Bio Guide Jun 08

Medical College of Georgia
Biological Safety Guide

June 2008
TABLE OF CONTENTS
List of Abbreviations .............................................................................. viii
Forward .................................................................................................. x
1 Introduction ......................................................................................... 1.1

1.1 Emergency Phone Numbers and Office of Environmental Health and Safety Contacts ...................1.1
1.2 Responsibilities .................................................................................................................................1-2

1.2.1 Department Chairperson and/or Center/Institute Director..............................................1-2
1.2.2 Principal Investigators, Clinical Directors and/or Instructional Course Directors ..........1-2
1.2.3 Research and MCG Clinical Personnel ..........................................................................1-5
1.2.4 Division of Environmental Health and Safety Biological Safety Office and Biosafety
Officer ............................................................................................................................1-5
1.2.5 MCG Institutional Biosafety Committee........................................................................1-7
1.2.6 The Medical College of Georgia (MCG)........................................................................1-8
1.2.6.1 Other Offices at MCG .................................................................................1-9
1.2.6.1.1 Facilities Operation & Maiantenance.......................................1-9
1.2.6.1.2 The Institutional Animal Care and Use Committee (IACUC)
and Division of Laboratory Animal Services (DLAS).............1-9
1.2.6.1.3 The Institutional Review Boards (IRBs) and the MCG Office
of Human Research Compliance (OHRP)................................1-10
1.2.6.1.4 Division of Sponsored Projects Administration (DSPA) .........1-11
1.2.6.1.5 Office of Institutional Audits and Compliance (OIAC) and the
Compliance Oversight Council (COC) ....................................1-11
1.2.6.1.6 Office of Technology Transfer and Economic Development
(OTTED)..................................................................................1-12
2 Biosafety Requirements ...................................................................... 2-1

2.1 Registration for the Use of Biological Materials...............................................................................2-1
2.1.1 Annual Affirmation of Biosafety Protocol Information...............................................2-2
2.1.2 Ammendments to Biosafety Protocols .......................................................................2-3
2.1.2.1 Adding New Personnel to or Removal of Personnel from a Biosafety
Protocol ...................................................................................................2-3
2.1.2.2 Amendments for New or Modified Biological Agents or Operations to a
Biosafety Protocol ...................................................................................2-4
2.1.2.3 Modifying the Locations or Equipment on a Biosafety Protocol.............2-4
2.1.2.4 Documenting New or Additional Project Titles Associated with a
Biosafety Protocol (or Multiple Biosafety Protocols)..............................2-5
2.2 Training Requirements....................................................................................................................2-6
2.3 Laboratory Establishment, Close-outs and Moves ........................................................................2-8

2.3.1 Incoming New Faculty to MCG ..................................................................................2-8
2.3.2 Laboratory Close-outs .................................................................................................2-9
2.3.3 Laboratory Moves .......................................................................................................2-10
2.4 Laboratory Record-keeping ............................................................................................................2-11
2.5 Paperwork Flow ..............................................................................................................................2-11

2.5.1 IBC Review/Approval Process for Biosafety Protocols (BSPs) and Amendments .....2-12
2.5.2 Verification that IBC has Received and approved Experiments, Agents, and Uses that
were Proposed in a Particular Sponsored Project, Clinical Protocol and/or Animal Use
Protocol to Other MCG Compliance Offices. .............................................................2-14
Medical College of Georgia Biosafety Guide-June 2008

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2.6 Biological Materials and Applications Which Must Be Documented on Biosafety Protocols ........2-17
2.6.1 Recombinant DNA Experiments .................................................................................2-17
2.6.1.1 Definition of Recombinant DNA ............................................................2-17
2.6.1.2 Definition of Exempt vs. Non-Exempt Recombinant DNA ....................2-18
2.6.1.3 Recombinant DNA Registration Requirements .......................................2-21
2.6.1.4 Human Gene Transfer/Therapy (HGT) ...................................................2-21
2.6.2 Human, Animal and/or Agricultural Pathogens and/or Toxins (or Materials
Potentially Contaminated with these Pathogens or Toxins) ........................................2-22
2.6.2.1 Toxins of Biological Origin.....................................................................2-23
2.6.2.2 Select Agents and Toxins ........................................................................2-23
2.6.2.2.1 Exclusions from Select Agents and Toxins Regulations..........2-26
2.6.2.2.2 Exemptions from Select Agents and Toxins Regulations for
Clinical/Diagnostic Laboratories .............................................2-27
2.6.2.2.3 Criteria for Handling Select Agent or Toxins Contaminated
Specimens for Diagnosis or Verification under the SAT
Exemption Clause ....................................................................2-28
2.6.3 Human Blood, Body Fluids, Cells, Tissues and Other Potentially Infectious
Material .......................................................................................................................2-29
2.6.3.1 Cultures of Cell Lines of Human and/or Primate Origin .........................2-31
2.6.3.1.1 Human Embryonic Stem (hES) Cells and Embryonic Germ
Cell Lines................................................................................2-32
2.6.4 Animals and the Use of Biological Materials in Animals ...........................................2-33

2.6.5 Insects/Arthropods ......................................................................................................2-34
2.7 Experiments Prohibited at MCG ....................................................................................................2-34
3 Risk Assessment .................................................................................. 3-1

3.1 Risk Assessment and Management Table .........................................................................................3-2
3.2 Considerations in a Risk Assessement ..............................................................................................3-3

3.2.1 Risk Groups: Hazardous Characteristics of a Biological Agent ....................................3-3
3.2.2 Routes of Exposure ........................................................................................................3-4
3.2.3 Operations with may be associated with additional risk of exposure .............................3-7
3.2.3.1 Zoonotic Disease Risks/Animal Research Risks ........................................3-7
3.2.3.1.1 Non-Human Primates................................................................3-7
3.2.3.1.1.1 Cercopithecine Herpes Virus (CHV-1); Herpes
B virus..................................................................3-8
3.2.3.1.1.2 Other Viral Infections associated with NHPs........3-9
3.2.3.1.1.3 Gastrointestinal Diseases associated with NHPs ...3-9
3.2.3.1.1.4 Bacterial Respiratory Diseases associated with
NHPs ....................................................................3-10
3.2.3.1.2 Birds.........................................................................................3-10
3.2.3.1.3 Dogs (Canines) .......................................................................3-11
3.2.3.1.4 Cats (Felines) ...........................................................................3-11
3.2.3.1.5 Rabbits (Lagomorphs) .............................................................3-12
3.2.3.1.6 Rodents (Mice, Rats, Hamsters, Gerbils, Guinea Pigs)............3-12
3.2.3.1.7 Ferrets ......................................................................................3-13
3.2.3.1.8 Pigs/Swine ...............................................................................3-13
3.2.3.1.9 Large Hooved Mammals (Cows, Horses, Sheep, Goats) .........3-14
3.2.4 Amount of Material Present ..........................................................................................3-15
3.2.5 Experience of Staff........................................................................................................3-15
3.2.6 Mode of Transmission...................................................................................................3-15
3.2.7 Environmental Stability.................................................................................................3-15
3.2.8 Institutional Public Image..............................................................................................3-15

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4 Risk Management: Biosafety Levels ................................................ 4-1
4.1 Laboratory Practices..........................................................................................................................4.4
4.1.1 Human Factors and Attitudes in Relation to Laboratory Accidents ...............................4.4
4.1.2 Practices Associated with Biosafety Levels 1 through 2+ Laboratories ........................4.4
4.1.2.1 Biosafety Level 1 (BSL-1/BL-1) ................................................................4.4
4.1.2.2 Biosafety Level 2 (BSL-2/BL-2). ..............................................................4-6
4.1.2.3 Biosafety Level 2+ (BSL2+) ......................................................................4-7
4.1.3 Practices Associated with Activities Involving Animals ...............................................4-8
4.1.3.1 Animal Biosafety Level 1 (ABSL-1/BL-1N)..............................................4-9
4.1.3.2 Animal Biosafety Level 2 (ABSL-2/BL-2N)..............................................4-11
4.1.4 Tissue/Cell and Microbiological Culture Practices ........................................................4-13
4.1.4.1 Tissue/Cell Culture Practices .....................................................................4-13
4.1.4.2 Microbiological Practices ...........................................................................4-15
4.1.4.3 Glass Ampules............................................................................................4-16
4.1.4.4 Cryovials/Cryopreservation in Liquid Nitrogen .........................................4-17
4.1.4.4 Vacuum-packed tubes/vials ........................................................................4-17
4.1.4.5 Embryonated eggs ......................................................................................4-17
4.1.5 Transport of Biohazards on Campus (between labs or buildings) ..................................4-18
4.1.6 Housekeeping .................................................................................................................4-18
4.1.6.1 Objectives of Housekeeping .......................................................................4-19
4.1.6.2 Scope ..........................................................................................................4-19
4.1.6.3 Assignment of Responsibilities...................................................................4-20
4.1.7 Standard Operating Procedure Development.................................................................4-20
4.2 Laboratory Equipment ......................................................................................................................4-21

4.2.1. Personal Protective Equipment (PPE) ...........................................................................4-21
4.2.1..1 Laboratory Clothing....................................................................................4-22
4.2.1.1.1 Gloves.......................................................................................4-23
4.2.1.1.2 Procedure for Removing Gloves...............................................4-24
4.2.1.1.3 Shoes.........................................................................................4-24
4.2.1.1.4 Gowns, Lab Coats, Jumpsuits, Aprons and Other Protective
Clothing.................................................................................4-24
4.2.1.1.5 Face and Eye Protection............................................................4-25
4.2.1.1.1.6 Respiratory Protection ...........................................................4-25
4.2.1.2 Selection of PPE .........................................................................................4-27
4.2.1.3 PPE Requirements Table .............................................................................4-28
4.2.2 Biological Safety Cabinets ..........................................................................................4-30
4.2.3 Laminar Flow Hoods/Clean Benches ..........................................................................4-32
4.2.4 Fume Hoods .................................................................................................................4-32
4.2.5 Centrifuges ...................................................................................................................4-32
4.2.6 Vacuum Line Chemical Traps and Filters ....................................................................4-33
4.2.7 Syringes and Needles ...................................................................................................4-34
4.2.8 Pipettes .........................................................................................................................4-35
4.2.9 Blenders, Mixers, Sonicators, and Cell Disruption Equipment ....................................4-36
4.2.10 Lyophilizers.................................................................................................................4-36
4.2.11 Microtome/Cryostat.....................................................................................................4-37
4.2.12 Fluorescence Activated Cell Sortiers (FACS) and FACS analyzers............................4-38
4.2.13 Miscellaneous Equipment (Waterbaths, Cold Storage, Shakers).................................4-39
4.3 Laboratory Facilities .........................................................................................................................4-40

4.3.1 Physical Separation of Laboratory Spaces from Non-Laboratory Spaces ......................4-41
4.3.2 Doors and Locks.............................................................................................................4-41
4.3.3 Laboratory and Biological Material Security .................................................................4-42
4.3.4 Windows ........................................................................................................................4-42
4.3.5 Floor Coverings, Walls and Ceilings.............................................................................4-42
4.3.6 Furnishings ....................................................................................................................4-42
4.3.7 Lab Benches/Cabinetry ................................................................................................4-43
4.3.8 Exhaust systems/Directional Airflow/HVAC...............................................................4-43
4.3.9 Plumbing/Eyewashes/Showers .....................................................................................4-43
4.3.10 Biosafety Cabinets.......................................................................................................4-44
4.3.11 Lighting, Air Ducts, Utility Pipes................................................................................4-44

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4.3.12 Additional Animal Facility Consdierations .................................................................4-44
5 Medical Surveillance Program ........................................................... 5-1

5.1 Tuberculosis (TB) Screening.............................................................................................................5-2
5.2 Immunizations ..................................................................................................................................5-3

5.2.1 Vaccinia Virus and Immunization Recommendations.............................................5-4
5.3 Respiratory Protection Program .......................................................................................................5-7
5.4 Medical Restrictions.........................................................................................................................5-7

5.4.1 Pregnancy .......................................................................................................................5-8
5.4.1.1 Reproductive Biological Hazards ................................................................5-8
5.4.2 Work with Animals ........................................................................................................5-9
5.4.3 Allergies .........................................................................................................................5-9
5.4.3.1 Allergies to Laboratory Animals (ALAs) ...................................................5-9
5.4.3.2 Latex Gloves and Related Allergies............................................................5-10
5.4.3.3 Antibiotic Allergies ....................................................................................5-10
5.4.3.4 Mold Allergies............................................................................................5-11
5.4.4 Other Restrictions.........................................................................................................5-12
5.5 Minors in Laboratories, Clinics or Other Hazardous Areas .............................................................5-12

5.6 Employee Serum Storage .................................................................................................................5-13
6 Accidents, Exposures, Spill Response ................................................ 6-1

6.1 Emergency Procedures for Exposure Incidents.................................................................................6-1
6.1.1 Percutaneous Injury ........................................................................................................6-1
6.1.2 Splash to Face.................................................................................................................6-1
6.1.3 Aerosol Exposure ...........................................................................................................6-1
6.2 Reporting Incident.............................................................................................................................6-1
6.3 Medical Assistance............................................................................................................................6-2
6.4 Spill Clean-up Procedures................................................................................................................6-3

6.4.1 Composition of a Basic Biohazard Spill Kit..................................................................6-3
6.4.2 Spills Occurring Outside of Laboratory Areas ..............................................................6-3
6.4.3 Spills Within Laboratory Areas .....................................................................................6-4
6.4.3.1 Biosafety Level 1 (BSL-1) Spills................................................................6-4
6.4.3.2 Biosafety Level 2 (BSL-2) Spills................................................................6-4
6.4.3.3 Blood Spills ................................................................................................6-5
6.4.3.4 Spill in a Biosafety Cabinet ........................................................................6-5
6.4.3.5 Centrifuge Spill...........................................................................................6-6
6.4.3.6 Spill of a Biohazardous Radioactive Material.............................................6-6
6.5 Investigation of Laboratory Accidents .............................................................................................6-7
7 Decontamination of Biologicals .......................................................... 7-1

7.1 Decontamination Methods ................................................................................................................7-1
7.1.1 Heat ................................................................................................................................7-1
7.1.1.1 Autoclave Use and Maintenance .................................................................7-1
7.1.2 Chemical Decontaminants .............................................................................................7-2
7.1.2.1 Liquid Chemical Decontaminants...............................................................7-2

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7.1.2.2 Vapors and Gases ........................................................................................7-3
7.1.3 Physical Methods ..........................................................................................................7-3
7.2 Characteristics of Chemical Decontaminants....................................................................................7-4

7.2.1 Properties of Some Common Decontaminants ...............................................................7-4
7.2.1.1 Chlorine/Hypochlorite ................................................................................7-4
7.2.1.2 Alcohols......................................................................................................7-5
7.2.1.3 Aldehydes (Formaldehyde, Glutaraldehyde) ..............................................7-5
7.2.1.4 Phenols .......................................................................................................7-5
7.2.1.5 Quaternary Ammonium Compounds (“Quats”)..........................................7-6
7.2.1.6 Iodine..........................................................................................................7-6
7.2.1.7 Peroxygens .................................................................................................7-6
7.2.2 Selecting Chemical Disinfectants ...................................................................................7-6
7.2.3 Characteristics of Some Liquid Disinfectants Table ......................................................7-7
7.3 Deactivation of Biological Toxins....................................................................................................7-10
7.4 Deactivation of Prions......................................................................................................................7-13
8 Waste Management ............................................................................ 8-1

8.1 Purpose of Biological Waste Management Program .......................................................................8-1
8.2 Definition of Biomedical Waste......................................................................................................8-1
8.3 Solid Biomedical Waste ..................................................................................................................8-3
8.3.1 Proper Solid Biohazardous Waste Disposal Procedures................................................8-4
8.3.2 Non-contaminated Glass ...............................................................................................8-6
8.3.3 Improper Solid Biohazardous Waste Disposal ..............................................................8-6
8.4 Pathological Wastes ........................................................................................................................8-7
8.5 Liquid Biological Wastes ................................................................................................................8-7
8.6 Chemotherapy Wastes.....................................................................................................................8-8
8.7 Mixed Waste ...................................................................................................................................8-8
8.7.1 Biological/Chemical Mixed Waste................................................................................8-8
8.7.2 Biological/Radioactive Mixed Wasete ..........................................................................8-9
8.8 Requisitions for New or Additional Biohazardous Waste Containers.............................................8-9
8.9 Requisition for Biohazardous Waste Pick-up..................................................................................8-9
9 Placard Signs and Labels ................................................................... 9-1

9.1 Door Placards....................................................................................................................................9-1
9.2 Labels and Color-Coding ..................................................................................................................9-2
9.3 Labeling Equipment Sent Out for Repair or Disposal.......................................................................9-2
10 Shipping .............................................................................................. 10-1

10.1 Training Requirements...................................................................................................................10-1
10.2 Shipping Overview..........................................................................................................................10-4
10.3 Shipment Type ...............................................................................................................................10-4

10.3.1 Unregulated Biological Materials............................................................................10-4
10.3.2 Infectious Substances ..............................................................................................10-7
10.3.2.1 Category A Infectious Substances ...........................................................10-7
10.3.2.1.1 Packaging...............................................................................10-7
10.3.2.1.2 Labeling .................................................................................10-10
10.3.2.2 Category B Infectious Substances............................................................10-10

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10.3.2.2.1 Packaging...............................................................................10-10
10.3.2.2.2 Labeling .................................................................................10-10
10.3.3 Patient Specimens....................................................................................................10-11
10.3.3.1 Packaging ................................................................................................10-11
10.3.3.2 Labeling...................................................................................................10-11
10.3.3.3 Dried Blood .............................................................................................10-11
10.3.4 Biological Products .................................................................................................10-11
10.3.5 Genetically Modified Organisms or Microorganisms..............................................10-12
10.3.5.1 Packaging ................................................................................................10-12
10.3.5.2 Labeling...................................................................................................10-12
10.3.6 Regulated Biomedical Waste...................................................................................10-12
10.4 Packaging Biological Material .......................................................................................................10-13

10.4.1 Triple Packaging......................................................................................................10-13
10.4.2 Other Packaging Requirements ...............................................................................10-14
10.4.2.1 Overpacks ................................................................................................10-14
10.4.2.2 Dry Ice .....................................................................................................10-14
10.4.2.2.1 Hazard Identification .............................................................10-14
10.4.2.2.2 Packaging Dry Ice..................................................................10-15
10.4.2.3 Liquid Nitrogen .......................................................................................10-17
10.4.2.4 Samples Preserved in Fixative .................................................................10-18
10.4.2.4.1 In Aqueous Formaldehyde Solutions <25% (Formalin).........10-18
10.4.2.4.2 In Aqueous Ethanol Solutions of 55-100%............................10-20
10.5 Shipper’s Declaration for Dangerous Goods..................................................................................10-21
10.6 Regulated Agents Which May Require Special Permits for Transfer ............................................10-25
10.6.1 .... CDC/USDA Select Agents & Toxins......................................................................10-25
10.6.2 .... Agricultural Pests, Pathogens and Biological Agents..............................................10-25
10.6.3 .... Agents or Vectors of Human Disease......................................................................10-26
10.6.4 .... Department of Commerce- Bureau of Industry and Security (BIS) ........................10-26
10.6.5 .... FDA Import Permits ................................................................................................10-28
10.6.6 .... Fish and Wildlife Service Permits ...........................................................................10-28
10.7 International Shipments................................................................................................................10-29

10.7.1 .... Exporting from the United States ............................................................................10-29
10.7.2 .... Importing into the United States..............................................................................10-29
10.8 Shipping Company Restrictions ...................................................................................................10-30

10.8.1 .... DHL.........................................................................................................................10-30
10.8.2 .... FedEx ......................................................................................................................10-30
10.8.3 .... United Parcel Service (UPS) ..................................................................................10-30
10.8.4 .... United States Postal Service (USPS) ......................................................................10-30
10.9 United States Postal Service Mailings..........................................................................................10-30

10.9.1 Mailing Category B Substances...............................................................................10-30
10.9.2 Mailing Exempt Human and Animal Specimens.....................................................10-31
10.9.3 Mailing Non-regulated Materials.............................................................................10-31
10.9.3.1 Non-Regulated Liquid Substance, Not Exceeding 50 ml.........................10-31
10.9.3.2 Non-Regulated Liquid Substance, Exceeding 50 ml................................10-31
10.9.3.3 Non-Regulated Dry Substance.................................................................10-31
10.10 Transport as Checked Airline Baggage .........................................................................................10-31
10.11 Transport in Ground Vehicles ......................................................................................................10-32

10.11.1 Passenger Automobiles............................................................................................10-32
10.11.2 Public Modes of Transportation (e.g., Shuttle busses) ............................................10-32
10.11.3 Courier Services ......................................................................................................10-32

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Appendicies.............................................................................................. A-1
Appendix A Biosafety Protocol Applications Forms...................................................................................A-1
Appendix B Classification of Human Etiologic Agents on the Basis of Hazard .........................................B-1
Appendix C Biosafety Levels......................................................................................................................C-1
Appendix D BL2+ Work Practices ..............................................................................................................D-1
Appendix E SOP Development Template ...................................................................................................E-1
Appendix F Human Gene Transfer Clinical Trials .....................................................................................F-1
Appendix G Sources of Contamination .......................................................................................................G-1
Appendix J Biosafety Notice for Decontamination of Equipment .............................................................J-1
Appendix K Shipping Labels.......................................................................................................................K-1
Appendix L Map of MCG Campus .............................................................................................................L-1
Acknowledgements

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LIST OF ABBREVIATIONS
AAV Adeno-associated Virus

ABSA American Biological Safety Association
AdV Adenovirus
ALA Allergy to Laboratory Animals
ANSI American National Standards Institute
APHIS (U.S.D.A) Animal & Plant Health Inspection Sevice
ARO Alternate Responsible Official
BBP Bloodborne Pathogen
BMBL Biosafety in Microbiological and Biomedical Laboratories (CDC/NIH Guidelines)
BSC Biosafety Cabinet (a.k.a “Tissue Culture Hood”)
BSO Biosafety Officer
BSP Biosafety Protocol
CCRI Chesapeake Research Review, Inc. (an MCG-authorized IRB)
CDC Centers for Disease Control
cDNA Complementary DNA
CFR (U.S.) Code of Federal Regulations
COC (MCG) Compliance Oversight Council
DLAS Division of Laboratory Animal Services
DOAS (Georgia State) Department of Administrative Services
DOC (U.S.) Department of Commerce
DOT (U.S.) Department of Transportation
DSPA (MCG) Division of Sponsored Project Administration (formerly, Contracts & Grants)
EHS Environmental Health and Safety
EPA (U.S.) Environmental Protection Agency
EPD (GA) Environmental Protection Division
FACS Fluorescence Activated Cell Sorting
FDA (U.S.) Food and Drug Administration
GeMCRIS Genetic Modification Clinical Research Information System (NIH/FDA)
HAC (MCG) Human Assurance Committee (MCG’s internal IRB)
IACUC Institutional Animal Care and Use Committee
IATA International Air Transport Association
IBC Institutional Biosafety Committee
ICAO International Civil Aviation Organization
IND (FDA) Investigational New Drug
IRB Institutional Review Board
LAS Laboratory Animal Services (see also: DLAS)
LD50 Lethal Dose 50 (i.e. dose of agent required to kill 50% of recipient subjects)
LFH Laminar Flow Hood
MCG Medical College of Georgia
MCGHI Medical College of Georgia Health Incorporated (the MCG hospital, a private entity, separate from MCG)
MCGRI Medical College of Georgia Research Institute
MLV Moloney Leukemia Virus (a mouse retrovirus)
MSDS Material Safety Data Sheet
NHP Non-human Primate
NIH National Institutes of Health
NIOSH National Institute for Occupational Safety and Health
NSF National Sanitation Foundation, International

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OBA (NIH) Office of Biotechnology Activities
OCGA Official Code of Georgia
OCIS (MCG) Office of Clinical Investigative Services
OHRP (MCG) Office of Human Research Protection
OIAC (MCG) Office of Internal Audits and Compliance
OSHA (U.S.) Occupational Safety and Health Administration
OTTED (MCG) Office of Technology Transfer and Economic Development (a.k.a. Intellectual Properties Office)
PI Principal Investigator
PPD Purified Protein Derivative (a.k.a. Mantoux) tuberculin sensitivity test (a common test for tuberculosis)
RAC Recombinant DNA Advisory Committee (to NIH/OBA)
RCL Replication Competent Lentivirus
RCR Replication Competent Retrovirus
rDNA Recombinant DNA
RO Responsible Official
SAE Serious Adverse Event
SAT Select Agents and Toxins
SOP Standard Operating Procedures
UN United Nations
USDA U.S. Department of Agriculture
VPR Vice President for Research
WHO World Health Organization

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FORWARD
This Biosafety Manual has been developed by the Division of Environmental Health and Safety Biosafety
Office and the Institutional Biosafety Committee (IBC) at the Medical College of Georgia (MCG). This
Guide is part of the MCG Biosafety Program instituted to accomplish the following goals:
To protect against exposures of personnel (MCG employees and students, community

members and visitors) to biological agents
To prevent environmental contamination
To provide an environment for high quality research while maintaining a safe workplace
To comply with applicable federal, state and local regulations and guidelines
To comply with Guidelines implemented by federal funding agencies and accepted by
MCG as a condition of funding eligibility
To create a secure laboratory environment to prevent unauthorized utilization of a
biological agent.
This Biosafety Manual provides university-wide safety guidelines, policies and procedures for the use,
possession, manipulation and transport of biological materials. Although the implementation of these
procedures is primarily the responsibility of the Principal Investigator (PI), its success depends largely on
the cooperative efforts of laboratory supervisors, employees and students. Please read the section on
responsibilities for additional information. Planning for and implementation of biological safety must be
part of every laboratory activity in which potentially biohazardous material is used.
Recommendations in this Biosafety Manual define a “standard of practice” that laboratories should follow.
In general, the possession, handling, manipulation and transport or biological agents, including:
Recombinant DNA molecules
Infectious or potentially infectious agents, including human or non-human primate derived
material, cultures and genetically modified cells
Microbial agents (e.g. viruses, bacteria, mycobacteria, rickettsia, yeast, fungi, prions,
parasites) or specimens which may be exposed to microbial agents
Toxins of biological origin
requires the use of various precautionary measures depending on the material(s), facilities, personnel and
their experience, and procedures involved. This manual will provide assistance in the evaluation,
containment and control of these biohazards. It is required that all parties involved and/or working with
these material be familiar with the contents of this manual, complete the required training, and that they
seek additional advice when necessary. The IBC Chairperson, IBC members, as well as the Biosafety
Officer (BSO), are available to assist in this endeavor.
This manual focuses on Biosafety Levels 1 and 2, as all MCG laboratories fall within these designations as
of May, 2008. A separate manual will be available for researchers working in Biosafety Level 3 research
laboratories which will be developed as part of establishing these facilities . No work with Biosafety Level
4 agents may be conducted at MCG.
We urge you to use the manual as a road map to compliance within your laboratory. Consult the sections
relevant to your research and apply the appropriate safety procedures. The Biosafety Office is available for
consultation if you have any question or concern with any aspect of the Biosafety Program at MCG. The
credo, “Think before you act,” and “If you do not know, ask,” are relevant to the use of this manual. If you
are unsure of a requirement or biosafety practice, please contact the Biosafety Office at x1-2663 or
BIOSAFETY@mcg.edu for assistance. We also would appreciate any feedback or comments that you may
have with the use of this manual, and will incorporate any suggestions in future versions.

x
1. INTRODUCTION
1.1 EMERGENCY PHONE NUMBERS AND SAFETY, COMPLIANCE
AND OPERATIONS CONTACTS
Emergency Telephone Numbers
Ambulance/Fire/Police x1-2911
Employee Health Office (FG-1174) x1-3418
Student Health Office (AF-1040) x1-3487
Biological/Chemical/Radiological Emergencies x1-2663 (Monday - Friday, 8:30 AM to 5:00 PM)
(After at other times contact Campus Police at x1-2911)
MCG Human Resources & Benefits (HS-1111) x1-3770
Georgia Department of Administrative Services 1-877-656-7475
Office of Environmental Health and Safety (EHS) Contacts (CI-1001)

http://www.mcg.edu/services/ehs/Index.html
Office of Environmental Health and Safety Main Office (CI-1001) x1-2663

Dr. Stacey Kraemer, Biosafety Officer x1-2663
Duane Perry, Environmental & Occupational Health Officer x1-2663
Kenneth Erondu, Chemical Safety Office x1-2663
Philip Maguire, Assistant Radiation Safety Officer x1-9826
Laboratory Animal Services (LAS) and Institutional Animal Care and Use
Committee Contacts
http://www.mcg.edu/research/animal/
LAS Main Office (CB-1901) x1-3421

Dr. Laura Tambrallo, LAS Director x1-0199
Dr. Nancy Rodriguez, LAS Assoc. Director x1-8575
Rennie Wolfe, LAS Asst. Director x1-2685
Dr. Patricia Charlton, LAS attending vet x1-0111
IACUC Office (CA-1094) x1-0198
Jenny Whitlock, IACUC compliance officer
Facility Management/Research Support & Compliance Office Telephone

Numbers
Facilities Management x1-2434

Environmental Services x1-2434
(housekeeping services, biowaste disposal/containers)
Laboratory Equipment Services (LES) (CB-0110) x1-6124
(lab equipment repair, biosafety cabinet certification)
Information Technology x1-4000
Telecommunication x1-3985
Public Safety (non-emergency) x1-2914
Key/I.D. Badging Office x1-6287
MCGHI Epidemiology x1-2224
Office of Technology Transfer and Economic Development (CA-2105) x1-9822
http://www.mcg.edu/research/techtransfer/
Office of Human Research Protection (OHRP) (CJ-2103) x1-1478
http://www.mcg.edu/research/ohrp
Medical College of Georgia Biosafety Guide- June 2008

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Office of Clinical Investigative Services (OCIS) (FF-100) x1-9680
http://www.mcg.edu/research/ohrp/
Division of Sponsored Project Administration(DSPA) Contacts (CJ-3301) x1-2592
http://www.mcg.edu/SPA/
Office of Institutional Audit and Compliance (HS-3135) x1-2661
http://www.mcg.edu/audits/
24-hour Compliance Hotline (for confidential reporting of compliance concerns) 1-800-576-6623
http://www.mcg.edu/audits/hotline.htm
Legal Affairs Office (AA-211) x1-4018
http://www.mcg.edu/services/legal/
1.2 ROLES AND RESPONSIBILITIES

1.2.1 Department Chairperson and/or Center/Institute Director
The Department Chairperson and/or Center/Institute Director bears overall responsibility for the
implementation and maintenance of safe practices and procedures in the department.
Department/Center/Institute Heads have the following responsibilities:
To ensure that prior to initiation of work, each Principal Investigator of a Research laboratory,
Clinical Director or each clinical laboratory, and/or Instructional Course Director within their
department which may expose MCG personnel, students, animals, the environment or the public
to biological material files a Biosafety Protocol (BSP) for review by the IBC through the
Biological Safety Officer and that approval has been granted prior to the initiation of the research.
To ensure that students, staff and faculty within their department/center/institute have had
instruction in safety procedures in research and teaching laboratories or field situations where
biological agents are used or collected.
To ensure that resources are made available to researchers, health care providers, laboratory and/or
clinic staff, instructors and/or students who require health screening and/or vaccination due to
potential risk of exposure to particular biological materials.
To assume responsibility for maintaining the appropriate Biosafety standards and documentation
of shared departmental facilities or delegate that responsibility to an appropriate faculty member
within the department.
To provide leadership in laboratory or clinic safety at the management level in the department or
institute.
1.2.2 Principal Investigators, Clinical Directors and/or Instructional Course

Directors
The Principal Investigator (PI), Clinical Director, and/or Instructional Course Director in which biological
material is used bears the ultimate responsibility and authority for assessing risks, establishing policies and
procedures, training personnel and maintaining the facility and equipment.
The principal investigator, clinical director, and/or instructional course director is responsible for:
Performing appropriate risk assessment of research projects. The level of detail should be
dependent on the hazard associated with the organism under study (e.g., an assessment of risk
associated with research on Risk Group 1 agents might reasonably be less detailed than a risk
assessment of a Risk Group 2 or unknown agents). Each evaluation should be completed before
work is undertaken and the project should be reassessed periodically as new data is obtained. The

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assessment should include an analysis of the risks posed by the particular organism under
investigation and of any specific research, clinical or teaching methods that may affect that risk
(e.g., procedures requiring highly concentrated amounts of microorganisms or inoculation of
laboratory animals). No human or animal pathogen should be studied without prior written
approval of the MCG Institutional Biosafety Committee (IBC). The procedures for handling
unclassified agents must also be reviewed by the MCG IBC, the Division of Environmental Health
and Safety (EHS), as well as the MCG Division of Laboratory Animal Resources (DLAS) if work
with animals is anticipated. The agents must be registered and information about these agents must
be provided to EHS.
Registering research work involving biological materials, particularly non-exempt recombinant

DNA and potential animal, human or plant pathogens with the Institutional Biosafety Committee
(IBC). This Biosafety Protocol (BSP) application must detail the nature of the proposed
experiments and an assessment of the levels of physical and biological containment required for
them as established by the NIH and CDC guidelines. The research described in BSP applications
must be accurate and complete description of the research projects in the laboratory and reflect the
agents, locations, operations, experiments, manipulations, personnel and safety measures that will
be employed in the laboratory. Changes in the protocols must be submitted as amendments to the
BSPs to the IBC prior to its initiation. PIs, Clinical Directors and/or Course Directors are
responsible for ensuring that no research is initiated by any laboratory personnel prior to review
and approval by the IBC.
Developing, establishing and implementing appropriate safety practices and procedures within
their laboratories prior to bringing new biological agents to campus and/or before initiating any
new research project (independent of funding status) to ensure safe operation and instructing
students and staff of potential hazards. This involves:
• Being knowledgeable in good laboratory practices and maintaining current knowledge of
new safety practices and/or equipment which may improve safety within the laboratory.
• Demonstrating a positive safety attitude.
• Making available to the laboratory staff copies of the written procedures that describe
potential biohazards, precautions, and actions to be taken in response to spills and
accidents to include decontamination procedures, and emergency procedures. These
procedures and other information addressing Biological Safety-related issues will be
produced in the form of a standard operating procedure (SOP) for the work. The SOP
will reside within the laboratory and be easily accessible for reference and provided to the
IBC for review as part of the Biosafety Protocol application.
• Maintaining up-to-date knowledge to changes in international federal, state, local

regulations and guidelines pertaining to biological materials, in consultation with the
Biosafety Office, and modifying the laboratory procedures to comply with these.
Approving research personnel to work in the laboratory and documenting that personnel are
competent to conduct the work. PIs, Clinical Directors and/or Course Directors are responsible
for the safety of personnel on their Biosafety Protocols (BSPs) and their actions. This includes:
• Providing laboratory staff with documented formal and informal instruction and training
in the practices and techniques required to ensure safety and in the procedures for dealing
with accidental spills, personnel contamination, and other laboratory accidents or
emergencies.
• Informing the laboratory staff of the risks involved with the biological agents in the
laboratory and the reasons and provisions for any precautionary medical practices (e.g.
physical examinations, serum collection, and vaccinations).

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• Making provisions for any precautionary medical practices, including occupational health
physical examinations, vaccinations and/or other medical surveillance of personnel when
required by the agents and nature of the experiments.
• Supervising and monitoring the performance of the staff to ensure that required safety
practices and techniques are employed.
• Ensuring the authorized staff complete the appropriate IBC-required training modules
and keeping these training records up-to-date.
Maintaining a liaison with the Biosafety Office. This includes:

• Amending and modifying Biosafety Protocols (BSPs) to reflect changes in agents,
personnel, locations, experimental application, operations and/or safety equipment.
• Reporting, in writing, any accident, potential exposure or exposure of personnel,

suspected illness, and/or release from containment of any biohazardous agents. Any
significant problems pertaining to the operation and implementation of containment
practices, procedures or facilities should also be reported to the Biosafety Office.
• Providing accurate information for compliance verification processes when requested.
Maintaining compliance with all Federal, State and/or local regulations related to possession, use,
transfer and/or disposal of Biohazardous materials. This would include the following regulations:
• Federal Select Agent regulations (42 CFR §73.3, 7 CFR §331.3 and 9 CFR §121.4) for
use, possession and transfer of Select Agents and Toxins
• U.S. Department of Transportation Regulations (49 CFR), U.S. Public Health Service (42
CFR §72) and IATA guidelines for shipping and/or transport of hazardous or etiological
materials
• U.S. Departments of State, Commerce and Treasury Regulations related to Export control
laws
• U.S. Department of Agriculture Animal and Plant Health Inspection Service Regulations
related to transportation, importation or exportation of animal or animal products,
genetically engineered organisms, plants or plant products and/or soil samples.
• U.S. Public Health Service importation and/or exportation requirements for Etiologic
Agents, any arthropod and/or other animal host or vector of human disease, including
unsterilized specimens of human and animal tissues (such as blood, body discharges,
fluids, excretions or similar material) containing an infectious or etiologic agent
• O.C.G.A. §31-12-13 and OSHA (29 CFR §1910.1030) standards for blood-borne
pathogen handling, medical surveillance, training and record-keeping
• Georgia EPA Solid Waste Management Laws Chapter 391-3-4-.15 for Biomedical Waste
Ensuring that the terms and conditions of NIH Grants Policy Statement are maintained within the
laboratory for all research projects (independent of funding status or sponsor). This includes
compliance with the NIH Guidelines for Research with Recombinant DNA Molecules, the
Occupational Health and Safety Administration (OSHA) standards included in 29 CFR Part 1910,
and other applicable safety guidelines, including those in the CDC/NIH publication Biosafety in
Microbiological and Biomedical Laboratories.

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1.2.3 Research and MCG Clinical Personnel and MCG Students
Research and MCG Clinical Personnel and MCG students are responsible for:
Completing requirements for approval to work in the laboratory and ensure that all work is
conducted in compliance with MCG, NIH, CDC, OSHA, state labor and waste management laws,
DOT and other applicable guidelines and regulations and MCG IBC policies prior to initiation of
any work with biological materials at MCG (See Section 2, Biosafety Requirements). Follow the
MCG Biological Safety Guide except where superseded by the MCG BSL-3 Manual.
Learning the standard operating procedures (SOPs) for the laboratory, the potential hazards of the
infectious agents in use and emergency procedures. Personnel are responsible for helping to
maintain the facility in good working condition and maintaining compliance with the laboratory
Biosafety Protocol and SOPs.
Maintaining their work areas neat and clean. All containers in which biological materials are
placed should be appropriately labeled with biohazard stickers. Biohazardous waste must be
disposed according to laboratory Standard Operating Procedures (SOPs).
Completing any medical surveillance requirements as required by the IBC on the Principal
Investigator’s Biosafety Protocol Agreement with the IBC prior to initiation of work with
biological materials at MCG.
Reporting to the Principal Investigator (PI), Clinical Directors and/or Course Directors any
medical restrictions, reportable illnesses, and any event that may be an exposure or result in the
creation of a potential hazard. Report all irregular conditions.
Bring to the attention of the PI, Clinical Director and/or Course Director any practice on the
laboratory SOPs that is impossible or impractical to maintain operations within the laboratory so
amendments to SOPs and Biosafety Protocols can be submitted which accurately reflect
laboratory operations or provide suggestions of new SOPs which should be added to the
laboratory SOP list to better facilitate communication of safety issues among the laboratory staff.
If inexperienced in handling human pathogens, tissue culture, recombinant DNA and/or

microorganisms, receiving additional training from the Principal Investigator and demonstrating
proficiency in these practices to the Principal Investigator prior to initiation.
Performing responsibilities assigned to them by the Principal Investigator, Clinical Director or

Instructional Course Director. The operation of the facility is the responsibility of the users;
therefore a number of tasks must be assigned. These tasks may include the following:
• Training of other staff members
• Autoclave maintenance and waste management
• Freezer, refrigerator, equipment maintenance
• Cleaning
• Vacuum trap and filter maintenance
• Maintenance of supplies, including personnel protective equipment
• Security of infectious agents; i.e. store infectious agents in a locked freezer in a locked
laboratory
1.2.4 Division of Environmental Health and Safety (EHS), Biological Safety Office
and Biosafety Officer (BSO)
MCG’s Biological Safety Office has responsibility for the daily administration of standards set by the MCG
Institutional Biosafety Committee (IBC) and acts as the agent of the committee in the implementation of
their standards. In addition, the Biosafety Office serves as a resource to researchers, administration,

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compliance and maintenance departments, as resources allow. Responsibilities include:
Providing information and consultation on operation of the laboratory to ensure compliance with
CDC, NIH, USDA, OSHA, DOT, EPA, state and local requirements to researchers, administrators
and other institutional compliance and maintenance offices.
Evaluation and inspection of laboratory facilities for work with infectious agents, recombinant
DNA and other potentially hazardous biological agents. This includes advising on safety
measures and equipment for new procedures that may be utilized to mitigate risks associated with
working with potentially hazardous materials.
Administration and maintenance of records for the Institutional Biosafety Committee, including
Biosafety Protocols, Personnel training records, meeting minutes and scheduling of IBC meetings.
Periodic review of rDNA experiments documented in the Biosafety Protocols to ensure

compliance with the NIH Guidelines for Research Involving Recombinant DNA Molecules (NIH
Guidelines)
Providing general biosafety training programs related to proper handling of biological materials
and maintenance of training records for compliance with federal, state and University
requirements (laboratory-, agent- and operation-specific training is the responsibility of the
Principal Investigator (see Section 1.2.2)
Providing advice and assistance in the event of large, high hazard or public biological material
spills.
Investigation of laboratory incidents, accidents, exposures, potential exposures, illnesses which

may have resulted from potential exposures to biological material in the laboratory, releases or
possible releases from containment of biological materials to ensure appropriate emergency
follow-up procedures have been followed. If alternate incident mitigation and/or management
procedures are required to circumvent future similar incidents, the Division of Environmental
Health and Safety will make recommendations to the appropriate safety committees to address
these.
Coordinating/consulting with other institutional safety and compliance offices to maintain

compliance and safety standards.
Serve as a point of contact and information for Facilities Maintenance, Security, IT, Public Safety,
internal and external responders related to safety with biological materials and research activities
within MCG facilities.
Identification and updating of areas of known and potential biohazards at MCG to the IBC on a
regular basis.
Within the Biosafety Office, the Biological Safety Officer (BSO) is the institutional biological safety
officer for recombinant DNA research, and required to maintain compliance with NIH Guidelines for
Research with Recombinant DNA Molecules (NIH Guidelines). NIH assigns the BSO the following roles
and responsibilities:
Periodic inspections to ensure that laboratory standards are rigorously followed;
Reporting to the Institutional Biosafety Committee and the institution any significant problems,
violations of the NIH Guidelines, and any significant research-related accidents or illnesses of
which the Biological Safety Officer becomes aware unless the Biological Safety Officer
determines that a report has already been filed by the Principal Investigator;
Developing emergency plans for handling accidental spills and personnel contamination and

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investigating laboratory accidents involving recombinant DNA research
Providing advice on laboratory security
Providing technical advice to Principal Investigators and the Institutional Biosafety Committee on
research safety procedures.
The BSO also currently serves as the Institutional Responsible Official (RO) while the Director of
Environmental Health and Safety currently serves as the Alternate Responsible Official (ARO) for
Compliance with the Federal Regulations on the Possession, Use, and Transfer of Select Agents and Toxins
as described in 42 CFR 73, 7 CFR 331 and 9 CFR 121. As such, the RO and ARO have the authority and
responsibility to act on behalf of the institution as dictated by these laws.
1.2.5 MCG Institutional Biological Safety Committee (IBC)

The MCG Institutional Biosafety Committee (IBC) serves to maintain institutional compliance with the
National Institutes of Health (NIH) Guidelines for Research Involving Recombinant DNA Molecules (NIH
Guidelines), the Centers for Disease Control (CDC) Biosafety in Microbiological and Biomedical
Laboratories (BMBL), and International, Federal, state and local regulations pertaining to the handling of
biological materials. The IBC shall also advise the President, Provost, Vice Presidents of Research and
Administration and the Director of the Division of Environmental Health and Safety (EHS) and/or other
pertinent offices on policy matters concerned with the protection of personnel from biohazardous agents
including both infectious organisms and allergens that may be present in either laboratory materials or the
environment. The IBC shall also recommend guidelines relating to procedures and facilities used at the
University, including such matters as safety training and health surveillance.
The IBC shall offer its counsel to all University personnel regarding matters of biological safety. The
President, Provost and/or Vice Presidents of Research or Administration may ask the Committee to inform
the community about developments in the general area of biological safety.
The IBC’s composition, roles and responsibilities adhere to those dictated in NIH Guidelines and the
membership can be viewed at: http://www.mcg.edu/services/ehs/biosafe/IBCmembers.pdf. As such, the
Committee is required to review applications for research involving recombinant DNA and biological
materials to determine whether the facilities, procedures, and practices meet the standards required by the
University and the NIH. It shall, in addition, have the responsibility to certify annually to the NIH that
such facilities, procedures, and practices, and the training and expertise of personnel meet NIH standards.
Meetings called for the purpose of such review and certification may be open to the public. Minutes of
these meetings shall be kept and made available for public inspection.
The IBC's responsibilities include:
Review applications and perform comprehensive risk assessments to determine the
appropriateness and adequacy of containment levels and safety measures proposed and/or used in
research, clinical duties and teaching.
Assess the adequacy of containment facilities for biological agents and rDNA molecules as
required by NIH or other funding or regulatory agencies. The IBC may down-grade or up-grade
containment levels as appropriate to address the risks associated with the proposed activities.
Assess for adequacy the facilities, procedures, practices, training and expertise of personnel
involved in the research, clinical duties and/or instructional activities.
When applications involve Select Agents, ensure that approval is granted only to those individuals
who meet the access requirements stated in Federal Regulations on the Possession, Use, and
Transfer of Select Agents and Toxins as described in 42 CFR 73, 7 CFR 331 and 9 CFR 121 and
other applicable Federal regulations.

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Develop with the Biological Safety Officer informational and training seminars and workshops on
biohazards for the MCG community.
Periodically review biohazardous research, clinical duties, and/or instructional activities being
conducted at MCG to ensure that the requirements of the University, funding sources, and
regulatory agencies are being fulfilled.
Recommend to MCG Administration appropriate sanctions for noncompliance with Biological

Safety standards, guidelines, or regulations.
Adopt emergency plans covering accidental spills and personnel contamination resulting from
biohazardous research.
When experiments involve rDNA in humans, the IBC will grant no approval and ensure that no
activities are conducted until NIH OBA Recombinant DNA Assurance Committee approval has
been obtained and compliance with Appendix M of NIH Guidelines can be ensured.
In addition, the IBC shall oversee the activities of the Biosafety Office in the sense that it shall:
review its objectives and performance goals;
monitor its progress in meeting those objectives and goals, and
recommend changes in the organization and activities of the Biosafety Office that the committee
may find desirable
The IBC shall meet regularly with the Director of EHS and the Biological Safety Officer to receive
progress reports and advise on specific safety issues as well as on general safety policy.
On matters of oversight that involve the evaluation of performance by the Biosafety Office, the Committee
may, at the discretion of the chair, meet in executive session. In such cases the Biological Safety Officer,
the Director EHS and any other EHS office representatives shall be excused from participation and voting.
1.2.6 The Medical College of Georgia (MCG)

The Medical College of Georgia and its administrative officers are ultimately responsible for the following:
Developing and maintaining appropriate policies regarding the conduct of potentially
biohazardous research, education, and service activities.
Developing mechanisms for ensuring adherence to Biological Safety policies.
Providing the resources necessary for the construction of safe research, clinical and teaching
facilities and for the implementation of the Biological Safety Program.
Providing adequate resources for IBC member training on biohazards and biological safety
procedures, including training programs and workshops.
Providing resources for appropriate medical surveillance measures to protect the health and safety
of employees.
Providing appropriate and sufficient legal protection for faculty and staff members who conduct
activities in compliance with appropriate regulations and guidelines.

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Coordinate efforts between institutional safety and compliance offices to ensure compliance with
International, Federal, State and Local regulations and guidelines for research and clinical care.
1.2.6.1 Other Offices and Committees at the Medical College of Georgia

(MCG)
1.2.6.1.1 Facilities Operations & Maintenance
The MCG Facility Operations and Maintenance Division is responsible for:
Ensuring that the design and construction of all laboratory and clinical facilities meet
the standards of containment to maintain compliance with Federal, State and Local
Regulations, NIH Guidelines for Research with Recombinant DNA Molecules, the
Centers for Disease Control Biosafety in Microbiological and Biomedical
Laboratories and the GA Board of Regents among other standards and their intended
purposes.
Making sure the mechanics and integrity of the facilities are maintained to ensure
proper protection of workers within the facilities and proper protection of the
environment from hazardous materials within the buildings.
Notifying the Biosafety Office, laboratory director and building manager any
planned maintenance of the facilities which would require even short suspension of
utility operations required to maintain safety in the laboratory. This may include
tests or repairs of HVAC, electrical, plumbing or vacuum systems.
Immediately reporting any possible failures in facility containment that may have
resulted in environmental release of biological materials or potential exposures of
any personnel to biohazardous materials to the Biosafety Office (x1-2663 or
BIOSAFETY@mcg.edu), as well as the building directors, departmental chairs and
researchers.
Reporting any issues of non-compliance that are noticed while within laboratories,
particularly if these may potentially place Facilities personnel at higher risk for
exposure (e.g. inappropriate waste disposal for sharps or biological material).
Because proper design and function of laboratory facilities is one of the key components
involved in management of risk within a laboratory, Facility Operations and Maintenance is
encouraged to consult and coordinate with the Biosafety Office during design of new or
renovated laboratory facilities and/or during construction or maintenance operations which
may require untrained personnel to enter areas containing biohazardous materials.
1.2.6.1.2 The Institutional Animal Care and Use Committee (IACUC)

and Division of Laboratory Animal Services (LAS)
To comply with federal laws and institutional policies governing the humane care and
treatment of laboratory animals, MCG requires that all use of invertebrate and vertebrate
animals for research, education, or for any other purpose be documented in an Animal Use
Protocol (AUP) which must be reviewed and approved by the Institutional Animal Care and
Use Committee (IACUC) prior to initiation of the work. The IACUC administrative office
specifically supports the IACUC compliance mission, which works closely with the
Division of Laboratory Animal Services (LAS), to address laboratory animal care and
welfare issues
The IACUC evaluates AUPs based on several criteria, most of which focus on the ethical

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and humane treatment and care of the animals, including:
Rationale and purpose of the proposed use of animals.
Justification of the species and number of animals requested. Whenever possible,
the number of animals requested should be justified statistically.
Availability or appropriateness of the use of less-invasive procedures, other
species, isolated organ preparation, cell or tissue culture, or computer simulation.
Adequacy of training and experience of personnel in the procedures used.
Unusual housing and husbandry requirements.
Appropriate sedation, analgesia, and anesthesia.
Unnecessary duplication of experiments.
Conduct of multiple major operative procedures.
Criteria and process for timely intervention, removal of animals from a study, or
euthanasia if painful or stressful outcomes are anticipated.
Postprocedure care.
Method of euthanasia or disposition of animal.
Safety of working environment for personnel.
Although the IBC’s oversight of the health and safety of the (human) researchers, the
community and the environment (including control of infection within the animal care
areas) may overlap with the IACUC’s concerns for animal welfare and caretaker safety,
each committee reviews research protocols from different perspectives. Therefore, PIs may
need to submit both AUP and BSP applications to the IACUC and IBC, respectively, in
order to receive both committees’ approvals to maintain compliance.
To address common concerns, the IACUC and IBC (as does LAS and Biosafety Office)
work in close collaboration with each other. IBC approval may be required by IACUC
before initiation of projects involving biohazardous materials or recombinant DNA in
conjunction with live animals or prior to completion of processing IACUC approvals.
1.2.6.1.3 The Institutional Review Boards (IRBs) and the MCG Office
of Human Research Compliance (OHRP)
The purpose of the MCG-approved Institutional Review Boards (IRBs) (i.e., the Human
Assurance Committee (HAC) or Chesapeake Research Review, Inc.) is to ensure the
principles outlined in the Department of Health and Human Services (DHHS) policies, the
Belmont Report, the Nuremberg Code and the Declaration of Helsinki are maintained in all
MCG investigations involving human subjects. These principles were established to
safeguard the rights and welfare of human subjects of research investigations, and to fulfill
the moral and legal obligations and commitments of the institution. The IRB administrative
office and compliance officers are located within the MCG Office of Human Research
Compliance (OHRP).
The IRBs evaluate Human Use/Clinical Protocols (i.e., HAC or CCRI applications) based
on several criteria, most of which focus on the ethical and legal obligations of MCG
toward human research subjects including:
The rights and welfare of the human research subjects involved
The appropriateness of the methods used to obtain informed consent from research
subjects
The risks to the human subjects and the potential benefits of the investigations to the
patients and mankind.
With the exception of protocols which may involve the delivery of recombinant DNA or
other biological materials (e.g., cells, tissues, toxins of biological origin), IBC reviews
focus more on the health and safety of the researchers, the community and the environment;

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therefore, IRB and Biosafety Protocol (BSP) applications and the information required of
each differs. Therefore, PIs may need to complete two separate applications and receive
both committees’ approvals to maintain compliance with both the human use and biosafety
requirements. Although the IRB and IBC reviews are independent of one another, some of
the issues (e.g. delivery/collection of biohazardous material or recombinant DNA into/from
human research subjects) overlap; therefore, the IRB and IBC (and consequently the Office
of Human Research Protection and Biosafety Office) work in close collaboration.
IBC approval may be required by OHRP before completion of IRB approvals for projects
involving biohazardous materials or recombinant DNA in conjunction with human research
subjects.
1.2.6.1.4 Division of Sponsored Projects Administration (DSPA)

The Division of Sponsored Program Administration (DSPA), formerly the Office of Grants
and Contracts, serves as the principal interface between the Medical College of Georgia and
external agencies providing sponsored program support, including the Medical College of
Georgia Research Institute (MCGRI).
As part of each application for sponsored funding, DSPA acts on behalf of the Vice
President of Research to ensure that each research project or contract complies with
Federal, State, funding agency and institutional standards. The routing form that
accompanies each application for funding serves as a way for DSPA to document that the
Principal Investigator verifies that all compliance requirements, including those related to
Biological Materials, have been completed. Any use of biological material needs to be
fully disclosed on the routing sheet(s), and any Biosafety protocol (BSP) Authorization
number(s) listed on the routing form must refer to the Biosafety Protocol(s) (BSPs) in
which the biological materials, operations, facilities, personnel and equipment which will
be used to accomplish the experiments described in the project application, have been fully
described, reviewed and approved by the IBC. As indicated on the routing form, you “may
be subject to criminal, civil or administrative penalties should any of the information
contained on this form or in the submitted application… be false, fictitious, or include
fraudulent statements.”
Prior to establishment of an account associated with a sponsored project or contract, DSPA
contacts the relevant compliance offices (OHRP, IACUC, Biosafety, Chemical Safety,
Radiation Safety) to request verification from each office regarding the compliance status
of the specific project. If compliance cannot be verified by any one of these offices for the
project in question, DSPA will not establish the account associated with the sponsored
project.
1.2.6.1.5 Office of Institutional Audits and Compliance (OIAC) and the

Compliance Oversight Council (COC)
The Office of Institutional Audit and Compliance (OIAC), formerly the Division of Internal
Audits, is responsible for the institution’s internal audit function and compliance oversight
activities. The OIAC maintain a Compliance Hotline to which any complaints or concerns
related to any compliance issues can be reported (confidentially, if desired). Compliance
hotline information can be accessed at the following URL:
http://www.mcg.edu/audits/hotline.htm
In addition to their financial audit responsibilities, OIAC administers training and addresses
questions regarding compliance with the Health Insurance Portability and Accountability
Act of 1996 (HIPAA), Family Educational Rights and Privacy Act of 1974 (FERPA), and

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other research and business ethical standards. The OIAC is also currently serving as a
central point of contact for compliance with the Departments of Commerce, State and
Treasury Export Control Regulations, which may limit access and shipping of materials
with potential commercial, military or economic value, including some biological materials.
Further information and contacts can be accessed at their web page:
http://www.mcg.edu/audits/index.htm
The Compliance Oversight Council (COC) is made up of member’s from MCG’s executive
management and is chaired by the Director of the Office of Institutional Audits and
Compliance. Their purpose is to coordinate and monitor, as necessary, other MCG campus
compliance efforts, collect annual compliance information and assemble an institutional
compliance report, coordinate and track the MCG/MCGHI/PPG compliance hotline issues.
Information on the COC can be found at:
http://www.mcg.edu/audits/coc.htm
1.2.6.1.6 Office of Technology Transfer and Economic Development

(OTTED)
The Office of Technology Transfer and Economic Development (OTTED) (often referred
to as the Intellectual Properties Office) is designed to help faculty, staff and student
researchers with their inventions by serving as a resource on matters related to intellectual
property. By identifying, evaluating and protecting novel ideas, the OTTED serves the
inventor but also MCG and the MCG Research Institute.
Because of its potential value, sharing of Intellectual Property among collaborators must be
documented in writing using a Material Transfer Agreement (MTA). The OTTED will
assist researchers in setting up MTAs to document shipment or receipt of Intellectual
properties. MTAs are not typically viewed by the Biosafety Office or the IBC.
Receipt of new biological materials on campus must be documented on the Principal
Investigator’s Biosafety Protocol (BSP) and approved by the Institutional Biosafety
Committee (IBC) prior to receipt on campus to ensure appropriate containment is available
for the risks that the agent(s) may pose. Therefore, receipt and documentation of any new
biological material on an MTA does not, by any means, substitute for documentation and
approval by the IBC for receipt of new biological materials.

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2. BIOSAFETY REQUIREMENTS
The following information describes the requirements for MCG researchers as defined by the MCG
Institutional Biosafety Committee (IBC) and the MCG Biosafety Office. It is the responsibility of each
Principal Investigator, Clinical Director and/or Instructional Course Director to ensure their workplaces are
in compliance.
2.1 REGISTRATION FOR THE USE OF BIOLOGICAL MATERIALS

All Principal Investigators working with biological materials are required to complete and submit a
Biosafety Protocol (BSP) Application prior to bringing new biological materials to campus or initiation of
the research independent of its funding status. Incoming Principal Investigators establishing a new
laboratory at MCG must submit the application prior to bringing biological materials to campus (see
Section 2.3.1, Incoming New Faculty to MCG). The BSP must be kept current to accurately reflect the
biological materials and their manipulation, personnel handling the material and the locations in which the
material may be handled or stored (see Section 2.1.2, Amendments to Biosafety Protocols).
A copy of the Biosafety Protocol Application form can be found in Appendix A and forms can be accessed
online at: http://www.mcg.edu/research/ibc/apps.htm. The risk assessment and mitigation methods associated
with the PI’s research project(s) must be performed and documented in this application (See Section 3, Risk
Assessment and Section 4, Risk Management for further information) to enable the Institutional Biosafety
Committee (IBC) to evaluate and perform a mandated independent comprehensive risk assessment. In
particular, BSPs require documentation of the following materials and operations:
Non-exempt recombinant DNA (as defined by the NIH Guidelines for Research with Recombinant
DNA), mammalian cells, tissues, organs or fluids requiring ≥BSL-2 containment
Risk Group ≥2 microbial agents (as described in the Centers of Disease Control publication
Biosafety in Microbiological and Biomedical Laboratories and the American Biosafety
Association)
Large scale (≥10 liters) cultures of biological materials
Biological materials delivered into animal or human subjects
Toxins of biological origin with LD50 ≤100μg/kg body weight
Shipping of biological materials, toxins of biological origin, materials on dry ice or liquid
nitrogen, or genetically modified organisms
The BSP application is a modular form and consists of several parts:
1. The Primary Application Form for Research Involving Biological Materials and/or
Recombinant DNA. This form is required for all applications and is intended to describe:
• the overall project aims
• the general nature of biological materials/recombinant DNA and manipulations of these
• the locations in which the biological materials/recombinant DNA may be located at any
moment in time
• personnel working on the project, and their experience levels (note: completion of all
current required training modules by all personnel and PIs on the BSP is required by the
IBC prior to release of IBC approvals)
2. Biosafety Schedule(s), as required to document and address the risks associated with each BSP’s
agents and research applications. The Biohazardous Material Compliance Worksheet on p. iii of
the Primary Application Form (see Appendix A) is intended as a guide to help assess which, if

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any, additional supplemental schedules may be required for completion of the BSP. These
schedules are as follows:
A: Non-exempt recombinant DNA

B: Handling and/or culture of mammalian cells, tissues, and/or organs requiring ≥ BSL-2
containment (including human or primate blood, tissues or fluids; cell, tissue or organ
cultures of human or non-human primate origin, or animal cells, tissues or organs which
have been potentially infected with Risk Group ≥2 agents
C: Research involving large-scale (≥10 liter) cultures
D: Research involving potentially infectious microbial agents (Risk Group ≥2) or specimens
exposed to these infectious microbial agents (excluding any material previously described
in either Schedule A or B)
E: The use of biological materials in association with live animals
F: Human gene transfer/therapy
G: HHS/USDA regulated Select Agents and Toxins
O: Use of biological agents and/or recombinant DNA in applications for sponsored funding
or IRB approval
3. A copy of the laboratory’s Standard Operating Procedures (SOPs). The risks associated with
the experiments involving the agents, operations, locations, personnel and environment described
in the forms above should be addressed by the mitigating measures in these SOPs. (Laboratory
SOPs should also be provided to incoming laboratory staff members and reviewed with the PI as a
training measure within the laboratory).
4. A copy of the laboratory’s Self-Audit form.
Once completed the above forms should be submitted to the Biosafety Office. Hard-copies can be
submitted via campus mail to: Biosafety Office, EHS, CI-1001 or electronic copies can be sent to
BIOSAFETY@mcg.edu.
Please note that all Biosafety Protocol applications and amendments are reviewed for compliance with the
annual training requirements as described in Section 2.2, Training Requirements, below, to comply with
specifications with the Georgia Board of Reagents (BOR), NIH Guidelines for Research Involving
Recombinant DNA Molecules (NIH Guidelines), the Centers of Disease Control and the Georgia State
Department of Labor. The Biosafety Office will also assist in updating information during periodic the
biological lab inspections.
2.1.1 Annual Affirmation of Biosafety Protocol Information

The Division of Environmental Health and Safety must maintain accurate information regarding the use of
biological materials (e.g., microorganisms, cell lines, human materials, animals, and toxins) by MCG
personnel. Approvals for BSPs typically issued by the IBC are for an indefinite period of time, as long as
the research, personnel, practices and locations have not changed. Therefore, the IBC requires all
Principal Investigators, Clinical Directors and/or Instructional Course Directors to annually review their
BSPs to ensure that the information provided continues to accurately reflect the current situation in their
laboratories, and submit a written affirmation to the IBC that their BSP(s) are up-to-date and accurate.
Protocol Renewal forms can be found at: http://www.mcg.edu/research/ibc/apps.htm and can be submitted via
campus mail to the Biosafety Office, Division of Environmental Health and Safety, CI-1001.

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2.1.2 Amendments to Biosafety Protocols
Amendments must be submitted to the Biosafety Office by the Principal Investigator, Clinical Director
and/or Instructional Course Director and approved prior to implementing any change(s) to IBC-approved
Biosafety Protocol. Documentation should be provided for changes in any of the following:
Addition or deletion of biological materials and/or non-exempt recombinant DNA
Changing the design, operations, scope or location of existing work, including alterations in:
• Any non-exempt recombinant DNA, including new vectors, inserts or operations
• Administration or exposures of new target cells, organs, organisms and/or animals
• Employing new techniques which may increase risk of exposure of personnel (see Risk
Assessment and Risk Management Sections 3 & 4 of this Biosafety Guide)
• Creation of a new transgenic strain of animal
• Embarking upon large-scale culture (≥10 liters)
• Changes in locations
• Changes in major equipment used (e.g., Biosafety cabinets, centrifuges)
Addition or deletion of any personnel to your list of authorized users who may be handling your
biological materials
Providing infectious agents to another investigator on or off campus
Arranging for visiting researchers and/or volunteers to work in your laboratory
Amendments must comprehensively describe the new proposed changes, and document any new risk
assessment(s) and proposed risk management methods, as appropriate. Amendments can be submitted by
altering the original Biosafety Protocol Application forms to include the new information and re-submitting
them to the Biosafety Office, or via a fully detailed email to the Biosafety Office. Any amendment which
involves an alteration in risk relative to the previously-approved BSP will be submitted to the IBC for
review and approval; simple amendments involving no alteration in risk (i.e., personnel changes, additions
of grant/contract/clinical protocol titles to previously-approved BSPs) will be handled administratively by
the Biosafety Office. All questions can be addressed to the Biosafety Office (x1-2663 or
BIOSAFETY@mcg.edu).
2.1.2.1 Adding New Personnel to or Removal of Personnel from a Biosafety

Protocol
Before a new employee, student, volunteer, or visitor may handle the biological material
documented on a Biosafety Protocol, their name must be added to the list of authorized personnel
on the BSP. PIs should first submit the name(s) of new personnel to the Biosafety Office
(BIOSAFETY@mcg.edu). In doing so, the PI takes responsibility for ensuring that all training and
laboratory entrance requirements listed in the laboratory SOPs, BSP document and Institutional
Biosafety Committee Authorization document have been completed by this new person before
authorizing them to work in the laboratory.
The Biosafety Office will verify that the new person has completed the IBC-required training
modules (as described in Section 2.2, Training, in addition to any special training requirements as
required by the IBC for the BSP and documented in the PI’s Institutional Biosafety Committee
Authorization document) before administratively adding their names to the list of authorized
personnel of the Biosafety Protocol.
As with all personnel listed in the BSP, the PI, Clinical Director and/or Instructional Course
Director is responsible for communicating the risks associated with the biological material and
procedures in the laboratory, as well as the laboratory-specific mitigation methods. Therefore, the
PI should review the laboratory standard operating procedures (SOPs), biosafety protocol(s) and
MCG Biosafety Guide with any new personnel, including emergency response procedures and
entrance requirements before allowing them to start work within the laboratory. The PI is also
responsible for ensuring proper supervision and training is provided to any new personnel until
proficiency in handling the materials and mastery of the techniques can be demonstrated.

2-3
New personnel should also be offered and provided employee health counseling and/or
vaccinations as documented in the Biosafety Protocol prior to initiation of work within the
laboratory.
Written notification should be provided to the Biosafety Office by the PI, Clinical Director or
Instructional Course Director if someone should be removed from their list of authorized
personnel upon their departure from the laboratory.
2.1.2.2 Amendments for New or Modified Biological Agents or Operations

to a Biosafety Protocol
Any changes in the Biological Agents to be utilized in the laboratory that are not covered in the
previously-approved BSP, must be documented to enable the comprehensive risk assessment by
the IBC and evaluation whether the documented mitigation methods are appropriate for the new
materials or operations.
In particular, any changes in non-exempt recombinant DNA materials or applications must be

documented and submitted to the full IBC for review to maintain strict compliance with NIH
Guidelines for Research with Recombinant DNA Molecules. This includes changes in inserts,
vectors or target cells/tissues/organisms or operations. This is a condition of NIH funding to the
entire institution and applies to all research at the institution, independent of its funding source.
Any change which might alter the risk of a BSP must also be documented. For instance, if the
infectious potential status of biological material handled differs from that previously documented
and approved in a BSP, an amendment must be submitted. For instance, if a previous BSP
included approval for handling blood samples obtained from non-infectious patients and the PI
wishes to initiate a study involving taking blood specimens from infectious patients (e.g., HIV+
patients), this change must be submitted and approved by the IBC prior to initiation.
Similarly, if new operation will be employed with the biological agent(s) which may involve
additional risk, this must also be documented. For instance, if a previous BSP included approval
for handling the biological agents in vitro, and the PI wishes to initiate a study involving in vivo
administration of the biological agent into animals or humans, this change must be submitted and
approved by the IBC prior to initiation. Another example of operational changes that may alter
the risk of a BSP includes utilizing new procedures with the biological which may involve
additional splash, spray or aerosol exposure risks (e.g., sonication, homogenization, FACS sorting,
etc.). See Section 3, Risk Assessment for further details.
BSP Amendments involving changes in agents or operations are reviewed and approved by the
IBC via the same procedures as the original BSP (see Section 2.5.1 for further details).
2.1.2.3 Modifying the Locations or Equipment on a Biosafety Protocol

If a PI’s biological agents will be handled or stored in locations other than those documented on
their IBC-approved BSPs, the new location(s) must be documented and approved by the IBC in
order to ensure that the containment is appropriate to handle the risks presented by the agents. A
laboratory assessment by the Biosafety Office will likely be required prior to approval of the
amendment.
Some common locations (such as core laboratories or common storage rooms) may have
previously been approved by the IBC for other BSPs involving similar agents/operations;

2-4
amendments to add these locations to a BSP are often approved administratively through the
Biosafety Office. However, any area proposed for handling or storage of biological materials
which has not been previously reviewed and approved by the IBC will need to be authorized by
the IBC before its use to ensure appropriate containment.
Because major equipment used to contain or handle biological agents, such as Biosafety Cabinets
or centrifuges, may not only impact the risk of the protocol, but may also require certification
documentation prior to use, such amendments must also be submitted to the Biosafety Office.
2.1.2.4 Documenting New or Additional Project Titles Associated with a

Biosafety Protocol (or Multiple Biosafety Protocols)
Biosafety Protocols (BSPs) are intended to document the risks and mitigations associated with
working with biological agents, including which agents, operations, personnel, locations and
major equipment to be utilized, independent of the funding status of the project. If a PI, Clinical
Director or Instructional Course Director wishes to initiate a new project which involves identical
agents, operations, personnel, locations, etc. as previously described on an IBC-approved BSP, the
IBC considers the new project “covered” by the terms of the previously-approved BSP.
This is often the case with many clinical research laboratories. If the study agents administered to
the patients are not biological in nature (i.e., they do not involve administration of rDNA, cells,
biological fluids, or toxins of biological origin), the infectious status of the patients are identical,
the operations involved are the same (e.g., serum preparation, aliquotting and shipping specimens
to sponsors), and the locations and personnel handling the materials are identical for multiple
clinical protocols, a single BSP may “cover” these multiple clinical protocols.
However, information may be required to “link” each of these new projects to an existing BSP to
enable the Biosafety Office to provide IBC verification information to DSPA, OHRP or IACUC,
as required (see Section 2.5.2 for more information about requirements for documentation of IBC
compliance to DSPA, OHRP or IACUC). To accomplish this “linkage”, the PI listed on the BSP
must provide verification that the information provided in his/her existing BSP fully “covers” the
new project. Also, by submitting this verification, the PI on the BSP signifies that he/she takes
responsibility in ensuring that the terms of the IBC Agreement document will be followed in the
accomplishment of the new project or protocol.
Verification can be accomplished via completion and submission of a Schedule O form (via email
or submission of a hard-copy, signed by the PI) or by providing the following information via
email to the Biosafety Office (BIOSAFETY@mcg.edu):
a. The PI name on the project/protocol if other than the PI name on the BSP (this
may be the case for student or post-doctoral fellowships accomplished under the
supervision of a laboratory director)
b. The BSP # which covers the new project/protocol
c. The BSP title (if available)
d. The BSP approval date
e. The project/protocol title
f. The project/protocol sponsor or funding agency
g. The anticipated funding period (until the next competitive renewal application)
h. The HAC or CCRI # (if verification to OHRP is required for IRB approval
processing)
The Biosafety Office also recognizes that some projects or protocols may be collaborative in
nature, and may involve handling of biological material under the oversight of two or more
different collaborating laboratory directors. As long as the BSPs dovetail to seamlessly document

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the agents, their handling and operations and transport between the laboratories, multiple BSPs
may be required to “cover” a single project or protocol. Again, the PIs on each of the BSPs
required to “cover” the project/protocol must provide documentation of verification of “coverage”.
Random audits will be performed to compare the experiments and agents described in the grants,
contracts, or protocol descriptions verified by the PIs with the contents of their IBC-approved
BSPs to ensure compliance. During these audits, the PIs will be requested to provide the
requested grant, contract or protocol information to the Biosafety Office for review.
2.2 TRAINING REQUIREMENTS

Successful completion of a range of biosafety training programs may be required prior to the initiation of
your work at MCG.
Please review the following table for information on the IBC-required training modules. The Biosafety
Office must be able to document completion of the required training modules listed in this table prior to
completion of any IBC-approval paperwork:
Intended for all You must satisfactorily Training Options

personnel who work complete the following
on MCG projects or training:
areas, who:
Work at MCG or on Basic Awareness Right-to- http://www.mcg.edu/services/ehs/chemsafe/RTKTraining.htm
MCG projects Know Training
Required once upon initial

hire
GA Board of Regents Requirements
Work with chemicals Chemical Specific Right-to- http://www.usg.edu/ehs/training/chemical/

Know Training
Required once prior to

initiation of work and
annually thereafter
Work with or may Hazardous Waste Awareness http://www.usg.edu/ehs/training/hazwaste/
produce hazardous waste Training
(including chemicals,
and blood and other Required once prior to
biological materials)
annually thereafter
Work with human or Bloodborne pathogens http://www.usg.edu/ehs/training/pathogens/
primate blood, tissues, training
fluids other potentially
infectious materials and Required once prior to
bloodborne pathogens
annually thereafter

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Are listed on the Biosafety for Basic Didactic initial training offered monthly. Contact EHS for
MCG IBC Training Requirements for all personnel listed on BSPs personnel lists of BSPs Research* next scheduled date/time.
and work with biological
materials in a basic An Initial Training session is Annual refresher training sessions may be offered in a
laboratory setting. didactic training format per request of the department
Includes Recombinant
required prior to initiation of (typically in conjunction with all EHS sections)
DNA, microbial agents work. OR
≤ RG2, cell culture & Online refresher training module may be accessed at:
biological materials with Annual Refresher required http://www.mcg.edu/services/ehs/biosafe/biosafe/training.htm
animals thereafter.
Are listed on BSPs that Biosafety for Clinical Didactic initial training offered monthly. Contact EHS for
pertain to simple clinical Research* next scheduled date/time.
protocols only. Involves
taking human blood, Annual refresher training sessions may be offered in a
tissues or fluid didactic training format per request of the department
specimens from patients
An Initial Training session is (typically in conjunction with all EHS sections)
who are not considered required prior to initiation of OR
infectious with any work. Online refresher training module may be accessed at:
agent ≥RG3, preparation http://www.mcg.edu/services/ehs/biosafe/biosafe/training.htm
of serum or plasma Annual Refresher required
samples, aliquotting of thereafter.
specimens and shipping
to off-site sponsors.
Does NOT cover the

delivery of any
biological materials
(recombinant DNA,
biotoxins, cells/tissues)
into patients or on-site
analysis or culture of
specimens.
Are responsible for Shipping of Hazardous Currently, a CD Training module developed by Saf-T-Pak
packaging, shipping, Materials can be obtained on loan from the Biosafety Office. Because
transporting, or of limited numbers available, please contact the office (x1-
Requirements
receiving Biohazards 2663 or BIOSAFETY@mcg.edu) to determine if one is

IATA/DOT
(infectious agents,
Required biennially (i.e. once available or to be placed on a waiting list.
biological toxins, human every 2 years)
clinical specimens,
animal diagnostic
specimens, dry ice,
liquid nitrogen,
genetically modified
organisms)
Work with agents or IBC-required need-specific Any special training requirements will be listed as a Special
Training
applications which, Condition of the IBC Authorization document. Assistance in

Special
training
based on the risk, the locating or obtaining the training can be obtained from the
IBC assesses a need for Biosafety Office (x1-2663 or BIOSAFETY@mcg.edu).
a specific training.
*Personnel listed on BSPs are required to take only one of these basic Biosafety classes, not both. The “Biosafety for
Clinical Research” is a slightly abbreviated, more clinically-oriented version of the more comprehensive “Biosafety for
Basic Research” class. The “Biosafety for Clinical Research” class is intended as an option only for those whose work is
not anticipated to ever involve recombinant DNA, culture of biological materials or higher risk agents or operations.

2-7
In addition to the above training modules, the IBC expects that the following training will be provided to
any person who is listed on a Biosafety Protocol prior to initiation of work within the laboratory.
Documentation of completion of the following training should be maintained by the Principal Investigator,
Clinical Director and/or Instructional Course Director:
May be in contact with Infection Control Training Currently, this training should be provided by the Clinical or
Standards
Infection
patients or clinical Laboratory Director and involve review of the specific

Control
Bloodborne Pathogens and

material in an MCG Tuberculosis Training Exposure Control Plan for the particular clinical area.
clinical setting
May be in contact with Laboratory-specific Training The Principal Investigator, Clinical director or Instructional
Training
Biological materials (includes review of SOPs, Course Director is responsible for educating all personnel in
specific
Lab-
BSPs, and hands-on training) the laboratory related to the laboratory-specific risk issues,
and mitigation methods. The PI or director is also
responsible for providing supervision training for all
personnel in the laboratory-specific operations.
2.3 LABORATORY ESTABLISHMENT, CLOSE-OUTS, AND MOVES

To ensure appropriate containment is maintained, safety measures are implemented and compliance with
all Federal, State, local regulations and guidelines for all biological materials at the Medical College of
Georgia have been met, documentation must be provided, reviewed and approved by the IBC prior to
moving any biological agents into any new facilities (which includes any materials which may be brought
to campus by incoming new faculty or materials which may be moved to new locations not previously
authorized as part of the Principal Investigator’s, Clinical Director’s or Instructional Course Director’s
IBC-approved Biosafety Protocol). In addition, to ensure that the biological materials are properly
removed and facilities are decontaminated prior to closing of a laboratory, PIs, Clinical Directors or
Instructional Course Directors must document compliance with appropriate close-out procedures listed
below.
2.3.1 Incoming New Faculty to MCG

New MCG researchers must receive authorization from the IBC prior to transfer of any biological material
to campus prior to shipment of this material. Initial applications to transfer biological agents to MCG
campus must, at minimum, document:
The biological agents to be transferred to MCG

The method of transfer (which must be in compliance with IATA/DOT standards, and any
required USDA or CDC permits for infectious materials must be obtained by the PI prior to
shipment. See Section 10, Shipping for further information)
The location(s) which these materials will be stored prior to the new faculty member’s laboratory
establishment
The MCG personnel responsible for the materials prior to arrival of the new faculty member (if
applicable).
The IBC will review the above to ensure containment issues have been addressed and compliance with
regulations, guidelines and policies are met, and will typically issue authorization only for the transfer of
the material. Prior to initiation of research in MCG facilities, the incoming faculty member is expected to:
Complete and submit a complete Biosafety Protocol Application to the Biosafety Office for IBC
review and approval (including documentation of all of the locations in which their biological
materials may be handled or stored and submission of the laboratory-specific Standard Operating
Procedures)

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Ensure all laboratory personnel listed on the BSP have completed the IBC training requirements
(see Section 2.2, Training for further details)
Have all laboratory facilities assessed by the Biosafety Office. Once the facilities have been
equipped for research, the PI should contact the Biosafety Office to make an appointment for a full
laboratory assessment (x1-2663 or BIOSAFETY@mcg.edu).
Incoming faculty members are encouraged to contact the Biosafety Office (706-721-2663 or
BIOSAFETY@mcg.edu) for assistance with these BSP submission processes as early as possible. The
Biosafety Office is more than happy to help “walk” new faculty members through the process and assist
them in obtaining any permits or authorizations required to transfer materials to campus.
New faculty members are advised that review and approval of BSPs involving non-exempt recombinant
DNA or any materials which may pose a higher risks than those previously approved at MCG may take
several (3-7) weeks due to requirements for full IBC review (the IBC meets once per month, and the
deadline for submission for Biosafety Protocols for review within that month is the first day of the month).
Because of these time constraints, the IBC recommends that Department chairs, Institute Directors, and/or
Departmental Managers notify the Biosafety Office of any incoming faculty members in their programs as
soon as possible after recruitment to initiate these application processes to enable the faculty member to
begin research as promptly as possible after arrival at MCG.
2.3.2 Laboratory Close-outs

Any faculty member who may be vacating a laboratory needs to ensure that all biological materials have
been removed from the laboratory and the laboratory has been decontaminated prior to departure. This is
one of the terms of the IBC Authorization agreement between the IBC and the PI.
Biological materials should be removed from the laboratory by one of the following methods:
Decontamination/deactivation and disposal of the biological material (as per the PI’s SOPs)
Disposal in the authorized Biohazard waste containers (as per the PI’s SOPs)
Transfer of the biological materials to another authorized user (or to another authorized location).
• If the material is to be transferred to another MCG laboratory, written documentation of
this transfer should be provided to the Biosafety Office (an email to
BIOSAFETY@mcg.edu is sufficient). If the materials are being transferred to a different
PI, the receiving PI should have authorization to possess this material in the new
locations prior to transfer. The receiving PI must also agree to take responsibility for the
new materials.
• If the material is to be transferred outside of MCG, this must be done in accordance with
IATA/DOT standards by personnel with documented training (since many biological
materials and dry ice are considered hazardous materials by the federal government).
Any transport, import and/or export permits must also be obtained by the PI prior to
transfer for shipping (see Section 10 for more details). Contact the Biosafety Office for
assistance (x1-2663 or BIOSAFETY@mcg.edu).
After removal of the biological materials from the laboratory, all surfaces and equipment must be
decontaminated using the appropriate disinfection procedures as documented in the laboratory standard
operating procedures prior to departure from the laboratory. Some equipment, such as biosafety cabinets,
may require additional decontamination methods, such as gaseous fumigation with paraformaldehyde or
vapor hydrogen peroxide (VHP), prior to removal from the laboratory. Contact the Laboratory Equipment
Services (LES) office (x1-6124) to arrange for decontamination of biosafety cabinets prior to moving.
After removal of all biological materials and decontamination of the laboratory and equipment, the
Biosafety Office must be notified to complete the “clearance” process.

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2.3.3 Laboratory Moves
If biological materials or equipment are to be transferred from one MCG laboratory to a new MCG
laboratory, the PI, Clinical Director and/or Instructional Course Director must:
Submit a BSP amendment request to the Biosafety office for IBC authorization of the new
locations for the biological materials prior to moving the materials (see Section 2.1.2,
Amendments to Biosafety Protocols for more details)
Transport the biological materials in a method which complies with their laboratory standard
operating procedures. Contact the Biosafety Office (x1-2663 or BIOSAFETY@mcg.edu) for
assistance and see below for special notes about transport considerations.
Arrange for a full laboratory assessment with the Biosafety Office (x1-2663 or
BIOSAFETY@mcg.edu) to ensure appropriate containment measures are in place in the new
laboratory prior to initiation of work in the new laboratory (this is usually a stipulation in the IBC
approval of any new location)
Complete the laboratory close-out procedures (as described above in 2.3.2, Laboratory Close-outs)
for the former laboratory
Standard Operating Procedures for biological materials typically require the biological material to be
transported outside of the laboratory:
By authorized personnel (i.e., those personnel listed on the BSP who are familiar with the risks
associated with the biological material, the emergency spill and exposure/release procedures)
Contained in a sealed, leakproof primary container inside a well-labeled sealed, leakproof, durable
secondary container.
However, transport of large amounts of biological materials in laboratory moves poses unique challenges.
First, laboratory moves are often accomplished with the help of non-authorized personnel (e.g.,
professional movers or personnel from the MCG’s Materials Management Office). PIs are reminded that
these personnel are not authorized to handle the biological materials because they have not been fully
educated in the risks, received appropriate medical evaluation/vaccinations and are not prepared for any
emergency procedures should spills or releases occur en route. Therefore, whenever possible, equipment
containing biological materials should be transported separately by authorized personnel. However, if this
is not feasible (due to the size/weight constraints of the equipment), the equipment containing the
biological materials should be securely sealed to prevent moving personnel from exposure (e.g.,
refrigerators and freezers should be locked and/or securely taped shut), the exterior of the equipment should
be decontaminated prior to the movers handling them and during the move, the movers and equipment
should be accompanied by at least one member of the authorized laboratory staff equipped with the
appropriate spill clean-up materials to ensure proper emergency procedures are followed should a
spill/release occur en route.
Secondly, biological materials are often transported in refrigerators, freezers and cryotanks during
laboratory moves to maintain the integrity of the samples. In preparation of moving, any glass, breakable
items or other hazardous materials (e.g., chemicals or radiological materials) should be removed from any
refrigerators or freezers. Biological materials should be contained in one or more forms for sealed, durable
containment within the refrigerator or freezer and secured to prevent spillage or scattering of biological
materials or samples within the freezer. Keep in mind: freezers and refrigerators require tipping to
transport on moving dollies; a good deal of tipping is often required to get large equipment into tight
spaces, such as elevators. For instance, sealed microcentrifuge tubes should be contained within boxes
with lids (not in open racks); plastic tubes should have tightly secured lids and should be in fitted
containers which would prevent movement during transport (or contained within ziplock bags). Filling any
open spaces within the freezers/refrigerators with packing materials will also prevent shifting of materials
during transport (foam rubber works well for this purpose). The refrigerator or freezer should be sealed
shut (locked or securely taped shut) and the exterior decontaminated prior to moving.
Because cryotanks cannot be completely sealed during transport to prevent leakage of liquid nitrogen if
tipped, and any vehicle transporting these materials on public roads may require special placarding for
DOT regulatory compliance, these items should not be transported via moving truck. Alternate methods of

2-10
moving this equipment should be discussed with the Biosafety Office prior to moving.
The need for gaseous decontamination of a Biosafety cabinet (BSC) must be evaluated by the Biosafety
office prior to moving. Should a BSC require decontamination, arrangements must be made in advance
with Laboratory Equipment Services (LES) (x1-6124) to perform this service before the BSC can be
moving the BSC.
2.4 LABORATORY RECORD-KEEPING

At least one copy of the Laboratory Biosafety Manual should be maintained within each laboratory for
training and reference of all laboratory personnel. This manual should include the following material:
• A copy of the MCG Biosafety Guide
• The Laboratory Biosafety Protocol(s), amendments and renewal documentation
• The Laboratory-specific Standard Operating Procedures (SOPs)
• Copies of documentation of training of all laboratory staff. This should include:
o Copies of certificates of completion of any BOR or IBC-required training modules
o Documentation of completion of IATA/DOT shipping training,
o Documentation that all authorized personnel listed on the BSP has received laboratory-
specific education and training from the PI or his/her designated representative.
Signed/dated statements should affirm that training included:
Laboratory-specific risk communication and training. This includes providing
references and a discussion of the risks associated with the materials within the
laboratory, any possible signs/symptoms which may suggest occupational
exposure has occurred and any incident/emergency procedures that may be
required.
Each staff member should verify that they have reviewed and understand the
materials documented within the Biosafety Guide, Laboratory Biosafety
Protocols and the Laboratory SOPs.
• Copies of signed documentation that each person authorized to work within the laboratory has
been offered Employee Health screening/counseling and vaccinations by the PI as appropriate for
the agents within the laboratory and as described in the laboratory SOPs and IBC Approval
documents.
• Records of any incident, exposure, possible exposure, spill, release from primary containment or
environmental exposure of any ≥Risk Group 2 biological agent (see Section 3, Risk Assessment
for further information on Risk Groups). This should include the date of the incident, the agents,
locations and personnel involved, a narrative description of the circumstances and outcome, the
dates when the Biosafety Office was informed and any follow-up measures taken (e.g., was health
care sought for those potentially exposed? are subsequent tests/health care follow-up measures
required and performed and when? were any SOPs altered to prevent similar incidents in the
future?)
2.5 PAPERWORK FLOW

There are two critical routine administrative processes that occur within the Biosafety Office:
1. Receipt and administration of new Biosafety Protocols (BSPs) or amendments, presentation of
these to the IBC for review and processing of IBC approvals.
2. Verification that IBC approval has been received for a particular project, clinical protocol or AUP
by a Principal Investigator to other campus compliance offices (DSPA, OHRP or IACUC).

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2.5.1 IBC Review/Approval Process for Biosafety Protocols and Amendments
Biosafety Protocols and amendments are first reviewed by the Biosafety Office to determine the most
appropriate review/approval track for the protocol. See Figure 2.5.1 for a diagram of the paperwork flow
for Biosafety protocol reviews, which describe the three review tracks available for BSPs:
1. All BSPs or amendments involving non-exempt recombinant DNA agents, or those involving
higher risk organisms (some RG2 and all RG3 organisms, and/or Select Agents) are required to be
reviewed by the full IBC during a convened meeting in which minutes are recorded as per NIH
and CDC Guidelines.
o The IBC meets monthly; typically on the third Wednesday of the month
o All BSPs and amendments received on or before the first day of the month will be
submitted to the IBC for review within that month.
o Delays on the order of approximately 3-7 weeks from the date of submission of a
complete BSP application and training documentation until IBC review approval
should therefore be anticipated
2. BSPs or amendments which do not involve recombinant DNA or involve only exempt
recombinant DNA are eligible for an expedited review and conditional approval by an IBC
subcommittee. Responses are requested from subcommittee members within 3 days of receipt of
the completed application or amendment.
3. Simple amendments of previously-IBC approved Biosafety protocols (BSPs) which do not involve
additional risk (e.g. an addition of a grant title/clinical protocol title to the protocol; most
personnel changes) are handled administratively within the Biosafety Office.
During the IBC meeting, the committee may vote to: approve the BSP/amendment without stipulations,
approve the BSP/amendment with stipulations, disapprove or table the proposed BSP/ amendment. The PI
will be notified of the IBC decision subsequent to the meeting.
If the BSP/amendment is approved with stipulations, the PI will be notified and will be given 2 months to
address these stipulations. The BSP/amendment is not considered finally approved until all stipulations
have been met. If the stipulations are not met within the 2 month time window after receipt of the
stipulations, the BSP/amendment application will be considered withdrawn and must be re-submitted for
further IBC reconsideration.

2-12
START
BSP submitted by PI to Biosafety Office
(DEADLINE: 1st day of month)
Involves YES
non-exempt rDNA?
NO
Involves higher risk RG ≥ 2

YES
agents and/or operations
and/or Select Agents/Toxins
NO
Is this a simple amendment to

YES previously approved BSP w/ no
add’l risk? (e.g. addition of grant
title, personnel changes)
NO
IBC Subcommittee
Review Recommended for Full Committee Review
(~3 working days)
Conditional Approval
(Note: this review will be followed by pro forma review by
the full IBC at next committee meeting for final approval)
Review During
Are there stipulations IBC Full
NO Approval
that PI must meet Committee
prior to approval?
Meeting
(3rd Wed. of month)
YES
Disapproval
Have the stipulations and training NO BSP Returned to PI

requirements of the IBC been met
within a 2 month period?
with no action
YES
END
1. PI can initiate research
2. BSO can verify IBC approval to DSPA,

OHRP, IACUC, etc. upon request
3. BSP Agreement Contract signed by BSO

and Chair of IBC
PI returns signed BSP Agreement Contract to the Biosafety Office
END
Figure 2.5.1. Paperwork Flow for Biosafety Protocol (BSP) Review by the Institutional Biosafety Committee (IBC)
and the Biosafety Office.
2.5.2 Verification that the IBC Has Reviewed and Approved Experiments, Agents
and Uses that Were Proposed in a Particular Sponsored Project, Clinical
Protocol or Animal Use Protocol to Other Compliance Offices
Part II of the NIH Grants Policy Statement, describes the terms and conditions of NIH Grant Awards
http://grants.nih.gov/grants/policy/nihgps_2003/NIHGPS_Part4.htm#_Toc54600062 . Among these are specific
requirements for institutions receiving NIH funds to meet Federal, State and local health and safety
standards and guidelines for all research at their institution. Grantee institutions are responsible for
meeting Federal, State, and local health and safety standards and for establishing and implementing
necessary measures to minimize their employees’ risk of injury or illness in activities related to NIH grants.
Among these are the NIH Guidelines for Research Involving Recombinant DNA Molecules (NIH
Guidelines) (April 2002 or latest revision), which apply to all research projects that involve recombinant
DNA and are conducted at or sponsored by an organization that receives NIH support for recombinant
DNA research. The IBC is required to review each proposed project for recombinant DNA experiments
and certify that the procedures, project, personnel, and facilities are adequate and in compliance with the
NIH Guidelines.
In addition to applicable Federal, State, and local laws and regulations, the following regulations must be
followed when developing and implementing health and safety operating procedures and practices for both
personnel and facilities:
29 CFR 1910.1030, Bloodborne pathogens; 29 CFR 1910.1450, Occupational exposure to hazardous

chemicals in laboratories; and other applicable occupational health and safety standards issued by the
Occupational Health and Safety Administration (OSHA) and included in 29 CFR Part 1910. These
regulations are available at http://www.osha.gov/comp-links.html.
The NIH recommends following these guidelines for use in developing and implementing health and
safety operating procedures and practices for both personnel and facilities. Therefore, these have been
adopted as part of MCG IBC Policies.:
• Biosafety in Microbiological and Biomedical Laboratories, CDC and NIH, HHS. This
publication is available at http://www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm
• Prudent Practices for Safety in Laboratories (1995), National Research Council, National
Academy Press, 500 Fifth Street, NW, Lockbox 285, Washington, DC 20055 (ISBN 0-309-
05229-7). This publication can be obtained by telephoning 800-624-8373. It also is available at
http://www.nap.edu/catalog/4911.html .
Although grantee organizations are not typically required to submit documented assurance of their
compliance with or implementation of the health and safety regulations and guidelines, if requested by the
awarding office, the grantee institutions should be able to provide evidence that applicable Federal, State,
and local health and safety standards have been considered and have been put into practice. NIH expects
the grantee institutions to provide safe working conditions for their employees and foster work
environments conducive to high-quality research.
Therefore to maintain these standards, and to ensure compliance with all Federal, State and Local laws, and
maintain appropriate documentation of compliance MCG compliance offices have a verification process for
all research involving hazardous materials, including biological hazards. To maintain compliance the
Biosafety Office is asked to verify that the specific agents, experiments, operations, locations, personnel
and equipment used in each sponsored project proposal, clinical protocol submission for IRB review,
and/or animal use protocol, have been reviewed and approved by the IBC prior to account establishment,
IRB or IACUC approval processing, respectively. See Figure 2.5.2 for guidance on when IBC approvals
need to be completed relative to verification to DSPA, OHRP or IACUC.
In order to facilitate communication between these offices, the Biosafety Office asks that you provide

2-14
verification and information required to “link” a particular biosafety protocol to a particular sponsored
project or clinical protocol. As previously described in Section 2.1.2.4, Documenting New or Additional
Project Titles Associated with a Biosafety Protocol (or Multiple Biosafety Protocols), this type of
amendment is handled administratively by the Biosafety Office and can be accomplished by submitting a
Biosafety Protocol Schedule O form http://www.mcg.edu/research/ibc/pdf/New/ScheduleO-DSPA.pdf or
providing written documentation (e-mail to BIOSAFETY@mcg.edu is fine) for the Biosafety Office
records from the PI on the BSP in which he/she verifies that the project/protocol information should be
“linked” to the Biosafety Protocol(s) (BSP) indicated:
i. The PI name on the project/protocol if other than the PI name on the BSP (this may be
the case for student or post-doctoral fellowships accomplished under the supervision of a
laboratory director)
j. The BSP # (or #s) which covers the new project/protocol
k. The BSP title(s) (if available)
l. The BSP approval date
m. The project/protocol title
n. The project/protocol sponsor or funding agency
o. The anticipated funding period (until the next competitive renewal application)
p. The HAC or CCRI # (if verification to OHRP is required for IRB approval processing)
Please see Section 2.1.2.4, Documenting New or Additional Project Titles Associated with a Biosafety
Protocol (or Multiple Biosafety Protocols) for further information.

2-15
START
Does your research involve the handling,

administration or potential exposure to potentially hazardous biological materials?
This includes:
• Human or non-human primate blood, tissues, and other bodily fluids or
specimens
• Recombinant DNA (including creation or cross-breeding of transgenic animals) Your research project
• Tissues or other material that possibly may contain infectious to humans, NO will not require an IBC-
animals, and/or plants approved Biosafety END
• Animals that have been handled in a room designated BSL-2 containment Protocol (BSP)
• Toxins of Biological Origin
• Shipping/transport of any tissues, fluids or organisms or materials derived from
living organisms or other hazardous materials used to preserve these materials
(such as dry ice, liquid nitrogen)
• Any cell, tissue, organ or microbial cultures ≥10 liters in volume
YES
When do you intend to initiate

research described in this project
You will need to submit a BSP to the Biosafety Office
ASAP
(including biological agents, locations, and receive IBC approval prior to initiation of your
operations, personnel which have not (e.g. for preliminary data, Independent experiments independent of other review committees.
yet received IBC approval? of sponsored funding)
Only after this project is funded and/or I

receive IRB or IACUC approval
1. Will you behandling biological materials in

any MCG facilities?
2. Will any of these biological materials be Are all biological
Will your research require handled by MCG employees? materials handled
Institutional Review Board 3. Will any of these biological materials be within the scope of
YES YES to ANY NO
(HAC or CCRI) approval? This shipped/transported outside of campus by your MCGHI or
would include any project using MCG employees? VA clinical duties
ANY patient specimens 4. Will your project involve the delivery of any (i.e. as per clinical
biological materials (e.g. recombinant DNA, standard-of-care)?
biotoxins (e.g. BOTOX), cells, tissues or
organs into any human subjects?
NO
YES
NO to ALL
Office of Human Research Protection (OHRP)

will request verification of IBC approval for this project
prior to completion of the IRB approval paperwork
subsequent to IRB review/approval.
You will need to complete the IBC review/approval process for this protocol and ensure that the Biosafety Office has
received documentation linking your IBC-approved BSP to this clinical protocol before OHRP can complete your
authorization documentation; however HAC and IBC reviews can take place concurrently.
Linkage of BSPs to clinical protocols can be accomplished via a Schedule O, which verifies that the protocol with the following information is
fully covered in the indicated BSP. Required information for OHRP verification include:
1. Title of Clinical Protocol

2. Sponsor of Clinical Protocol (if any)
3. PI name (if different than that on the BSP)
4. HAC or CCRI #
5. Projected project period
6. BSP#, date of approval
Note: the Biosafety Office will be performing random audits of PIs research to make sure the information in their BSPs and Clinical Protocols
Will your research involve introduction of biological reconcile.
materials (cells, tissues, biotoxins, recombinant
DNA) into vertebrate animals?
Institutional Animal Care and Use Committee (IACUC)
YES to
Will your research involve the introduction of will request verification of IBC approval for this project
recombinant DNA into vertebrate animals (including ANY prior to completion of the IACUC approval paperwork
the creation of new trangenic/knock-out straings by subsequent to IACUC review/approval.
cross-breeding Tg lines strains with strains of
different genetic backgrounds?
You will need to complete the IBC review/approval process for these experiments before IACUC can
complete your AUP authorization documentation; however IACUC and IBC reviews can take
place concurrently.
NO to all As a consultant on the IACUC, the BSO reads all AUPs, therefore the BSO will automatically verify documentatiion of all
experiments of concern have been documented prior to approval as part of the IACUC review process.
Division of Sponsored Program Administration (SPA)

will request verification that IBC approval for this project
prior to establishment of any account associated with this project.
You will need to complete the IBC review/approval process for the experiments described in this project and ensure that the
Biosafety Office has received documentation linking your IBC-approved BSP to this project before SPA can set up any account
associated with this project. Note: submission of BSPs can take place after grant submissions as long as the IBC approval
Is your research sponsored (and process has been completed “just in time” for the receipt of funding notification by SPA.
therefore will require an account be YES
established by the
Linkage of BSPs to a sponsored project can be accomplished via a Schedule O, which verifies that the experiments/agents/locations described within this
Division of Sponsored Project
specific project are fully covered in the indicated BSP information. Required information to verify IBC approval to DSPA include:
Administration (DSPA)?
1. Title of the Project
2. Sponsor of the Project
3. PI Name (if different than that on the BSP)
4. Funding period (date of next competitive renewal period)
NO 5. BSP#, date of approval.
Note: the Biosafety Office will be performing random audits of PIs research to make sure the information documented in their BSPs and grants/contract
reconcile.
END
Figure 2.5.2 IBC Verification Paperwork Flow
2.6 BIOLOGICAL MATERIALS AND APPLICATIONS WHICH MUST
BE DOCUMENTED ON BIOSAFETY PROTOCOLS
2.6.1 Recombinant DNA Experiments
As a condition for receipt of NIH funding, MCG must ensure that all such research conducted at or
sponsored by the institution, irrespective of the source of funding, shall comply with the NIH Guidelines
for Research Involving Recombinant DNA Molecules. The NIH Guidelines for Research Involving
Recombinant DNA Molecules (NIH Guidelines) is available via the NIH Office of Biotechnology Activities
(NIH OBA) web page:
http://www4.od.nih.gov/oba/rac/guidelines/guidelines.html.
On behalf of the institution, the MCG IBC is required by NIH to review all recombinant DNA research
conducted at or sponsored by the institution and perform a comprehensive risk assessment of containment
levels, facilities, procedures, practices, training and expertise of personnel for proposed research to ensure
compliance with the NIH Guidelines.
In addition, the IBC is expected to periodically review recombinant DNA research conducted at the
institution and report any significant problems with or violations of the NIH Guidelines and any significant
research-related accidents or illnesses to the appropriate institutional official and NIH/OBA within 30
calendar days, unless the Institutional Biosafety Committee determines that a report has already been filed
by the Principal Investigator (Section IV, NIH Guidelines:
http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Guidelines_Apr_02.htm#_Toc7261578). Reporting of
adverse events specifically associated with Human Gene Transfer protocols must be accomplished even
faster than 30 days (see Appendix M, NIH Guidelines for details:
http://www4.od.nih.gov/oba/rac/guidelines_02/Appendix_M.htm). The MCG Institutional Biosafety
Committee (IBC) therefore requires registration and/or approval is required prior to the initiation of all
recombinant DNA experiments.
2.6.1.1 Definition of Recombinant DNA

The NIH Guidelines (Section 1-B) defines recombinant DNA molecules as either:
i) molecules that are constructed outside living cells by joining natural or synthetic
DNA segments to DNA molecules that can replicate in a living cell, or
ii) molecules that result from the replication of those described in (i) above.
NIH Guidelines also specifies that synthetic DNA segments which are likely to yield a
potentially harmful polynucleotide or polypeptide (e.g., a toxin or a pharmacologically active
agent) are considered as equivalent to their natural DNA counterpart. If the synthetic DNA
segment is not expressed in vivo as a biologically active polynucleotide or polypeptide
product, it is exempt from the NIH Guidelines.
Therefore, according to NIH Guidelines, the following are technically not considered
“recombinant DNA”:
Isolation of genomic DNA or RNA from natural sources
PCR amplification of genomic DNA or cDNA from non-cloned templates (e.g. for
genotyping purposes), as long as the resultant amplicons are not subsequently cloned
Chemically synthesized oligonuleotides such as those used in some siRNA/shRNA or
DNA sequencing applications. Please note, methods of producing siRNA or shRNA
other than chemical synthesis of oligonucleotides typically involve recombinant DNA
templates and/or cloning techniques and are therefore not exempt from NIH Guidelines
(e.g. production via in vitro transcription, expression of siRNA from an expression
plasmid or viral vector, or expression of a PCR-derived siRNA expression cassette)

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2.6.1.2 Definition of Exempt vs. Non-Exempt Recombinant DNA
NIH categorizes recombinant DNA experiments into six categories, as described in Sections IIIA
through F of NIH Guidelines for Research with Recombinant DNA Molecules. Five of these
categories (described in Sections IIIA through E of NIH Guidelines) refer to research that the NIH
considers “non-exempt” from the requirements for notification and/or approval of internal
institutional review committee(s) (i.e. the IBC and/or IRB) and/or external agencies (NIH Office
of Biotechnology Activities (OBA), the Recombinant DNA Advisory Committee (RAC) and/or
the NIH Director). Although the NIH does not explicitly require notification or approval for
recombinant DNA experiments which fall into a sixth (“exempt rDNA”) category (as described in
Section III-F of NIH Guidelines) prior to initiation of the experiment, the MCG IBC asks
researchers register and/or discuss their non-exempt rDNA experiments with the Biosafety Office
in an effort to avoid accidental misclassification of rDNA experiments.
The factors that distinguish non-exempt rDNA research vs. exempt rDNA research involve one or
a combination of the following, any one of which may alter the risk of the overall research
proposal:
Vectors
Typically, vectors that are contain large regions or are derived from genomes or
episomes of Risk Group ≥ 2 agents (i.e. infectious material or material capable
of infection of human cells) are considered “non-exempt” unless/until proven
incapable of infection or transmitting harmful effects upon target cells (e.g. viral
vectors—including those that are designed to be replication-incompetent and/or
are commercially available. Please note that viral vectors are still infectious,
even if they are designed to be replication-incompetent!)
Inserts
Some inserts, by themselves, will relegate a recombinant DNA protocol to the
non-exempt category, particularly those which:
o confer drug resistance trait to microorganisms that are not known to
acquire the trait naturally if such acquisition could compromise the use
of the drug to control disease agents in humans, veterinary medicine, or
agriculture (See Section III-A of NIH Guidelines for further
information:
http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Guidelines_Apr_0
2.htm#_Toc7261559)
o are derived from genomes or episomes of RG ≥ 2 organisms (i.e.
pathogens) or Select Agents (http://www.cdc.gov/od/sap/docs/salist.pdf)
which may transmit a harmful effect upon an intended or unintended
target cell.
o are biological toxins with LD50 < 100 μg/kg body weight. Those
toxins with LD50 < 100 ng/kg body weight (e.g. botulinum toxins,
tetanus toxin, diphtheria toxin, and Shigella dysenteriae neurotoxin)
require NIH/OBA approval; those with LD50s ≥100 ng/kg but < 100
μg/kg may require NIH/OBA registration prior to initiation (e.g.
Staphylococcus aureus alpha toxin, Staphylococcus aureus beta toxin,
ricin, Pseudomonas aeruginosa exotoxin A, Bordetella pertussis toxin,
the lethal factor of Bacillus anthracis, the Pasteurella pestis murine
toxins, the oxygen-labile hemolysins such as streptolysin O, certain
neurotoxins present in snake venoms and other venoms, cholera toxin,
the heat labile toxins of Escherichia coli, Klebsiella, and other related
proteins that may be identified by neutralization with an antiserum

2-18
monospecific for cholera toxin, and the heat stable toxins of
Escherichia coli and of Yersinia enterocolitica) See Appendix F NIH
Guidelines and the CDC/USDA Select Agent Program
(http://www.cdc.gov/od/sap/final_rule.htm) for further information.
In addition, expression of some inserts in non-exempt vectors may alter the risk
associated with the research proposal. For instance, the biological effect of
over- or under-expression of DNA inserts in non-exempt vectors which
modulate any of the following biologically active molecules may require special
additional consideration by the IBC during the comprehensive risk assessment
process:
o Oncogenes or proto-oncogenes
o Anti-apoptotic molecules
o Immunomodulatory molecules
o Toxins
Volumes
Large-scale culture volumes ≥10 liters require special operations as per NIH
Guidelines (see Appendix K, NIH Guidelines).
Applications/ Targets
Delivery of rDNA into the following targets may relegate a recombinant DNA
protocol to non-exempt status
o Risk Group ≥ 2 microorganisms (i.e. potential pathogens)
o Human subjects (i.e. Human Gene Transfer/Therapy—see Section
2.6.1.4, below for additional information)
o Live Animals.
Introduction of recombinant DNA into animals (e.g. injection
of rDNA or transplantation of genetically engineered cells)
may fall into the non-exempt rDNA categories as per NIH
Guidelines.
Creation of a new strain of Transgenic (Tg) and/or Knock-out
(KO) animals via:
• Standard Tg/KO techniques (rDNA delivery into ES
cells or blastocysts)
• Cross-breeding 1+ strain of Tg/KO animals with
another strain of different genetic background, thus
creating a new strain.
However, the purchase or transfer of existing transgenic or
KO rodent strains (e.g. from commercial suppliers, such as
Jackson, Harlan or Charles River Laboratories or from other
IBC-authorized investigators) is exempt from the NIH
Guidelines under Section III-F (see Appendix C-VI, NIH
Guidelines) if these animals are appropriately contained at
BSL-1.
The following guideline, reproduced in the Biosafety Protocol Application Form is intended to
further assist researchers in properly classifying their recombinant DNA experiments (see Table
2.6.1.2). Please note that experiments 1-3 listed in Table 2.6.1.2 under “Experiments which must
be registered with the IBC and approved prior to initiation” require registrations/approvals of
external agencies (the NIH Director, RAC, and/or NIH/OBA), so IBC processing of these
protocols will require the most time to complete. Please contact the Biosafety Office x1-2663 or
BIOSAFETY@mcg.edu for any additional assistance:

2-19
“Non-exempt” Recombinant DNA “Exempt”
Experiments which must be registered with the Experiments that require registration with the Recombinant DNA experiments that do not require
IBC and approved prior to initiation: IBC simultaneous with initiation: IBC approval:
1. Deliberate transfer of a drug trait to a 1. Experiments using as vectors < 2/3 of the genome 1. rDNA containing less than 1/2 of an eukaryotic
microorganism not known to acquire it naturally of a eukaryotic virus, demonstrated to be free of viral genome propagated in cell culture (with the
(if it could compromise the use of the drug to helper virus or complementing helper virus exception of expression of DNA from Risk Group4
control disease agents in humans, animals or components. 2, 3, 4 or restricted agents);
agriculture)2. (Note: this would likely exclude
most subcloning procedures using antibiotic 2. rDNA work involving E. coli K12 derivatives, S.
selectable markers in E. coli K12 derivatives) 2. Transgenic or knockout rodent experiments for cerevisiae, and B. subtilis host-vector systems (with
which BSL-14 containment is appropriate (NOTE: the exception of expression of DNA from Risk
2. Cloning of DNA encoding toxic molecules lethal the purchase of transgenic rodents for BSL-1 Group 2, 3, 44 or restricted agents).
to vertebrates at an LD50 of <100 µg/kg body experiments falls into the “exempt” category;
weight2,3. however, breeding of these animals with different
strains of mice is considered non-exempt rDNA
3. Human gene transfer/therapy experiments2; experiments).
4. Cloning using human or animal pathogens as

host-vector systems.
5. Cloning of DNA from all Risk Group4 2, 3, 4 or

restricted human or animal pathogens (including
HIV and related viruses and viruses capable of
infecting human cells).
6. Experiments using more than 2/3 of the genome

of infectious animal or plant viruses or defective
viruses grown in the presence of helper virus or
complementing helper virus components.
7. Recombinant DNA experiments involving whole

animals, including transgenic or knockout rodent
experiments requiring BSL24 containment; or
transplantation of genetically engineered cells
into animals.
8. Large scale DNA projects (≥10 liter cultures at

any moment in time).
1
For further explanations/details, please see: http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Guidelines_Apr_02.htm
2
Please note that these experiments may require approval of NIH Director, NIH’s Office of Biotechnology Activities (OBA) approvals or registration
and/or the Recombinant DNA Advisory Committee (RAC) review. Please contact the Biosafety Office (x1-2663) as soon as possible if your
experiments fall into these categories for assistance.
3
LD50 for many toxins can be viewed on commercially available Material Safety Data Sheets (MSDSs), e.g.
http://www.sigmaaldrich.com/Area_of_Interest/The_Americas/United_States.html or contact the Biosafety Office x1-2663 if you need assistance.
4
See the following links for assistance in Risk Group classification and recommended Biosafety Level usage:
• American Biosafety Association Risk Group Guide: http://www.absa.org/XriskgroupsX/index.html
• Public Health Agency of Canada MSDSs: http://www.phac-aspc.gc.ca/msds-ftss/
• NIH Guidelines for Recombinant DNA Research: http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Guidelines_Apr_02.htm
• CDC’s Biosafety in Microbiological and Biomedical Laboratories (BMBL): http://www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm
Table 2.6.1.2 Simplified Guideline for Classifying Recombinant DNA Experiments according to NIH
Guidelines for Research with Recombinant DNA Molecules. MCG IBC requests that all recombinant
DNA experiments be registered with the Biosafety Office prior to initiation to avoid potential mis-
classification errors. (Table based on that from Yale University’s Biosafety Manual)

2-20
2.6.1.3 Recombinant DNA Registration Requirements
Principal Investigators and/or Lab Supervisors must submit or amend Biosafety Protocols:
before bringing new non-exempt recombinant DNA materials to campus which had not
already been approved by the IBC
before initiating any new non-exempt recombinant DNA work.
New submissions of Biosafety Protocol describing new work must be submitted by the PI using
the Biosafety Protocol Application forms, which are available in Appendix A of the Biosafety
Guide and are also available online on the MCG Biosafety Office web site
(http://www.mcg.edu/research/ibc/apps.htm).
Amendments must be submitted by the PI fully describing the new work if the scope of the work
has changed from that approved by the IBC. Changing the scope of the work would include
changes in: vectors or inserts, administration or exposures of new target materials or animals,
creation of a new transgenic strain or species, or embarking upon large-scale culture (≥10 liters),
changes in locations, personnel or major equipment (see Section 2.1.2 regarding BSP amendment
procedures). Amendments can be submitted by altering the original Biosafety Protocol
Application forms to include the new information and re-submission to the Biosafety Office, or
via a fully detailed email. All questions can be addressed to the Biosafety Office (x1-2663 or
BIOSAFETY@mcg.edu).
2.6.1.4 Human Gene Transfer/Therapy (HGT)
Proposed clinical trials involving the deliberate transfer of recombinant DNA, or DNA or RNA
derived from recombinant DNA, into human research participants are defined as human gene
transfer experiments. Human gene transfer for therapeutic purposes is referred to as human gene
therapy.
Human gene transfer experiments require registration and approval from at least two campus
safety committees (the IBC and the IRB) as well as review by federal agencies (the NIH Office of
Biotechnology’s (NIH/OBA’s) Recombinant DNA Advisory Committee (RAC) and the Federal
Drug Administration (FDA)) before enrollment of the first subject. These requirements are
described in Section III-C and Appendix M of NIH Guidelines
(http://www4.od.nih.gov/oba/rac/guidelines_02/Appendix_M.htm).
In addition to Appendix M of NIH Guidelines, the NIH has provided reference information for
researchers involved in Human Gene Transfer Research on their web page. Principal Investigators
should review these materials and implement the guidance in preparation of protocol applications.
The NIH Guidance for the development of Informed Consent documents specifically related to
Gene Transfer Research can be found at: http://www4.od.nih.gov/oba/rac/ic/. NIH has also
provided answers to Frequently Asked Question (FAQ) about the NIH Review Process for Human
Gene Transfer Trials at: http://www4.od.nih.gov/oba/RAC/RAC_FAQs.htm .
The NIH/FDA Genetic Modification Clinical Research Information System (GeMCRIS) maintains
a comprehensive information resource and analytical tool for scientists, research participants,
sponsors, institutional oversight committees, federal officials, and others with an interest in human
gene transfer research. GeMCRIS allows public users to access basic reports about human gene
transfer trials registered with the NIH and to develop specific queries based on their own
information needs. GeMCRIS information can be accessed via
http://www4.od.nih.gov/oba/rac/GeMCRIS/GeMCRIS.htm .
The IBC is required by the terms of NIH Guidelines to ensure compliance with all aspects of
Appendix M, which includes review of materials which are more typically considered the domain

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of the Institutional Review Board (IRB). Therefore, in addition to the standard BSP Application
forms and protocol-specific SOPs, PIs of human gene transfer protocols must include the Clinical
Protocol materials, Investigator’s Brochure, the Informed Consent Document, Recombinant DNA
Advisory Committee (RAC) submission information and documentation of review/responses from
the RAC, and any pertinent IND information, including preliminary animal or human studies and
SAE report information in their application for IBC review and approval.
After IBC review and approval, a record of the IBC review and comments will be provided to the
Institutional Review Board (IRB) for consideration during their review and a copy will be
provided to the Office of Human Research Protection (OHRP) for their files.
Copies of any adverse event reports must be provided to the sponsor, the IRB, the NIH/OBA and
IBC within the time frames described in Appendix M of NIH Guidelines. Human gene transfer
protocols must be reviewed by the IBC at least annually. Therefore, any updated information
related to the protocol must be provided to the Biosafety Office no later than 11 months after the
IBC approval anniversary.
2.6.2 Human, Animal and/or Agricultural Pathogens or Toxins (or Materials

Potentially Contaminated with these Pathogens or Toxins)
Special considerations may be required prior to possession and/or transfer of any known human, animal
and/or agricultural pathogens or toxins or materials which may be potentially contaminated with these
pathogens or toxins to MCG campus to comply with CDC and/or USDA regulations. This may include
laboratory inspections for compliance with Biosafety standards and permits for transfer of these agents.
The Biosafety Office can assist researchers in determining whether special inspections and/or permits may
be required.
The MCG Institutional Biosafety Committee (IBC), the Institutional Animal Use and Care Committee
(IACUC) and the Division of Laboratory Animal Services (LAS) must approve all experiments involving
the introduction of infectious agents or potentially hazardous biological materials into animals prior to
initiation.
All researchers working with etiologic agents (Risk Group 2 or higher) must receive training in both
biosafety and the microbiological procedures that will be utilized for the experiment. Biosafety training
sessions for new staff and faculty are provided through the Division of Environmental Health and Safety on
a monthly basis (call x1-2663, BIOSAFETY@mcg.edu or check EHS web site for the next scheduled training
date), The Principal Investigator is responsible for ensuring that all researchers are trained in the
appropriate procedures and techniques used in the laboratory.
Consultation in Risk Group classification of biological agents can be sought through the Biosafety Office
(x1-2663 or BIOSAFETY@mcg.edu). Information can also be found at the following URLs:
CDC BMBL: http://www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm
NIH Guidelines: http://www4.od.nih.gov/oba/rac/guidelines/guidelines.html
ABSA Risk Group Classification Database: http://www.absa.org/XriskgroupsX/index.html
Public Health of Canada Biological MSDSs: http://www.phac-aspc.gc.ca/msds-ftss/
Work with Risk Group 3 agents (or those requiring Biosafety Level 3 containment) and/or Select Agents or
Toxins will require additional registration and training requirements by the PI and laboratory staff as
specified in the BSL-3 Laboratory Manual. Contact the Biosafety Office for additional information. Work
with Risk Group 4 agents or experiments requiring Biosafety Level 4 containment are not permitted at
MCG.

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2.6.2.1 Toxins of Biological Origin
Because of the risks posed by the acute toxicity of some biological toxins, the IBC will require
documentation of Standard Operating Procedures (SOPs) to ensure safety handling and disposal
procedures for any toxin of biological origin with an LD50 ≤ 100 µg/kg body weight as part of the
Principal Investigator’s Biosafety Protocol (BSP). These should incorporate the risk assessment
and management considerations as described in Appendix I of the CDC/NIH publication Biosafety
in Microbiological and Biomedical Laboratories (BMBL)
http://www.cdc.gov/od/ohs/biosfty/bmbl5/sections/AppendixI.pdf .
These SOP considerations should include:

• Special training should be provided to all personnel related to the unique risk
characteristics of toxins and particular emergency spill and/or exposure procedures
should be provided
• Special procedures and protective equipment required to prevent exposure to any
dry/freeze-dried toxins, since the potential for electrostatic dispersal of concentrated
forms of toxin pose uniquely high risks. Respiratory protective measures must be
considered.
• The containment practices and equipment required to reduce the risks of accidental
exposure by direct contamination of mouth, eyes or other mucous membranes by
inadvertent aerosol generation; and by needle-sticks or other accidents that may
compromise the normal barrier of the skin while working with solutions of biotoxins.
When working with toxins that pose direct percutaneous hazards, special care must be
taken to select gloves that are impervious to the toxin and the diluents or solvents
employed.
• Restricted access to designated isolated rooms while toxins are in use. The room should
be clearly posted: “Toxins in Use—Authorized Personnel Only.” while toxin work is on-
going and unrelated and nonessential work should be restricted from areas where stock
solutions of toxin or organisms producing toxin are used.
• Special deactivation/decontamination procedures may be required for surfaces and
liquids contaminated with the toxins. Toxins may not be deactivated by typical
disinfectants used against pathogens, and therefore, special procedures should be
documented and followed for deactivation and disposal (see Section 7.3 for further
information and guidance on deactivation of biological toxins).
• Additional security measures which may be required to limit access to the stored
biological toxins.
2.6.2.2 Select Agents and Toxins (SATs)

A subset among human, animal and/or agricultural pathogens and toxins are the Select Agents and
Toxins (SATs). Select agents/toxins are agents that the U.S. Department of Health and Human
Services (HHS) and/or the Department of Agriculture/Animal & Plant Health Inspection Service
(USDA/APHIS) considers to have the potential to pose a severe threat to human health, animal or
plant health, or to animal or plant products (See Table 2.6.2.2 for a list of these Select Agents of
Toxins or http://www.selectagent.gov/agentToxinList.htm).
The SAT regulations pertain not only to the intact agents or toxins, but also several genetic
elements or recombinant nucleic acids from these agents and recombinant Select Agent organisms.
The HHS and USDA originally enacted interim regulations governing the possession, use, and/or
transfer of these agents on December 13, 2002, and subsequently published their final rules on
March 18th, 2005 and these can be viewed here:

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42 CFR Parts 72 and 73 Office of Inspector General 42 CFR Part 1003: Possession,
Use, and Transfer of Select Agents and Toxins; Final Rule:
http://www.selectagent.gov/resources/42_cfr_73_final_rule.pdf
7 CFR Part 331 and 9 CFR Part 121: Agricultural Bioterrorism Protection Act of 2002;
Possession, Use, and Transfer of Biological Agents and Toxins; Final Rule:
http://www.selectagent.gov/resources/APHISFinalRule.pdf
The National Select Agent Registry has established a URL that serves as an informational center,
as well as a source of all registration materials, and forms. http://www.selectagent.gov/

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Among these requirements of these regulations is that any entity (defined by HHS/CDC and
USDA/APHIS as “any government agency (Federal, State, or local), university, corporation,
company, partnership, society, association, firm, sole proprietorship, or other legal entity”)
wishing to use, possess, or transfer SATs must register with either the Centers of Disease Control
(CDC) or USDA/APHIS. This registration process requires the following:
Designation of a Responsible Official (RO) for the entity, which is the person
“designated by an entity to act on its behalf…” and that “…this individual must have the
authority and control to ensure compliance with the regulations in this part”. Currently,
the MCG RO is the Biosafety Officer. An Alternate Responsible Official (ARO) may
also be designated by the entity and who is similarly required to ensure compliance with
the regulations. Currently, the MCG ARO is the Directory of Environmental Health and
Safety.
Approval of the HHS or USDA Secretary or Administrator based on a Security Risk
Assessment (SRA) by the U.S. Attorney General for any individual or entity who may
potentially have access or control over any Select Agent or Toxin. Note: an application
for an individual may be denied or a certificate of registration revoked or suspended if an
individual is reasonably suspected by any Federal law enforcement or intelligence agency
of:
o Committing a crime specified in 18 U.S.C. 2332b(g)(5),
o Knowing involvement with an organization that engages in domestic or
international terrorism (as defined in 18 U.S.C. 2331) or with any other
organization that engages in intentional crimes of violence, or
o Being an agent of a foreign power (as defined in 50 U.S.C. 1801)
Identification of the particular physical location in which these SATs may be present, and
the associated comprehensive documentation must be provided and approved by the CDC
and/or USDA for the specific locations, agents and experiments proposed for use in these
facilities:
o Safety plans
o Security plans
o Incident/Emergency response plans
o Precise Inventories and all records of transfer and use.
Agent- and Use-specific training of all who may have access must be documented and
approved, and must include drills and exercises.
Notifications of any theft, loss, or unaccounted samples, releases from containment and
possible exposures must be made to the CDC and/or USDA, and may require further
Federal investigation and cooperation.
Any individual who intends to possess, use or transfer any Select Agent or Toxin to MCG campus
which does not qualify for any of the exclusions and exemptions listed below in Sections 2.6.2.2.1
and 2.6.2.2.2, must contact the Biosafety Office (x1-2663 or BIOSAFETY@mcg.edu) immediately
to discuss registration procedures and the adequacy of the facilities to secure the agents or toxins
in question, and the measures that will need to be taken to qualify for registration. They must
refer to the BSL-3/Select Agent and Toxin Manual and receive additional training as required by
Federal law.
2.6.2.2.1 Exclusions from Select Agents and Toxins Regulations
HHS/USDA select agents or toxins that meet any of the following specific criteria are
excluded from the Select Agent of Toxin Regulations as described in 42 CFR §73.3 (d) &
(e), 7 CFR §331.3 (d) & (e) and 9 CFR §121.4 (d) & (e):
1. Any HHS/USDA select agent or toxin that is in its naturally occurring

environment provided the select agent or toxin has not been intentionally
introduced, cultivated, collected, or otherwise extracted from its natural source.

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2. Non-viable HHS/USDA select agents or nonfunctional HHS/USDA toxins.
3. HHS/USDA toxins under the control of a principal investigator, treating
physician or veterinarian, or commercial manufacturer or distributor, if the
aggregate amount does not, at any time, exceed the following amounts:
Abrin 100 mg
Botulinum neurotoxins 0.5 mg
Clostridium perfringens epsilon 100 mg
toxin
Conotoxins 100 mg
Diacetoxyscirpenol 1,000 mg
Ricin 100 mg
Saxitoxin 100 mg
Shiga-like ribosome inactivating 100 mg
proteins (i.e. verotoxins)
Shigatoxin 100 mg
Staphylococcal enterotoxins 5.0 mg
Tetrodotoxin 100 mg
T–2 toxin 1,000 mg
The regulations in 42 CFR Part 73 and 9 CFR Part 121 also include a procedure by which
an attenuated strain of a select biological agent or toxin that does not pose a severe threat
to public health and safety, animal health, or animal products may be excluded from the
list of select biological agents and toxins upon application and approval of the HHS or
USDA (See 42 CFR §73.3 (e), 7 CFR § 331.3 (e) and 9 CFR §121.4 (e).
Based upon consultations with subject matter experts and a review of relevant published
studies and information provided by the entities requesting the exclusions, the HHS and
USDA have determined that some attenuated strains are not subject to the requirements
of 42 CFR §73, 7 CFR §331, and 9 CFR§121 if used in basic or applied research, as
positive controls, for diagnostic assay development, or for the development of vaccines
and therapeutics. A list of excluded attenuated strains of SATs can be viewed at:
http://www.selectagent.gov/exclusions.htm . However, please note that an individual or
entity that possesses, uses, or transfers an excluded attenuated strain will be subject to the
regulations if there is any reintroduction of factor(s) associated with virulence or other
manipulations that modify the attenuation such that virulence is restored or enhanced.
All Select Agents Toxins and their uses must still be documented on the associated
Biosafety Protocol even if exclusion levels are not met or exceeded. Because of the very
specific requirements to maintain exclusion criteria, the IBC will likely require additional
documentation or standard operating procedures (SOPs) for possession use and/or
transfer of any excluded Select Agent or Toxin. This may include testing to prove
attenuated/non-viable agents/toxins are, indeed, attenuated/non-viable upon receipt and
maintaining additional security on any room or refrigerator or freezer which may contain
the SAT, and/or maintaining an accurate log book to document the amount of Select
Agent Toxin received/used in the laboratory to document the aggregate amounts in the
laboratory at any moment in time.
2.6.2.2.2 Exemptions from Select Agents and Toxins Regulations for

Clinical/Diagnostic Laboratories
As described in 42 CFR §73.5, 7 CFR §331.5 and 9 CFR §121.5, provisions have been
made specifically for clinical and/or diagnostic laboratories or other entities that possess,

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use, or transfer an HHS/USDA select agent or toxin under certain specified conditions
when the SAT is:
Contained in a specimen presented for diagnosis or verification (see below for
conditions)
Contained in a specimen presented for proficiency testing
A product that is, bears or contains SATs that is cleared, approved, licensed, or
registered under any of the following laws, insofar as its use meets the
requirements of such laws:
• The Federal Food, Drug, and Cosmetic Act (21 U.S.C. 301 et seq.),
• Section 351 of the Public Health Service Act pertaining to
biological products (42 U.S.C. 262)
• The Act commonly known as the Virus-Serum-Toxin Act (21
U.S.C. 151–159)
• The Federal Insecticide, Fungicide, and Rodenticide Act (7 U.S.C.
136 et seq.)
An agent or product for which the HHS Secretary or USDA Administrator has
granted a special exemption as per their application/approval process
(CDC/USDA Form 5). These special circumstances may include an
investigational/experimental product that is, bears, or contains a select agent or
toxin
• when such product is being used in an investigation authorized
under any Federal Act and additional regulation under this part is
not necessary to protect public, animal or plant health and safety or
protect animal or plant product safety.
• being used, possessed or transferred to allow for timely participation
of the individual or entity in response to a domestic or foreign
public health emergency, agricultural emergencies or outbreaks, or
in endemic areas.
Is a diagnostic reagent or vaccine that is, bears, or contains USDA Veterinary
Service Select agents or Toxins that are produced at USDA diagnostic facilities.
The Biosafety Officer/Responsible Official must be contacted immediately (x1-2663)

when any material containing a Select Agent or Toxin is identified or is likely to be
transferred to campus (even if it may qualify for an exemption or exclusion) to
ensure that the strict compliance standards with these Federal laws are maintained.
2.6.2.2.3 Criteria for Handling Select Agents or Toxins Contaminated

Specimens in Clinical/Diagnostic Laboratories under the
Exemption Clause
Please note, if a SAT is contained in a specimen presented for diagnosis or verification,

please be aware that the following conditions must be met:
1. The SAT needs to be transferred or destroyed on-site within 7 calendar
days in accordance with specific guidance in the Federal laws.
2. The SAT must be secured against theft, loss or release prior to transfer
or destruction.
3. With the exception of certain agents which have requirements for
immediate reporting to the CDC/USDA (see below), once the
diagnosis/identification of SATs in any specimens must be reported to
the CDC and/or APHIS with seven (7) calendar days after
identification and other appropriate authorities when required by law.

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The following agents must be reported to the CDC immediately by
telephone, facsimile, or e-mail, followed by submission of
CDC/USDA Form 4:
• Ebola viruses
• Lassa fever virus
• Marburg virus
• Variola major virus (Smallpox virus)
• Variola minor (Alastrim)
• Yersinia pestis
• South American Haemorrhagic Fever viruses (Junin,
Machupo, Sabia, Flexal, Guanarito)
The following agents must be reported to the USDA immediately

by telephone, facsimile, or e-mail, followed by submission of
CDC/USDA Form 4:
• African horse sickness virus
• African swine fever virus
• Avian influenza virus (highly pathogenic)
• Bovine spongiform encephalopathy agent
• Classical swine fever virus
• Foot-and-mouth disease virus
• Newcastle disease virus (velogenic)
• Rinderpest virus
• Swine vesicular disease virus
2.6.3 Human Blood, Body Fluids, Cells, Tissues and Other Potentially Infectious
Materials
The Occupational Safety and Health Administration (OSHA) created the Occupational Exposure to
Bloodborne Pathogens (BBP) Standard, 29 CFR Part 1910.1030 (Bloodborne Pathogens Standard) to
minimize or eliminate exposure to infectious agents that may be present in any human blood, tissues or
certain body fluids (bloodborne pathogens). Georgia does not have an OSHA-approved state plan for
occupational safety and health. Consequently, while federal OSHA requirements do apply to most private
employers in the state, they do not extend to public employers such as government agencies. However,
Georgia Code §31-12-13 requires public employers to adopt a bloodborne pathogen standard governing
occupational exposure of public employees to blood and other potentially infectious materials. This
standard must be "at least as prescriptive" as the OSHA standard. Lastly, as described in Section 2.6.1,
one of the terms and conditions for acceptance of NIH Grant awards that any grantee institution adheres to
the OSHA Bloodborne Pathogen Standard 29 CFR 1910.1030.
http://grants.nih.gov/grants/policy/nihgps_2003/NIHGPS_Part4.htm#_Toc54600062. For these reasons, it
is MCG’s IBC policy to adhere to the OSHA standards for Bloodborne Pathogens.
The Bloodborne Pathogens Standard applies to all employers having employees that are “occupationally
exposed” to human blood, materials which may have been exposed to human blood or other potentially
infectious materials. An employee is considered occupationally exposed if there is “reasonably anticipated
skin, eye, mucous membrane, or parenteral [via injection, infusion, cut exposure, or transdermal methods]
contact with human blood or other potentially infectious materials in the performance of an employee’s
duties.” An individual is also considered occupationally exposed even if they do not have direct contact
with blood or other potentially infectious material, if the employee uses equipment that is used to process
or store blood, other potentially infectious materials or bloodborne pathogens.

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Potentially infectious materials other than human blood may include:
Human cell, tissue or organ cultures
Any unfixed human tissue or organ, other than intact skin, from a human being (living or dead)
Human body fluids, except urine, feces, saliva or tears unless contaminated with blood. This
includes:
• semen
• vaginal secretions
• cerebrospinal fluid
• synovial fluid
• pleural fluid
• pericardial fluid
• peritoneal fluid
• amniotic fluid
• saliva in dental procedures
All body fluids in situations where it is difficult or impossible to differentiate between body fluids
or is contaminated with blood.
HIV-containing cell, tissue or organ cultures
HIV- or HBV-containing culture medium or other solutions
Blood, organs, or other tissues from experimental animals infected with HIV or HBV or other
bloodborne pathogens.
OSHA has determined that occupational exposure to human blood, tissues and body fluids poses a
significant health risk because these may contain bloodborne pathogens such as:
• Human Immunodeficiency Virus (HIV)
• Hepatitis B virus (HBV)
• Hepatitis C virus
• Hepatitis D virus
• Plasmodium species
• Treponema species
• Babesia species
• Borrelia species
• Brucella species
• Leptospira species
• Francisella species
• Streptobacillus moniliformis
• Colorado Tick Fever viruses
• Arboviruses
• Spirillum minus
• Creutzfeldt-Jakob virus
• Human T-lymphotropic Virus Type I
• Hemorrhagic Fever viruses
All occupationally exposed employees are required by OSHA to attend a Bloodborne Pathogens training
session prior to beginning work and annually thereafter. Employees must receive training when they are
first assigned to tasks where occupational exposure to bloodborne pathogens may occur. The training must
be repeated annually, and must be provided at no cost to the employees and during work hours.
Employees must receive additional training when changes occur that affect the employees' occupational
exposure, such as task or procedural modifications or the institution of new tasks or procedures. This
additional training only needs to address new exposures created.
Training must include:

• making accessible a copy of the regulatory text of the standard and explanation of its contents
• general discussion on bloodborne diseases and their transmission

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• an explanation of the exposure control plan and how the employee can obtain a copy of the written
plan
• how to recognize tasks and other activities that may involve exposure to blood and other
potentially infectious materials
• use and limitations of methods for preventing or reducing exposure, including engineering and
work practice controls and personal protective equipment
• information on the hepatitis B vaccine
• response to emergencies involving blood or other potentially infectious materials
• how to handle exposure incidents
• the post-exposure evaluation and follow-up program
• information about signs/labels/color-coding related to bloodborne pathogens
Due to the importance of information regarding occupational exposure to potentially life-threatening

bloodborne pathogens, employees must have the opportunity to ask questions and receive answers from a
knowledgeable trainer. Laboratory and production facility workers must receive additional specialized
initial training.
Consult the Bloodborne Pathogen Training Manual for Clinical and Laboratory Personnel for additional
information on the exposure control plan, training requirements, work practices, housekeeping, engineering
controls, personal protective equipment, signs/label requirements, Hepatitis B vaccination, emergency
actions, exposure incident procedures, post-exposure evaluation and follow-up, and recordkeeping. Contact
the Biosafety Office at 706-721-2663 or BIOSAFETY@mcg.edu for assistance with exposure determination
and for training information.
2.6.3.1 Cultures of Cell lines of Human and/or Primate Origin

Human cell lines are commonly used in biomedical research, yet appropriate biosafety
requirements for handling human cell lines are often subject to debate within the scientific
community. While human blood, most body fluids, unfixed human tissues and organs were
clearly included within the scope and application of the OSHA Bloodborne pathogen standard (29
CFR Part 1910.1030), the inclusion of human cell lines was ambiguous.
In 1994, OSHA issued a letter of interpretation related to the applicability of the BBP Standard
towards human cell lines:
http://www.osha.gov/pls/oshaweb/owadisp.show_document?p_table=INTERPRETATIONS&p_id
=21519.
According to the interpretation, human cell lines are considered to be potentially infectious and
within the scope of the BBP Standard unless the specific cell line has been characterized to be free
of hepatitis viruses, HIV, Epstein-Barr virus, papilloma viruses and other recognized bloodborne
pathogens. In alignment with this interpretation, the American Type Culture Collection (ATCC)
recommends that all human cell lines be accorded the same level of biosafety consideration as a
line known to carry HIV: http://www.atcc.org/TechnicalInfo/faqCellBiology.cfm#Q53
Moreover, Appendix H of the Fifth Edition of the CDC publication, Biosafety in Microbiological
and Biomedical Laboratories, recommends that human and other primate cells should be handled
using Biosafety Level 2 (BSL2) practices and containment
http://www.cdc.gov/od/ohs/biosfty/bmbl5/APPENDIX%20H.pdf .
Tumorigenic human cell lines also present a potential hazard as the result of self-inoculation. In
fact, there has been at least one reported case of development of a tumor resulting from an
accidental needlestick (Gugel EA and Sanders ME. (1986) Needle-stick transmission of human

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colonic adenocarcinoma. N Engl J Med. 315(23):1487; PMID: 3785302). Because of the extra
protection that the laboratory worker’s functioning immune system affords him/her in case of
accidental needlestick or exposure, laboratory workers should never handle autologous cells or
tissues.
In consideration of the aforementioned regulatory interpretation and consensus guidelines and

other factors, the MCG Institutional Biosafety Committee considers that all cell and organ
cultures of human or primate origin, including well-established cell lines should be handled
in accordance with the OSHA Bloodborne Pathogens Standard and under Biosafety Level 2
(BSL-2) containment.
2.6.3.1.1 Human Embryonic Stem (hES) Cell and Embryonic Germ Cell
Lines
On August 9, 2001, at 9:00 p.m. EDT, President George W. Bush announced his decision
to allow Federal funds to be used for research on existing human embryonic stem cell
lines as long as prior to his announcement
1. the derivation process (which commences with the removal of the inner cell
mass from the blastocyst) had already been initiated and
2. the embryo from which the stem cell line was derived no longer had the
possibility of development as a human being.
In addition, the President established the following criteria that must be met:
1. The stem cells must have been derived from an embryo that was created for
reproductive purposes;
2. The embryo was no longer needed for these purposes;
3. Informed consent must have been obtained for the donation of the embryo;
4. No financial inducements were provided for donation of the embryo.
In order to facilitate research using human embryonic stem cells, the NIH created the
Human Embryonic Stem Cell Registry, which lists the human embryonic stem cell
lines—at varying stages of development—that meet the eligibility criteria. A list of the
embryonic stem cell lines which meet the President’s criteria and are therefore eligible
for federal funding can be viewed at: http://stemcells.nih.gov/research/registry/ .
No federal funds may be used, either directly or indirectly, to support research on human
embryonic stem cell lines that do not meet the criteria established by President Bush on
August 9, 2001. Thus, research on lines (or their derivatives) not listed on the NIH
Human Embryonic Stem Cell Registry may not be supported by federal funds. This
includes considerations to strictly prevent any indirect costs, or F&A costs, to be utilized
to support any research with any unallowable human embryonic stem cell lines. Strict
compliance with cost allocation methodologies described in the circular, including the
Cost Accounting Standards, must prevent the shifting of unallowable stem cell research
costs to federally sponsored programs. A properly documented F&A proposal utilized in
the establishment of F&A rates should demonstrate that none of the costs of unallowable
stem cell research or other unallowable activities have been shifted to federally sponsored
activities. Generally, this means that separate facilities, supplies and equipment must be
maintained for research with any unallowable hES lines to prevent inadvertent use of
Federal indirect funds for the support of unallowable hES line research. See:
http://stemcells.nih.gov/info/faqs.asp for more information.
MCG hES research should also adhere to Guidelines for Human Embryonic Stem Cell
Research developed by the National Research Council of the National Academies of
Sciences. http://www.nap.edu/catalog.php?record_id=11278 . While MCG does not currently
support an Institutional Embryonic Stem Cell Research Oversight Committee (ESCRO),

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as recommended by the Guidelines, the ethics and compliance oversight will still be
monitored by the IRB, IBC and IACUC. Much of the recommendations in these
Guidelines relate to ensuring the source and consent of the donors and/or recipients of
these cells (i.e. issues generally under review by the Institutional Review Board);
however, please note the Guidelines specifically recommends against the following
applications for hES research:
• Research involving in vitro culture of any intact human embryo, regardless of
derivation method, for longer than 14 days or until formation of the primitive
streak begins, whichever occurs first.
• Research in which hES cells are introduced into nonhuman primate blastocysts
or in which any ES cells are introduced into human blastocysts.
• No animal into which hES cells have been introduced at any stage of
development should be allowed to breed.
Research use of human embryonic germ cells derived from fetal tissue with Federal funds
requires review of compliance with the NIH Guidelines for Research Involving Human
Pluripotent Stem Cells. The review process is described at:
http://grants.nih.gov/grants/guide/notice-files/NOT-OD-02-049.html .
2.6.4 Animals and the use of Biological Materials in Animals

All research experiments involving animals must be conducted in accordance with the associated MCG
Institutional Animal Care and Use Committee (IACUC) approved animal use protocol (AUP). If the
animals are infected, infectious or pose a higher risk for human infection such that the animals or their
tissues require Biosafety Level-2 containment or higher (see Section 4, Risk Management for further
information), research with these animals will require IBC authorization, as well. This would include
working with non-human primates (NHP), their tissues and cells.
Because of the evolutionary similarities between humans and non-human primates, NHPs are at higher risk
of carrying bloodborne pathogens infectious to humans. In addition, most species of macaque monkeys are
at high risk of being carriers of Cercopithecine Herpes Virus 1 (CHV-1, Herpes B virus) (see Section
3.2.3.1, Zoonotic Disease Risks/Animal Research Risks for further information). For these reasons, a
minimum of BSL-2 containment is recommended for handling of NHP species, their tissues and cells by
the CDC BMBL, Appendix H (http://www.cdc.gov/od/ohs/biosfty/bmbl5/sections/AppendixH.pdf). Materials
and practices must be documented on a BSP Application, and all personnel who may be at risk of exposure
to NHP materials should receive BBP training and preventative measures (see Section 2.6.3 for further
information).
Animal research that involves the introduction of biological hazards (e.g., agents which require ≥BSL-2
containment or recombinant DNA) must have an approved Biosafety Protocol (BSP) to reflect these uses
and fully describe and disclose the use of these biological agents/recombinant DNA in their AUP and BSP
applications. The Biosafety Office must approve work with pathogens or recombinant DNA in animals
(including transgenic animals) prior to initiation. Biosafety Protocol Application forms can be found at:
http://www.mcg.edu/research/ibc/apps.htm or please contact the Biosafety Office (x1-2663 or
BIOSAFETY@mcg.edu) for assistance.
Once approved by the IBC, the MCG Division of Laboratory Animal Resources Center (LAS) will be
contacted prior to initiation to ensure that a safety protocol have been established and appropriate facilities
are obtained for these experiments. Contact LAS at 706-721-3421 for additional information.

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2.6.5 Insects/Arthropods
Insects present unique challenges to containment which are not encountered with either microbial
pathogens or biological toxins. In addition, because some arthropods may serve as vectors for disease
transmission, special containment considerations, including facilities (insectaries) and practices specifically
designed to prevent accidental escape of arthropods, are crucial for health and environmental safety. With
this in mind, the American Committee of Medical Entomology (ACME), a subcommittee of the American
Society of Tropical Medicine and Hygiene (ASTMH) have drafted Arthropod Containment Guidelines
(ACG) and related materials which should be considered during the risk assessment process:
http://www.astmh.org/SIC/acme.cfm.
Currently, MCG has no areas appropriately configured as insectaries to handle any insects which could
serve as disease vectors. This limits research projects involving live insects only to those involving insects
which do generally not serve as disease vectors (e.g., Drosophila melanogaster (i.e., fruit flies)), which
may or may not carry exempt (RG-1) recombinant DNA. Even with these experiments, special precautions
should be taken to prevent accidental escape/release of laboratory insects, particularly those which have
been genetically engineered, to prevent environmental contamination.
2.7 EXPERIMENTS PROHIBITED AT MCG

The following experiments are prohibited at MCG:
Experiments using agents or organism that require containment facilities and/or equipment which
are not available at the Medical College of Georgia.
Experiments using any organism or agent that is prohibited by any federal or state agency from
importation into Georgia or for which the required permits/authorizations have not been received.
Experiments involving recombinant DNA manipulation of plants or plant pathogens
Research with venomous insects or animals
Experiments involving purposeful environmental release of any biological agent and/or
recombinant DNA materials.
Experiments with agents or organisms which are not permitted as a term of maintaining our federal
fundability status (e.g., human embryonic stem cells which are not eligible for federal funding)
Experiments which violate the ethical standards laid out by the National Institutes of Health and/or
the National Research Council of the National Academies of Sciences (e.g., see Section 2.6.3.1.1).

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3. RISK ASSESSMENT
Responsibility for biosafety exists at all levels and is shared throughout the University. The President,
Provost and Vice President of Research acknowledge the institution’s role in providing a safe workplace
and have given the Institutional Biosafety Committee (IBC) and Biosafety Office the authority to
administer the campus biosafety program. The Institutional Biosafety Committee (IBC) establishes
policies for the safe use of biohazards and for compliance with all applicable regulations and guidelines to
secure the safety of the university, personnel, research assets, the environment and the surrounding
community. The Biosafety Office acts as the agent of the IBC in ensuring compliance with these policies,
disseminating pertinent information; consulting with faculty, staff, students and visitors, and monitoring for
non-compliance.
The researchers, clinicians, students and technicians who perform work with biohazards are perhaps the
most important component of the biosafety program, as they must incorporate the biosafety requirements
and safety precautions into all facets of their work.
The Principal Investigator is ultimately responsible for safety within the laboratory (See Section 1.2.2,
Responsibilities, Principal Investigator). An integral part of this responsibility is to conduct a review of
proposed work to identify potential hazards (risk assessment) and to adopt appropriate safety procedures
before initiation of the experiments (risk management). The PI must also monitor the work to ensure that
the safety procedures are being utilized by the staff and assess whether any improvements should be made
based on logistics of the experiments or additional safety concerns.
Certain experiments require advanced registration and Biological Safety Committee approval prior to
initiation (See Section 2).
A risk assessment/risk management matrix has been prepared to illustrate key elements of the process (see
below). Relevant sections providing additional details are indicated within the matrix. Information on the
routes of exposure is included at the end of this section.
The five P’s of risk assessment and risk management are:

Pathogen – hazardous biological agent.
Procedures – proposed experimental manipulations and safe work practices.
Personnel – appropriate training and skills.
Protective equipment – protective clothing and safety equipment.
Place – laboratory design.
Consider the five P’s in each facet of laboratory work. Properly conducted, risk assessment can help
prevent exposure to biohazards and minimize the potential for laboratory acquired infection. Remember
that prior planning prevents poor performance from both a biosafety and research integrity standpoints..
After reading this section and relevant sections of the Biological Safety Manual contact the Biosafety
Office at x1-2663 or BIOSAFETY@mcg.edu for help applying the principles of risk assessment and risk
management to experimental procedures.

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3.1 RISK ASSESSMENT AND MANAGEMENT TABLE
Risk Assessment Risk Management
Pathogen • Agent classification (See Appendix B) • Registration – See Section 2 .
• Routes of infection/exposure Biosafety Office
• Infectious disease process Biological Safety Committee .
• Virulence, pathogenicity, quantity, State of Georgia- certain infectious agents
concentration, incidence in community, USDA – restricted agents or Select Agents/Toxins .
presence of vectors CDC – Select Agents/Toxins
FDA/NIH - human gene therapy
• Ensure strain identity, attenuation, replication competency
(safety tests & documentation)
Procedures • Aerosol risk: sonicating, centrifuging, • Written set of standard operating procedures (SOPs) with
homogenizing, blending, shaking, etc. safety practices incorporated
• Percutaneous risk: needles, syringes, • Adherence to basic biosafety principles
glass Pasteur pipettes, scalpels, cryostat • Label labs, areas, and equipment housing BL2 or higher
blade/knife, etc. agents
• Splash/splatter risk: pipetting, microbial • Conduct lab inspections to review practices and containment
loop, etc. equipment
• Transport/shipping of agents outside of • Use trial experiments with non-infectious material to test new
laboratory procedures/equipment
Personnel • Host immunity • Safety training

Neoplastic disease . • Prior work experience with biohazards
Infection . • Demonstrated proficiency with techniques
Immunosuppressive therapy • Prompt reporting of all exposure incidents, near misses, as
Age, race, sex, pregnancy well as signs and symptoms of related disease to PI and
Surgery (splenectomy, Employee Health & Biosafety Office
gastrectomy) • Investigation/review of incidents/spills, etc. to prevent future
Diabetes, Lupus occurrence
• Immunization
• Post-exposure prophylaxis
• Serum banking
• Attitude toward safety
• Comfort
• Open wounds, non-intact skin, eczema,
dermatitis
Protective Protection (containment) for: • Personal protective equipment (PPE):

Equipment Respirators – HEPA, N-99, N-95, etc.
• Aerosols – respirable size particles Surgical masks (for droplets)
<5µm Face (eye, nose, mouth) protection – mask and safety
• Droplets/splatter glasses, or chin length face shield .
• Sharps Solid front gown or lab coat
Gloves
• Biological safety cabinets
• Centrifuge safety buckets/rotors
• Plexiglass shielding, glove box
Place – • Risk group/biosafety level requirements • Basic lab – door, sink, surfaces easily cleaned, eyewash,
Laboratory • Aerosol risk screens on windows that open
facility • Restricted access • Labels
• Containment laboratory with directional airflow

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3.2 CONSIDERATIONS IN A RISK ASSESSMENT
In general, a risk assessment includes considerations of the following three items for all of the following
factors listed below :
a. Identification of the potential hazards in an experiment.
b. Characterization of the relative potential for the hazards to become an issue.
c. Characterization of the potential severity of the consequences if the hazard exposure did occur (to
personnel, community, environment, institutional image)
3.2.1 Risk Groups: Hazardous Characteristics of a Biological Agent

The principal hazardous characteristics of an agent are:
Its capability to infect and cause disease or pernicious response in a susceptible human or animal
host, which includes the following considerations about the agent:
• Infective dose
• Attenuation
• Allergenicity
• Physiological activity
• Oncogenicity
Is virulence as measured by the severity of disease that results from infection, which includes the
following considerations:
• Pathogenicity
• Attenuation
• Genetic modifications (toxic effects, oncogenecity, allergenicity, physiological activity)
The availability of preventive measures and effective treatments for the disease, which includes
the following considerations about the agents:
• Are there vaccinations or treatment modalities available against the agent?
• Is this a drug resistant strain?
The World Health Organization (WHO), NIH Guidelines for Research with Recombinant DNA Molecules,
and CDC/NIH publication Biosafety in Microbiological and Biomedical Laboratories (BMBL) established
a risk-group (RG) classification system based on the risks that the agent, alone, presents to the health of
healthy human adults based on an assessment of the characteristics described above. See Table 3.2.1 for
a general description of these risk groups. The risk group of an agent should be only one factor to be
considered in association with mode of transmission, procedural protocols, amount of material present,
experience of staff, and other factors in determining the appropriate biosafety containment level (BSLs)
which the work will be conducted.
Keep in mind that since these guidelines are based on the effects on healthy human adults, and do not
account for individual health considerations, such as allergies, pregnancy, breast feeding, medication
effects, a compromised immune system (due to illnesses or medical treatments such as steroids or
chemotherapy) or other illnesses which may make individuals more susceptible to agents. In addition, the
potential for differential effects of these agent in the immature systems of minors are also not considered in
these guidelines. Therefore, the guidelines represent only a starting point for an biological agent’s risk
assessments. Other known health considerations should also be factored in when performing a
comprehensive risk assessment. Also, for this reason, for their own safety, any individual with special
health concerns is strongly encouraged to discuss these with the Principal Investigator, Clinical Director or
Instructional Course Director prior to initiation of work within the laboratory. In turn, PIs, Clinical

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Directors or Instructions Course Directors should offer the opportunity for the individual to seek Employee
Health Counseling through the Employee Health Office to discuss their potential special risks (See Section
5, Medical Surveillance Program, for more information).
Additional guidance in determining the appropriate Risk Group for microbial agents can be found at:
NIH Guidelines, Appendix B
(http://www4.od.nih.gov/oba/rac/guidelines_02/APPENDIX_B.htm)
American Biological Safety Association Risk Group Database:
http://www.absa.org/XriskgroupsX/index.html
CDC/NIH Biosafety in Microbiological and Biomedical Laboratories (BMBL), particularly
Section VIII: http://www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm
Public Health of Canada MSDSs for biological agents: http://www.phac-aspc.gc.ca/msds-ftss/
Table 3.2.1 NIH Guidelines (2002) Definitions of Risk Groups
Risk Group Description of Risk of Agents

(RG)
Classification
RG 1 Agents that are not associated with disease in health human adults
RG 2 Agents that are associated with human disease which is rarely

serious and for which preventive or therapeutic interventions are
often available.
RG 3 Agents that are associated with serious or lethal human disease for
which preventive or therapeutic interventions may be available
(high individual risk but low community risk).
RG 4 Agents that are likely to cause serious or lethal human disease for
which preventive or therapeutic interventions are not usually
available (high individual risk and high community risk).
3.2.2 Routes of Exposure

In order for microbiological agents to cause disease, they must:
1. Enter or invade the body in sufficient numbers. Routes of entry include oral, respiratory, and
parenteral, via mucous membrane and/or via animal contacts (bites, scratches). See the table
below for additional information on routes of exposure or contact the Biosafety Office at x1-2663
or BIOSAFETY@mcg.edu.
Any risk assessment process should consider the possible unique routes of entry presented during
the course of the proposed experiment in addition to the risks associated with potential spread of
the infectious organism outside of the laboratory should a researcher be inadvertently infected.
Please note: these infectious routes of entry may differ. For instance, although a parasitic
infection may normally require a specific arthropod vector to transmit the infectious agents, and
the risk of having the arthropod vector within the laboratory may be extremely low, the researcher
must also consider that a parasitic infection may arise in the laboratory after accidental parenteral
introduction of the organism (via a needlestick, for instance).
It is difficult to determine a minimum infectious dose when discussing biohazards. The same
dose of a pathogen may produce no disease symptoms in one individual but may cause serious or
even fatal disease in another. There are microorganisms for which it is thought one organism
entering the body is sufficient to invade and promote the disease process; Mycobacterium

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tuberculosis, the bacteria that causes tuberculosis or Coxiella burnetii, the causative agent of Q
fever are examples. For many pathogens, 10 to 100 or more organisms must enter the body to
cause infection leading to disease.
2. Once inside the body, microbial agents must colonize in a hospitable area and establish an
infection in the host body cells, tissues and/or organs
3. Lastly, the microorganisms must overcome the host body’s natural defense mechanisms and/or
mutate or adapt to body changes to persevere within the host organism.
Other factors contribute to an individual’s susceptibility to the disease process. These include age,
immunological state, occupation, physical and geographic environment and predisposing
conditions (such as alcoholism and other drug abuse, pregnancy and diseases such as diabetes).
Therefore all factors must be considered in a comprehensive risk assessment process. When performing
risk assessments using recombinant DNA materials, all of the above factors must also be considered, in
addition to issues more specific to recombinant DNA, including the recombinant agents’ ability to replicate
once established inside the host, its ability to spread horizontally (to others or to unintended regions of the
host’s body ), and/or the ability to transfer recombinant DNA vertically via germ cells into future
generations.
Once the risks are assessed, appropriate mitigation/management measures can be developed which
specifically address the risks posed by the agents and how these will be used (See Section 4, Risk
Management, for further information).

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Routes of Transmission for Infectious Agents

3-6
3.2.3 Operations which may be associated with additional risk of exposure
The following laboratory operations are associated with additional risk for aerosol, splash and/or spray
production. Therefore, additional risk considerations must be factored in during the risk assessment, and
these activities must be documented on all Biosafety Protocols for the IBC to review.
• Blowing out pipettes • Homogenizing

• Cell sorters (FACS) • Blending
• Shaking , vortexing or stirring tubes • Grinding
• Opening lyophilized cultures • Cell disruption with French press
• Opening snap top tubes • Intranasal inoculation of animals
• Breakage of culture containers • Cage cleaning, changing animal
• Flaming loops or slides bedding
• Pulling needles out of septums • Harvesting infected material from
• Filling a syringe animals, eggs, and other virology
• Placing liquids under pressure procedures
• Pouring liquids • Necropsies of infected animal
• Centrifugation
• Sonication
3.2.3.1 Zoonotic Disease Risks/Animal Research Risks

Use of animals and animal tissues may pose additional inherent risks due to the somewhat
uncontrollable nature of animals as well as the potential risk of exposure to zoonotic diseases,
which are diseases which are communicable from animals to humans and in many cases, visa
versa, under natural conditions. Some animals are natural carriers of infections which may be
associated with mild symptoms in animals, and therefore difficult to diagnose; however, some of
these diseases can be quite serious or lethal in humans. Several of these illnesses may pose a
particular hazard to pregnant women or their developing fetuses (Q-fever, toxoplasmosis);
therefore pregnant workers should consider discussing their special risks and protective measures
with a physician. Familiarization of the potential symptoms of the zoonotic diseases which may
be carried by the research animals used in the laboratory should be part of the laboratory-specific
training/education measures provided to all personnel working with the materials and should be
communicated to any health care provider, should any exposures or symptoms present. Protective
measures should always be addressed in each laboratory’s standard operating procedures (SOPs)
to mitigate the risk of these diseases. A recommended initial reference for information on
zoonotic diseases and their associated pathogens is the interactive tables developed by the Air
Force Public Health Services: http://phsource.us/PH/ZD/DiseasesTable.htm.
Additional risks associated with working with animals may include those related simply with the
logistics for working in animal facilities. Heavy and large equipment is often used in these
facilities, such as cage racks or changing stations, and the metal caging can often become bent in
the day-to-day activities in the animal laboratory. These additional risks, including risks for crush
injuries or lifting hazards must also be considered by the Facility Director.
3.2.3.1.1 Non-human primates

Because of primates evolutionary similarities to humans, many of the diseases that affect
one species can affect the other; albeit, often with different symptoms. Care and
consideration must not only be given to the protection of the workers while working with
NHP, but also to protecting the NHP colonies to infections brought in my human

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caregivers and researchers, which can quickly devastate a primate colony. A minimal
BSL-2 containment facilities and safety measures should be followed when working with
NHP and materials derived from NHP.
3.2.3.1.1.1 Cercopithecine Herpes Virus (CHV-1); Herpes B virus

Most species of macaque monkeys (rhesus, cynomolgus, pigtail) can carry a
virus known as B virus, Herpesvirus simiae or Cercopithecine Herpes Virus I
(CHV-1). CHV-1 is to primates what human herpes simplex virus-1 (i.e. the
cold sore virus) or -2 (i.e. genital herpes) are to humans. In monkeys, it is a
relatively mild disease; most of the time, one cannot tell whether a monkey is
infected by sight. The virus tends to "lie dormant" in the body, but it can cause
"breakouts" with occasional mucous membrane or genital lesions, especially
during periods of high stress. It's during the "breakouts" that monkeys can shed
the virus and potentially expose humans.
Unfortunately, in humans CHV-1 has a mortality rate somewhere in the

neighborhood of 64-80% if left untreated. Infection in humans is rare, but most
cases have involved contact with nonhuman primates. Immediate diagnosis and
treatment with antiviral drugs has saved lives.
Since B virus has a high prevalence in conventional primate colonies, and

animals can carry the disease without showing symptoms, it is best to assume
that all primates are infected. Body fluids such as saliva and urine present the
highest risk to workers. Caretakers are at special risk, since routine care and
cage washing involves frequent contact with sharp edges on contaminated cages
and production of aerosols from spraying animal waste.
In laboratories, all materials of non-human primates should be handled under

very strict BSL-2 containment. Activities such as pipetting, opening containers
and centrifuging urine, blood or mucosal swabs can present risk of
contamination.
CHV-1 is fairly stable outside of the body (estimates suggest it can remain
viable for up to 7 days at 37°C and weeks at 4°C). However, it can be
inactivated by 70% ethanol, freshly diluted bleach (1:5), heating (50-60 C for
>30 min), acidic pH's and/or detergent solutions.
Personnel working with NHP will be required to attend a special NHP training
class offered by DLAS Veterinarians. Use of protective clothing, eye protection
and respiratory protection are essential when working with NHP or their
materials. Consistent caution and respect for the monkey is always a good idea.
In addition, if you ever spot what looks like a cold sore on a monkey, be sure to
contact a DLAS veterinarian before handling the animal or its cage. Any even
potential exposure should be assessed and treated by a medical professional
immediately. See incident response guidance and contact DLAS Veterinarians
(x1-3421) and/or the Biosafety Office (x1-2663 or BIOSAFETY@mcg.edu) for
assistance.
Further information about CHV-1 and measures for prevention, please see the
following guidance:
Public Health Service of Canada MSDS on CHV-1: http://www.phac-

aspc.gc.ca/msds-ftss/msds81e.html
Cohen JI, Davenport DS, Stewart JA, Deitchman S, Hilliard JK,

Chapman LE and the B Virus Working Group (2002).

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Recommendations for prevention of and therapy for exposure to B
virus (cercopithecine herpesvirus 1). Clin Infect Dis. 35(10):1191-203.
(A copy of this paper can be downloaded from MCG computers from
the following URL:
http://web.ebscohost.com/ehost/detail?vid=3&hid=5&sid=da4a1eb6-
61c4-475e-b22a-a5d1b9e3058d%40sessionmgr103 )
3.2.3.1.1.2 Other viral infections associated with NHP
Nonhuman primates can develop measles, usually following exposure to

infected humans. Workers should be vaccinated against measles. Workers with
measles should not work with nonhuman primates until they have recovered.
Clinical signs in the animals are similar to those in humans, including a red rash
on the face and hands. Transmission is usually by direct contact. More
information on measles and measles virus can be viewed at: http://www.phac-
Influenza virus can be transmitted between humans and nonhuman primates.

Signs of disease for both humans and animals include fever and upper
respiratory signs such as coughing, sneezing and nasoocular secretions. The
disease is potentially fatal in immunocompromised individuals. Transmission is
by aerosol. Prevention is achieved through the use of protective clothing and
respiratory protection. More information about influenza can be found at the
CDC informational URL: http://www.cdc.gov/flu/ .
Several pox viruses, including monkeypox and Yaba virus can cause painful
nodular lesions in the skin of humans and nonhuman primates. Transmission is
by direct contact, although infected tissues could present a risk to laboratory
workers. Prevention of disease is through use of protective clothing. More
information on monkey pox can be seen at:
http://www.cdc.gov/ncidod/monkeypox/
Nonhuman primates may be carriers of filoviruses

(http://www.cdc.gov/Ncidod/dvrd/spb/mnpages/dispages/filoviruses.htm) such
as Ebola virus (http://www.phac-aspc.gc.ca/msds-ftss/msds53e.html). These
viruses cause an acute hemorrhagic fever in humans and nonhuman primates
that is often fatal. Flood and body fluids are potential sources of the virus. Use
of protective clothing and eyewear is preventative. Animals are screened upon
entry into the country for filoviruses.
3.2.3.1.1.3 Gastrointestinal Diseases associated with NHPs
Bacterial agents such as Salmonella (http://www.phac-aspc.gc.ca/msds-

ftss/msds135e.html), Shigella (http://www.phac-aspc.gc.ca/msds-
ftss/msds139e.html) and intestinal parasites such as Giardia lamblia
(http://www.phac-aspc.gc.ca/msds-ftss/msds71e.html), Entamoeba histolytica
(http://www.phac-aspc.gc.ca/msds-ftss/msds58e.html), and Balantidium coli
(http://www.phac-aspc.gc.ca/msds-ftss/msds15e.html) can be carried by many
species of nonhuman primates. Feces are the primary source of infections.
Gloves, a laboratory coat and a respiratory mask should be worn when handling
fecal samples or sanitizing caging. Symptoms of these diseases include diarrhea
with or without blood, vomiting, and fever in both humans and nonhuman
primates. Medical attention should be sought immediately.

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3.2.3.1.1.4 Bacterial Respiratory Diseases associated with NHPs
Streptococcus pneumonia can cause an acute bacterial pneumonia in humans

and nonhuman primates. Infected animals and lung or sputum samples can
present a risk to workers. Sudden onset of fever, malaise and coughing are
symptoms of bacterial pneumonia. Medical attention should be sought
immediately.
Tuberculosis caused by Mycobacterium tuberculosis (http://www.phac-

aspc.gc.ca/msds-ftss/msds103e.html) infection causes an extremely rapidly fatal
pneumonia in most Old World primate species, and chronic pneumonia in
humans. Typically, infected humans present a much greater risk to the animals
than animals do to humans. It would be unusual for the disease to be transmitted
to humans, unless the animal is undergoing surgery or pathologic examination.
Infected tissue samples can also present a risk to laboratory workers. Prevention
of disease includes routine use of respiratory protection and protective clothing
when working with tissues or when coming into close proximity to animals.
Animals and human handlers should be screened every 6 months for disease.
Animals should also be quarantined and screened on entry into the facility.
3.2.3.1.2 Birds
Birds carry diseases such as psittacosis and an avian form of tuberculosis. Only
inspected, properly quarantined birds should be used in research studies or teaching
demonstrations. Mycological fecal contamination is also frequent. The causative agents
for some common avian transmitted diseases are described below and the following links
describe some of the potential illnesses associated with birds. Contact the Biosafety
Office or Division of Laboratory Animals Services for further information.
• Chlamydia psittaci: http://www.phac-aspc.gc.ca/msds-ftss/msds31e.html

• Histoplasma capsulatum: http://www.phac-aspc.gc.ca/msds-ftss/msds82e.html
• Cryptococcus neoformans: http://www.phac-aspc.gc.ca/msds-ftss/msds47e.html
• Mycobacterium avium: http://www.phac-aspc.gc.ca/msds-ftss/msds102e.html
Transmission to humans occurs by exposure via the inhalation route for the fungal
infections (Histoplasma, Cryptococcus) due to inhaling spores. Contact with tissues
through cuts or scratches may also pose a risk. Another route of exposure may be surface
contact while handling avian fecal specimens.
Those at risk include investigators, animal caretakers, laboratory personnel, or others who
routinely handle birds, their tissues, and feces. Scratches or cuts involving birds or
injuries from objects contaminated with body fluids or feces from birds require
immediate first aid and medical attention.
Gloves, masks and a laboratory coat (or other dedicated protective clothing such as a
scrub suit) should be worn when working with birds. In some cases protective eye wear is
also indicated. Do not eat, drink, or apply cosmetics while working in an aviary, and
always wash your hands after handling birds. Remember that unfixed tissues, body fluids,
and other materials derived from birds may also pose a risk. Guano (feces), hair and
feathers may also exacerbate allergies.

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3.2.3.1.3 Dogs (Canines)
Dogs used in research have been vaccinated against rabies. However, it may be prudent
to consider prophylactic immunization.
Bites and scratches may also pose serious problems through trauma and/or bacterial
infection. Dogs may also have enteric bacteria such as Salmonella released in the feces so
cage washers and any personnel who must clean bedding should wash hands with a
disinfectant hand soap before leaving the facility. Dogs, like most mammals, can shed fur
so anyone with allergies to fur, dander or animal bedding should wear personal protective
clothing to minimize discomfort. Dogs may also carry biting insects, such as fleas, so
personal protective equipment may also be used in this instance as well.
The following links describe some of the potential illnesses associated with dogs and may
be found on-line:
• Brucella canis: http://www.phac-aspc.gc.ca/msds-ftss/msds23e.html

• Campylobacter jejuni: http://www.phac-aspc.gc.ca/msds-ftss/msds29e.html
• Rabies: http://www.phac-aspc.gc.ca/msds-ftss/msds124e.html
scrub suit) should be worn when working with dogs. In some cases protective eye wear is
also indicated. Do not eat, drink, or apply cosmetics while working in an animal use area,
and always wash you hands after handling dogs. Remember that unfixed tissues, body
fluids, and other materials derived from dogs may also pose a risk.
Bites or scratches involving dogs or injuries from objects contaminated with body fluids
from dogs require immediate first aid and medical attention.
3.2.3.1.4 Cats (Felines)

Bites and scratches may also pose serious problems through trauma and/or bacterial
infection. Cats may also release microorganisms such as Salmonella and parasites such as
Toxoplasma in the feces so cage washers and any personnel who must clean bedding
should wash hands with a disinfectant hand soap before leaving the facility. Cats, like
most mammals, can shed fur so anyone with allergies to fur, dander or animal bedding
should wear personal protective clothing to minimize discomfort. Cats may also carry
biting insects, such as fleas, so personal protective equipment may also be used in this
instance as well.
The following links describe some of the potential illnesses associated with cats and may
be found on-line:
• Bartonella henselae (Cat Scratch Fever):

http://www.nlm.nih.gov/medlineplus/ency/article/001614.htm
• Rabies: http://www.phac-aspc.gc.ca/msds-ftss/msds124e.html
• Toxoplasma gondii: http://www.phac-aspc.gc.ca/msds-ftss/msds153e.html
Bites or scratches involving these species or injuries from objects contaminated with
body fluids from cats require immediate first aid and medical attention. Pregnant women
need to be aware that toxoplasmosis, caused by infection with Toxoplasma gondii can
cause problems with pregnancy, including abortion and should address these risks with
the Employee Health Office, Division of Laboratory Animal Services or Biosafety
Office, as desired.

3-11
scrub suit) should be worn when working with cats. In some cases protective eye wear is
also indicated. Do not eat, drink, or apply cosmetics while working in an animal use area,
and always wash you hands after handling cats. Remember that unfixed tissues, blood,
serum, urine, and other materials derived from cats may also pose a risk.
3.2.3.1.5 Rabbits (Lagomorphs)

Rabbits raised in "clean" facilities are considered relatively free of zoonotic diseases
unless they have been experimentally or accidentally infected with a human pathogen.
However, those working with rabbits should also be aware of possible allergic reactions.
These allergic reactions are often associated with cage cleaning due to the dust hazards of
bedding and surface contact with urine and other waste materials.
The following links describe some of the potential zoonotic illnesses which may be
associated with wild rabbits and may be found on-line:
• Bordetella: http://www.phac-aspc.gc.ca/msds-ftss/msds19e.html
• Pasteurella : http://www.phac-aspc.gc.ca/msds-ftss/msds117e.html
• Tularemia (Francisella tularensis): http://www.phac-aspc.gc.ca/msds-
ftss/msds68e.html
Bites or scratches involving these rabbits or injuries from objects contaminated with body
fluids from rabbits require immediate first aid and medical attention.
scrub suit) should be worn when working with rabbits. In some cases protective eye wear
is also indicated. Do not eat, drink, or apply cosmetics while working in an animal use
area, and always wash you hands after handling rabbits. Remember that unfixed tissues,
blood, serum, urine and other materials derived from rabbits may also pose a risk.
Bedding, hair and fur may also exacerbate allergies.
3.2.3.1.6 Rodents (mice, rats, hamsters, gerbils, guinea pigs)

Rodents raised in "clean" facilities are considered relatively free of zoonotic diseases
unless they have been experimentally or accidentally infected with a human pathogen.
Note: those working with rodents should also be aware of possible allergic reactions.
These allergic reactions are often associated with cage cleaning due to the dust hazards of
bedding and surface contact with rodent urine proteins.
Wild rodents pose additional concerns. Wild-caught animals may act as carriers for such
viruses as hantavirus and lymphocytic choriomeningitis (LCMV) depending on where
they were captured. Additionally, each rodent species may harbor their own range of
bacterial diseases, such as tularemia and plague. These animals may also have biting
insect vectors which can act as a potential carrier of disease (mouse to human
transmission).
The following links describe some of the potential zoonotic illnesses associated with
rodents and may be found on-line:
• Lymphocytic Choriomeningitis Virus (LCMV) : http://www.phac-

• Hantaviruses (Hemorrhagic fever with renal syndrome or Korean hemorrhagic
fever): http://www.phac-aspc.gc.ca/msds-ftss/msds74e.html
• Tularemia (Francisella tularensis): http://www.phac-aspc.gc.ca/msds-
ftss/msds68e.html

3-12
• Plague (Yersinia pestis): http://www.phac-aspc.gc.ca/msds-ftss/msds169e.html
Bites or scratches involving these rodents or injuries from objects contaminated with
body fluids from rodents require immediate first aid and medical attention.
3.2.3.1.7 Ferrets
Commercially-raised laboratory ferrets are typically free of infections that could pose a
risk to humans. Disease development from typical exposure to laboratory ferrets is not
recognized as a significant public health risk. Risk of rabies is minor due to pre-arrival
and on-site conditioning of ferrets. Zoonotic agents that can be transmitted by ferrets
include:
• Salmonella spp. (http://www.phac-aspc.gc.ca/msds-ftss/msds135e.html )

• Campylobacter spp. (http://www.phac-aspc.gc.ca/msds-ftss/msds29e.html )
• Cryptosporidia (http://www.phac-aspc.gc.ca/msds-ftss/msds48e.html )
• Giardia spp. (http://www.phac-aspc.gc.ca/msds-ftss/msds71e.html )
• Leptospira (http://www.phac-aspc.gc.ca/msds-ftss/msds95e.html )
• Influenza (http://www.cdc.gov/flu/about/disease.htm )
Ferrets are very susceptible to influenza viruses and have served for years as an animal
model in the laboratory. In ferrets flu is characterized by sneezing, fever, lethargy,
mucoserous nasal discharge, conjunctivitis and photophobia. The course of the influenza
infection usually lasts less than a week. The disease can be severe in young ferrets.
Human cases of influenza have occurred from contamination by aerosols from infected
ferrets. Similarly ferrets can be infected by humans shedding the virus.
Ferrets should not be allowed to roam freely, and their feces should be discarded in a
hygienic manner. They also share parasites with dogs and cats (Toxocara, Dipylidium) as
well as dermatophytosis (Microsporum canis, T. mentagrophytes).
Risk of exposure of workers to zoonotic materials may result from the following
activities: changing animals from dirty (exposed to animals or their wastes) to clean
cages; handling animals for injections, surgery, etc.; handling dirty (exposed to animals
or their wastes) animal room supplies; interacting with people who have entered animal
areas and have not changed clothes or showered; and eating and touching the face with
contaminated hands (exposed to animals or their wastes). There is a moderate risk of
injury from ferrets bites or scratches. Bites or scratches or other potential exposures or
injuries involving ferrets or objects contaminated with body fluids from ferrets require
immediate first aid and medical attention
3.2.3.1.8 Pigs/Swine
Swine harbor a range of parasites and diseases that can be transmitted to humans. These
include:
• Trichinosis (http://www.phac-aspc.gc.ca/msds-ftss/msds155e.html)
• Cysticercosis
(http://www.cdc.gov/ncidod/dpd/parasites/cysticercosis/default.htm)
• Brucellosis (http://www.phac-aspc.gc.ca/msds-ftss/msds23e.html)
• Salmonella spp. (http://www.phac-aspc.gc.ca/msds-ftss/msds132e.html and
http://www.phac-aspc.gc.ca/msds-ftss/msds135e.html )
• Pathogenic E. coli (http://www.phac-aspc.gc.ca/msds-ftss/msds23e.html).

3-13
Pigs are also known to host large concentrations of parasitic ascarid worms
(http://www.phac-aspc.gc.ca/msds-ftss/msds10e.html) in their digestive tract.
Pigs can be susceptible to pneumonia, usually caused by weather. Pigs have small lungs
in relation to body size; for this reason, bronchitis or pneumonia can kill a pig quickly.
Pigs can be aggressive and pig-induced injuries are relatively common in areas where
pigs are reared or where they form part of the wild or feral fauna. Their relatively large
size and weight also pose a physical hazard for animal care givers and researchers.
scrub suit) should be worn when working with swine. In some cases protective eye wear
is also indicated. Do not eat, drink, or apply cosmetics while working in an animal use
area, and always wash you hands after working with animals. Remember that unfixed
tissues, blood, serum, urine and other materials derived from swine may also pose a risk.
Bedding, hay, dust and hair may also exacerbate allergies. Bites or scratches involving
swine or injuries from objects contaminated with body fluids from swine require
immediate first aid and medical attention
3.2.3.1.9 Large Hooved Mammals (Cows, Horses, Sheep, Goats)

The size of hooved mammals pose additional concerns for researchers, due to the
physical hazards of weight and strength of the animal. Hooved mammals may resist
handling and may require multiple workers to administer medication or other functions.
With regard to pathogens, sheep are known to shed a rickettsia, Coxiella burnetii, that is
the causative agent for Q-Fever. Ruminants and pigs may harbor their own range of
bacterial pathogens and parasites, such as Salmonella, Campylobacter and
Cryptosporidium. Skin conditions, such as Erysipelas and Orf may result after contact
with pigs and sheep and goats, respectively. In addition, these animals may carry biting
insect vectors who can act as a potential carrier of disease.
The following links describe some of the potential illnesses associated with hooved
mammals, farm animals, and may be found on-line:
• Coxiella burnetii/ Q-fever (http://www.phac-aspc.gc.ca/msds-

ftss/msds43e.html)
• Salmonellosis (http://www.phac-aspc.gc.ca/msds-ftss/msds132e.html and
http://www.phac-aspc.gc.ca/msds-ftss/msds135e.html )
• Orf (“Sore Mouth” infection) http://www.cdc.gov/ncidod/dvrd/orf_virus/
• Pasteurella Pneumonia (http://www.phac-aspc.gc.ca/msds-ftss/msds117e.html )
• Campylobacter jejuni (http://www.phac-aspc.gc.ca/msds-ftss/msds29e.html)
• Brucella spp./Brucelosis (http://www.phac-aspc.gc.ca/msds-ftss/msds23e.html)
Bites or scratches involving these species or injuries from objects contaminated with
body fluids from hooved mammals require immediate first aid and medical
attention.
scrub suit) should be worn when working with hooved mammals. In some cases
protective eye wear is also indicated. Do not eat, drink, or apply cosmetics while working
in an animal use area, and always wash you hands after working with hooved mammals.
Remember that serum, urine and other materials derived from hooved mammals may also
pose a risk. Bedding, hay, dust and hair may also exacerbate allergies.

3-14
3.2.4 Amount of Material Present
High quantities of infectious material pose a greater risk of spread than small quantities. Large volumes of
cultures require additional considerations of the risk of spills, splashes, and laboratory logistics. These
considerations should be factored into a comprehensive risk assessment.
Cultures of recombinant materials in volumes exceeding 10 liters have specific biosafety standards which
should be followed, as described in Appendix K of NIH Guidelines
(http://www4.od.nih.gov/oba/rac/guidelines_02/Appendix_K.htm)
3.2.5 Experience of Staff

MCG personnel receive basic biosafety training during the IBC-required initial training class and annual
refresher training modules. However, specialized training in laboratory-specific operations is the
responsibility of the Principal Investigator.
If a laboratory is utilizing technology which no one, including the PI, has previous experience in (e.g.
recombinant viral vector systems or use of new equipment, such as a French Press), additional training
should be sought by the Principal Investigator before embarking on these experiments. Often the Biosafety
Office can be of assistance in providing additional training, or identifying appropriate trainers for the
laboratory. Contact the Biosafety Office (x1-2663 or BIOSAFETY@mcg.edu) for assistance.
3.2.6 Mode of Transmission

Infectious diseases may spread between people via different modes. Some, like the diseases associated
with the most common bloodborne pathogens (HIV, Hepatitis B & C) require contact with blood via open
wounds, needlestick or sexual transmission. This makes these diseases inherently more difficult to spread
than those transmitted via an airborne route of transmission, such as influenza. The extent of risk to the
public health and environment may factor in to the natural modes of transmission of diseases which may
result from inadvertent infection of individuals.
3.2.7 Environmental Stability

Because of their resistance to common disinfectant measures and their heat stability, biological agents such
as endospore-forming species of bacteria (e.g., strains of Bacillus, Clostridium), and cyst-forming
protozoan parasites pose an additional risk than agents which are more easily decontaminated. See Section
7 on for further information on decontamination.
3.2.8 Institutional Public Image

Some biological agents may present additional considerations based on the perceived risk that others may
assess who are outside of the laboratory or the university. A good example of this might be a laboratory
who wishes to work with the avirulent Sterne strain of Bacillus anthracis. Although this strain is
commonly used as a vaccination in animals, and may be available through veterinary suppliers, and poses
very little risk of infection in healthy adults, if someone became aware that a laboratory works with
anthrax, a higher levels of risk may be perceived than actually exists. Anticipation and addressing these
sorts of concerns prior to use of these agents may be necessary in order to maintain the institution’s public
image.

3-15
4. RISK MANAGEMENT: BIOSAFETY LEVELS
Management of the risks associated with research involving biological materials is accomplished via a
combination of Practices, Safety Equipment and Facilities. Biosafety Levels (BSLs) refer to the level of
containment which is required and appropriate to contain the risk as assessed as described in Section 3, Risk
Assessment. The CDC, WHO and NIH have established standards for four biosafety containment levels (BSLs)
for work with all biohazardous materials in the publication Biosafety in Microbiological and Biomedical
Laboratories (BMBL) http://www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm. These standards are reflected
(although not always identical) to the four biosafety containment levels (BLs) described for work with
recombinant DNA materials in Appendix G of the NIH Guidelines for Research with Recombinant DNA
Molecules (“NIH Guidelines”) http://www4.od.nih.gov/oba/rac/guidelines/guidelines.html. Both publications provide
general descriptions of the combination of microbiological practices, laboratory facilities, and safety equipment
as well as their recommended uses in order to contain biological agents which may be infectious to humans, or
impact the environment.
Because working with hazardous materials in animals imposes additional risks to the laboratory worker or the
environment and laboratory logistics, there are also standards for four biocontainment levels for research with
biohazardous materials and animals or ABSLs (Section V of BMBL for all biohazardous material
http://www.cdc.gov/od/ohs/biosfty/bmbl5/sections/SectionV-VertebrateAnimalBiosafetyLevelCriteria.pdf ) or
BL-Ns (in Appendix Q of NIH Guidelines for work with recombinant materials
http://www4.od.nih.gov/oba/rac/guidelines_02/Appendix_Q.htm ). Similarly, the American Committee of
Medical Entomology of the American Society of Tropical Medicine and Hygiene has also developed arthropod
containment levels (ACL1-4) for research with arthropods (http://www.astmh.org/SIC/acme.cfm).
NIH Guidelines also has developed specific biosafety containment levels for large scale cultures (>10 liters)
BL-Large Scale in Appendix K (http://www4.od.nih.gov/oba/rac/guidelines_02/Appendix_K.htm).
These Biosafety containment level standards are intended as a guideline for management of the risks as
determined during the risk assessment. However, this is not a prescriptive, formulaic process. Mitigation and
management methods should always provide prudent measures to contain the materials and protect exposure of
the users to the materials based on the risks assessed. Therefore, since the risks in each laboratory are unique to
the laboratory, the Standard Operating Procedures (SOPs) should be specific for the laboratory, agents,
operations and practices within the laboratory. Protective measures to block the routes of transmission, based
on the operations involved and locations must be reviewed (See Figure 4, below for suggestions of appropriate
mitigation methods to prevent exposure via the common routes of transmission within the laboratory). Keep in
mind that on occasion, a risk may be present which may require an additional or alternative protective measure
(e.g., development of a special practice or procedure, use of special equipment or restriction to a facility) to
protect personnel and the environment from the biological agents proposed for use. These may go beyond those
explicitly stated in the BMBL or NIH Guideline standards; however, it is the IBC’s responsibility (as per NIH
Guidelines) to ensure the assessed risks involved in a proposed experiment have been reduced to an acceptable
level based on the mandated comprehensive risk assessment. Therefore, the IBC may stipulate special
containment measures as a condition of receiving approval.
Below is a summary of practices, equipment and facility requirements for agents assigned to basic biosafety
levels 1–4 (BL 1–4) (Table 4). Additional information on biosafety levels may be found in Appendix C of this
as well as in the BMBL at: http://www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm or NIH Guidelines at:
Only work at biosafety levels 1-2 is currently permitted at MCG. Most laboratories at the Medical College of
Georgia work under BSL-2 containment levels. Work at BSL-3 containment levels will require special
considerations, training and the standards will be documented in the MCG BSL-3 Biosafety Guide. No
Biosafety level 4 work is allowed at MCG.

4-1
Table 4. Summary of Recommended Biosafety Levels for Infectious Agents
Biosafety Agents Practices Safety Equipment Facilities (Secondary

Level (Primary Barriers) Barriers)
RG1; Not known to Standard Microbiological None required. Door required on lab
cause disease in Practices. Handwash sink must be
1 healthy adults. available
RG2; Associated BSL-1 practice plus: Class I or II BSCs or other BSL-1 plus: Autoclave or
with human disease, • Limited access to lab physical containment devices other method for
hazards are auto- • Biohazard warning used for all manipulations of decontamination available.
inoculation, signs posted agents that cause splashes or
ingestion, mucous • Sharps precautions aerosols of infectious
membrane • Biosafety manual materials;
2 exposure. defining any needed
waste & surface PPE: laboratory coats;
decontamination or gloves; face protection as
medical surveillance needed.
policies.
RG3; Indigenous or BSL-2 practice plus: Class II BCSs used for all BSL-2 plus:
exotic agents with • Restricted access manipulations of agents (no • Physical separation from
potential for aerosol • Decontamination of all open benchwork) access corridors
transmission; waste before leaving • Self-closing, double-
disease may have facility PPE: back-closing gowns or door access;
serious or lethal • Decontamination of lab jump suits, shoe covers, • Air exhausted from
3
consequences. clothing before gloves; respiratory protection facility, not recirculated
laundering; as needed. • Negative airflow must be
• Baseline serum maintained into
sampling as laboratory
appropriate • Sealed penetrations
RG4; Dangerous or BSL-3 practices plus: All procedures conducted in BSL-3 plus:
exotic agents which • Clothing change Class III BSCs or Class I or II • Separate building or
pose high risk of before entering; BSCs in combination with isolated zone;
life-threatening • Shower on exit full-body, air-supplied, • Dedicated
disease, aerosol- • All material positive pressure personnel supply/exhaust, vacuum,
transmitted lab decontaminated on exit suit. and decon systems;
4 infections; or related from facility. • Double-redundancy of
agents with facility systems
unknown risk of • Other requirements
transmission. outlined in BMBL.
Adapted from the Office of Health and Safety, Centers for Disease Control and Prevention & Yale University

4-2
Figure 4. Protection for the Routes of Transmission
Route of Transmission Protection

Mucous Membranes Achieve Face Protection:
Through mucous membranes or the eyes Wearing full-face shield or safety glasses and
nose or mouth (splash, splatter). surgical mask
Working in a Biosafety cabinet or behind
protective shields
Following good microbiological practices.
Ingestion Prevent exposure via ingestion, by:

Mouth pipetting, eating, drinking, smoking in Not eating, drinking, chewing gum, smoking
the lab. in lab
Mechanical pipettors
No storage of food items or utensils in lab
Following good microbiological practices
Inhalation Avoid exposure to aerosols by:

Breathing in respirable sized aerosols Working in a Biosafety Cabinet
(<5µm), centrifuge leaks, spills, & Using sealed rotors or safety caps for
aerosol-generating procedures such as centrifuge buckets
pipetting, homogenizing, etc. Safety containment equipment
HEPA filtered respirator
Following good microbiological practices.
Percutaneous Avoid percutaneous exposures by:
Through intact or non-intact Using extreme precautions with sharps

skin via needlestick, puncture Using needleless, retractable or safety sharp
with a contaminated sharp systems
object, animal scratch, bite, Disposing sharps immediately in rigid
through wounds, abrasions, or leakproof sharps container placed near site of
use
Using animal restraints
Using cut/bite resistant gloves
Using sleeve covers, water proof bandages
over any open wounds
Using double gloves
Using good work practices
Contact (indirect transmission) Prevent indirect exposure by:
Via mucous membranes or non-intact Decontamination of work surfaces
skin from hands that have been in Maintaining good personal hygiene (avoid
contact with a contaminated surface touching your face or other unprotected skin
(i.e. benches, phones, computers, with glove or non-gloved hands)
equipment handles) or by failure to Avoid touching non-contaminated areas (e.g.
wash hands after working. phones) with contaminated hands.
Not applying cosmetics within the laboratory.
Removing gloves and wash hands before
exiting laboratory
Frequent glove changing

4-3
4.1 LABORATORY PRACTICES
In this section, an attempt has been made to provide information regarding hazards involved with certain
laboratory practices and methods for preventing them. Prevention is an important element to biohazard control,
and it is recommended that anyone working in a laboratory read this section carefully. It is the responsibility of
the Principal Investigator, Clinical Director and/or Instructional Course Designer or their designees to establish
their laboratory-specific Standard Operating Procedures to address the risks within each laboratory. These
should be documented as part of the Biosafety Protocol (BSP) Application process and these documents should
provide a basis to begin laboratory-specific safety training for all laboratory personnel prior to initiation of
work.
4.1.1 Human Factors and Attitudes in Relation to Laboratory Accidents
For the purpose of safety, an attitude can be defined as an accumulation of information and experience that
predisposes an individual to certain behavior. Human factors and attitudes result in tendencies on the part of the
individual to react in a positive or negative fashion to a situation, a person or an objective. Laboratory
supervisors and Principal Investigators should understand the importance of attitudes and human factors in their
own efforts to control biohazards in their laboratory. Some observations that may be of help to supervisors are
listed below:
The lack of accident perception ability is often a significant factor in laboratory accidents.
Inflexibility of work habits, that tend to preclude last minute modification when an accident
situation is recognized, plays a part in the causation of some laboratory accidents.
Working at an abnormal rate of speed is a significant causal factor.
Intentional violations of regulations are a frequent cause of accidents. This is termed excessive
risk taking.
The performance of routine procedures such as diluting and plating cultures is the most frequent
task being performed at the time of laboratory accidents.
Working when one is very tired is more likely to create a higher potential for accidents.
Working at a well-organized and uncrowded laboratory bench will help in the prevention of lab
accidents.
Working alone or without supervision in a laboratory with higher hazard agents should be
discouraged.
Each employee working with biohazardous agents must be constantly aware of the importance of the proper
attitude in preventing accidents in the laboratory.
4.1.2 Practices Associated with Biosafety Levels 1 through 2+ Laboratories
4.1.2.1 Biosafety Level 1 (BSL-1/BL-1)

As defined in:
Section IV of the CDC BMBL:
http://www.cdc.gov/od/ohs/biosfty/bmbl5/sections/SectionIV-LaboratoryBiosafetyLevelCriteria.pdf
Appendix G-II-A for recombinant DNA BL-1:
http://www4.od.nih.gov/oba/rac/guidelines_02/Appendix_G.htm#_Toc7246561

4-4
Keep laboratory door closed when experiments are in progress. There should be floor-to-
ceiling physical separation from “laboratory areas” and “non-laboratory areas”. Therefore,
any doors leading into non-laboratory areas, such as offices or hallways must be kept shut
when experiments are in progress.
Use procedures that minimize aerosols. For example, pipette gently along the sides of tubes
to prevent aerosol production. Open vacuum pressured containers/vials carefully using
protective covers to avoid exposures to any aerosols that may arise from the back-pressure
that may result.
Eating, drinking, smoking, handling contact lenses, applying cosmetics, and storing food for
human consumption must not be permitted in laboratory areas. Food must be stored outside
the laboratory area in cabinets or refrigerators designated and used for this purpose.
Wear appropriate PPE:

Gloves must be worn to protect hands from exposure to hazardous materials. Glove
selection should be based on an appropriate risk assessment. Alternatives to latex
gloves should be available.
Laboratory gowns or coats should be worn over street clothes.
Protective eyewear should be worn when conducting procedures that have the
potential to create splashes of microorganisms or other hazardous materials. Persons
who wear contact lenses in laboratories should also wear eye protection.
Gloves should be changed when contaminated, their integrity have been compromised and/or
when otherwise necessary. Periodic glove-changing is recommended. Be aware that some
petroleum-based hand moisturizers can impact the integrity of latex gloves and should be
avoided.
Gloves should be removed and hands washed after completing experimental procedures and
before leaving the laboratory. Hand washing protocols must be rigorously followed. Soap
should be used and hands washed for approximately 20 seconds prior to rinsing with warm
water.
Dispose of used gloves with other contaminated laboratory waste after removal. Do not wash,
save and/or reuse disposable gloves.
Do not mouth pipette. Use mechanical pipetting devices.
Avoid using hypodermic needles, scalpels, blades, glass or other sharp items whenever
possible. Consider alternatives or devices with safety features, for instance retractable
blades, or substitute plastic for glass whenever feasible.
Disinfect work surfaces daily and immediately after a spill with an appropriate disinfectant (as
documented in the laboratory SOPs). Use of taped-down benchkote paper is discouraged,
since this is often not changed daily.
Decontaminate all biological wastes before discarding. Decontaminate other contaminated

materials before washing, reusing, or discarding.
For off-site decontamination, package contaminated materials in closed, durable, leakproof

containers before leaving the laboratory. See Section 4.1.5 for more information about
appropriate transport procedures.
Ensure that all containers which contain biological waste are labeled with the biohazard
symbol. This includes intermediary beakers or bins used on bench-tops to contain pipette
tips or small microcentrifuge tubes which may have been used with biological materials.

4-5
Control insect and rodent infestations.
Keep areas neat and clean.
The laboratory supervisor must ensure that laboratory personnel receive appropriate training
regarding their duties, the necessary precautions to prevent exposures, and exposure
evaluation procedures.
Personnel must receive annual updates or additional training when procedural or policy
changes occur. Personal health status may impact an individual’s susceptibility to infection,
ability to receive immunizations or prophylactic interventions. Therefore, all laboratory
personnel and particularly women of child-bearing age should be provided with information
regarding immune competence and conditions that may predispose them to infection.
Individuals having these conditions should be encouraged to self-identify to the institution’s
healthcare provider for appropriate counseling and guidance.
4.1.2.2 Biosafety Level 2 (BSL-2/BL-2)

As defined in:
Section IV of the CDC BMBL:
http://www.cdc.gov/od/ohs/biosfty/bmbl5/sections/SectionIV-LaboratoryBiosafetyLevelCriteria.pdf
Appendix G-II-B for recombinant DNA BL-2:
http://www4.od.nih.gov/oba/rac/guidelines_02/Appendix_G.htm#_Toc7246566
Biosafety Level 2 containment practices are the same as those listed for Biosafety Level 1, with the
following additions:
Keep laboratory door closed and facilities must be locked after work hours. There should be
floor-to-ceiling physical separation from “laboratory areas” and “non-laboratory areas”.
Therefore, any doors leading into non-laboratory areas, such as offices or hallways must be
kept shut.
Only persons who have been informed of the research and its risks should be permitted to
enter BSL-2 areas.
Keep plants and animals not used in BSL-2 experiment out of the laboratory.
Wear appropriate PPE (as documented in the laboratory SOPs):

Open wounds should be covered with occlusive bandages.
Gloves must be worn to protect hands from exposure to hazardous materials. Glove
selection should be based on an appropriate risk assessment. Alternatives to latex
gloves should be available. Double-gloving may be necessary for some procedures.
Laboratory gowns or coats must be worn within the laboratory and not removed from
the laboratory. These should either be disposable or laundered by the institution—
not taken home.
Protective eyewear should be worn when conducting procedures that have the
potential to create splashes of microorganisms or other hazardous materials. Persons
who wear contact lenses in laboratories should also wear eye protection.
Eye, face and respiratory protection should be used in rooms containing infected
animals as determined by the risk assessment.
Use biological safety cabinets or similar containment equipment to contain aerosol-producing

equipment.
Maintain a biological spill kit within the laboratory.

4-6
Vacuum aspirator lines need to be properly maintained to prevent overflow. It is
recommended that vacuum traps never be allowed to collect liquids past half-full to enable
full decontamination prior to disposal. Vacuum traps outside of biosafety cabinets should be
placed in secondary containment sufficient in size to contain the contents of the trap, should
the trap implode under vacuum pressure. Vacuum lines should be protected with High
Efficiency Particulate Air (HEPA) filters, or their equivalent. Filters must be replaced as
needed.
Laboratory equipment should be routinely decontaminated by staff properly trained and

equipped to work with the material, after spills, splashes, or other potential contamination.
Equipment must be fully decontaminated before repair, maintenance, or removal from the
laboratory.
Spills involving infectious materials must be contained, decontaminated, and cleaned up by

staff properly trained and equipped to work with infectious material.
Report spills, accidents, potential exposures, near misses and disease symptoms related to
laboratory acquired infection to the PI and the Biosafety Office. Additional reporting
requirements may be required to comply with Human Resources and/or Campus Safety
Committee requirements (See Section 6 for further information).
Laboratory personnel must be provided medical surveillance and offered appropriate

immunizations for agents handled or potentially present in the laboratory. Some agents may
require the collection and storage of serum samples from at-risk personnel prior to initiation
of work within the laboratory. These measures must be documented in the laboratory
Biosafety Manual and SOPs.
A laboratory-specific biosafety manual must be prepared and adopted as policy. The biosafety
manual must be available and accessible to all personnel within the laboratory.
The laboratory supervisor must ensure that laboratory personnel demonstrate proficiency in
standard and special microbiological practices before working with BSL-2 agents.
4.1.2.3 Biosafety Level 2+ (BSL-2+)

Biosafety level 2+ (BSL-2+) is the designation utilized for those biohazard experiments that require
practices that are more stringent than standard BSL-2 procedures. It is not strictly defined by the CDC
or NIH, however, generally, BSL-3 practices are mandated in a space designed for BSL-2 work.
The BSL-2+ laboratory should be self-contained with all equipment required for the experiment
located within the laboratory. A biohazard door sign listing the agent in use, emergency contact, and
entry requirements is posted on the door while BSL-2+ work is in progress and the laboratory director
should control access to the laboratory and restricts access to persons whose presence is required for
program or support purposes. The laboratory doors should be kept closed when experiments are in
progress. Access is especially restricted to those involved in the experiment. When work is completed
and equipment has been decontaminated, the sign can be removed and the laboratory is returned to
standard BSL-2 use.
Special precautions should be taken to limit the handling of any contaminated sharp items, including
needles and syringes, slides, pipettes, capillary tubes, and scalpels. The use of additional personal
protective equipment should also be assessed based on the risks of the agents and operations within the
laboratory.

4-7
Laboratory and support personnel in BSL-2+ laboratories should receive appropriate training on the
potential hazards associated with the work involved and the necessary precautions to prevent
exposures. Personnel must demonstrate proficiency in safe microbiological practices and techniques.
Special health counseling measures, and possibly baseline serum samples should be collected as
appropriate and stored for all laboratory and other at-risk personnel.
Under BSL-2+ containment, all manipulations of biological material are conducted in a class II
biological safety cabinet and secondary containment is utilized for centrifugation and other potential
aerosol generating procedures. All potentially contaminated waste materials from the BSL-2+
laboratory should be decontaminated prior to disposal in the Stericycle Biohazard Waste boxes.
Additional requirements for work at BSL-2+ are listed in Appendix D. Please consult the Biosafety
Office prior to initiating any work at BSL-2+.
4.1.3 Practices Associated with Activities Involving Animals

Laboratory animal facilities are a special type of laboratory. Although many of the practices followed under
ABSL (or BL-N) containment levels are often comparable to those used in vitro, the animal room can present
unique problems since the activities of the animals themselves can present unique hazards not found in standard
microbiological laboratories. Animals may generate aerosols, they may bite and scratch, and they may be
infected with a zoonotic agent. Necropsies of infected animals also present a risk of exposure to personnel.
The co-application of Biosafety Levels and the Animal Biosafety Levels are determined by a protocol driven
risk assessment in order to appropriate contain the risks of the agents and animals used in an experiment.
The Institutional Biosafety Committee works in close conjunction with the Institutional Animal Care and Use
Committee (IACUC) and the veterinarians in the Division of Laboratory Animals Services (DLAS) in order to
ensure that the safety of human researchers and caretakers, the health of the animal colonies and humane
treatment of research animals are ensured. These considerations not only impact the safety, but the integrity
and ethics of the science occurring in the facilities. Considerations are given to the proper handling of animals
which may have been experimentally infected as part of a research protocol, those which may naturally harbor
zoonotic infectious agents (e.g., non-human primates), or those which may inadvertently have been infected
with animal pathogens which may pose hazards for colony maintenance and integrity.
Animal Biosafety Levels as defined in the CDC BMBL (Section V) and the NIH Guidelines (Appendix Q)
presuppose that laboratory animal facilities, operational practices, and quality of animal care meet applicable
standards and regulations as described in:
Institute of Laboratory Animal Research (ILAR), Commission on Life Sciences, National
Research Council’s Guide for the Care and Use of Laboratory Animals:
http://www.nap.edu/catalog.php?record_id=5140
U.S. Federal Laboratory Animal Welfare Regulations (9 C.F.R.)
http://www.access.gpo.gov/nara/cfr/waisidx_07/9cfrv1_07.html
Training videos are available through the American Biosafety Association (ABSA) web site to provide
guidance to animal handlers at various Animal Biosafety Levels http://www.absa.org/restraining.html .
In addition to the animal biosafety levels described in the CDC BMBL and NIH Guidelines, the USDA has
developed facility parameters and work practices for handling agents of agriculture significance. These are
described in Appendix D in the CDC BMBL http://www.cdc.gov/od/ohs/biosfty/bmbl5/sections/AppendixD.pdf.
USDA requirements are unique to agriculture because of the necessity to protect the environment from
pathogens of economic or environmental impact. The importation, possession or use of the following agents are
either prohibited or restricted by law or by USDA regulations or administrative policies. These agents of
agricultural significance include:
• African horse sickness virus • Lumpy skin disease virus

• Louping ill virus • Akabane virus
• African swine fever virus

4-8
• Malignant catarrhal fever virus (exotic • Classical swine fever virus
strains or alcelaphine herpesvirus type 1) • Swine vesicular disease virus
• Avian influenza virus (highly pathogenic) • Coccidioides immitis
• Menangle virus • Teschen disease virus
• Bacillus anthracis • Cochliomyia hominivorax (Screwworm)
• Mycobacterium bovis • Theileria annulata
• Besnoitia besnoiti • Coxiella burnetti (Q fever)
• Mycoplasma agalactiae • Theileria lawrencei
• Bluetongue virus (exotic) • Ephemeral fever virus
• Mycoplasma mycoides subsp. mycoides, • Theileria bovis
(small colony type) • Ehrlichia (Cowdria) ruminantium
• Borna disease virus (heartwater)
• Mycoplasma capricolum • Theileria hirci
• Bovine infectious petechial fever agent • Eastern equine encephalitis virus
• Nairobi sheep disease virus (Ganjam • Trypanosoma brucei
virus) • Foot and mouth disease virus
• Bovine spongiform encephalopathy prion • Trypanosoma congolense
• Newcastle disease virus (velogenic • Francisella tularensis
strains)
• Trypanosoma equiperdum (dourine)
• Brucella abortus
• Goat pox
• Nipah virus
• Trypanosoma evansi
• Brucella melitensis
• Hemorrhagic disease of rabbits virus
• Peste des petits ruminants virus (plague of
• Trypanosoma vivax
small ruminants)
• Hendra virus
• Brucella suis
• Venezuelan equine encephalomyelitis
• Rift Valley fever virus
virus
• Burkholderia mallei/Pseudomonas mallei
• Histoplasma (Zymonema) farciminosum
(Glanders)
• Vesicular exanthema virus
• Rinderpest virus
• Infectious salmon anemia virus
• Burkholderia pseudomallei
• Vesicular stomatitis virus (exotic)
• Sheep pox virus
Japanese encephalitis virus
• Camelpox virus
• Wesselsbron disease virus
• Spring Viremia of Carp virus
4.1.3.1 Animal Biosafety Level 1 (ABSL-1, BL-1N)

As defined in:
Section V of the CDC BMBL:
http://www.cdc.gov/od/ohs/biosfty/bmbl5/sections/SectionV-
VertebrateAnimalBiosafetyLevelCriteria.pdf
Appendix Q-II-A for recombinant DNA BL-1N:
http://www4.od.nih.gov/oba/rac/guidelines_02/Appendix_Q.htm#_Toc7256318
The animal facility director establishes and enforces policies, procedures, and protocols
for institutional policies and emergency situations.
Each institute must ensure that research staff and animal handler safety and health
concerns are addressed as part of the animal protocol review.
Prior to beginning a study animal protocols must also be reviewed and approved by the
Institutional Animal Care and Use Committee (IACUC) and the Institutional Biosafety
Committee.
Personnel must have specific training in animal facility procedures and must be

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supervised by an individual with adequate knowledge of potential hazards and
experimental animal procedures.
A safety manual specific to the animal facility is prepared or adopted in consultation with
the animal facility director and appropriate safety professionals.
The safety manual must be available and accessible. Personnel are advised of potential
hazards and are required to read and follow instructions on practices and procedures.
Appropriate medical surveillance program is in place, as determined by risk assessment.

The need for an animal allergy prevention program should be considered.
Facility supervisors should ensure that medical staff is informed of potential occupational
hazards within the animal facility, to include those associated with research, animal
husbandry duties, animal care and manipulations.
Personal health status may impact an individual’s susceptibility to infection, ability to

receive immunizations or prophylactic interventions. Therefore, all personnel and
particularly women of child-bearing age should be provided information regarding
immune competence and conditions that may predispose them to infection. Individuals
having these conditions should be encouraged to self-identify to the institution’s
healthcare provider for appropriate counseling and guidance.
Personnel using respirators must be enrolled in an appropriately constituted respiratory

protection program.
A sign incorporating safety information must be posted at the entrance to the areas where
infectious materials and/or animals are housed or are manipulated.
The sign must include the animal biosafety level, general occupational health
requirements, personal protective equipment requirements, the supervisor’s name (or
other responsible personnel), telephone number, and required procedures for entering and
exiting the animal areas. Identification of specific infectious agents is recommended
when more than one agent is being used within an animal room.
Advance consideration should be given to emergency and disaster recovery plans, as a

contingency for man-made or natural disasters to prevent exposure of work personnel,
accidental release/escape of animals or environmental contamination.
Protective laboratory coats, gowns, or uniforms are recommended to prevent

contamination of personal clothing. These should not be worn outside of the facility, and
non-disposable PPE must be decontaminated and cleaned by the institution—not taken
home to launder.
Open wounds should be covered with occlusive bandages.
Gloves are worn to prevent skin contact with contaminated, infectious and hazardous
materials, and when handling animals.
Gloves and personal protective equipment should be removed in a manner that minimizes
transfer of infectious materials outside of the areas where infectious materials and/or
animals are housed or are manipulated.
Persons must wash their hands after removing gloves, and before leaving the areas where
infectious materials and/or animals are housed or are manipulated.

4-10
Eye and face and respiratory protection should be used in rooms containing infected
animals, as dictated by the risk assessment.
When applicable, laboratory supervisors should adopt improved engineering and work
practice controls that reduce the risk of sharps injuries. Precautions, including those listed
below, must always be taken with sharp items. These include:
o Needles and syringes or other sharp instruments are limited to use in the animal
facility when there is no alternative for such procedures as parenteral injection,
blood collection, or aspiration of fluids from laboratory animals and diaphragm
bottles.
o Disposable needles must not be bent, sheared, broken, recapped, removed from
disposable syringes, or otherwise manipulated by hand before disposal.
o Used disposable needles must be carefully placed in puncture-resistant
containers used for sharps disposal. Sharps containers should be located as close
to the work site as possible.
o Non-disposable sharps must be placed in a hard-walled container for transport to
a processing area for decontamination, preferably by autoclaving.
o Broken glassware must not be handled directly. Instead, it must be removed
using a brush and dustpan, tongs, or forceps. Plasticware should be substituted
for glassware whenever possible.
Equipment containing sharp edges and corners should be avoided.
4.1.3.2 Animal Biosafety Level 2 (ABSL-2, BL-2N)

As defined in:
Section V of the CDC BMBL:
http://www.cdc.gov/od/ohs/biosfty/bmbl5/sections/SectionV-
VertebrateAnimalBiosafetyLevelCriteria.pdf
Appendix Q-II-B for recombinant DNA BL-2N:
http://www4.od.nih.gov/oba/rac/guidelines_02/Appendix_Q.htm#_Toc7256323
In addition to the practices required by ABSL-1 containment, ABSL-2 requires that:
Access to the animal facility is restricted.
Personnel must have specific training in animal facility procedures, the handling of
infected animals and the manipulation of pathogenic agents.
Personnel entering the facility must be supervised by individuals with adequate

knowledge of potential hazards, microbiological agents, animal manipulations and
husbandry procedures.
Procedures involving the manipulation of infectious materials, or where aerosols or

splashes may be created, including animal necropsies, and injections of animals, should
be conducted in BSCs or by use of other physical containment equipment.
Procedures involving a high potential for generating aerosols should be conducted within
a biosafety cabinet or other physical containment device.
When a procedure cannot be performed within a biosafety cabinet, a combination of

personal protective equipment and other containment devices must be used.
Consideration should be given to the use of restraint devices and practices that reduce the
risk of exposure during animal manipulations (e.g., physical restraint devices, chemical
restraint medications, etc).

4-11
Materials to be decontaminated outside of the immediate areas where infectious materials
and/or animals are housed or are manipulated must be placed in a durable, leak proof,
covered container and secured for transport. The outer surface of the container is
disinfected prior to moving materials. The transport container must contain a universal
biohazard label.
Equipment, cages, and racks should be handled in a manner that minimizes

contamination of other areas. These should be periodically inspected to ensure that sharp
edges are not present which may present a laboratory hazard.
Equipment must be decontaminated before repair, maintenance, or removal from the

areas where infectious materials and/or animals are housed or are manipulated.
Spills involving infectious materials must be contained, decontaminated, and cleaned up

by staff properly trained and equipped to work with infectious material.
Incidents that may result in exposure to infectious materials must be immediately

evaluated and treated according to procedures described in the safety manual. All such
incidents must be reported to the animal facility supervisor and the Biosafety Officer.
Medical evaluation, surveillance, and treatment should be provided as appropriate and
records maintained.
Persons having contact with the NHP should assess risk of mucous membrane exposure
and wear appropriate protective equipment (e.g., masks, goggles, faceshields, etc.) as
needed.
Respiratory protection is worn based upon risk assessment.
Sink traps are filled with water, and/or appropriate liquid to prevent the migration of
vermin and gases.

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4.1.4 Tissue, Cell and Microbiological Culture Practices
4.1.4.1 Tissue/Cell Culture Practices

Adhering to appropriate tissue culture techniques is intended to contain the risks of the biological
agents used in an experiment, but also intends to improve the integrity of the science. The
following measures are general guidelines for improved safety while performing cell culture
techniques within a biosafety cabinet (BSC, also commonly called a “tissue culture hood”).
Wear long sleeved gowns with knit cuffs and long gloves when working in the biosafety cabinet.
Maintain a clean lab coat reserved solely for cell culture work. Cover any open wounds with
occlusive bandages prior to donning PPE.
Avoid causing unnecessary air disturbances in and around the BSC. Avoid moving ones
hands in and out of the biosafety cabinet or sweeping side-to-side motions. Avoid talking
during culture manipulations as aerosols may be drawn into the work area. BSCs should
be placed in low traffic areas away from doors to prevent disturbance of the air curtain
protecting the containment and sterility in the BSC.
Place all reagents and supplies inside the Biosafety cabinet before the experiment to
reduce the disturbance of air within the BSC during use.
Work from a clean to a dirty

side. Place all unused (clean)
materials and reagents on one
side of the work surface, and
waste containers on the other
(dirty) side.
Do not over-crowd your

biosafety cabinets, which may
interfere with proper laminar CLEAN DIRTY/WASTE
air flow required for BSC
function.
Do not block the grates in the front or the back of the BSC which would prevent proper
laminar air flow required to maintain containment and sterility within the BSC. Work
approximately 4 inches back from the front of the BSC work area.
Allow the BSC to run for fifteen (15) minutes prior to use and fifteen (15) minutes after
use to purge the air within the BSC of any contaminants.
Clean all work surfaces, interior vertical surfaces and face shields before and after use
with an appropriate disinfectant.
Do not use open flames inside the BSC. Heat currents generated from the flame may
interfere with the laminar airflow of the BSC. In addition, the heat can
damage the HEPA filters or the adhesives in the filter units or other
components of the BSC. This can impact the warranty or UL ratings
of the BSC. Lastly, flames pose a high flammability risk when used in
the vicinity of alcohols, which are often used in tissue culture
situations. Alternative devices or measures should be utilized, such as
glass bead sterilizers, microincinerators, use of disposable instruments Glass bead sterilizer
Microincinerator
or having multiple packets of sterile instruments on-hand. If no appropriate

substitute can be found for a flame, only the use on flame-on-demand devices, such
as properly-used touch-plate microburners will be permitted. Gas and air balance
Touch-plate
microburner

4-13
should be set in the microburner such that only a small pilot light is visible most of the
time; flame is only produced after depressing the plunger when needed.
Do not use volatile chemicals in unducted Class IIA2 Biosafety Cabinets. After HEPA
filtration (which will not filter out chemicals), these BSC typically recirculate 70% of the
air and exhaust 30% air from the BSC into the room. If volatile chemicals are used
within the BSC, recirculation will serve only to concentrate the chemicals within the
BSC, and pose a spark hazard; in addition, the chemicals will be inappropriately
exhausted into the room posing health risks. Minute amounts of volatile chemicals can
only be used in BSCs which are equipped with ducted exhaust ventilation—either via a
thimble/canopy connection (a “ducted” Class IIA2 BSC); while somewhat larger amounts
of volatile chemicals can be used in hard-ducted Class IIB2 BSCs. Please be aware, most
BSCs on the MCG campus are unducted Class IIA2 types and therefore inappropriate for
the use of volatile chemicals; please contact the Biosafety Office (x1-2663 or
BIOSAFETY@mcg.edu ) before using volatile chemicals in the BSC if you are not sure
of what type of BSC you possess.
Radiological materials should not be used in unducted Class IIA2 Biosafety Cabinets
unless protected using a charcoal filter box around the experiment. Contact the Radiation
Safety Office for further information about charcoal protection devices for use in
unducted BSCs. Radiological materials should not be used in these BSCs for similar
reasons that volatile chemicals should not be used (see above); in addition, radiological
material which adheres to dust particles may also contaminate the HEPA filters and lead
to inadvertent exposures of BSC maintenance and certification personnel. Minute
amounts of radiological materials can only be used in BSCs which are equipped with
ducted exhaust ventilation—either via a thimble/canopy connection (a “ducted” Class
IIA2 BSC); while somewhat larger amounts can be used in hard-ducted Class IIB2 BSCs.
Please be aware, most BSCs on the MCG campus are unducted Class IIA2 types and
therefore inappropriate for the use of radiological materials without additional protective
measures.
Liquid wastes should be collected in order to decontaminate using an appropriate method

which should be described in the laboratory SOPs. Typically, liquid wastes are collected
in vacuum aspirators into which some disinfectant has been placed. To ensure
appropriate decontamination, subsequent disinfection measures should be followed prior
to disposal.
Do not allow vacuum traps to become overfull (recommended not greater than
half-full). This not only prevent liquids from being inadvertently drawn into the
vacuum line, but will allow for full decontamination of the liquid wastes prior to
disposal
HEPA filters or equivalents should be placed in the vacuum lines of any

laboratory requiring BSL-2 containment or higher.
In-line HEPA vacuum filters
Do not leave pipettes in the ends of the vacuum aspirator hoses. After use,
remove them from the hose and place in disinfection tray/container prior to
disposal. Leaving pipettes within the hoses only presents additional exposure or

4-14
contamination risks.
Rinse vacuum tubing with disinfectant after use. This will prevent backflow of
contaminated liquids within the vacuum line and subsequent contamination.
If the vacuum traps are outside of the Biosafety Cabinet, place in sufficient
secondary containment to hold the volume of liquid which may be spilled if
implosion of the vacuum flask should accidentally occur.
Glassware/plasticware and other contaminated items should be disinfected or autoclaved

before washing, reuse or disposal. Glassware should be thoroughly cleaned and rinsed,
by washing repeatedly with tap water and distilled water.
Place dirty pipettes, tips and tubes in a decontamination tray or

container in which disinfectant has been placed on the “dirty
side” of the BSC rather than move hands in and out of the BSC
to dispose of these. This will avoid disrupting the protective air Pipette/decontamination tray
curtain when hands are removed from the BSC. Discard empty tubes immediately into
the disinfection tray or similar containment device; after the experiment, drain the
disinfectant from the plastic wastes then dispose of the wastes in the biohazard waste
containers..
Keep open tubes parallel to the airflow.
After transferring inoculums, always recap vials.
Do not place tubes on work surface.
Pipette gently along the sides of tubes to prevent production of aerosols
Photo courtesy of Yale Biosafety
Spray and aerosol produced by

expelling liquids from a pipette
Work with one specimen at a time; recap before going to the next.
Autoclave verification should be performed routinely.
If a problem with contamination develops please contact the Biosafety Office (x1-2663 or
BIOSAFETY@mcg.edu) for further assistance.
4.1.4.2 Microbiological Practices

In the absence of definite accidents or obvious spillage, it is not certain that the opening of plates,
tubes and bottles of other microorganisms has caused laboratory infection. However, it is probable
that among the highly infective agents some infections have occurred by this means. Particular
care is required when opening plates, tubes, or bottles containing fungi, for this operation may
release a large number of spores. Such cultures should be manipulated in a biological safety
cabinet.

4-15
To assure a homogenous suspension that will provide a representative sample, liquid cultures are
agitated before a sample is taken. Vigorous shaking will create a heavy aerosol. A swirling action
will generate homogenous suspension with a minimum of aerosol. When a liquid culture is re-
suspended, a few minutes should elapse prior to opening the container to reduce the aerosol.
The insertion of a sterile, hot wire loop or needle into a liquid or slant culture can cause splattering
and release of an aerosol. To minimize the aerosol production, the loop should be allowed to cool
in the air or be cooled by touching it to the inside of the container or to the agar surface where no
growth is evident prior to contact with the culture of colony.
Placing an inoculating loop, wire or needle directly into a flame after use can also cause
splattering and release of an aerosol. Following use of inoculating loop or needle, it is preferable
to sterilize the instrument in an electric or gas incinerator specifically designed for this purpose
rather than heating in an open flame. These small incinerators have a shield to contain any
material that may spatter from the loop or needle. Disposable inoculating loops are also
commercially available. Rather than decontaminating them immediately after use with heat, they
are discarded first into a disinfectant solution.
The practice of streaking an inoculum on rough agar results in aerosol production created by the
vibrating loop or needle. This generally does not occur if the operation is performed on smooth
agar. It is good safety practice to discard all rough agar poured plates that are intended for
streaking purposes with a wire loop.
Water arising from syneresis in Petri dish cultures usually contains viable microorganisms and
forms a film between the rim and lid of the inverted plate. Aerosols are dispersed when opening
the plate breaks this film. Vented plastic Petri dishes, where the lid touches the rim at only three
points, are less likely to pose this hazard. The risk may also be minimized by using properly dried
plates, but even these (when incubated anaerobically) are likely to be wet after removal from an
anaerobic jar. Filter papers fitted into the lids reduce, but do not prevent dispersal. If plates are
obviously wet, they should be opened in the biological safety cabinet.
Less obvious is the release of aerosols when screw-capped bottles or plugged tubes are opened.
This happens when a film of contaminated liquid, which may collect between the rim and the
liner, is broken during removal of the closure. The practice of removing cotton plugs or other
closures from flasks, bottles, centrifuge tubes, etc., immediately following shaking or
centrifugation can generate aerosols and cause environmental contamination. The technique of
shaking tissue cultures with glass beads to release viruses can create a virus-laden aerosol.
Removal of wet closures, which can occur if the flask or centrifuge tube is not held in an upright
position, is also hazardous. In addition, when using the centrifuge, there may be a small amount
of foaming and the closures may become slightly moistened. Because of these possibilities, it is
good safety practice to open all liquid cultures of infectious or hazardous material in a biological
safety cabinet wearing gloves and a long sleeved laboratory garment.
Dried, infectious culture material may also collect at or near the rim or neck of culture tubes/flasks
and may be dispersed into the air when disturbed. Containers of dry powdered hazardous
materials should be opened in a biological safety cabinet.
4.1.4.3 Glass Ampules

When a sealed ampule containing a lyophilized or liquid culture is opened, an aerosol may be
created. Aerosol creation should be prevented or minimized; opening of ampules should be done
in biological safety cabinets. When recovering the contents of an ampule, care should be taken not
to cut the gloves or hands or disperse broken glass into eyes, face, or laboratory environment. In
addition, the biological product itself should not be contaminated with foreign organisms or with
disinfectants. To accomplish this, work in a biological safety cabinet and wear gloves. Nick the
ampule with a file near the neck. Wrap the ampule in disinfectant wetted cotton. Snap the ampule

4-16
open at the nick, being sure to hold the ampule upright. Alternatively, at the file mark on the neck
of the ampule, apply a hot wire or rod to develop a crack. Then wrap the ampule in disinfected
wetted cotton, and snap it open. Discard cotton and ampule tip into disinfectant. The contents of
the ampule are reconstituted by slowly adding fluid to avoid aerosolizing the dried material. Mix
contents without bubbling, and withdraw the contents into a fresh container. Some researchers
may desire to use commercially available ampules prescored for easy opening. However, there is
the possibility to consider that this may weaken the ampule and cause it to break during handling
and storage. Ampules of liquid cultures are opened in a similar way.
Ensure that all hazardous fluid cultures or viable powdered infectious materials in glass vessels are
transported, incubated, and stored in easily handled, nonbreakable leakproof secondary containers
that are large enough to contain all the fluid or powder in case of leakage or breakage of the glass
vessel. The secondary container must be labeled with a biohazard label bearing the name of the
infectious material.
4.1.4.4 Cryovials and Cryopreservation in Liquid Nitrogen

There is a foreseeable risk that when samples that have been stored in liquid nitrogen are removed
from the liquid phase and warmed they might explode. Clearly there is potential for significant
physical injury to anyone nearby and in many cases there might also be an associated infection
risk. Therefore, all personnel working with liquid nitrogen storage vessels or near them should be
made aware of the explosion and infection risks and appropriate control measures should be taken
to protect personnel from these risks.
When plastics are placed in liquid nitrogen they become brittle and shrink due to the effects of the
extremely low temperatures. This applies even to those plastics used for cryogenic vials sold
specifically for storage of samples in liquid nitrogen. While the use of tubes with internal threads
and a gaskets will lower the risk of leakage, it is virtually impossible to achieve a leakproof vial
once the specimen is placed in liquid phase liquid nitrogen. Inevitably, some seepage in to the vial
will occur. When the vial is removed from liquid nitrogen for thawing, the quick warming and
expansion of air within the vials can cause the tube to explode.
Therefore, cryopreserved materials should be stored in the vapor phase of liquid nitrogen in a
cryotank. If it is placed in the liquid phase, commercially available plastic tubing may be sealed
around the cryovial like a sausage skin. Special caution should be taken when removing cryovials
from liquid nitrogen storage, and should include cryoprotective gloves to protect against liquid
nitrogen burning, and full eye and face protection. Cryovials should be placed into a secondary
container with a closed lid and into a Biosafety Cabinet as soon as possible to further protect the
handler should the vial explode.
4.1.4.5 Vacuum Packed Tubes/Vials

When a vacuum-sealed tube or vial (e.g., Vacutainer blood tubes or septum bottles) are opened,
the lyophilized material or liquid culture may become aersolized due to the sudden influx of air
within the tube/vial. Protective measures to protect personnel from exposures to these aerosols
should be incorporated into the laboratory Standard Operating Procedures. Ideally, opening of
vacuum-sealed tubes/vials should be done in biological safety cabinets. Covering the lid or plug
with an disinfectant-soaked gauze pad will also provide a barrier against exposures to the
aerosolized material that may be generated upon opening. Commercial “cap” devices are also
available to limit the spread of aerosols generated when opening tubes or vials.
4.1.4.6 Embryonated Eggs

Harvesting cultures from embryonated eggs is a hazardous procedure and leads to heavy surface
contamination of the egg trays, shells, the environment, and the hands of the operator. It is

4-17
essential that operations of this type be conducted in a biological safety cabinet. A suitable
disinfectant should be at hand and used frequently.
4.1.5 Transport of Biological Material on Campus (between Labs or Buildings)

To prevent exposure to non-authorized personnel (with unknown health status) and potential environmental
release and contamination, biological materials which must be transported between laboratory areas by
authorized personnel only through common hallways or walkways must be properly packaged, contained
and labeled. Here are the guidelines which should be followed:
Biological materials must be contained inside two leakproof containers prior to removal from the
laboratory. The IBC standard for intramural transport is:
• Inside a sealed, leakproof primary container. A container is considered “leakproof” if it
can be filled with water, turned upside down without holding the lid in place and leakage
does not occur.
• The primary container must be placed inside a sealed, leakproof, durable secondary
container. The IBC standard for “durable” is puncture-resistant. Ziplock bags are not
considered “durable” by this standard. Sealed 15 ml or 50 ml plastic centrifuge tubes or
“burpable” household plastic storage containers (e.g., Tupperware®, Rubbermaid® or
similar brands) are considered “durable.
• Absorbent material (e.g., paper towels) must be placed between the primary and
secondary containers suitable for the volume transported.
• A biohazard sticker and label must be affixed on the outside of the secondary container
with agent name, lab address and emergency contact phone number.
Utilize plastic containers whenever feasible. Avoid glass. If glass primary containers must be used,
place containers within a sealed rigid plastic container with absorbent and padding to cushion vials
during transport.
The outside of the primary container should be decontaminated before placing into the secondary
container. The outside of the secondary container should be decontaminated before leaving the
laboratory.
Biological materials may not be transported through non-MCG areas (e.g., the MCGHI hospital or
Augusta Veterans Administration Hospital) without prior permission from these entities. Live
animals must not be transported through non-MCG areas. Consider that the risk of transporting
potentially infectious materials through hospitals is higher because of the poorer health status of
individuals within the hospital.
Transport of biological materials in a private vehicle is discouraged. Keep in mind that transport
of hazardous materials (which may include biological material, dry ice and liquid nitrogen) is
against the terms of most private automobile insurance policies. Check with your insurer.
Any extramural transport of biological materials must comply with the IATA/DOT shipping
standards as described in Section 10 of the MCG Biosafety Guide.
4.1.6 Housekeeping
Well-defined housekeeping procedures and schedules are essential in reducing the risks associated with
working with pathogenic agents and in protecting the integrity of the research program. This is particularly
true in the laboratory operating under less than total containment concepts and in all areas used for the

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housing of animals, whether or not they have been intentionally infected. A well conceived and well
executed housekeeping program limits physical clutter that could distract the attention and interfere with
the activities of laboratory personnel at a critical moment in a potentially hazardous procedure, provides a
work area that will not in itself be a source of physical injury or contamination, and provides an area that
promotes the efficient use of decontaminates in the event of inadvertent release of an etiologic agent. Less
immediately evident are the benefits of establishing among personnel of widely varying levels of education
some concepts of the nature and sources of contamination.
4.1.6.1 Objectives of Housekeeping

The objectives of housekeeping in the laboratory are to:
Provide an orderly work area conducive to the accomplishment of the research program.
Provide work areas devoid of physical hazards.
Provide a clean work area with background contamination ideally held to a zero level but
more realistically to a level such that extraordinary measures in sterile techniques are not
required to maintain integrity of the biological systems under study.
Prevent the accumulation of materials from current and past experiments that constitute a
hazard to laboratory personnel.
Prevent the creation of aerosols of hazardous materials as a result of the housekeeping

procedures used.
Procedures developed in the area of housekeeping should be based on the highest level of risk to
which the personnel and integrity of the experiments will be subject. Such an approach avoids the
confusion of multiple practices and retraining of personnel. The primary function, then, of routine
housekeeping procedures is to prevent the accumulation of organic debris that may:
Harbor microorganisms potentially a threat to the integrity of the biological systems

under investigation.
Enhance the survival of microorganisms inadvertently released in experimental

procedures.
Retard penetration of decontaminants.
Be transferable from one area to another on clothing and shoes.
With sufficient buildup, become a biohazard as a consequence of secondary

aerosolization by personnel and air movement
Cause allergenic sensitization of personnel (e.g., to animal dander).
Housekeeping in animal care units has the same primary function as that stated for the laboratory
and should, in addition, be as meticulously carried out in quarantine and conditioning areas as in
areas used to house experimentally infected animals. No other area in the laboratory has the
constant potential for creation of significant quantities of contaminated organic debris than do
animal care facilities.
4.1.6.2 Scope of Housekeeping

In all laboratories, efforts to achieve total decontamination and to conduct a major cleanup of the
biological materials are normally undertaken at relatively long time intervals. Routine

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housekeeping must be relied on to provide a work area free of significant sources of background
contamination. The provision of such a work area is not simply a matter of indicating in a general
way what has to be done, who will do it, and how often. The supervisor must view each task
critically in terms of the potential biohazard involved, decide on a detailed procedure for its
accomplishment, and provide instructions to laboratory personnel in a manner that minimizes the
opportunity for misunderstanding.
The following list outlines a portion of the terms requiring critical review by the laboratory
supervisor. It is not intended to be complete but is presented as an example of the detailed manner
in which housekeeping in the laboratory complex must be viewed.
• Aisles • Hallways
• Eyewashes • Supply Storage
• Lab Entry and Exit Ways • Deep Freezer Chests
• Bench Tops • Incubators
• Floors • Waste Accumulations
• Lab Equipment Cleanup • Dry Ice Chests
• Biological Safety Cabinets • Insect and Rodent Control
• Glassware • Work Surfaces
• Refrigerators • Instruments
• Cold Rooms
4.1.6.3 Assignment of Responsibilities

Housekeeping in the laboratory is one avenue that leads to safely accomplishing the research
program. It is important that housekeeping tasks be assigned to personnel who are knowledgeable
of the research environment. The recommended approach to housekeeping is the assignment of
housekeeping tasks to the research teams on an individual basis for their immediate work areas
and on a cooperative basis for areas of common usage. Similarly, animal caretaker personnel
should be responsible for housekeeping in animal care areas. The laboratory supervisor must
determine the frequency with which the individual and cooperative housekeeping chores need be
accomplished. The supervisor should provide schedules and perform frequent inspection to assure
compliance. This approach assures that research work flow patterns will not be interrupted by a
contracted cleanup crew; delicate laboratory equipment will be handled only by those most
knowledgeable of its particular requirements; and the location of concentrated biological
preparations, as well as contaminated equipment used in their preparation and application, will be
known.
4.1.7 Standard Operating Procedure (SOP) Development

Central to the concept of risk assessment and management is the development of laboratory-specific
Standard Operating Procedures (SOPs). These function within the laboratory to train all personnel on the
“rules of the laboratory”, and provide easy future reference for all staff members. These SOPs also serve as
documentation to the Institutional Biosafety Committee that appropriate practices are in place to contain
the biological risks posed in a particular experiment. All mitigation methods specific to the agents,
locations and operations which are required to contain their risks should be documented.
At minimum, laboratory SOPs should address the following:

Risk communication information. The risks of the agents and operations which may present
possible routes of entry should be documented. Symptoms of any illnesses which may arise due to
inadvertent exposure to the materials should also be documented, and instructions provided to
personnel about communication of this information to any healthcare provider, should they
symptoms present (even if the personnel was not knowingly exposed to the materials).
Prophylactic measures (e.g., vaccinations) and possible post-exposure health measures should be

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documented.
Emergency Response Procedures. The procedures documenting the response of the personnel and
supervisors should an incident, accident, exposure, spill, release or equipment failure involving
biological materials should be documented. This includes spill response, centrifuge tube
breakages, exposure responses, reporting requirements, etc.
Personal Protective Equipment (PPE) and Protective Equipment. The typical PPE to be worn by
the personnel in the laboratory and any special PPE and/or equipment which must be used during
particular agents (e.g., animal or biotoxin handling) or during particular operations which may
present additional splash, spray or aerosol risks (see Section 3.2.3 for further information) must be
documented and followed by personnel.
Disinfection/Decontamination/Disposal Procedures. Documentation should be provided to

personnel related to the appropriate disinfection, decontamination agents and procedures for all
surfaces (e.g., benchtops, stainless steel surfaces of equipment), equipment, liquid wastes, solid
wastes, spills and instruments. Disposal procedures for sharps, solid waste, liquid waste and
pathological waste (e.g., animal carcasses or anything identifiable as a body part) should also be
documented. If autoclaves are used as a method of decontamination, autoclave maintenance and
certification procedures and records should be documented. Any special deactivation procedures
for biological toxins must also be included in the SOPs.
Standard Prudent Practices for Laboratories (e.g., no eating, drinking, gum chewing, mouth
pipetting, hand-washing) as well as those particular to the PIs laboratory (e.g., closing the doors to
inner laboratories).
Storage and Access Control measures. The procedures the laboratory is expected to follow to
ensure no unauthorized personnel accesses the biological materials.
Transport/shipping procedures which should be followed by all personnel removing biological

agents from the laboratory to transport to another MCG laboratory or shipping. This includes
training requirements which may be required by these personnel.
Any training requirements of personnel.
A template has been developed to help assist researchers in starting to development of their laboratory
SOPs: http://www.mcg.edu/services/ehs/biosafe/bslsoptemplate.doc . This form is not intended to be
comprehensive; PIs are encouraged to modify this form as appropriate for their laboratories. New or
amended SOPs should be submitted to the Biosafety Office as part of the IBC review/approval process.
The Biosafety Office encourages researchers to seek assistance from the biosafety office in SOP
development; contact the office (x1-2663 or BIOSAFETY@MCG.EDU).
4.2 LABORATORY EQUIPMENT

Along with practices and facilities, proper use of laboratory equipment provides containment for work with
biological materials. Equipment can refer to Personal Protective Equipment (PPE), as well as operational
equipment, which are often used for the dual purposes of maintaining scientific integrity as well as
maintaining safety. Laboratory staff should be trained by the Principal Investigator, Laboratory Director or
Instructional Course Director in the proper use of laboratory equipment.
4.2.1 Personal Protective Equipment (PPE)

Multidisciplinary research conducted in MCG laboratories requires that personal protective equipment
(protective clothing and safety apparatus/equipment) be used to protect the researcher from contact with
infectious, toxic and corrosive agents, excessive heat, cold, fire and other physical hazards. Suitable

4-21
Personal Protective Equipment (PPE) also protects the experiment from contamination. The extent and
kind of clothing and equipment to be selected for any particular activity depends upon the research
operations and levels of risk associated with the research. While PPE is an important component of any
biological safety program, PPE is used with the understanding that PPE serves as a second line of defense.
Good laboratory techniques, procedures and appropriate laboratory equipment are the primary barriers
against potential exposure to hazardous agents.
For additional information you are urged to consult the Biosafety Office (x1-2663 or
BIOSAFETY@mcg.edu). In the event the Biosafety Office does not have a listing of the kind of protective
devices you are seeking, efforts will be made to acquire the information needed.
4.2.1.1 Laboratory Clothing

A commonly used PPE item within the laboratory is special clothing. Both reusable and
disposable clothing is available. Whichever is used, it must be durable, designed to provide
protection and prevent exposure of the skin to harmful agents, as well as be compatible with the
methods of decontamination employed.
Laboratory clothing serves to protect the wearer, the experiment, and environment against
contamination. If proper precautions are not taken, contaminated clothing may carry infectious
materials outside the laboratory and into other work areas, cafeterias, or the home. Infectious
agents can remain viable on cotton and wool fabrics and be disseminated from these fabrics.
Some additional points:

Overt exposure to agents at all level of risk should be followed by immediate
decontamination of the PPE and change into clean PPE to protect the worker, the
experiments and the environment.
Provisions should be made for PPE to be provided to visitors and maintenance or security
personnel, if applicable.
PPE worn within the laboratory should not be worn outside the facility to the library,
cafeteria, or other places accessible to the public.
Personnel should be encouraged to use disposable facial tissues instead of personal

handkerchiefs.
PPE should be placed in an appropriately designated area or container for storage,

washing, decontamination or disposal.
All PPE should be decontaminated before being sent to the laundry or discarded. Treat
contaminated areas of PPE with an appropriate disinfectant. Lab coats with extensive
contamination may by placed in a biohazard bag and autoclaved.
Do not take PPE home to launder; select a laundry service that follows universal
precautions.
Change PPE as soon as feasible whenever it is compromised, soiled or torn.
Wear appropriate sizes and keep an adequate supply of PPE available in the laboratory.
Wash hands whenever PPE is removed.
Do not touch door handles, elevator buttons, telephones, computers or other clean
surfaces or items with gloved hands.

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Wear closed-toe shoes and long pants to guard against skin contamination or chemical
exposure. Do not wear sandals or shorts in the laboratory.
4.2.1.1.1 Gloves
Gloves should be comfortable and of sufficient length to prevent exposure of the wrist
and forearm. Depending upon intended use, the composition and design of the glove may
vary to provide the desired level of flexibility, strength, impermeability, and resistance to
penetration by sharp objects, as well as protection against heat and cold. Quality
assurance is an important consideration.
No one glove can be expected to be satisfactory for all intended uses. Gloves may be
fabricated of cloth, leather, natural and synthetic rubbers, or plastics. New formulations
of synthetic rubber and plastic continue to be developed as research makes varied and
changing demands on the protective capabilities of gloves. Changing applications lead to
improved capabilities of impermeability, strength, flexibility, tactile sense and control.
Within even the modest laboratory, the glove applications may be such that no less than
four or five types of protective gloves need to be stocked and used.
Disposable (single use) gloves provide a barrier between infectious agents and the skin.
Glove use is a basic precept of preventing infectious agent transmission. Breaks in the
skin barrier of the hand (damaged cuticles, scrapes, micro-cuts, dermatitis, etc.) are
common.
Gloves shall be removed and hands washed before exiting the laboratory. Use the one
glove method, or an appropriate secondary container, when transporting materials
through common use areas.
The MCG Division of Environmental Health and Safety (EHS) can provide information
on gloves needed for various tasks, such as working with animals, dry ice, heat, acids,
etc. Consult the Division of EHS with details of your work to receive further information
about the type and availability of gloves that will best meet your requirements.
Considerations for the selection and use of gloves:

Gloves are not 100% leakproof; change gloves periodically and when soiled and
always wash hands after removing gloves or other PPE.
Gloves will not prevent needle sticks or other puncture injuries.
Check gloves for visible tears before use.
Avoid wetting examination gloves as water or disinfectants will encourage

wicking and leaking which may lead to exposure.
Do not reuse examination gloves; discard contaminated gloves in a biohazard

bag immediately after use.
Double glove or use household utility gloves when cleaning spills. Household
utility gloves may be decontaminated and reused (replace when compromised.)
Latex allergies among laboratory and clinic personnel are common and can
become extremely severe (see Section 5.4.3.2, Latex Gloves and Related
Allergies. Alternatives to latex should be considered.

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4.2.1.1.2 Procedure for Removing Gloves
Gloves should be the last item of PPE removed as part of the de-gowning procedure. The
exterior of the gloves should be considered contaminated and exposure to skin should be
avoided. Grip the outside of one glove at wrist with the other gloved hand, pull glove off
and gather in palm of gloved hand. Place index or middle finger of the ungloved hand on
wrist of gloved hand, slide finger under the glove opening and pull glove off inside out.
When removing PPE, remove lab coat or solid front gown first, then remove gloves
aseptically), remove face protection last to avoid touching your face with contaminated
hands. If wearing double gloves, remove outer gloves before removing lab coat or solid
front gown.
4.2.1.1.3 Shoes
Shoes worn in the laboratory must have closed-toes. Special protective shoes (e.g., steel-
toed shoes) may be required for certain work activities (e.g., when working in areas
where there is a danger of foot injuries due to falling or rolling objects, or objects
piercing the sole, and where feet are exposed to electrical hazards). When working with
infectious agents it is advisable to wear shoe covers over street shoes; shoe covers should
be removed and decontaminated (autoclaved) before disposal. For work in tissue culture
laboratories it may be necessary to change from street shoes to specific laboratory shoes
for protection of cultures from contamination.
In certain animal facilities, the MCG Division of Laboratory Animal Services (LAS)
requires personnel to wear overshoes to protect the animals in containment areas.
Similarly, people who work with animals and do cage washing are required to wear
protective shoes. All personnel working in LAS facilities follow these and other
Standard Operating Procedures established by MCG Division of Laboratory Animal
Services.
4.2.1.1.4 Gowns, Lab Coats, Jumpsuits, Aprons and Other

Protective Clothing
Gowns, lab coats and jumpsuits protect the wearer’s clothing and skin from
contamination. As with all PPE, the type of clothing needed depends on the task being
performed and the degree of exposure anticipated.
Solid front wrap-around clothing offers better protection than pull-over type clothing or
clothing with front closures. Lab coats are not 100% leakproof; change PPE when soiled,
and always wash your hands after removing any PPE. Lab coats or other protective
clothing will also not provide protection against needle sticks or other punctures.
Spills and splashes occur most often in the chest or lap area. The contaminated surface
must be touched during removal of a front closing jacket or lab coat. The contaminated
portion often ends up in the wearer’s face during removal of pullover clothing. Many

4-24
workers prefer not to button up front closing jackets, which leaves street clothing
exposed. If front closing jackets must be worn, strict measures shall be implemented to
assure the clothing is closed at all times when performing procedures or tasks that may
cause exposure.
Long-sleeved garments with snug fitting cuffs are preferred over open or short sleeves.
Snug fitting cuffs prevent splashes, splatters and aerosols from making contact with
exposed skin on the lower arms. Longer single-use gloves can be pulled over snug fitting
cuffs to seal out any infectious materials.
Plastic, vinyl or rubber aprons are usually worn over other protective clothing when extra
protection is desired. Aprons are necessary for protection against liquids spilling or
splashing on clothing. It is recommended that appropriate aprons be worn to protect
against the potential harmful effects of liquid waste. Aprons may also be used to provide
protection from steam and hot water in locations such as animal handling facilities,
autoclave rooms and laboratory glasswashing rooms.
4.2.1.1.5 Face and Eye Protection

Protection of the face and eyes is of prime importance in laboratories due to the potential
for foreign material, both liquid and solid, to splash on the head, face and eyes or contact
lenses. A variety of face shields, head covers/hoods, protective goggles, and lenses are
available from safety supply houses. The selection is dependent upon materials of
construction, fit, comfort, and compatibility with the work and the overall facial area
requiring protection.
Some of the considerations for selection and use of face and eye protection are indicated
below:
Face shields and hoods protect the face and the neck from flying particles and
sprays of hazardous material; however, they do not provide basic eye
protection against impacting objects.
Shields should cover the entire face, permit tilting back to clean the face if
desired, and be easily removed in the event of an accident.
If an eye hazard exists in a particular operation or experiment, the soundest

safety policy would be to require that eye or face protection, or both, be worn
at all times by all persons entering or working in the laboratory.
Contact lenses do not provide eye protection; in fact, they may present more
risk to the eyes by holding hazardous materials in contact with the eye for a
longer period of time until they can be removed. It is recommended that
contact lenses not be worn when working around chemicals, fumes, and other
hazardous material and dust particles since these items may become trapped in
the space between the contact lens and the cornea. When contact lenses are
worn, eye protection, such as tight fitting goggles, must be worn.
4.2.1.1.6 Respiratory Protection

Protection of the respiratory system is a major concern of any biological safety program
because infectious organisms can readily enter the human body through the respiratory
tract. The possibility of this occurring depends on the type and infectious dose of the
particular organism. For some, as few as one to ten organisms, when inhaled, may cause
infection. Particles with an effective aerodynamic diameter of between 0.5 and 5.0 µm
(the respirable fraction) are most effective at penetration and retention in the deep

4-25
pulmonary spaces. Particles larger than 5 micrometers are generally trapped in the upper
respiratory tract and eventually cleared or swallowed.
Engineering controls, such as the use of biological safety cabinets, should always be
considered as a first line of defense against respiratory infection when working with
infectious organisms. Respirators should only be considered as a second line of defense
after feasible engineering controls have been put into place and additional controls are
still needed.
Respirators vary in design, application, and protective capability. Respirators can be

placed into two categories:
air purifying
supplied air
By far, the most commonly used respirators in laboratories for protection against
biological materials are air purifying respirators. These protect by purifying the existing
breathing air through a filter (for particulates) or cartridge (for gases and vapors), or both.
Standard air purifying respirators used at MCG are half-mask, full face, or (battery)
powered air purifying respirators (PAPR). These rely on the proper cartridge selection to
filter out the contaminant.
Dust masks that have been approved by the National Institute of Occupational Safety and
Health (NIOSH) are also considered to be air purifying respirators. Approved dust masks
will have one of the following designations – N95, N99, N100, R95, R99, R100, P95,
P99, or P100. The selection of N-, R-, and P-series filters depends on the presence or
absence of oil particles, as follows:
If no oil particles are present in the work environment, use a filter of any
series (i.e., N-, R-, or P-series).
If oil particles (e.g., lubricants, cutting fluids, glycerine, etc.) are present,
use an R- or P-series filter. Note: N-series filters cannot be used if oil
particles are present.
If oil particles are present and the filter is to be used for more than one work
shift, use only a P-series filter.
Note: To help you remember the filter series, use the following guide:
N for Not resistant to oil,
R for Resistant to oil
P for oil Proof
Selection of filter efficiency (i.e., 95%, 99%, or 99.97%) depends on how much filter
leakage is acceptable. Higher filter efficiency means lower filter leakage. The NIOSH
Guide to the Selection and Use of Particulate Respirators Certified under 42 CFR 84 can
be found at: http://www.cdc.gov/niosh/userguid.html . OSHA also has an online
electronic tool for assisting in the proper selection of appropriate respirators:
http://www.osha.gov/SLTC/etools/respiratory/respirator_selection.html .
Federal OSHA regulations (29 CFR 1910.134) require initial and annual training and fit-
testing, and well as medical surveillance of all respirator wearers. If a respirator is to be
used in the course of research with biological materials, the MCG IBC will require
adherence to these OSHA standards:
http://www.osha.gov/pls/oshaweb/owadisp.show_document?p_id=12716&p_table=STA
NDARD .
Please make sure that the MCG Office of Environmental Health and Occupational Safety
(EHOS) is notified whenever the use of a respirator is being considered. Proper selection
of cartridges and respirators is very important and should not be made without input from

4-26
the EHOS. In addition, the EHOS office will assist in guiding researchers through the
evaluation process and provide the required training and fit testing. The EHOS Office
can be contacted at x1-2663. The Employee Health Office must also be notified so that
medical evaluation/surveillance and clearance can be issued prior to wearing the
respirator.
4.2.1.2 Selection of PPE

Use the following PPE to minimize exposure via mucous membrane OR non-intact skin:
For face protection, wear safety glasses and a mask, or a chin length face shield whenever
splashing, splattering or droplets may be anticipated (any work with liquids on the open
bench). An impact resistant face shield should be used when operating the autoclave or
when working with cryogenic materials. Impact resistant face shields will protect the
user’s face against splatters of hot liquids or broken glass or plastic fragments.
Gloves and a lab coat are worn to protect the skin and clothing from contact with
potentially infectious materials. Wear gloves that are long enough to extend over the
sleeves of the lab coat and cover wrists so no bare skin is exposed. Consider double
gloving when working with cultures of infectious agents or handling spills. Thicker
household utility gloves can be worn for cleaning blood or BSL-2 spills. Utility gloves
can be decontaminated and reused until the integrity of the glove is compromised.
Temperature resistant gloves should be worn to protect hands from physical damage
when working with very hot (autoclave) or cold (liquid nitrogen tank, -70°C freezer)
materials.
Sleeve covers may be worn over lab coat and gown sleeves to provide protection to the
sleeves and wrists from contamination when working in the biological safety cabinet.
Disposable sleeve covers have tight fitting grips at both ends.
Waterproof bandages are worn to cover any wounds or non-intact skin before gloving. It
is preferred to double glove when skin is damaged or non-intact. Inform your supervisor
of any severe skin conditions or wounds. Avoid working with Risk Group 2 or higher
potentially infectious materials if non-intact skin cannot be adequately covered.
Solid front gowns provide more protection to clothing and skin than lab coats. Solid
front gowns are worn for high hazard infectious agent work. The tight fitting cuffs of the
gown help to minimize wrist contamination.
Impervious lab coats, gowns or aprons are worn when heavy contamination or soiling is
likely.
Head covers are worn to protect the hair and scalp from splatter or droplets when
working with heavy contamination or when contact with the head is likely. When
choosing a head cover make sure it is impervious to liquids (some head covers are not
impervious).
Shoe covers are worn over the shoes to protect shoes from contamination when working
in heavily contaminated areas (such as large spills, crime scenes, morgues, cadaver
dissection areas, surgical operation areas).
Gowns, head and shoe covers also help keep contaminants from entering the sterile area
in clean rooms, surgical suites and barrier animal facilities.

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Use the following PPE to minimize exposure via cuts, slices, or scratches:
Kevlar gloves and sleeves are cut resistant and will help guard against slices, scratches or
cuts, but will not prevent direct puncture or needlestick injuries. Steel mesh gloves also
protect against slices, cuts, and scratches but will not eliminate punctures. Neoprene and
other abrasive resistant gloves are cut resistant, but significantly reduce dexterity.
Use the following PPE to minimize exposure via aerosols:

HEPA filtered respirators (air purifying or powered air purifying) are worn to prevent
exposure to potentially infectious aerosols when cleaning spills of concentrated infectious
material or responding to centrifuge incidents. Employees who wear a respirator must
enroll in the MCG Respiratory Protection Program through the EHOS (x1-2663) before
using a respirator.
4.2.1.3 PPE Requirements Table

Please see Table 4.2.1.4 for a guidance table for PPE

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Table 4.2.1.4. PPE Requirements Table
PPE Biosafety Level 1 Biosafety Level 2 Biosafety Level 2+/3

Gloves Recommended to prevent skin or clothing Required Required
contact with biological materials. Note: work
that may involve radioactive materials or
chemicals will require the use of a lab coat and
gloves
Lab Coat Recommended to prevent skin or clothing Required Solid-front protective clothing such as back fastening
contact with biological materials. Note: work gown with tight fitting cuffs must be worn to protect
that may involve radioactive materials or street clothing, scrubs and/or skin from contact with
chemicals will require the use of a lab coat and infectious agents.
gloves
Face Protection Wear protective eyewear and surgical mask or Face protection is not required when performing all
chin-length face shield whenever splashing, work inside a biosafety cabinet (BSC). However if
splattering or spraying is anticipated to prevent there is a potential for splashing, such as from a
contact with mucous membranes of the eyes, nose dropped container during transport, face protection
and mouth. Researchers may choose to augment must be worn.
eye protection by performing experiments behind a
protective splash shield
Respiratory The MCG IBC may recommend respiratory The use of respiratory protective equipment such as a
Protection protection on a case-by-case basis if the potential powered air purifying respirator (PAPR) will be
for aerosol generation is high and alternate recommended or required by the MCG IBC and/or the
containment devices, such as a biosafety cabinet, Biosafety Office on a case-by-case basis. The use of
cannot be used PAPRs is required for response and cleanup of a BSL-
2+ or BSL-3 spill. All those who may wear a
respirator must be enrolled in the MCG Respiratory
Protection Program.
Other Other PPE, such as Tyvek coveralls, booties, sleeve Other PPE, such as Tyvek coveralls, booties, sleeve
guards, plastic aprons, and household rubber gloves guards, plastic aprons, and household rubber gloves
will be recommended on a case-by-case basis. will be recommended on a case-by-case basis.
Generally, additional protective clothing is required Generally, additional protective clothing is required
whenever there is a high potential for splashing of whenever there is a high potential for splashing of
potentially infectious materials, such as organ potentially infectious materials, such as organ
harvesting or large spill response and clean-up. harvesting or large spill response and clean-up.

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4.2.2 Biological Safety Cabinets (BSCs)
The function of a BSC is dependent upon high
efficiency particulate air (HEPA) filters within the
plenum of the unit. The overall basis for HEPA
filter function relies on the combination of multiple
physical forces: inertial impaction (direct blocking
of the particulates via the filter medium), diffusion
(Brownian motion), Interception (“straining”) and
electrostatic forces. Larger particulates are
typically more efficiently blocked by interception
and impaction; whereas smaller particles tend to be
more efficiently retained by diffusion or
electrostatic forces.
Together, these forces combine to enable these

HEPA filters to remove a minimum of 99.97% of
particles 0.3 µm in size. The overall efficiency of
the HEPA filter is greater for removal of particles
that are larger or smaller than 0.3 µm (see Figure,
right, to review the relationship between HEPA filter efficiency relative to particle size). However, HEPA
filters will not filter out gases and vapors.
HEPA filters have a limited life span (approximately 5-7 years) and are relatively fragile; they can be easily
dislodged or damaged during moves or repairs of the unit. For this reason, Biosafety Cabinet function must
be certified at least annually or after any move or repair. The Division of Environmental Health and Safety
and the Laboratory Equipment Service (LES) office jointly run a program that monitors the annual
performance of biological safety cabinets at least annually. Contact the EHOS office (x1-2663) or
Laboratory Equipment Services (x1-6124) to request certification of a new, moved or repaired BSC. The
BSC certification program conforms to guidelines established by the National Institutes of Health (NIH)
and the Centers for Disease Control and Prevention (CDC) in their publication Primary Containment for
Biohazards: Selection, Installation and Use of Biological Safety Cabinets
http://www.cdc.gov/od/ohs/biosfty/bsc/bsc.htm and the Occupational Safety and Health Administration’s
(OSHA) Bloodborne Pathogens Standard.
Biological safety cabinets (BSCs) are categorized by

Classes:
Class I units provide personnel protection only
Class II units provide both personnel and product
protection
Class III are glove-box units which provide
personnel protection and optional product
protection
The specifications for BSC function and types are defined
by NSF ANSI Standard 49.
Most BSCs at MCG are Class II units, which means, when

used properly, they provide a clean work environment for research or patient care activities while offering
personnel and environmental protection. The BSC provides primary containment for infectious materials.
The efficacy of BSCs depends upon the behavior of the operator and the orientation of the unit in the
facility. Class II BSCs fall into four types (A1, A2, B1 and B2), the most common of which is the Class
IIA2 BSC at MCG. Class IIA2 BSCs have the following characteristics:
100 feet per minute inward air face velocity

70% recirculated air, 30% exhausted (thru HEPA) (see airflow diagram, right)
Potentially contaminated ducts and plenums are under negative pressure or surrounded by negative
pressure ducts and plenums

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Class IIA2 cabinets can be either “ducted” or “unducted”. The overwhelming
30%
majority of BSCs in MCG facilities are unducted Class IIA2 BSC.
After HEPA filtration, the exhaust from a Class IIA2 Biosafety Cabinet can
be expelled into the room, or it can be exhausted to an external area
connected via a thimble or canopy connector through the buildings external
exhaust system (i.e. a “ducted” Class IIA2 BSC).
The great advantage of having a “ducted” Class IIA2 BSC is that it may be 70%
used with minute quantities of volatile toxic chemicals and tracer amounts
of radionuclides, since the air from the BSC is exhausted through properly
functioning exhaust canopies. Radionuclides and volatile chemicals may
not be used in “unducted” Class IIA2 BSCs unless charcoal filter
containment units are placed within the unit to capture any potentially
volatilized radionuclides prior to circulation within the BSC. Contact the
Radiation Safety Office for further information about these charcoal
devices.
Keep in mind: if the BSC is ducted using a thimble connection, because this is not an air-tight
connection, if failure were to occur in the building exhaust system during use, the BSC would still
function to filter particulates, but will expel the air exhaust into the room and may pose a hazard if
radionuclides or volatile chemicals are in use.
MCG also has a small handful of Class IIB2 BSCs in select areas on campus, which have the following
characteristics:
100 feet per minute inward air face velocity
100% of the air is exhausted, no air recirculation occurs.
Potentially contaminated plenums are under negative pressure or surrounded by negative pressure
ducts and plenums.
They are hard-connected to the building’s external exhaust system (i.e., they are always “ducted”)
The typical exhaust requirements for these units are significantly higher than those of Class IIA2
units (~700-1200 CFM at 2” w.g.)
Because the function of a Class IIB2 BSC is intricately connected to the function of the building external
exhaust system, any failure in the building exhaust system will compromise the function of a Class IIB2
BSC. Therefore, audible or visual alarm systems must be installed to alert any users in fluctuations in the
building exhaust system. However, because exhausts from Class IIB2 units are connected via air-tight
connection and exhausted externally from the building and no recirculation of air occurs within the BSC,
there are fewer restrictions for volatile chemicals or radionuclide use in these BSCs.
BSCs isolate biohazards from personnel by confining the biohazardous material in the unit, primarily
through the use of directional laminar airflow. The BSC removes aerosolized biohazardous material by
moving air through high efficiency particulate air (HEPA) filters. The intake air coming from the front of
the unit is drawn into the front grate of the BSC filtered through a HEPA filter before entering the BSC
work area. Exhaust air also passes through a HEPA filter. Aerosols generated in the work area of the BSC
are contained within the BSC.
Operating Procedures for Class II Biological Safety Cabinet (also see Section 4.1.4.1 for more information
on Tissue/Cell Culture Practices):
If used, turn off UV light within the unit and switch on fluorescent light and blower.
Disinfect all interior surfaces with 70% ethanol or suitable disinfectant.
Place items required for procedure into cabinet; do not obstruct grills.
Allow the unit to run approximately 15 minutes prior to use to purge contaminants from work
area.
Keep materials at least 4 inches inside work area.
Work should proceed from clean to contaminated areas.
After procedure, allow cabinet to run approximately 15 minutes before removing materials to

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purge contaminants from work area prior to shutting off motor.
Wipe down all work surfaces with suitable disinfectant (as described in the laboratory SOPs)
Turn off fluorescent light and blower if desired.
Many BSCs are equipped with germicidal ultraviolet (UV) lamps. Time of exposure, distance, presence of
dust or debris and UV lamp intensity contribute to the germicidal effect of the UV lamp. The visible blue-
violet glow of the UV lamp does not indicate there is germicidal effect; the efficacy of the UV lamp may
decrease as the lamp grows older. The UV lamp needs to be cleaned periodically to remove dust. UV
lamps may damage eyes, skin, and laboratory equipment. UV lamps should be turned off while the room is
occupied. For these reasons, the Biosafety Office discourages the use of UV lamps inside of BSCs due to
the potential damage which may result from UV lamp use, and the arguable reliability of UV
decontamination over time. For further information related to the use of UV lamps in BSCs, please see the
following references:
Burgener, J (2006) Position Paper on the Use of Ultraviolet Lights in Biological Safety Cabinets.
Applied Biosafety 11(4): 228-230.
Meecham, P.J. and Wilson, C. (2006) Use of Ultraviolet Lights in Biological Safety Cabinets: A
Contrarian View. Applied Biosafety 11(4): 222-227.
4.2.3 Laminar Flow Hoods/Clean Benches

Laminar Flow Hoods (a.k.a. Clean Benches) differ from
Biosafety Cabinets in that they offer only product protection, not
personnel protection. They function by blowing HEPA filtered
air through the metal baffle in the back of the unit directly across
the work surface toward the user (see airflow diagram, right).
While this provides a sterile work surface, this may increase the
exposure risk of the individual using the unit to the materials on
the work surface. Therefore, LFHs should not be used for work
with hazardous materials. These function well for media
preparation or similarly low hazard operations which may
require sterile conditions.
4.2.4 Fume Hoods

Unlike Biosafety Cabinets or Laminar Flow
Hoods, chemical fume hoods offer personnel
protection to the user, but no product or
environmental protection. Air drawn from the
front of the unit passes directly across the work
surface of the fume hood, thereby potentially
compromising the sterility of any materials
exposed on the work surface (see airflow
diagram, right). In addition, the exhaust air from
the fume hood is not filtered in any way before
expelling the air via the building’s exhaust systems. This may result in environmental contamination, and
potential exposure of personnel who may be near the exhaust outtakes. However, typically fume hood
exhausts are expelled at high velocity through an exhaust stack, reducing the latter risk. Fume hoods
should be used for working with hazardous materials, such as biological toxins; however, not if sterility is
needed.
4.2.5 Centrifuges
All centrifugation of Risk Group 2 agents or higher shall be done using centrifuge safety buckets with
safety caps or in sealed centrifuge tubes in sealed rotors. If a small centrifuge is used and centrifuge safety

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cups are not available, the centrifuge should be operated in the biological safety cabinet.
Each person operating a centrifuge should be trained on proper

operating procedures. Keep a log book detailing operation records
for centrifuges and rotors to assist in determining service
requirements.
The following procedures for centrifugation are recommended safety

measures to consider while using biological materials in centrifuges :
Examine tubes and bottles for cracks or stress marks before
using them.
Fill and decant all centrifuge tubes and bottles within the
biological safety cabinet. Centrifuge safety caps and buckets
Never overfill centrifuge tubes as leakage may occur when tubes are filled to capacity. The
maximum for centrifuge tubes is 3/4 full.
Always cap tubes before spinning.
Place all tubes in safety buckets with safety caps or in sealed rotors. Inspect the "O" ring seal of
the safety bucket and the inside of safety buckets or rotors. Correct rough walls caused by erosion
or adhering of matter and remove debris from the rubber cushions.
Wipe exterior of tubes or bottles with disinfectant prior to loading into rotor or safety bucket.
Wipe the exterior of the rotor or safety buckets before removal from Biosafety Cabinet.
Never exceed safe rotor speed.
Stop the centrifuge immediately if an unusual condition (noise or vibration) begins.
Wait five to fifteen minutes after the run before opening the centrifuge. This will allow aerosols to
settle in the event of a breakdown in containment.
Decontaminate safety carriers or rotors and centrifuge interior after each use.
Open safety buckets or rotors in a biological safety cabinet. If the rotor does not fit in the
biological safety cabinet, use the fume hood.
If construction of the centrifuge permits, the centrifuge chamber is to be connected to a vacuum
pump with a HEPA filter installed between the centrifuge and the vacuum pump.
4.2.6 Vacuum Line Chemical Traps and Filters

Considerations and Limitations of Vacuum Line Chemical Traps and Filters:
Liquid wastes should be collected in order to decontaminate using an appropriate method which
should be described in the laboratory SOPs. Typically, liquid wastes are collected in vacuum
aspirators into which some full-strength disinfectant has been placed.
When setting up a vacuum aspirator system, make sure the tubing inside the vacuum flasks extend
far below the vacuum arm of the flask to prevent liquid wastes from being drawn through the
flask’s vacuum arm and contaminating the vacuum line (see, e.g., “Collection flask” in figure
below).

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Flexible vacuum tubing or Tygon® tubing used in aspirator systems should have walls thick
enough to withstand the vacuum without collapsing, cracking or leaking. Periodically inspect and
replace vacuum tubing which has become cracked over time.
To ensure appropriate decontamination, subsequent disinfection measures should be followed
prior to disposal.
Do no allow vacuum traps to become overfull (recommended not greater than half-full). This not
only prevent liquids from inadvertently drawn into the vacuum line, but will allow for full
decontamination of the liquid wastes prior to disposal
HEPA filters or equivalents should be placed in the vacuum lines of any laboratory requiring BSL-
2 containment or higher.
Vacuum line filters shall be examined and replaced if clogged or if liquid makes contact with the
filter. Used filters shall be discarded in the medical waste stream.
Do not leave pipettes in the ends of the vacuum aspirator hoses. After use, remove them from the
hose and place in disinfection tray/container prior to disposal. Leaving pipettes within the hoses
only presents additional exposure or contamination risks.
Rinse vacuum tubing with disinfectant after use. This will prevent backflow of contaminated
liquids within the vacuum line and subsequent contamination.
If the vacuum traps are outside of the Biosafety Cabinet, place in sufficient secondary containment
to hold the volume of liquid which may be spilled if implosion of the vacuum flask should
accidentally occur
4.2.7 Syringes and Needles

The hypodermic needle is a dangerous instrument. To lessen the chance of accidental injection, aerosol
generation, or spills, the use of syringes should be avoided when alternate methods are available. For
example, use a blunt needle or cannula on the syringe for oral or intranasal inoculations and never use a
syringe and needle as a substitute for a pipette in making dilutions.
The following practices are recommended for hypodermic needles and syringes when used for parenteral
injections:
Use the syringe and needle in a biological safety cabinet only and avoid quick and unnecessary
movements of the hand holding the syringe.
Examine glass syringes for chips and cracks, and needles for barbs and plugs. This should be
done prior to sterilization before use. Use needle-locking syringes only, and be sure that the needle
is locked securely into the barrel. Replace glass
syringes with plastic disposable syringes
whenever possible.
Whenever possible use safer needle systems.
These might include retractable needle systems or
shielded needle systems (see right).
Wear latex or nitrile gloves for all manipulations
with needles and syringes.
Fill the syringe carefully to minimize air bubbles and frothing of the inoculum.
Expel excess air, liquid and bubbles from a syringe vertically into a cotton pledget moistened with
an appropriate disinfectant, or into a small bottle of sterile cotton.
Do not use the syringe to forcefully expel a stream of infectious fluid into an open vial for the
purpose of mixing. Mixing with a syringe is condoned only if the tip of the syringe is held below
the surface of the fluid in the tube.
If syringes are filled from test tubes, take care not to contaminate the hub of the needle, as this
may result in the transfer of infectious material to the fingers.
When removing a syringe and needle from a rubber-stoppered bottle, wrap the needle and stopper
in a cotton pledget moistened with an appropriate disinfectant. If there is concern of the
disinfectant contaminating sensitive experimental materials, a sterile pledget may be used and
immediately discarded into a biohazard bag.
When inoculating animals, position the hand that is holding the animal “behind” the needle or use

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a pair of forceps to hold the animal in order to avoid puncture wounds.
Be sure the animal is properly restrained prior to the inoculation and be on the alert for any
unexpected movements of the animal.
Before and after injection of an animal, swab the injection site with
an appropriate antiseptic.
Discard syringes into an authorized sharps container, which should
always be kept near the site of use. DO NOT bend, shear, recap or
otherwise manipulate the needle. If recapping is unavoidable, use a
one handed method. DO NOT discard syringes into biohazard bags.
4.2.8 Pipettes
The following is excerpted from Laboratory Safety, Principles and Practices 2nd Ed., ASM Press.:
Never suction or pipette by mouth; always use some type of pipetting aid when pipetting
infectious materials. Preferably, all activities should be confined to a biosafety cabinet.
Mouth pipetting should be prohibited even with mouth pipetting devices that use an hydrophobic
membrane filter that does not require fingers to touch the mouthpiece. This reusable pipetting
device requires storage on the bench or other location between usage, which can result in
contamination on the end piece that inserts into the mouth.
Pipetting of toxic chemicals should be performed in a chemical fume hood.
Infectious or toxic materials should never be forcefully expelled from a
pipette. Mark-to-mark pipettes are preferable to other types because they do
not require expulsion of the last drop.
Infectious or toxic fluids should never be mixed by bubbling air from a pipette
through the fluid.
Infectious or toxic fluids should never be mixed by
alternate suction and expulsion through a pipette.
Gently discharge from a pipette as close as possible to
the fluid or agar level, and the contents should be
allowed to run down the wall of the tube or bottle
whenever possible, not dropped from a height.
Pipettes used for transferring infectious or toxic
materials should always be plugged with cotton, even when safety pipetting aids are used.
Avoid accidentally dropping infectious or toxic material from the pipette onto the work surface.
Place a disinfectant dampened towel or other absorbent material on the work surface, and
autoclave before discard or reuse. Plastic backed bench paper is suitable for this purpose.
Contaminated pipettes should be placed horizontally into a
pan or tray containing enough suitable disinfectant, such as
hypochlorite, to allow complete immersion of the pipettes.
Pipettes should not be placed vertically in a cylinder that,
because of its height, must be placed on the floor outside the
biosafety cabinet. Removing contaminated pipettes from
the biosafety cabinet and placing them vertically in a
cylinder provides opportunity for dripping from the pipette
onto the floor, or the rim of the cylinder, thereby creating an aerosol, and the top of the pipettes
often protrude above the level of disinfectant.
Place discard pans for used pipettes within the biosafety cabinet.
After suitable contact time, excess disinfectant can be carefully poured down the sink. The pan
and pipettes can be autoclaved together, and replaced by a clean pan with fresh disinfectant.

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4.2.9 Blenders, Mixers, Sonicators and Cell Disruption Equipment
Hazardous aerosols are created by most laboratory
operations involving blending, mixing, stirring,
grinding or disrupting biohazardous materials. Even the
use of a mortar and pestle can be a hazardous
operation. Other devices that may produce aerosols are
ball mills, colloid mills, jet mills, tissue grinders,
magnetic mixers, stirrers, sonic cleaning
devices/sonicators, homogenizers, ultrasonic cell
disintegrators, French Presses, and shakers.
Adequate decontamination is essential prior to sonic

cleaning due to possible aerosol generation. Wherever
sonicators are used in the cleaning process; such as in
dishwashers, animal cage washers, etc.; all items
should be sterilized prior to cleaning.
The laboratory practices generally required when using equipment that may generate aerosols with
biohazardous materials are as follows:
Operate blending, cell disruption, and grinding equipment in a biological safety cabinet.
Use safety blenders designed to prevent leakage from the rotor bearing at the bottom of the bowl.
In the absence of a leakproof rotor, inspect the rotor for leakage prior to operation. A preliminary
test run with sterile water, saline, or methylene blue solution is recommended prior to use.
If the blender is used with infectious material place a towel moistened with an appropriate
disinfectant over the top of the blender. Sterilize the device and residual contents promptly after
use.
Glass blender bowls are undesirable for use with infectious material because of the potential for
glass bowls to break.
Blender bowls sometimes require supplemental cooling to prevent destruction of the bearings and
to minimize thermal effects on the product.
Before opening the safety blender bowl, permit the blender to rest for at least one minute to allow
settling of the aerosol cloud.
Grinding of infected tissues or materials with any open device is best done within a biological
safety cabinet.
4.2.10 Lyophilizers
Specimens snap-frozen in ampules are dried on a vacuum manifold or in a chamber-type drier at low
negative pressure. If the glass neck of the ampule is sealed off while the ampoule is still under vacuum, it
may cause implosion, either during the sealing or later when the evacuated ampule is being opened. To
avoid this, after drying is completed, and before sealing is done, bring the pressure within the ampule back
to normal by gradually introducing dry nitrogen, avoiding turbulent disturbance of the dry product.
The narrow or constricted neck of the ampoule is contaminated if the specimen is allowed to run down the
wall of the neck during filling. Subsequently, when the ampule is sealed with a torch, the dried material on
the wall becomes charred or partially decomposed; residues of this material may adversely affect the dried
material when it is reconstituted. To avoid this, a syringe with a long cannula or a Pasteur-type pipette
should be used to fill the vial. Do not allow the delivery end of the cannula or pipette to touch the neck of
the vial.
All ampules used for freeze-drying of cultures, toxins, or other biohazardous material should be fabricated
of Pyrex-type glass. This type of glass requires a high-temperature torch using an air-gas or oxygen-gas
mixture for sealing. These hard glass ampoules are much less apt to form gas bubbles that burst inwardly
during sealing under vacuum than the soft glass ampoules and are more resistant to breakage during

4-36
handling and storage.
The filling of ampules and vials with infectious specimens, the subsequent freeze-drying, and sealing or
closing of ampules and vials in the preparation of dry infectious specimens should be performed in a
biological safety cabinet. The same is true for the preparation of ampules and vials containing liquid
specimens not subject to freeze-drying.
Safety precautions to be taken will depend on the agents, equipment, and containment available. Therefore,
before initiating this procedure, the principal investigator should work out the protocol for each machine in
consultation with the Biological Safety Officer. All persons using the procedure must then follow the
protocol.
4.2.11 Microtome/Cryostat
Due to the very sharp blade and the nature of the materials used with the
microtome/cryostat, training is essential in the use of the equipment and in the
hazards of the materials used with the equipment. Users should be informed of
the need to prevent cuts and scrapes as well as protect the eyes, nose, mouth and
skin from exposure to the materials being used.
New personnel must be trained in the proper use and maintenance of the
equipment, and demonstrate proficiency prior to use.
If using human tissue, microtome/cryostat users are required to attend

Bloodborne Pathogens training. Fixatives take time to penetrate tissue; the
fixatives may not inactivate pathogens deep in the tissue. Freezing and drying
do not inactivate most pathogens, so, as with fixative use, the pathogens that may
be present in the tissue should be considered capable of causing infection.
Microtome/cryostat users will likely need to also attend Chemical Safety Laboratory Personnel training due
to the fixatives and dyes used in histology.
When purchasing new units the available safety features should be taken into
consideration prior to deciding on a manufacturer or model. Some available
safety features are:
Auto-decontamination cycle
Easy blade release for installing and changing blades.
Retractable knife/blade to permit safe entry into chamber for
cleaning, retrieving specimens, etc.
Disposable blades.
Never retrieve samples, change blades, or clean equipment by hand with the
blade in place; always use appropriate engineering controls (i.e. forceps,
tweezers, dissecting probes, and small brushes).
Things to remember when using and maintaining microtomes/cryostats:

Always keep hands away from blades.
Use extreme caution when aligning blocks, the blocks may be close to the blades. If available,
make sure block holder is in locked position when loading/aligning blocks.
Use knife-edge protectors/guards. Do not leave knife-edges that may extend beyond microtome
knife holder unprotected.
Keep blocks wet when in the microtome to minimize airborne shavings during slicing.
Use brushes to clean/brush equipment.
Use engineering controls such as forceps when removing or changing the blade.
Dislodge stuck blocks using mechanical means such as forceps and/or dissecting probes.

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Wear appropriate PPE such as a lab coat or gown, mask, safety glasses or goggles, surgical grade
Kevlar gloves that provide dexterity and cut protection, and examination gloves to protect against
biohazards.
When changing blades, wear stainless steel mesh gloves to provide additional protection from cuts
and scrapes.
Avoid freezing propellants that are under pressure as they may cause splattering or droplets of
infectious materials.
Decontaminate equipment on a regular schedule using an appropriate disinfectant.
Consider trimmings and sections of tissue as contaminated and discard in the appropriate waste
stream.
Do not move or transport microtome with knife in position.
Do not leave knives out of containers when not in use.
Do not leave motorized microtomes running unattended.
4.2.12 Fluorescence Activated Cell Sorters (FACS) and FACS analyzers

The following is excerpted from Schmid, I, Lambert, C. Ambrozak D and Perfetto SP (2007) Standard
Safety Practices for Sorting of Unfixed Cells. Current Protocols in Cytometry 3.6.1-3.6.20. John Wiley &
Sons, Inc. :
In 1994 the International Society of Analytical Cytology (ISAC) first recognized the need to formulate
safety guidelines for sorting and analysis of unfixed cells to provide laboratories with recommendations for
practices to reduce the potential for biohazard exposure of instrument operators. These standards are
periodically updated by ISAC, most recently in 2007 (See: http://www.isac-
net.org/media/Biosafety_sorting_2007.pdf ).
Biological particles 0.1 μm to 60 μm in size (e.g., aerosols) have been found to be important in the spread
of infectious diseases. Submicrometer particles formed through dehydration of small droplets (droplet
nuclei) can contain inorganic material, organic material, or infectious agents and may stay suspended in air
for prolonged periods of time. During inhalation, larger particles are deposited mainly into the nasal
passages, 3- to 7-μm particles into the tracheal area and pharynx, and ≤3-μm particles into the lungs of the
exposed individual. Droplets that fall out of suspension in air will land on surfaces, and pathogens they may
contain can then be transmitted by exposure to broken skin or mucous membranes, or by ingestion.
Consequently, protection of all laboratory workers from exposure is critical, in particular during high-risk
procedures such as droplet-based cell sorting using instruments with high system pressures.
Jet-in-air technology utilized for cell sorting involves a liquid stream carrying the cells through a nozzle
vibrating at high frequency. At a given distance from the nozzle orifice the stream is broken into individual
droplets. These droplets are then passed between high voltage plates. Droplets containing cells of interest
with parameters preselected by the operator are electrostatically charged and
deflected into sort sample receptacles. Overall droplet size depends on the
instrument operating pressure and the size of the nozzle orifice and its
vibration frequency. High-speed cell sorters utilize higher system pressures
and sort frequencies and thus produce more smaller droplets compared to
older instruments designed for lowspeed separations. All sorters also
generate microdroplets, i.e., satellite droplets, 3 to 7 μm. Owing to the high
fluid pressure produced in high-speed cell sorters large amounts of
secondary aerosols of various and undefined droplet sizes can occur during
instrument failures, for instance, when a partial clog in the nozzle causes a
deflection in the fluid stream that is hitting a hard surface, e.g., the waste
Aerosol produced after deflection from the waste
catcher. This figure shows the amount and position of catcher. Droplets larger than 80 μm constitute the majority of droplets
aerosols produced after deflecting from the side of the
waste catcher. (Photo from: Perfetto, SP, Ambrozak generated during sorting and settle quickly out of the atmosphere; smaller
DR, Koup RA, and Roederer M (2003) Measuring
Containment of Viable Infectious Cell Sorting in droplets, however, may be aerosolized, particularly when they are elevated
High-Velocity Cell Sorters. Cytometry Part A
52A:122–130.) by air currents. Because of the potential health risk to sorter operators and

4-38
the environment if aerosols escape into the room, aerosol containment of a sorter, whether free standing or
enclosed in a biological safety cabinet, must be verified routinely using appropriate testing methods, such
as the use of highly fluorescent Glo-Germ® (5-µm melamine copolymer resin beads in a 5-ml volume of
ethanol) under the same conditions as the cell sort.
Although FACS analyzers are typically a more contained unit that the FACS sorter, it is important to note
that some functional FACS analysis measurements on cells (e.g., evaluation of calcium flux or membrane
potential, certain apoptosis assays, cytokine assays, or live DNA or RNA staining) preclude cell fixation,
and when performed on jet-in-air flow cytometers, can also expose operators or bystanders to potentially
hazardous aerosols and sample splashes. Therefore, the safety practices outlined here apply whenever
unfixed samples are run through a jet-in-air flow cytometer or a cell sorter that combines a flow cell with
jet-in-air sorting.
When sorting any infectious or hazardous material, even if it is classified as BSL-2, it is critical to
understand that droplet-based sorting procedures are considered BSL-3 practices.
It is therefore recommended that viable, unfixed samples that are potentially infectious be sorted at a
minimum on a sorter which has been tested for aerosol containment located in a modified BSL-2 facility
using practices and containment equipment recommended for BSL-3 by the CDC. However, because of the
increased hazard of a sudden quick release of large amounts of fluid or aerosols into the environment, it is
highly recommended that high-speed sorting be performed in a BSL-3 laboratory facility under complete
BSL-3 containment.
All risks associated with materials being used or presented for sorting or analysis in any MCG FACS
facility should be fully disclosed to the facility manager before use and documented on the PI’s Biosafety
Protocol for risk assessment. Currently, the MCG core FACS facilities do not offer facilities capable of
adequately containing BSL-2 materials when FACS sorting.
For further information on FACS Biosafety, please contact the Biosafety Office (x1-2663 or
BIOSAFETY@mcg.edu) or the FACS core facility director. Also, you may wish to review the following
references:
Schmid I, Lambert C, Ambrozak D, Marti GE, Moss DM and Perfetto SP (2007) International
Society for Analytical Cytology Biosafety Standard for Sorting of Unfixed Cells. Cytometry
Part A 71A: 414-437.
Schmid I, Lambert C Ambrozak D and Perfetto SP (2007) Standard Safety Practices for Sorting of
Unfixed Cells. Current Protocols in Cytometry 3.6.1-3.6.20. John Wiley & Sons, Inc.
Perfetto SP, Ambrozak DR, Koup RA, Roederer M (2003) Measuring Containment of Viable
Infectious Cell Sorting in High-Velocity Cell Sorters. Cytometry Part A 52A:122–130.
4.2.13 Miscellaneous Equipment (Waterbaths, Cold Storage, Shakers, etc.)

Water baths and Warburg baths used to inactivate, incubate, or test infectious substances should contain a
disinfectant. For cold water baths, 70% propylene glycol is recommended.
Sodium azide should not be used as a bacteriostatic. It creates a serious
explosion hazard.
Whenever possible, the use of dry heat/incubator blocks and dry incubators
should be considered instead of water baths since this may reduce the risks
of water intrusion upon the samples and contamination of water in baths or
shakers. Dry heat blocks come with modular adapters designed to fit tubes
of multiple sizes as well as rectangular 12-, 24- or 96-well plates.
Deep freezer, liquid nitrogen, and dry ice chests as well as refrigerators should be checked, cleaned out
periodically to remove any broken ampules, tubes, etc. containing infectious material, and decontaminated.

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Use rubber gloves and respiratory protection during this cleaning. All infectious or toxic material stored in
refrigerators or deep freezers should be properly labeled. If equipment failure occurs, immediate action to
prevent specimen loss and contamination of equipment and facilities may be required. Security measures
should be commensurate with the hazards.
The degree of hazard represented by contaminated liquid nitrogen reservoirs will be largely dependent
upon the infectious potential of the stored microorganisms, their stability in liquid nitrogen, and their ability
to survive in the airborne state. Investigations suggest that storing tissue culture cell lines in containers
other than sealed glass ampoules might result in potential inter-contamination among cell lines stored in a
common liquid nitrogen repository.
Care must be exercised in the use of membrane filters to obtain sterile filtrates of infectious materials.
Because of the fragility of the membrane and other factors, such filtrates cannot be handled as non-
infectious until culture or other tests have proved their sterility.
Shaking machines should be examined carefully for potential breakage of flasks or other containers being
shaken. Screw-capped durable plastic or heavy walled glass flasks should be used. These should be
securely fastened to the shaker platform. An additional precaution would be to enclose the flask in a plastic
bag with or without an absorbent material.
No person should work alone while performing any extremely hazardous operation.
4.3 LABORATORY FACILITIES

Laboratory Facility design and function is the third element, along with practices and equipment, which
provides appropriate containment for work with biological materials. The unique design and construction
of laboratory facilities contributes to the laboratory workers' protection, provides a barrier to protect
persons outside the laboratory, and protects persons or animals in the community from infectious agents
that may be accidentally released from the laboratory. Keep in mind, conversion of non-laboratory spaces
into laboratory spaces is not always possible without significant monetary investment, since the design,
ventilation requirements and furnishings within the non-laboratory areas may not comply with those
required by the Board of Reagents or other safety standards.
Laboratory directors are responsible for providing facilities commensurate with the laboratory's function
and the recommended biosafety level for the agents being manipulated. The recommended secondary
barrier(s) will depend on the risk of transmission of specific agents. For example, the exposure risks for
most laboratory work in BSL-1 and BSL-2 facilities will be direct contact with the agents, or inadvertent
contact exposures through contaminated work environments. Secondary barriers in these laboratories may
include separation of the laboratory work area from public access, availability of a decontamination facility
(e.g., autoclave), and hand washing facilities.
When the risk of infection by exposure to an infectious aerosol is present, higher levels of primary
containment and multiple secondary barriers may become necessary to prevent infectious agents from
escaping into the environment. Such design features include specialized ventilation systems to ensure
directional air flow, air treatment systems to decontaminate or remove agents from exhaust air, controlled
access zones, airlocks as laboratory entrances, or separate buildings or modules to isolate the laboratory.
Design engineers for laboratories may refer to specific ventilation recommendations as found in the
ASHRAE Laboratory Design Guide published by the American Society of Heating, Refrigerating, and Air-
Conditioning Engineers (ASHRAE).
The Georgia Board of Reagents has issued general laboratory design criteria to which all facilities designed
and built in the University Systems of Georgia system must adhere. However, additional considerations,
based on the risk assessments of the biological, chemical and radiological materials agents to be used in the

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facility and anticipated operations should be incorporated into any laboratory construction or renovation
project during the design phase of the project.
Animal facilities, in particular, present significant design and construction challenges. Small disruptions
(loud or unusual noises, odors, illumination, etc.) can greatly impact animal research operations. From a
biosafety perspective, special considerations should be made in determining the locations of these facilities,
as well as the equipment and mechanical, electrical and plumbing (MEP) features in and around these
facilities. These design issues can be particularly challenging for architects and engineers unfamiliar with
these issues.
In addition, considerations should be made during design to ensure that the facility can be properly
maintained without disruption of operations which may lead to inadvertent hazardous material exposure of
maintenance personnel, laboratory personnel, ongoing research activities (including research animals), the
environment or the surrounding community. Proper facility operation and maintenance is key to
maintaining containment in any laboratory.
Some general biosafety considerations for design and operations of facilities are discussed below.
4.3.1 Physical Separation of Laboratory Spaces from Non-Laboratory Spaces

A physical barrier must exist which distinguishes “laboratory areas” from “non-laboratory areas”.
Basically, a room is either considered a laboratory or it is not. Laboratory areas are where hazardous
materials may be handled. “Non-laboratory” areas may include offices, hallway areas, kitchens, or library,
where unauthorized personnel (i.e., those who have not been properly informed of the hazards and provided
precautionary medical screening) may be present and in which activities not permitted within the laboratory
(e.g., food or drink consumption/storage, storage of utensils for food/drink consumption, application of
cosmetics, gum chewing, etc.) are permitted.
All laboratories require floor-to-ceiling physical separation between these two areas. Cubicles, study
carrels and/or artificially demarcated areas (e.g., taped-off areas) physically located within a laboratory do
not satisfy the physical separation criteria. Therefore, all restrictions within laboratories (e.g., food/drink
consumption and storage restrictions) apply to all desks physically located within laboratories which do not
have floor-to-ceiling separation from the laboratory space. During laboratory design, accommodation for
the needs of the laboratory research staff, particularly for separated areas for storage and consumption of
food or drink, should be considered.
Also, consider that gloves must be removed and hands must be washed before exiting the laboratory areas,
so hand-washing facilities near the exit doors is a convenient feature that should be considered during
design of facilities.
4.3.2 Doors and Locks

Doors leading from laboratories into non-laboratory areas (e.g., offices, hallways) should be kept closed
after entering or exiting from the laboratory to maintain the physical barrier between laboratory and non-
laboratory areas. Doors to hallways, offices or other “non-laboratory” area (as discussed in Section 4.3.1)
should be kept closed. If doors open inward into laboratory spaces, consideration should be made to
include a window in the door to prevent collisions into personnel handling biological (or other) hazardous
materials. Ventilation grilles are not permitted for laboratory doors.
Laboratory doors in BSL-2 facilities should be self-closing and have locks to provide security for/from the
materials stored and handled within the facility. Animal facility doors which open to the exterior should
not only be self-closing, but also self-locking. Doors to areas where infectious materials and/or animals are
housed should open inward and should be kept closed when experimental animals are present. These doors
should never be propped open. Doors to cubicles inside of animal rooms may open outward or slide
horizontally or vertically.

4-41
4.3.3 Laboratory and Biological Material Security
Measures should be taken to secure biological materials within the laboratory to prevent inadvertent
exposure of personnel who have not been informed of the risks and received appropriate health
screening/vaccinations, theft or inadvertent release. Whenever possible, accommodations for storage of
higher risk agents (≥ Risk Group 2 and biological toxins) should be made within the most secure area of the
PI’s laboratory rather than in common or shared storage rooms.
If materials must be stored in common/shared facilities, additional measures to secure the materials may be
required, including freezer/cryotank locks or lockboxes within refrigerators or freezers.
Security for any Select Agent or Toxin (See Section 2.6.2.2) is even more critical and is required by
Federal Law.
4.3.4 Windows
Laboratory windows that open to the exterior are strongly discouraged. However, if a laboratory does have
windows that open to the exterior, they must be fitted with screens.
In animal facilities, external windows are not recommended, since the presence of windows may impact
facility security. Exterior windows in animal facilities should therefore be assessed by security personnel.
However, if windows are present, they must be resistant to breakage, and where possible, sealed.
4.3.5 Floor Coverings, Walls and Ceilings

Laboratory facilities in which biological materials are housed should be designed to be easily cleaned and
decontaminated. Since organic material, including cloth and unvarnished wood, and porous materials are
virtually impossible to properly disinfect, the use of these materials in construction or furnishing biological
laboratories is discouraged. Carpets and rugs in laboratories are not permitted in any laboratory.
In animal facilities, all interior surfaces (walls, floors and ceilings) need to be assessed for cleanability.
These surfaces need to be water-resistant, and floors must be slip-resistant, impervious to liquids and
resistant to chemicals. Penetrations in floors, walls and ceiling surfaces should be sealed, to include
openings around ducts, doors and door frames, to facilitate pest control and proper cleaning. Ceilings
should be smooth, impervious to moisture, able to withstand cleaning with detergents and chemical
disinfectants and be free of imperfect junctions.
4.3.6 Furnishings
All furnishings within any laboratory must be capable of supporting anticipated work loads and uses and
should be easily cleaned and disinfected. Since organic material, including cloth or unvarnished wood, and
porous materials, such as unsealed cement, are virtually impossible to properly disinfect, the use of these
materials in construction or furnishing biological laboratories is discouraged.
Chairs must be covered with a non-porous material that can be easily cleaned and decontaminated with an
appropriate disinfectant. Sharp edges and corners should be avoided.
Spaces between benches, cabinets, and equipment should be accessible for cleaning. Storage of cardboard
boxes, particularly on the floor, is discouraged because this may impede cleaning and disinfection efforts.
In addition, cardboard may harbor insects or may beome moldy (particularly when used in damp areas,
such as cold rooms).

4-42
In BSL-2 and ABSL-2 facilities, a method for decontaminating all laboratory wastes should be available in
the facility (e.g., autoclave, chemical disinfection, incineration, or other validated decontamination
method). No plants or animals which are not associated with the experiments in the laboratory are
permitted in BSL-2 laboratories.
4.3.7 Lab Benches/Cabinetry

Again, because they must be durable and cleanable, bench tops and cabinets in all laboratories must be
impervious to water and resistant to heat, organic solvents, acids, alkalis, and other chemicals. Spaces
between benches, cabinets, and equipment should be accessible for cleaning.
4.3.8 Exhaust Systems/Directional Airflow/HVAC

Although there are no specific requirements on ventilation systems at BSL-1 or -2 facilities, the CDC/NIH
recommends that in planning new facilities, considerations should be made in the mechanical ventilation
systems to provide an inward flow of air without recirculation to spaces outside of the laboratory. This is
often required by the Chemical Safety regulations, that are also often present in MCG laboratories.
In addition, any specific ventilation requirements related to the proposed use or equipment in the laboratory
should also be addressed during laboratory design (e.g., compressed gas tanks, particularly tanks containing
compressed cryogenic liquids, are permitted only in well-ventilated areas due to the potential asphyxiation
risk associated with the rapid displacement of oxygen within an enclosed spaces).
Hazardous components of exhausted laboratory air can potentially contaminate the environment or
inadvertently expose personnel (particularly maintenance personnel, who are more likely to be working on
the ventilation systems on rooftops, etc.). This should also be considered when designing the exhaust
system of these facilities. Stacked exhaust systems which expel the discharged air at high velocities and/or
HEPA filtration should be considered based on the risks present.
Animal facilities have stricter requirements for ventilation to comply with the Guide for Care and Use of
Laboratory Animals. No recirculation of exhaust air should occur in any animal facility and should be
discharged to the outside without being recirculated to other rooms. While it is recommended that all
animal rooms have inward directional airflow compared to adjoining hallways or rooms; this is required for
compliance with ≥ABSL-2 standards. Ducted exhaust air ventilation systems are also an ABSL-2 standard.
Ventilation system design should consider the heat and high moisture load produced during the cleaning of
animal rooms, and especially the cage washing areas.
4.3.9 Plumbing/Eyewashes/Showers
Handwash sinks should be present in each laboratory room to enable personnel to wash their hands just
prior to exiting from the facility. In BSL-1 and BSL-2 containment laboratories, the sink may have a
manual, hands-free or automated operation. The location of the handwash sink should be near the exit
door, and additional sinks for hand-washing should be located in other appropriate locations within the
facility, as well. For instance, if an animal facility has segregated areas where infectious materials and/or
animals are housed or manipulated, a sink must also be available for hand washing at the exit from each
segregated area.
Emergency safety showers are required in all animal facilities and within a 10 second walking distance
from any location within an MCG laboratory, since hazardous chemicals are often present in the
laboratories, in addition to biological materials.
BSL-2 laboratories and all animal facilities must also have eyewashes. MCG uses only plumbed twin-
stream (binocular) eyewashes to enable compliance with ANSI standards for eyewashing. Contact the
EHOS department at x1-2663 for recommendations of makes and models of eyewashes. Eyewashes should

4-43
be flushed out regularly (weekly is recommended) by laboratory staff to prevent the growth of
microorganisms within the water lines, which could expose personnel to further mucous membrane damage
during emergency eyewash use.
Sink or floor drain traps are filled with water, and/or appropriate liquid to prevent the migration of vermin
and gases into the laboratories. If floor drains are provided, the traps are filled with water, and/or
appropriate disinfectant to prevent the migration of vermin and gases.
4.3.10 Biosafety Cabinets

BSCs must be installed so that fluctuations of the room air supply and exhaust do not interfere with proper
operations. BSCs should be located away from doors, windows that can be opened, heavily traveled
laboratory areas, and other possible airflow disruptions.
HEPA filtered exhaust air from a Class II BSC can be safely re-circulated back into the laboratory
environment if the cabinet is tested and certified at least annually and operated according to manufacturer’s
recommendations.
BSCs can also be connected to the laboratory exhaust system by either a thimble (canopy) connection (e.g.,
a Class IIA2 BSC) or a direct (hard) connection (e.g., a Class IIB2 BSC). Provisions to assure proper
safety cabinet performance and air system operation must be made and verified. All BSCs should be used
according to manufacturer’s recommendation, to protect the worker and avoid creating a hazardous
environment from volatile chemicals and gases. The choice of the most appropriate Class II BSC/exhaust
system for the intended uses in the facility will likely hinge on anticipated use of volatile chemicals and
radiological materials within the BSC. If the BSC is unducted, no work with radiological or volatile
chemicals is permitted in the BSC; minute amounts are permitted in thimble-connected Class IIA2 BSCs;
small amounts are permitted in hard-ducted Class IIB2 BSCs. However, laboratory designers are cautioned
to consider the differences in the mechanical, electrical and HVAC demands of the different BSCs. Class
IIB2 BSCs require significantly more static air pressure and energy to function than thimble-connected
Class IIA2 BSCs. In addition, because the exhaust of the Class IIB2 BSCs is intricately linked to the
HVAC exhaust system to which it is connected, they may be more prone to failure if exhaust fluctuations
occur. Accurate specifications should be obtained and incorporated prior to final design.
4.3.11 Lighting, Air Ducts, Utility Pipes

Adequate illumination is required for all activities in the laboratory. Care should be taken to avoid
reflections and glare that could impede vision.
To facilitate cleaning efforts, all internal facility appurtenances, such as light fixtures, air ducts, and utility
pipes, should be arranged to minimize horizontal surface areas to facilitate cleaning and minimize the
accumulation of debris or fomites. This is a requirement of animal facility design.
Air ducts should not be located over lab benches, but should be positioned over aisles so airflow readings
may be taken periodically to monitor the laboratory’s HVAC performance and air balance.
4.3.12 Additional Animal Facility Considerations

Laboratory animal facilities pose unique challenges to balance biosafety and animal care and use issues. In
general laboratory animal facilities requirements can be found in the Guide for the Care and Use of
Laboratory Animals and Laboratory Animal Welfare Regulations; however, assessments should also be
made based on biosafety, occupational health and safety risks to address potential hazards associated with
the conduct of laboratory animal research.

4-44
Facilities for laboratory animals used in studies of infectious or non-infectious disease should be physically
separate from other activities such as animal production and quarantine, clinical laboratories, and especially
from facilities providing patient care. Traffic flow that will minimize the risk of cross contamination
should be incorporated into the facility design. Animal areas should be as restricted as possible. Special
containment equipment or facility design may be required as determined by appropriate risk assessment.
In addition, provisions should be made for animal-specific operations. At ABSL-1, cages may be washed
manually or preferably in a mechanical cage washer. At ABSL-2 containment, the cages should first be
autoclaved or otherwise decontaminated prior to washing using a mechanical cage washer. The mechanical
cage washer should have a final rinse temperature of at least 180°F. The cage wash area should also be
designed to accommodate the use of high pressure spray systems, humidity, strong chemical disinfectants
and 180°F water temperatures, during the cage/equipment cleaning process.

4-45
5. MEDICAL SURVEILLANCE PROGRAM
All personnel who are authorized to work with biological materials in the execution of MCG’s research or
educational missions are required to have access to Employee/Occupational health services.
The purpose of an Occupational Health program is to conduct periodic health assessments of personnel
handling biological materials with particular attention devoted to factors or conditions associated with a
particular biological agent a given individual might handle. For a particular person, the medical
surveillance program might call for any of a number of precautionary measures, including immunizations, a
periodic physical examination and collection of a serum specimen. Each laboratory should have the
requirements for Employee Health Screening and vaccinations as part of their Standard Operating
Procedures. The purpose of the medical surveillance program is to:
recommend appropriate medical precautions to be followed, and
do periodic reassessment of employees to determine if medical conditions associated with

employment are prevalent and, if so, to undertake definitive measures to alleviate them.
The extent of medical surveillance for a given employee will vary greatly and be dependent upon:
the nature of the research project in which involved,

the biological agents to which directly or potentially exposed, and
certain additional factors relating to the current or previous health status of the individual.
The CDC BMBL standard for Biosafety Level 2 places the responsibility on the laboratory supervisor to
provide Employee Health services and education pertaining to their health risks while working as indicated:
“The laboratory supervisor must ensure that laboratory personnel receive appropriate training regarding
their duties, the necessary precautions to prevent exposures, and exposure evaluation procedures. Personnel
must receive annual updates or additional training when procedural or policy changes occur. Personal
health status may impact an individual’s susceptibility to infection, ability to receive immunizations or
prophylactic interventions. Therefore, all laboratory personnel and particularly women of child-bearing age
should be provided with information regarding immune competence and conditions that may predispose
them to infection. Individuals having these conditions should be encouraged to self-identify to the
institution’s healthcare provider for appropriate counseling and guidance.” (CDC BMBL, Biosafety Level
2, A. Standard Microbiological Practices, 11:
http://www.cdc.gov/od/ohs/biosfty/bmbl5/sections/SectionIV-LaboratoryBiosafetyLevelCriteria.pdf )
Employee Health services are also a requirement of the OSHA Bloodborne pathogen standard
(http://www.osha.gov/pls/oshaweb/owadisp.show_document?p_table=STANDARDS&p_id=10051) 29
CFR 1910.1030 and maintain health records on all employees with potential occupational exposure to
Bloodborne pathogens or other potentially infectious material.
MCG policies for seeking any appropriate health counseling and vaccination differ based on whether the
laboratory personnel is an employee or a non-employee (which would include any student who is not an
MCG employee). MCG contracts with the MCG Health, Inc. for Occupational Health services for MCG
employees. A medical surveillance program should be offered by the employer to personnel engaged in
biological research, which are conducted by the Employee Health office (x1-3418) located at room 1174
FG Building, which is at 1515 Pope Avenue (see map, below). Non-MCG employee students who may be
working with biological materials in the course of the education should seek health counseling and
surveillance services with the Student Health Office (x1-3487) which is at 1040 AF (Pavilion II) Building
off Laney Walker Blvd (see map, below). It is beholden upon MCG to offer any personnel who are not
MCG employees (volunteers, visitors) who may be working with biological materials to further the MCG
research or educational mission while at MCG the appropriate Occupational Health services prior to
authorizing them to work within the laboratory or clinic, or subsequent to an incident, exposure or accident

5-1
involving biological materials. It is the responsibility of the Principal Investigator, Clinic Director and/or
Instructional Course Director to map out how these personnel will receive the appropriate health
counseling, vaccination and post-exposure follow-up health care as part of the laboratory SOPs
Employee Health screening and medical surveillance should be provided without charge for any MCG
employee whose job may result in potential exposure. As MCG has no university-wide health program,
these costs are typically incurred at the departmental level. Individuals seeking Employee Health Services
should be provided by the authorized Employee Health IDR for services prior to their appointment.
5.1 TUBERCULOSIS (TB) SCREENING

Personnel who face occupational exposure to Tuberculosis (TB) should be enrolled in the University’s
Tuberculosis Exposure Control Plan. The Occupational Safety and Health Administration has identified
workers from the following areas as potentially exposed:
Laboratories that may handle M. tuberculosis or be exposed to patients or specimens from patients
with tuberculosis.
Healthcare facilities
Long term care facilities
Correctional facilities
Homeless shelters
Substance abuse treatment facilities
New employees at risk must be tested for TB exposure by a tuberculin skin test (PPD) at time of hire
(within 2 weeks of start date) to establish a baseline. All employees at risk must be PPD tested on an

5-2
annual basis. Students who may encounter TB exposure as part of their educational operations should
follow similar practices with the Student Health Services.
MCG personnel who have been exposed to active TB cases in the course of accomplishing their MCG-
research or educational duties must report the incident and undergo an initial baseline TB test at time of
exposure and a follow up test at 3 months post exposure. Please contact Employee Health (x1-3418) or
Student Health (x1-3487) to arrange for PPD testing. Contact the Biosafety Office in the Office of
Environmental Health and Safety at x1-2663 or BIOSAFETY@mcg.edu for information on TB training.
In addition, any personnel working with non-human primates should be carefully monitored for potential
exposures to tuberculosis. A TB infection causes an extremely rapidly fatal pneumonia in most Old World
primate species which can devastate an entire NHP colony, while it typically manifests as chronic
pneumonia in humans. Typically, infected humans present a much greater risk to the animals than animals
do to humans. It is unusual for the disease to be transmitted to humans, unless the animal is undergoing
surgery or pathologic examination. Infected tissue samples can also present a risk to laboratory workers.
Prevention of disease includes routine use of respiratory protection and protective clothing when working
with tissues or when coming into close proximity to animals. Animals and human handlers should be
screened every 6 months for disease. Animals should also be quarantined and screened on entry into the
facility.
5.2 IMMUNIZATIONS
In certain situations, personnel engaged in particular research or educational activities would be immunized
with appropriate vaccines, such as rabies, rubella and measles. Further information about adult vaccines
can be found at the following URL: http://www.cdc.gov/vaccines/vpd-vac/adult-vpd.htm and the
recommended schedule for adult vaccinations and boosters can be found at the following file:
http://www.cdc.gov/vaccines/recs/schedules/downloads/adult/06-07/adult-schedule.pdf . Vaccines not
commonly available will be obtained, whenever possible, for those engaged in specific research with
potential exposure to the agent in question.
Vaccine Recommendations
Rabies Vaccine Recommended for all personnel entering laboratories or animal

facilities with rabies vaccination entrance requirements.
Hepatitis B Vaccine Recommended for persons working with patients in a health care
setting or persons working with human blood, body fluids, tissues or
cell lines derived from human materials.
MMR (Measles, Mumps, Rubella) Recommended for persons working with patients in a health care
Td (Tetanus, diphtheria) or TDap (Tetanus, Recommended for persons working with patients in a health care
Diphtheria, Pertussis) setting or persons working with human blood, body fluids, tissues or
Current tetanus vaccination is required of all those who work with

laboratory animals, including researchers and animal caretakers. In
addition, anyone whose job responsibilities associated with an animal
facility may place them at risk of exposure to tetanus.
Influenza Recommended for persons working with patients in a health care
Varicella Zoster Vaccine Recommended for persons who have not previously had varicella
zoster infection (i.e. chicken pox and/or shingles) who work with
patients in a health care setting or persons working with human blood,

5-3
body fluids, tissues or cell lines derived from human materials.
Vaccinia (Smallpox) Vaccine Prior to working with vaccinia, employees are required to receive an
independent medical counseling session from the Employee Health
regarding vaccinia immunization to discuss the risks and benefits of
vaccinia vaccination. See 5.2.1, below, for further information.
Contact the Biosafety Office (x1-2663 or BIOSAFETY@mcg.edu) to
make arrangements.
In cases where infected animals are not housed in filter-top cages or

other primary containment devices, vaccination shall be required for
room entry.
Arboviruses: Eastern and Western Equine Prior to working with arboviruses, employees are required to receive a
Encephalitis Vaccines, Japanese Encephalitis medical evaluation and counseling from Employee Health regarding
Vaccine, Venezuelan Equine Encephalitis possible immunization.
Vaccine, Yellow Fever Vaccine, Rift Valley
Fever Vaccine
Lyme Disease Vaccine Recommended for persons working with the Lyme Disease agent or
vectors in research laboratories, with animals, or in fieldwork.
Other vaccines such as Salmonella typhi To be determined by the Employee Health Physician.
(Typhoid),
In some cases, appropriate follow-up serum samples will be collected at periodic intervals to measure
vaccine-induced antibodies when indicated.
5.2.1 Vaccinia Virus Use and Immunization Recommendations

Recombinant vaccinia and other pox viruses are useful microbiological research tools for expression of
exogenous proteins in a variety of cultured cell types. However, their use is not without risk to laboratory
personnel. Naturally or experimentally infected laboratory animals are a potential source of infection to
exposed unvaccinated laboratory personnel. Genetically engineered recombinant vaccinia viruses pose an
additional potential risk to laboratory personnel, through direct contact or contact with clinical materials
from infected volunteers or animals. Infectious vaccinia virus particles may be present in lesion fluids or
crusts, respiratory secretions, or tissues of infected hosts and could lead to secondary infections of other
individuals who may come in contact with these materials, such as family members of the laboratory
personnel. Ingestion, parenteral inoculation, and droplet or aerosol exposure of mucous membranes or
broken skin with infectious fluids or tissues, are the primary hazards to laboratory and animal care
personnel. Serious ocular infections have been known to occur even in vaccinated individuals, see:
Lewis et al.(2006) Ocular vaccinia infection in a laboratory worker, Philadelphia, 2004. Emerging
Infectious Diseases. 12(1): 134-7
Ruben FL & Lane JM. (1970) Ocular Vaccinia. Archives of Ophthalmology 84: 45-48.
Because of these risks, the IBC will expect that all manipulations of vaccinia strains be performed at BSL-2
containment in a biosafety cabinet. If work must be performed outside of a biosafety cabinet (e.g. animal
surgery, microscopy), the following personal protective equipment must be used:
gloves
lab coat
eye and mucous membrane protection [safety glasses or goggles which are ANSI certified and
surgical mask or face shield].

5-4
Multiple strains of vaccinia virus exist with varying levels of virulence for humans and animals. Depending
on the strain used, vaccinia virus presents varying levels of health risk to laboratory personnel. Strains that
are highly attenuated are typically unable to replicate or replicate poorly in human cells. On the other hand,
non-highly attenuated strains of vaccinia have the ability to replicate in human cells and thus pose a risk to
humans. Risks include localized skin infections and more severe, disseminated reactions to which
immunocompromized individuals may be more susceptible. However, vaccination against vaccinia poses
certain health risks to the individuals, and there are several contraindications for vaccination. The risks and
benefits of vaccination and exposure to vaccinia in the laboratory sett to the laboratory personnel and those
in close contact to the laboratory personnel should be discussed in a health counseling session with the
Employee Health Office.
The following information and recommendations are based on national guidelines issued by the CDC in
Vaccinia (Smallpox) Vaccine Recommendations of the Advisory Committee on Immunization Practices.
MMWR, Recommendations and Reports, 50, 2001
(http://www.cdc.gov/mmwr/preview/mmwrhtml/rr5010a1.htm ):
Because of the inherent risks in the vaccination process, vaccination will likely not be recommended by
the IBC for those working with the following highly attenuated strains, depending on the risk
assessment of the Biosafety Protocol:
Highly Attenuated Strain Biosafety Derived from:
Level *
MVA 2 Vaccinia virus (Ankara)
NYVAC 1 Vaccinia virus (Copenhagen)
TROVAC 1 Fowlpox virus
ALVAC 1 Canarypox virus
*Keep in mind: the eventual recommended biosafety level may increase depending on the presence and
characteristics of a foreign protein expressed by a recombinant vaccinia virus or other aspects of the
proposed experiment, as assessed by the IBC during its review process.
These recommendations were based upon the following considerations:

Laboratory personnel who work with highly attenuated strains of vaccinia virus (e.g., MVA
and NYVAC) or who work with the Avipoxvirus strains ALVAC and TROVAC do not
require routine vaccinia vaccination.
The Occupational Safety Health Board of NIH no longer requires vaccinia vaccination for
personnel manipulating MVA or NYVAC in laboratories using only those strains.
The Recombinant DNA Advisory Committee of the NIH reduced the biosafety level of
NYVAC, TROVAC and ALVAC to level 1 based on accumulated attenuation data and
biological properties of these strains.
Although there is no formal surveillance system in place, there have not been any reports of
laboratory-acquired infection resulting from exposure to any of the above highly attenuated
strains or recombinant vaccines derived from these strains in the literature or to the CDC.
Appropriate biosafety guidelines and infection control procedures should always be observed
when working with viral material even if vaccination is not indicated.
The IBC is likely to highly recommend vaccinia vaccination to laboratory workers who directly
handle cultures or animals infected with non-highly attenuated vaccinia virus strains, recombinant
vaccinia viruses derived from non-highly attenuated vaccinia strains, or other orthopox viruses that can
infect humans. Although recommended, the IBC will not require the vaccination, particularly considering
the number of contraindications associated with vaccinations. However, the risks of working with vaccinia
without vaccination should be fully communicated to the laboratory staff members. Anyone who has a

5-5
contraindication for vaccination (including: eczema, atopic dermatitis, immunosuppression and pregnancy,
or having household contacts with any of these conditions) should also consider these contraindications for
work with non-highly attenuated strains of vaccinia or orthopox viruses capable of human cell infection.
Non-highly attenuated strains WR (Western Reserve, mouse neuroadapted derivative)

NYCBOH (strain used in vaccinia vaccine)
Copenhagen
Temple of Heaven
Lister
Other orthopox viruses Cowpox, Monkeypox
The vaccine, Dryvax®, is a live virus preparation of vaccinia virus manufactured by Wyeth and
distributed by the CDC. It is administered with a bifurcated needle using a multi-puncture
technique.
Vaccinia immunization results in high seroconversion. The resulting immunity should provide
protection to recipients against infections resulting from uncontrolled, inadvertent inoculation by
unusual routes (e.g., the eye) with a substantial dose of virus of higher or unknown pathogenicity.
Individuals with certain conditions are more likely to experience severe side effects or
complications from the vaccine and should not be vaccinated. Contraindications include eczema,
atopic dermatitis, immunosuppression and pregnancy, or having household contacts with any of
these conditions.
Revaccination every 10 years is recommended for people working with non-highly attenuated
vaccinia strains; more frequent revaccination may be required for more virulent orthopox viruses.
Laboratory personnel not directly handling cultures of vaccinia or animals infected with vaccinia,
but working in the same lab where non-highly attenuated strains are being used should be offered
medical screening for potential contraindications to vaccinia exposure.
Other health-care workers (such as physicians and nurses) whose contact with these viruses is
limited to contaminated materials (for example, dressings), but who adhere to appropriate
infection control measures, are probably at lower risk for inadvertent infection than laboratory
workers. However, because a theoretical risk of infection exists, vaccination may be considered
for this group.
A summary of published case reports of laboratory-acquired vaccinia virus infections is available
in:
Byers K (2005) Biosafety Tips-Biosafety Issues in Laboratory Experiments Using
Vaccinia Viral Vectors. Applied Biosafety, 10(2): 118-122
Based on these guidelines, laboratory personnel for whom vaccination is recommended must receive
mandatory confidential medical counseling before beginning work with the virus. These individuals
must be counseled on the risks and benefits of the vaccine and medically screened for contraindications to
vaccinia exposure or vaccination. During the counseling session, the risks and benefits of vaccinia
immunization will be discussed and concerns or questions will be addressed. Following medical
consultation the individual will either: sign a vaccinia vaccination consent form and receive the vaccine.
(This will require follow-up visits to monitor the vaccination site), OR sign a declination form.
IBC approval may occur before all personnel listed on the Request for Vaccinia Vaccination form has been
counseled and vaccinated. It is the Principal Investigator's responsibility to ensure that all personnel
working directly with vaccinia have received counseling. Evidence of medical counseling must be
documented in the laboratory biosafety manual.
Additional information about the Smallpox vaccine is available through the Centers for Disease Control
and Prevention web site:
Smallpox Fact Sheet: http://www.bt.cdc.gov/agent/smallpox/vaccination/facts.asp
Smallpox Vaccine: What You Need to Know:
http://www.bt.cdc.gov/agent/smallpox/vaccination/vaccine.asp

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5.3 RESPIRATORY PROTECTION PROGRAM
As described in Section 4.2.1.2, Federal OSHA regulations (29 CFR 1910.134) require initial and annual
training and fit-testing, and well as medical surveillance of all respirator wearers
(http://www.osha.gov/pls/oshaweb/owadisp.show_document?p_id=12716&p_table=STANDARD). The
purpose of a medical evaluation program is to determine if employees can tolerate the physiological burden
associated with respirator use, including: the burden imposed by the respirator itself (e.g., its weight and
breathing resistance during both normal operation and under conditions of filter, canister, or cartridge
overload); musculoskeletal stress (e.g., when the respirator to be worn is a SCBA); limitations on auditory,
visual, and olfactory sensations; and isolation from the workplace environment. Since certain jobs and
workplace conditions in which a respirator is used can also impose a physiological burden on the user, the
medical evaluation must also consider the following factors: type and weight of the respirator to be worn;
duration and frequency of respirator use; expected physical work effort; use of protective clothing and
equipment to be worn; and temperature and humidity extremes that may be encountered. This information
must be provided to the physican or other licensed health care professional (PLHCP) before the PLHCP
makes a recommendation regarding an employee's ability to use a respirator.
The medical evaluation is designed to identify general medical conditions that place employees who use
respirators at risk of serious medical consequences. Medical conditions known to compromise an
employee's ability to tolerate respirator-, job-, and workplace-related physiological stress include:
cardiovascular and respiratory diseases (e.g., a history of high blood pressure, angina, heart attack, cardiac
arrhythmias, stroke, asthma, chronic bronchitis, emphysema); reduced pulmonary function caused by other
factors (e.g., smoking or prior exposure to respiratory hazards); neurological or musculoskeletal disorders
(e.g., ringing in the ears, epilepsy, lower back pain); impaired sensory function (e.g., perforated ear drums,
reduced or absent ability to smell); and psychological disorders (e.g., claustrophobia and severe anxiety).
If a respirator is to be used in the course of research with biological materials, the MCG IBC will require
adherence to these OSHA standards as a stipulation of IBC approval, since these represent prudent
precautionary safety measures.
Please contact the MCG Office of Environmental Health and Occupational Safety (EHOS) (x1-2663)
whenever the use of a respirator is being considered. EHOS staff will assist users in:
Proper selection of cartridges and respirators
Guiding researchers in the process to meet the appropriate medical evaluation requirements (which
typically involves documenting annual respirator medical evaluation through the Employee Health
office)
Providing the required annual training and respirator fit testing.
5.4 MEDICAL RESTRICTIONS

As previously described in Section 3.2.1 Risk Groups: Hazardous Characterizations of a Biological agent,
all risk group and containment recommendations presented in the CDC/NIH BMBL, the NIH Guidelines,
the WHO Biosafety Guidelines, etc.,. are based on the risks to healthy human adults. These
recommendations do not account for individual health considerations, such as allergies, pregnancy, breast
feeding, medication effects, a compromised immune system (due to illnesses or medical treatments such as
steroids or chemotherapy) or other illnesses which may make individuals more susceptible to agents.
For this reason, for their own safety, any individual with special health concerns is strongly encouraged to
discuss these with the Principal Investigator, Clinical Director or Instructional Course Director prior to
initiation of work within the laboratory. In turn, PIs, Clinical Directors or Instructions Course Directors
should offer the opportunity for the individual to seek Employee Health Counseling through the Employee
Health Office to discuss their potential individual special risks.

5-7
5.4.1 Pregnancy
It is recognized that exposure to certain infectious agents may adversely affect a fetus during pregnancy if
the mother is infected with the agent. Therefore, if pregnancy is possible while you are working in an
infectious disease laboratory or laboratory engaged in work with infectious agents you should consult your
Principal Investigator or supervisor. The Employee Health or Student Health Offices should also be made
available for answering questions regarding the potential harm from the biological agents present within
your laboratory.
Women who are pregnant or become pregnant are encouraged to inform their supervisors, Principal
Investigators, Clinic Directors and/or Instructional Course Directors, who should, in turn, encourage them
to seek appropriate health counseling through the Employee or Student Health Offices. Personnel are
urged to discuss exposure issues with their supervisors or principal investigators regarding associated risks
of research being conducted and pregnancy. Employee/Student Health will give advice about precautions
that might be necessary.
The Employee Health and Student Health Offices are resources for pregnant women to ask about any
questions or concerns they may have regarding risks in their work environment. The Employee Health and
Student Health Offices may require additional information about the agents and on-going operations within
the laboratory beyond what the laboratory personnel is able to offer.. The Employee or Student Health
Office may need to discuss these matters with the Principal Investigator, Clinic Director or Instructional
Course Director, or they may contact the Biosafety Office (x1-2663 or BIOSAFETY@mcg.edu) to discuss
the agents and operations documented in the laboratory’s Biosafety Protocol. The Employee Health or
Student Health Office may also act as a liaison between pregnant laboratory personnel and their respective
supervisors or principal investigators. Please contact the Employee Health (x1-3418) or Student Health
Offices (x1-3487) for further information on reproductive and fetal pathogens.
5.4.1.1 Reproductive Biological Hazards

The Employee Health Physician will offer confidential counseling to any woman or man of
childbearing age working with reproductive pathogens or other potentially infectious materials.
Reproductive biological hazards include, but are not limited to the following:
Cytomegalovirus (CMV)
Hepatitis B virus (HBV)
Hepatitis E virus
Human Immunodeficiency virus (HIV)
Human parvovirus B19
Rubella (German Measles)
Lymphocytic Choriomeningitis virus
Toxoplasma gondii (Toxoplasmosis)
Listeria monocytogenes
Varicella-zoster virus (chicken pox)
Coxiella burnetii (Q fever)
Vaccinia virus
Whenever necessary, Employee Health along with the Biosafety Office will offer an opportunity
to review work procedures in the lab to ensure that potential exposure is minimized.
Consideration for reassignment to other tasks that do not involve exposure to the reproductive
hazard (generally with actual pathogens, not necessarily for only other potentially infectious
materials such as blood or body fluids) should be given. Also, Principal Investigators actively
working with reproductive hazards should explain these risks at the time of hire.

5-8
5.4.2 Work with Animals
Occupational Health programs play an important part of both animal use regulations, as well as biosafety
guidelines. Special hazards exist for workers who are exposed to animals, and therefore guidance is
provided by the Institute for Laboratory Animal Research (ILAR) Commission on Life Sciences, National
Research Council related to Occupational Health issues in:
Occupational Health and Safety in the Care of Research Animals

http://www.nap.edu/catalog.php?record_id=4988
Occupational Health and Safety in the Care and Use of Nonhuman Primates
http://www.nap.edu/catalog.php?record_id=10713#toc.
5.4.3 Allergies
5.4.3.1 Allergies to Laboratory Animals (ALAs)

Allergic reaction to animals is among the most common condition that adversely affects worker
health. The estimated prevalence of allergic symptoms among workers exposed to animals is from
10% to 40%. Workers who are continually exposed to animal allergen tend to have progressively
more frequent and severe symptoms, and an estimated 10% develop asthma. Studies have shown
that about 50% of those with symptoms will eventually stop working with animals permanently or
temporarily because of the discomfort involved in ALA.
Initial allergy symptoms are usually:

Runny nose (allergic rhinitis),
Itchy eyes (allergic conjunctivitis)
Rashes (contact urticaria, atopy).
Symptoms usually evolve over a period of 1-2 years and may lead to acute anaphylaxis in a small
number of patients.
Hence, it is critical that all workers seek to minimize their exposure to animal allergens and that
Principal Investigators and Laboratory supervisors discuss these risks with their laboratory
workers. Supervisors are also responsible to ensure that appropriate operating procedures have
been established to prevent undue exposure of workers to animal allergens and provide Employee
Health/Student Health screening for workers who may be developing hypersensitivity to assess
their risks of further work with animals. These risks should also be communicated to others in the
laboratory to inform them of the potential consequences of exposure to their fellow laboratory
staff member to prevent inadvertent exposure.
In rodents, the allergen protein is of urinary origin while in rabbits, the primary allergen is
contained in the fur and dander and to a lesser degree in the saliva and urine. In guinea pigs, urine
is the main allergen with dander, fur, and saliva contributing. The major cat allergen is produced
in oil glands of the skin and coats the hair shafts. It is also present in saliva. Exposure to birds
can cause rhinitis and asthma symptoms. Multiple bird proteins have been identified as allergens
and can be found in serum and fecal droppings that contain serum. Fish proteins can be an
inhalation allergen for those who are sensitized.
Prudent efforts to prevent allergen exposure and reduce the frequency of sensitization in animal
workers require strict work practices and consistent use of PPE. Housing animals in filter-top
cages, working in well-ventilated areas, and using ventilated hoods for soiled bedding disposal
will minimize exposure to animal allergens.
The work area must be maintained clean to prevent inhalant and contact exposure. Procedures
should be adopted that minimize release of airborne materials, including bedding dust and

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antibiotic aerosols, and the contamination of hands, arms, body and face. Workers should adopt
the use of PPE during each and every animal contact or allergen exposure.
Of particular importance is wearing a face mask to reduce inhalation and hand-to-face spread of
allergens and covering all exposed skin (i.e. gloves, lab coat, sleeve protectors) to prevent allergen
contact. In some cases, respirators may be recommended; in which case the employee must be
enrolled in MCG’s respirator program, and undergo annual fit-testing as per OSHA standards (29
CFR 1910.134).
It is also important that once animal procedures are complete, all contaminated PPE and clothing
are removed and properly disposed of to prevent repeated exposure while performing subsequent
duties. Contact your supervisor, LAS supervisors or biosafety further information regarding PPE.
5.4.3.2 Latex Gloves and Related Allergies

Allergic reactions to natural rubber latex have been increasing
since 1987, when the Center for Disease Control recommended
the use of universal precautions to protect against potentially
infectious materials, bloodborne pathogens and HIV. Increased
glove demand also resulted in higher levels of allergens due to
changes in the manufacturing process. In additional to skin
contact with the latex allergens, inhalation is another potential
route of exposure. Latex proteins may be released into the air
along with the powders used to lubricate the interior of the
glove.
In June 1997, the National Institute of Occupational Safety and

Health (NIOSH) issued an alert, “Preventing Allergic
Reactions to Latex in the Workplace” (publication number
DHHS (NIOSH) 97-135). The full text of this publication is
available at the NIOSH web site,
http://www.cdc.gov/niosh/topics/latex/.
NIOSH studies indicate that 8-12% of healthcare workers

regularly exposed to latex are sensitized, compared to 1-6% of
the general population. Latex exposure symptoms include skin
rash and inflammation, respiratory irritation, asthma and
shock. The amount of exposure needed to sensitize an
individual to natural rubber latex is not known, but when
exposures are reduced, sensitization decreases.
NIOSH recommends the following actions to reduce exposure
to latex:
If latex gloves must be used, choose reduced-protein, powder-free latex gloves.

Whenever possible, substitute another glove material (for instance, nitrile gloves)
Wash hands with mild soap and water after removing latex gloves
5.4.3.3. Antibiotic Allergies

Allergic reactions have been described to a large number of medicines, and those against
antibiotics is one of the most common of these. Reactions to antibiotics can range from a rash or
hives starting a few days after exposure to sudden onset of rashes, difficulty breathing, stomach
upset and anaphylaxis soon after exposure. Because of the potential severity of these reactions,
any personnel with known allergies to antibiotics should discuss their personal health risks in
working in a laboratory with the Employee Health or Student Health Offices.

5-10
In the laboratory setting, antibiotic allergies may impact the risks in two ways:
1. Should exposures occur to biological materials for which the medical treatment modality
is administration of an antibiotic against which the laboratory personnel is allergic, an
alternate post-exposure treatment plan should be made prior to exposure in conjunction
with the Principal Investigator, Clinical Director and/or Instructional Course Director.
These supervisors should be also be made aware of the potential for allergic reaction, so
this can be communicated to health care providers in an emergency situation
2. Biomedical laboratories often use antibiotics for research purposes. For instance,
antibiotic selection is often used during culture operations in both microbiological and
mammalian cell/tissue culture settings. The risks of exposure of allergic personnel to the
antibiotic should be communicated to the Principal Investigator, Clinic Director and/or
Class Instruction to:
a. Enable development of any operating practices which would help limit the
exposure any allergic personnel to the antibiotic
b. Enable communication of the exposure risks to others in the laboratory to
consider the exposure risks to the allergic personnel.
5.4.3.4 Mold Allergies

Some people are very sensitive to molds. For these people, exposure to molds can cause symptoms
such as nasal stuffiness, eye irritation, wheezing, or skin irritation. Some people, such as those
with serious allergies to molds, may have more severe reactions. Severe reactions may occur
among workers exposed to large amounts of molds in occupational settings... Severe reactions
may include fever and shortness of breath. Some people with chronic lung illnesses, such as
obstructive lung disease, may develop mold infections in their lungs.
Molds can be found almost anywhere; they can grow on virtually any substance, providing
moisture is present. There are molds that can grow on wood, paper, carpet, and foods. There is no
practical way to eliminate all mold and mold spores in the indoor environment; the way to control
indoor mold growth is to control moisture and humidity which may provide ideal conditions for
mold growth. In buildings, conditions which can favor mold growth can include roof and
plumbing leaks or floods, condensation, and excess humidity. In large buildings, mold and
mildew are commonly found on the exterior wall surfaces of corner rooms in heating climate
locations. In laboratories, mold is often found in areas where moist conditions are present and
condensation is likely to occur such as cold rooms, inside refrigerators, or in thawed freezers or
decommissioned warm rooms-- particularly when organic materials (like cardboard) are placed in
these areas.
Because of the potential health consequences to personnel with mold allergies, care should be
taken by all MCG personnel (i.e., not just laboratory staff), to take measures to prevent mold
growth in MCG facilities. Prudent preventative measures include:
Not storing organic materials which can serve as growth medium to mold in moist
atmospheres. Do not store cardboard boxes in cold rooms, freezers or refrigerators.
Inspect the building for signs of mold, moisture, leaks, or spills routinely. Clean up
mold and eliminate sources of moisture as soon as possible.
Fix the source of water problems or leaks to prevent mold growth.
Reduce indoor humidity (to 30-60% ) to decrease mold growth by: venting
bathrooms, dryers, and other moisture-generating sources to the outside; using air
conditioners and de-humidifiers; increasing ventilation; and using exhaust fans
whenever steam-producing operations occur (such as glassware or cage washing or
shower facilities).
Clean and dry any damp or wet building materials, floors/carpeting and furnishings
within 24-48 hours to prevent mold growth.
Clean mold off hard surfaces with water and detergent, and dry completely.
Absorbent materials such as ceiling tiles, that are moldy, may need to be replaced.

5-11
Prevent condensation: Reduce the potential for condensation on cold surfaces (i.e.,
windows, piping, exterior walls, roof, or floors) by adding insulation.
In areas where there is a perpetual moisture problem, do not install carpeting (i.e., by
drinking fountains, by sinks, or on concrete floors with leaks or frequent
condensation).
Further information about mold in the indoor environment, please see the EPA Mold Resources
web page, at: http://www.epa.gov/mold/moldresources.html.
5.4.4 Other Restrictions

Restrictions or recommendations will be made on an individual basis after discussion with the Employee
Health Physician and the employee's personal medical doctor. Examples of conditions that might warrant
special precautions are HIV infection, immunosuppressive conditions and drug therapy that suppresses the
immune system (e.g. steroid therapy). Therefore, if you are suffering from any of the above conditions, you
must inform your physician and the Employee Health or Student Health Offices about the situation.
5.5 MINORS IN LABORATORIES AND OTHER HAZARDOUS AREAS

All risk group and containment recommendations presented in standard Biosafety Guidelines (CDC/NIH
BMBL, the NIH Guidelines, the WHO Biosafety Guidelines, etc.), are based on the risks to healthy human
adults and do not account for differential effects on immature bodies/systems. Persons under the age of 18
are prohibited from entering laboratory areas or other areas where hazardous materials or conditions may
be present except when such entry is a pre-authorized, escorted, scheduled open house tour. During these
times, no work with Risk Group ≥2 organisms or recombinant DNA will be permitted and the facility
should be decontaminated to prevent inadvertent exposure to minors to any residual hazardous material
while visiting.
Because of their biological, social, and economic characteristics, young workers have unique and
substantial risks for work-related injuries and illnesses. For this reason, both the Federal and State
Departments of Labor has issued special requirements for work of minors under the age of 18 years old
http://www.dol.gov/dol/allcfr/Title_29/Part_570/toc.htm. These laws specifically generally restrict minors
under the age of 18 from working in hazardous areas
http://www.dol.gov/dol/topic/youthlabor/hazardousjobs.htm, and specifically restrict any minor under the
age of 18 from working in areas where this is exposure to radiation or storage/manufacture of explosives.
CDC/NIOSH has also issued some recommendations for protecting the health and safety of minors while in
occupational settings (see: http://www.cdc.gov/niosh/topics/youth/ and
http://www.cdc.gov/niosh/docs/NIOSHRecsDOLHaz/default.html).
By definition, any laboratory is a potentially hazardous area, whether these hazards are presented by
biological materials, chemicals, or radioactive substances. Few studies exist which adequately characterize
potential differential health consequences of exposure of minors to these hazards. While the Biosafety
Office recognizes that paid or volunteer employment within a laboratory can be valuable learning
experience, minors under the age of 18 years should be restricted from working in these areas, unless
provisions can be made and followed to address the following concerns:
Federal law explicitly prohibits any minor under the age of 18 from working in areas where
exposure to radiation may occur or where explosive materials may be stored or used. This would
limit the MCG laboratories in which a minor would be permitted to enter.
Risk communication and the comprehension of those risks and mitigation methods is a crucial
safety measure within the biomedical laboratory. This may require a certain level of education
background and maturity to understand. General biosafety training modules assume a base level
of comprehension of biological and chemical concepts, and the Biosafety Office would not be

5-12
equipped to provide special training sufficient to compensate for the lack of education and
experience.
Signed liability waivers, normally obtained from parents and guardians to permit minors to enter
laboratories require that the parent or guardian be fully informed of the risks faced by their child in
these areas. Again, the amount of education/training required to bring parents or guardians of
minors who wish to work in a laboratory may greatly vary, and cannot be guaranteed.
Adequate supervision of the minor worker within the laboratory would have to be ensured. This
means all activities engaged by the minor within the laboratory would have to be monitored at all
times, which requires the physical proximity (i.e., within the supervisor’s line-of-sight) and
attentiveness of the supervisor at all times while the minor is within the laboratory. This level of
supervision is difficult to guarantee within the busy working laboratory.
Appropriate occupational health screening and vaccinations would be required of all personnel
within the laboratory. Some vaccinations require multiple administrations over time, and
completion of the vaccination series may not be possible within the time frame for the minor’s
employment/volunteer opportunity. In addition, special health care provisions would have to be
made for the minor in case of incident or potential exposure or illnesses which may have resulted
from a potential exposure. If the minor is in the laboratory on a volunteer basis, and is not an
MCG student, special provisions with the Employee Health and Student Health Offices may be
required to address the minor’s occupational health issues.
5.6 EMPLOYEE SERUM STORAGE

Many infections do not result in an overt disease condition. Such infections are detected by development
of antibodies to the agent in question. Therefore, Employee Health may need to establish a program for
persons engaged in BL3 research or working with non-human primates, which includes collection of pre-
assignment serum. If an illness occurs which may be related to the agent the person is working with,
additional serum samples may need to be collected.

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6. ACCIDENTS, EXPOSURES, SPILL RESPONSE
Laboratory-specific SOPs should address any emergency response procedures, including those required if
an accident, exposure, potential exposure, an illness which may have resulted from a possible laboratory
exposure, release from primary containment or environmental contamination of any biologically hazardous
material. Guidelines for the development of these SOPs are discussed below.
6.1 EMERGENCY PROCEDURES FOR EXPOSURE INCIDENTS

An "exposure incident" is specific contact (eye, mouth, other mucous membrane, respiratory tract via
inhalation, non-intact skin, or parenteral) with potentially infectious materials that results from the
performance of an employee's duties or commission of the person’s responsibilities in conjunction with the
research or educational mission of MCG. A person who sustains a known or potential exposure incident
must remove their gloves and treat the affected area immediately by following the appropriate exposure
incident response below.
6.1.1 Percutaneous Injury

Wash the affected area with antiseptic soap and warm water for 15 minutes.
6.1.2 Splash to Face

Flush affected area in eyewash continuously for 15 minutes.
6.1.3 Aerosol Exposure

Do not inhale and immediately leave room. Remove Personal Protective Equipment (PPE) carefully. When
removing PPE make sure to turn the exposed areas inward. Wash hands well with soap and water. Post a
spill sign on the laboratory entry doors to prevent others from entering. The laboratory should remain
evacuated for at least 30 minutes to allow for the droplets to settle and/or aerosols to be purged by the air
exchange rate within the laboratory.
The PI must clear the laboratory before re-entry and spill clean-up to commence. For extensive BL2
contamination (i.e. an incident involving a centrifuge) or incidents involving BL2+ or BL3 agents, the
Biosafety Office must be notified immediately (x1-2663) and will assume responsibility, in conjunction
with the PI, to clear the laboratory for re-entry.
6.2 REPORTING INCIDENT

The employee must report the incident to his/her supervisor. Because the health and safety issues of the
injured personnel is of primary importance, if the injury is emergent, the supervisor should take (or make
arrangements for the injured person to be taken) to the nearest emergency room and reporting requirements
should be completed post-hoc.
The supervisor should take measures to ensure that additional personnel are restricted from areas to prevent
further inadvertent exposures. Any incident, accident, exposure, possible exposure, illness which may have
resulted from exposure, releases from primary containment or environmental contamination involving
biological materials which occurred in the course of accomplishing the research and/or educational
missions of the university should be reported as soon as possible to the MCG Biosafety Office (x1-2663 or
BIOSAFETY@mcg.edu). It is the responsibility of the Biosafety Office to assist Principal Investigators,
Clinical Directors and/or Instructional Course Directors in completion of any funding agency-specific,
public health or environmental safety reporting requirements and to review the incident with the supervisor

6-1
and the injured person to discuss whether alterations in the laboratory’s or institution’s Standard Operating
Procedures would be prudent to prevent future similar occurrences.
Injured personnel and their supervisors should also keep in mind that the IBC and Biosafety Office are
primarily concerned with ensuring that the appropriate prevention, treatment and post-exposure follow-up
measures are implemented to ensure the health and safety of the exposed personnel and the environment
and prevention of future incidents. Other MCG offices may require the injured person and/or their
supervisor to report incidents, accidents or exposures for other purposes, such as completion of GA State
Workman’s Compensation requirements and/or to maintain campus safety statistics. Therefore, it is
recommended that PIs, Clinic Directors and Instructional Course Directors review these requirements and
incorporate these additional reporting requirements in any laboratory SOP to assist laboratory personnel in
completion of all reporting requirements. Please see the following MCG Policies and Procedures for
guidance:
For MCG Employees:

MCG Policy & Procedure 1.4.17: http://www.mcg.edu/policies/1417.html
For Persons who are not MCG employees (including students):
MCG Policy & Procedure 3.0.03: http://www.mcg.edu/policies/3003.html
For quick reference, here is some contact information and URLs of some reporting forms that may
be required as part of development of laboratory reporting SOPs for compliance with these other
MCG policies. (These are provided merely to assist PIs, Clinic Directors and Instructional Course
Designers in the development of their post-exposure follow-up laboratory SOPs):
• GA State Department of Administrative Services (DOAS):
o Phone number: 1-877-656-7475
o Pertinent forms may include:
• WC-1 form Employer's First Report of Injury
http://sbwc.georgia.gov/vgn/images/portal/cit_1210/2/61/13139736wc001.pdf
• DOAS “Incident Notice Only” form:
http://doas.georgia.gov/vgn/images/portal/cit_1210/4/47/27136091IncidentReportO
nly.pdf
• MCG Human Resources Benefits Section

o Phone number: x1-3770
o Fax: x1-1572
o Location: HS-1111, Annex Building
o Pertinent forms may include:
• MCG Human Resources Employee’s Report of Accident/Injury
form (MC-419) http://www.mcg.edu/hrforms/pdf/WCReportAccidentInjury.pdf
• MCG Public Safety Office x1-2911
6.3 MEDICAL ASSISTANCE

In non-emergent cases, employees are urged to follow the medical assistance instructions provided to them
by the DOAS and their health care provider after they have received proper available first aid at site of
exposure. For certain exposures such as non-human primate bites, scratches or splashes, tick or insect bites,
or exposure to infectious agents, the employee will be advised to be evaluated by the Employee Health
Office. In situations when Employee Health is not available or if more extensive treatment is required, the
employee will likely be referred to the nearest Emergency Room, and asked to follow up with visit(s) with
the Employee Health Office (see Section 5, Medical Surveillance for a map showing nearby health facility
locations).
Personnel who have been potentially exposed on the job will be provided with post-exposure evaluation
and follow-up at no cost to employees who experience "exposure incidents". The post-exposure monitoring

6-2
periods are dependent on the type of exposure. This time period is related to the various incubation periods
of the infectious agents.
Employees can obtain copies of their Employee Health records by contacting the MCGHI Employee Health
Office at FG-1174 (x1-3418). MCG must retain medical records for your duration of employment plus 30
years.
6.4 SPILL CLEAN-UP PROCEDURES

This section is intended to outline the basic procedures for dealing with some of the biological spills that
may be encountered in a research laboratory. All lab personnel should refer to the relevant spill response
procedures before initiating their experiments.
6.4.1 Composition of a Basic Biohazard Spill Kit

Microbiological and biomedical research laboratories should prepare and maintain a biological spill kit. A
spill kit is an essential safety item for labs working with agents which require Biosafety Level 2 or higher
containment and for groups working with large volumes (> 1 liter) under Biosafety Level 1 containment.
The following items should be included in the spill kit:
Concentrated household bleach (< 1 year old)

A spray bottle for making 10% bleach solutions
Forceps, disposable broom/dust pan, and/or other mechanical devices for handling sharps or
removing solid objects within the spill
Paper towels or other suitable absorbent (diapers, disposable shop towels)
Biohazard bags for the collection of contaminated spill clean-up items
Utility gloves and medical examination gloves
Face protection (eye wear and mask, or full face shield)
Signage for warning others to avoid the area
Additional personal protective equipment, such as a disposable cover-gown or disposable booties may also
be recommended equipment in some spill clean-up situations
Although household bleach is recommended as a standard disinfectant in the spill kit, other suitable
disinfectants may be used provided the disinfectant is effective against the agents in use at the appropriate
dilutions and contact time. Disinfectants should be registered with the Environmental Protection
Agency as tuberculocidal for compliance with the Occupational Health and Safety Administration
Bloodborne Pathogens Standard (29CFR 1910.1030).
Representatives from the Biosafety Office are available if you have any questions regarding biological spill
response procedures or decontamination (x1-2663 or BIOSAFETY@mcg.edu).
6.4.2 Spills Occurring Outside of Laboratory Areas

Because laboratory facilities are designed to contain hazards within the confines of the laboratory, where
all who are likely to encounter the material have knowledge of the risks, any spill outside of the laboratory
area poses a particular risk for exposure of the general public or environmental contamination. Therefore,
the procedures for addressing a spill which occurs outside the confines of a laboratory are:
Attend to any injuries or exposures

Alert others to avoid the area to prevent contamination of additional personnel and environment
Contact the Biosafety Office (x1-2663) immediately to assist in spill clean-up. If after hours,
contact Public Safety Office (x1-2911), who has the 24/7 on-call numbers for EHS staff members.

6-3
6.4.3 Spills within the Laboratory Areas
Because laboratory facilities are designed to contain hazards within the confines of the laboratory, spills
within the laboratory are generally not as potentially problematic as those which occur outside the
laboratory. Of course, the overall risk will depend on the agents, operations and personnel involved in the
spill and clean-up measures.
6.4.3.1 Biosafety Level 1 (BSL1) Spills

Immediate Procedures
• Notify others in the area, to prevent contamination of additional personnel and
environment.
• Remove any contaminated clothing and wash exposed skin with soap and water.
Clean-up of BSL1 Spill

• Wearing gloves, lab coat, and face protection, cover spill with paper towels, pour
concentrated disinfectant around the spill allowing it to mix with spilled material.
Allow suitable contact time, at least 15 min.
• Pick up any pieces of broken glass with forceps or other mechanical device(s)
(not with your hands!) and place in a sharps container.
• Discard all disposable materials used to clean up the spill into a biohazard bag.
• Wash hands with soap and water.
6.4.3.2 Biosafety Level 2 (BSL2) Spills

• Avoid inhaling airborne material, while quickly leaving the room. Notify others to
leave.
• Close door, and post with a warning sign.
• Remove contaminated clothing, turning exposed areas inward, and place in a
biohazard bag.
• Wash all exposed skin with soap and water.
• Inform Supervisor, and, if assistance is needed, consult the Biosafety Office (x1-
2663).
Clean-up of BL2 Spill

• Allow aerosols to disperse for at least 30 minutes before reentering the laboratory.
Assemble clean-up materials (disinfectant, paper towels, biohazard bags, and
forceps).
• Put on protective clothing (lab coat, face protection, utility gloves, and booties if
necessary).
• Depending on the nature of the spill, it may be advisable to wear a HEPA filtered
respirator instead of a surgical mask.
• Cover the area with disinfectant-soaked towels, and then carefully pour
disinfectant around the spill. Avoid enlarging the contaminated area. Use more
concentrated disinfectant as it is diluted by the spill. Allow at least a 30 minute
contact time.
• Pick up any sharp objects with forceps or other mechanical devices (not your
hands!) and discard in a sharps container.
• Soak up the disinfectant and spill using mechanical means, such as an
autoclavable broom and dustpan, since there may be sharps under the paper
towels, and place the materials into a sharps container. Smaller pieces of glass

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may be collected with cotton or paper towels held with forceps. If no sharps were
involved in the spill discard the materials into an autoclave bag.
• Wipe surrounding areas (where the spill may have splashed) with disinfectant.
• Spray the area with freshly prepared 10% household bleach solution and allow to
air-dry (or wipe down with disinfectant-soaked towels after a 30-minute contact
time).
• Place all contaminated paper towels and any contaminated protective clothing into
a biohazard bag and autoclave or dispose in the appropriate Stericycle waste box.
• Wash hands and exposed skin areas with soap and water.
6.4.3.3 Blood Spills

For blood or other material with a high organic content and low concentration of infectious
microorganisms:
• Wear gloves, eye protection, and a lab coat.
• Absorb blood with paper towels and place in a biohazard bag. Collect any sharp
objects with forceps or other mechanical device and place in a sharps container.
• Using a detergent solution, clean the spill site of all visible blood.
• Spray the spill site with 10% household bleach and allow to air-dry for 30
minutes.
• After the 30 minute contact time, wipe the area down with disinfectant-soaked
paper towels.
• Discard all disposable materials used to decontaminate the spill and any
contaminated personal protective equipment into a biohazard bag.
• Wash your hands with soap and water.
6.4.3.4 Spill in a Biological Safety Cabinet

• Leave the biological safety cabinet blower on and begin cleanup immediately.
• While wearing PPE (gloves and gown) cover the spill area with paper towels or
disinfectant soaked paper towels. Do not place your head in the cabinet to clean
the spill, keep your face behind the viewscreen.
• If necessary, flood the work surface as well as the drain pans and catch basins
below the work surface, with disinfectant. Be sure the drain valve is closed before
flooding the area under the work surface.
• Wipe cabinet walls, work surfaces, and inside the viewscreen with disinfectant.
• Lift the front exhaust grill and work surface; wipe all surfaces with disinfectant.
Be sure no paper towels or soiled debris are blown into the area under the spill
tray
• If the work surface, as well as drain pans and catch basins under the work surface,
have been flooded with disinfectant soak up the disinfectant in the work surface.
Place a container under the drain valve and drain the disinfectant under the work
surface into the container.
• Wipe the areas under the work surface to remove residual disinfectant.
• Wash hands and exposed skin with soap and water.
• Autoclave all cleanup materials and protective clothing.
• Notify your PI or supervisor.
• If the spill overflows the drain pan/catch basin under the wok surface into the
interior of the biological safety cabinet notify the Biosafety Office (x1-2663). A
more extensive decontamination of the biological safety cabinet may be required.

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6.4.3.5 Centrifuge Spill
• Always use sealed safety safety-cap sealed buckets or sealed rotors with O-rings.
Examine O-ring and replace if worn, cracking or missing. Check tubes and bottles
for cracks and deformities before each use.
• Wait five minutes before opening the centrifuge following the end of a run with
potentially hazardous biological material if using safety caps or sealed rotors. If a
spill is identified after the centrifuge lid is opened, carefully close the lid and
evacuate the laboratory and close the laboratory door. Remain out of the
laboratory for at least 30 minutes. Post a sign on the laboratory door indicating
there is a biohazard spill and do not enter.
• Remove any contaminated protective clothing and place into a biohazard bag.
Wash hands and any exposed skin surfaces with soap and water.
• Notify your supervisor and the Biosafety Office (x1-2663 or
BIOSAFETY@mcg.edu).
After 30 minutes...
• Enter the lab with personal protective equipment and spill cleanup materials. Full-
face protection, lab coat and utility gloves should be worn.
• Transfer rotors and buckets to a biological safety cabinet. Immerse rotor/buckets
in 70% ethanol or a non-corrosive disinfectant effective against the agent in use.
Allow at least a one hour contact time. Intact tubes may be wiped down and
placed into a new container. Handle any broken glass with forceps and discard
into a sharps container.
• Carefully retrieve any broken glass from inside the centrifuge using forceps and
discard into a sharps container. Smaller pieces of glass may be collected with
cotton or paper towels held with forceps. Carefully wipe the inside of the
centrifuge with disinfectant.
• Spray the inside of the centrifuge with disinfectant and allow to air dry. If bleach
is used, follow by wiping with 70% ethanol to remove any corrosive residues.
• Place contaminated items and disposable personal protective equipment in an
autoclave bag and autoclave.
6.4.3.6 Spill of a Biohazardous Radioactive Material

A biohazardous spill involving radioactive material requires emergency procedures that are
different from the procedures used for either material alone. Use procedures that protect you from
the radiochemical while you disinfect the biological material.
Before any clean up, consider the type of radionuclide, characteristics of the microorganism, and
the volume of the spill. Contact the MCG Radiation Safety Office (x1-2663) for isotope cleanup
procedures.
• Avoid inhaling airborne material, while quickly leaving the room. Notify others to
leave.
• Close door, and post a warning sign.
• Remove contaminated clothing, turning exposed areas inward, and place in a
biohazard bag labeled with a radioactive materials label or a radioactive waste
container labeled with a biohazard label.
• Wash all exposed skin with soap and water; follow with a three-minute water
rinse.

6-6
• Inform supervisor and Radiation Safety Office of spill, and monitor all exposed
personnel for radiation. If assistance is needed in handling the microorganism,
contact the Biosafety Office at x1-2663.
Clean-Up of a Biohazardous Radioactive Material

• Allow aerosols to disperse for at least 30 minutes before reentering the laboratory.
Assemble clean-up materials (disinfectant, autoclavable containers, forceps,
towel, and sponges), and confirm with the Radiation Safety representative that it
is safe to enter the lab.
• Put on protective clothing (gown, surgical mask, gloves, and shoe covers).
Depending on the nature of the spill, it may be advisable to wear a HEPA-filtered
respirator instead of a surgical mask.
• Cover the area with disinfectant-soaked towels, and carefully pour disinfectant
around the spill. Avoid enlarging the contaminated area. Use more concentrated
disinfectant as it is diluted by the spill. Allow at least 30 minutes contact time.
Do Not use bleach solutions on iodinated material, radioiodine gas may

be released. Instead, use an alternative disinfectant such as an iodophor
or phenolic.
• Handle any sharp objects with forceps. Wipe surrounding areas, where the spill
may have splashed, with disinfectant.
• Soak up the disinfectant and spill, and place the biologically decontaminated
waste, along with all protective clothing contaminated with radioactive materials,
into an approved radioactive waste container and label it according to Radiation
Safety Guidelines.
• Contaminated protective clothing must also be biologically decontaminated prior
to disposal as radioactive waste.
Do Not autoclave the waste unless this action is approved by the

Radiation Safety Officer. If waste can not be autoclaved, add additional
disinfectant to ensure biological decontamination of all the materials.
• Wash hands and exposed skin areas with soap and water; monitor personnel and
spill area for residual radioactive contamination.
• If skin contamination is found, repeat decontamination procedures under the
direction of the Radiation Safety Officer.
• If the spill area has residual activity, determine if it is fixed or removable and
handle accordingly.
• Discarding items contaminated with radioactive materials:
• Place the contaminated item(s) on absorbent paper.
• Spray disinfectant (freshly prepared 10% household bleach) on the
contaminated areas and allow 20 minute contact time.
• Wrap the item(s) inside the paper and dispose of as radioactive
waste.
6.5 INVESTIGATION OF LABORATORY ACCIDENTS

The Division of Environmental Health and Safety (EHS), in cooperation with the principal investigator and
his or her staff, will conduct the necessary investigation of a laboratory accident. The goal of the
investigation is the prevention of similar accidents as well as obtaining information concerning the
circumstances and number of employees who have been exposed to the agent in question. In addition,

6-7
EHS, in consultation with the Employee Health and Human Resources might institute further steps to
monitor the health of those who may have been exposed to the agent in question.
It should be emphasized that the reporting of accidents to the principal investigator or laboratory supervisor
is the responsibility of the employee who has the accident. The principal investigator or the laboratory
supervisor should then report to the Biosafety Office (x1-2663 or BIOSAFETY@mcg.edu).
We encourage principal investigators and employees to also report incidents that did not result in an
exposure (“near misses”) to the Biosafety Office. Evaluation of near misses can lead to alternative work
practices and implementation of engineering controls to minimize future incidents.
Whenever an injury involves a sharp and human material (body fluid, tissue, cell line, etc.) the Biosafety
Office must perform an investigation to determine if a safe sharps device is available to prevent future
occurrences of the injury. If safe sharps devices are available, they must be evaluated by the biosafety
office in conjunction with the Group or Department. The incident must also be recorded on the
University’s Sharps Injury Log, maintained by the Human Resources Office. The confidential log will
include the type and brand of device involved in the incident; the Department or work area where the
exposure incident occurred; and an explanation of how the incident occurred.

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7. DECONTAMINATION OF BIOLOGICALS
7.1 DECONTAMINATION METHODS

Decontamination methods fall into three main categories: heat, application of chemical decontaminants
(including vapors and gases) and physical methods (such as filtration or irradiation).
7.1.1 Heat
The application of heat, either moist or dry, is recommended as the most effective method of sterilization.
Steam at 121°C under pressure in the autoclave is the most convenient method of rapidly achieving sterility
under ordinary circumstances. Dry heat at 160°C to 170°C for periods of two to four hours is suitable for
destruction of viable agents on an impermeable non-organic material such as glass, but is not reliable in
even shallow layers of organic or inorganic material that can act as insulation. Incineration is another use
of heat for decontamination. Incineration serves as an efficient means of disposal for human and animal
pathological wastes.
7.1.1.1 Autoclave Use and Maintenance

Moist heat causes the denaturation of proteins at lower temperatures and shorter times than dry
heat. One of the most effective physical decontamination controls is steam sterilization
(autoclave) which generates moisture and high temperature pressurized steam within a sealed
chamber. Autoclaves can sterilize all items that are heat stable. In gravity autoclaves, a cycle of
250°F (121°C) at 15 to 18 pounds per square inch (psi) of pressure for one hour may be required
for decontamination. In the newer vacuum autoclaves, decontamination may require a cycle of
270°F (132°C) at 27 to 30 psi for 45 minutes.
The hazards of handling hot solids and liquids are reasonably familiar. Laboratory personnel
should be cautioned that steam under pressure, such as that found in autoclaves, could be a source
of scalding jets if the equipment is misused. When preparing to use an autoclave, check the door
seal gaskets each time to ensure these are intact prior to using the autoclave. Also, before use,
check to make sure the drain at the bottom of the autoclave is unobstructed. Prepare to autoclave
loads of manageable size only. Do not overfill autoclaves.
Fluids treated by steam under pressure may be superheated if removed from the sterilizer too soon
after treatment; which may cause a sudden and violent boiling of contents from the containers that
can splash scalding liquids onto personnel handling the containers. Therefore slow exhaust cycles
should be used to autoclave liquids. In addition, bottles with liquids should allow for the liquid
expansion during autoclaving; “head room” should be left in the vessels before autoclaving.
Similarly, to avoid over-pressurization will occur inside of vessels with liquids if gas is not able to
freely enter/exit the bottle during the cycle. Therefore, only loosely fit covers or caps on these
vessels prior to autoclaving.
For solid materials, keep in mind that steam must be in contact with the materials to efficiently
sterilize them. Therefore autoclave bags should be left partially opened and/or some additional
water should be placed on the inside of the bag prior to autoclaving.
Because of the air and liquid exchange inside of bags or vessels, other hazardous materials should
not be included in autoclave loads. Mixed waste—either chemicals (such as phenol: chloroform)
or radiological material mixed with biological materials should never be autoclaved due to the
chemical and radiological hazards present.
In addition, some materials should never be autoclaved. Nitrocellulose materials (tubes/filters),

for instance, can explode under autoclave temperatures. Make sure any plastic materials that are
autoclaved are guaranteed “autoclavable” by the manufacturer. Most plastics will melt inside the

7-1
autoclave and produce irritating odors.
Always place materials to be autoclaved inside of a metal or autoclavable plastic autoclave tray to
prevent spillage of agar or melted plastics into the bottom of the autoclave. (Note: most plastics
are not autoclavable unless specially formulated, so check the manufacturer’s specifications to
ensure your plastics are autoclavable!). Melted and re-solidified agar or plastic plugs the drain at
the bottom of the autoclave, which prevents proper function of the autoclave and often requires
maintenance to repair.
Wear closed toe shoes, pants, lab coat, face shield and long sleeved insulated gloves when
operating an autoclave. A heavy, rubberized insulated apron is further recommended for those
who autoclave frequently. Allow time for loads to cool before removing them from autoclaves
after a run. Take proper precautions when first opening the door; first crack the seal on the door
sufficiently to allow the initial burst of steam to escape, then leave the door open about ½ inch to
vent the autoclave for about 5-10 before fully opening the door.
All autoclaves should be on a preventative maintenance program and certified regularly to ensure
proper function. Heat-sensitive autoclave tape can be used to ensure that an autoclave
got warm; however, this is insufficient to tell the user that the If the autoclave is
appropriate temperature and pressure were maintained over a sufficient Prior to not functioning
period of time to provide full decontamination. Please note: MCG autoclaving correctly and spores
does not have a campus-wide autoclave certification and maintenance germinate
program. Responsibility for certifying and maintaining autoclaves falls
to the owner—the department or Principal Investigator who owns the
autoclave. Autoclaves should be certified using bioindicators for steam
autoclaves. Typically, these are vials containing spores of Geobacillus
stearothermophilus in growth media with an colored indicator. To certify
that an autoclave is functioning properly, a vial is placed in the middle of
a typical autoclave load (attaching a string to the vial prior to placement
allows for later retrieval). The autoclave load is run as usual, and the vial
is retrieved from the load after autoclaving. The vial is cultured to
determine whether the spores are capable of germinating, which would Bioindicators
cause a color change in the indicator. Germination of the spores would
indicate that the autoclave is not decontaminating the loads sufficiently, and maintenance should
be performed on the autoclave.
If you experience any problems or unusual occurrences during autoclave use, please report these
to your supervisor and/or building/department manager to enable them to contact the autoclave
maintenance provider. For autoclaves attached to “house” steam lines, ensure that steam pressure
is sufficient to operate the autoclave prior to contacting external repair offices. Insufficient steam
pressure should be reported to the Facilities Management Office (x1-2434) for repair.
7.1.2 Chemical Decontaminants

In general, chemical decontaminants find their most practical use in surface decontamination and, at
sufficient concentration, as decontaminants of liquid wastes for final disposal in sanitary sewer systems.
Further information about the characteristics of the specific chemicals used for disinfection can be found in
Section 7.2.
7.1.2.1 Liquid Chemical Decontaminants

There are many misconceptions concerning the use of liquid decontaminants. This is due largely
to a characteristic capacity of such liquids to perform dramatically in the test tube and to fail
miserably in a practical situation. Such failures often occur because proper consideration was not
given to such factors as temperature, contact time, pH, the presence and state of dispersion,
penetrability and reactivity of organic material at the site of application. Small variations in the

7-2
above factors may make large differences in the effectiveness of decontamination. For this reason
even when used under highly favorable conditions, complete reliance should not be placed on
liquid decontaminants when the end result must be sterility.
There are many liquid decontaminants available under a wide variety of trade names. In general,
these can be categorized as halogens, acids and alkalis, heavy metal salts, quaternary ammonium
compounds, phenols, aldehydes, ketones, alcohols, and amines. Unfortunately, the more active
the decontaminant, the more likely it will posses undesirable characteristics such as corrosivity. In
addition, some of the chemical disinfectants will require disposal as chemical waste after
disinfection.
None is equally useful or effective under all conditions for all infectious agents. Particular care
should be observed when handling concentrated stock solutions of disinfectants. Personnel
assigned to the task of making up use-concentrations from stock solutions must be informed of the
potential hazards and trained in the safe procedures to follow and appropriate personal protective
equipment to use as well as the toxicity associated with ocular, skin and respiratory exposure.
7.1.2.2 Vapors and Gases

A variety of vapors and gases possess decontamination properties. The most useful of these are
formaldehyde, vapor-phase hydrogen peroxide (VHP), chlorine dioxide and ethylene oxide. When
these can be employed in a closed system and under controlled conditions of temperature and
humidity, excellent decontamination can result. Vapor and gas decontaminants are primarily
useful in decontaminating biological safety cabinets and associated air-handling systems and air
filters; bulky or stationary equipment that resists penetration by liquid surface decontaminants;
instruments and optics that may be damaged by other decontamination methods; and rooms,
buildings and associated air-handling systems.
Only those who have received appropriate training in vapor and gas decontamination should
attempt these operations. They should avoid inhalation of vapors of any of these vapors of gasses.
Stock containers of these products should be capable of confining these vapors and should be kept
in properly ventilated chemical storage areas. In preparing use-dilutions and when applying them,
personnel should control the operations to prevent exposure of others and wear respiratory
protection as necessary.
Mutagenic potential has been attributed to ethylene oxide; toxic and hypersensitivity effects are
well-documented for formaldehyde. Ethylene oxide use is very limited and is generally used in
surgical and clinical areas.
Use of vapor and gas disinfection methods is monitored closely by the MCG Division of
Environmental Health and Safety. Please contact EHS at x1-2663 for information regarding the
exposure monitoring program.
7.1.3 Physical Methods

Alternative measures for decontamination which rely on physical properties of decontaminants are often
used as precautionary measures, such as HEPA filtrations on BSC or exhaust systems, rather than a primary
mode of disinfection. The function of HEPA filters was discussed earlier, in Section 4.2.2, Biosafety
Cabinets, and will not be repeated here. Most HEPA filters remove a minimum of 99.97% of particulates
from air.
Gamma-wave irradiation is another method used for decontamination; however, this often requires a high-
energy irradiator, currently making it impractical as an everyday method for decontamination of most
materials at MCG.

7-3
UV irradiation is a somewhat controversial method of disinfection for surfaces. While it is an effective
measure, time of exposure, distance, presence of dust or debris and UV lamp intensity can all affect the
germicidal effect of the UV lamp. Users should recognize that the visible blue-violet glow of the UV lamp
does not indicate there is germicidal effect. The UV lamp needs to be cleaned periodically to remove dust.
UV lamps may damage eyes, skin, and laboratory equipment and shut off while the room is occupied.
Because users tend to over-rely on UV irradiation for surface decontamination in lieu of thorough chemical
surface decontamination and the compounding factors which may impede proper decontamination using
UV irradiation, the Biosafety Office discourages the use of UV lamps. For further information related to
the use of UV lamps for surface decontamination, please see the following references:
Burgener, J (2006) Position Paper on the Use of Ultraviolet Lights in Biological Safety Cabinets.
Applied Biosafety 11(4): 228-230.
Meecham, P.J. and Wilson, C. (2006) Use of Ultraviolet Lights in Biological Safety Cabinets: A
Contrarian View. Applied Biosafety 11(4): 222-227.
7.2 CHARACTERISTICS OF CHEMICAL DECONTAMINANTS

Chemicals with decontaminant properties are, for the most part, available as powders, crystals, or liquid
concentrates. These may be added to water for application as surface decontaminants, and some, when
added in sufficient quantity, find use as decontaminants of bulk liquid wastes. Chemical decontaminants
that are gaseous at room temperature are useful as space-penetrating decontaminants. Others become gases
at elevated temperatures and can act as either aqueous surface or gaseous space-penetrating
decontaminants.
Inactivation of microorganisms by chemical decontaminants may occur in one or more of the following
ways:
Coagulation and denaturation of proteins
Lysis
Binding to enzymes or inactivation of an essential enzyme by oxidation, binding, or
destruction of enzyme substrate.
The relative resistance to the action of chemical decontaminants may be altered substantially by such
factors as: concentration of active ingredient, duration of contact, pH, temperature, humidity, and presence
of extrinsic organic matter. Depending on how these factors are manipulated, the degree of success
achieved with chemical decontaminants may range from minimal inactivation of target microorganisms to
an indicated sterility within the limits of sensitivity of the assay system employed. Ineffectiveness of a
decontaminant is due primarily to the failure of the decontaminant to contact the microorganisms rather
than failure of the decontaminant to act. If an item is placed in a liquid decontaminant, tiny bubbles are
visible on the surface of the item. The area under the bubbles is dry and microorganisms in these dry areas
will not be affected by the decontaminant. If there are spots of grease, rust or dirt on the item,
microorganisms under these protective coatings will not be contacted by the decontaminant. Scrubbing an
item when immersed in a decontaminant is helpful. A decontaminant should have, and most do have,
incorporated surface-active agents.
7.2.1 Properties of Some Common Decontaminants
7.2.1.1 Chlorine/Hypochlorites
This halogen is a universal decontaminant active against a broad spectrum of microorganisms,
including bacterial spores. Chlorine combines with protein and rapidly decreases in concentration
in the presence of protein. Free available chlorine is the active element. Chlorine solutions must
be prepared frequently because of its instability in water.

7-4
Sodium hypochlorite is usually used as a base for chlorine decontaminants. An excellent
decontaminant can be prepared from household or laundry bleach. These bleaches usually contain
5.25%, or 52,500 ppm, available hypochlorite. If diluted 1 to 10, the resulting solution will
contain 0.525% or 5,250 ppm of available hypochlorite. The addition of a 0.7% (v/v) of nonionic
detergent can improve the disinfection properties.
Because hypochlorite is a strong oxidizing agent, it can be corrosive to metals. Therefore, caution
should be taken to prevent etching of stainless surfaces cleaned with bleach solutions.
7.2.1.2 Alcohols
Ethyl or isopropyl alcohol in a concentration of 70-85% by weight is often used; however, both
lose effectiveness at concentrations below 50% and above 90%. Alcohols denature proteins and
are somewhat slow in germicidal action. However, alcohols are effective decontaminants against
lipid-containing viruses. A contact time of ten minutes is generally employed in efficacy tests with
disinfectants. Due to the high evaporation rate of alcohols, repeated applications may be required
to achieve the required ten-minute contact time for decontamination. Because of this, the OSHA
Bloodborne Pathogens Standard does not recognize alcohol as an effective decontaminant for
surfaces.
Isopropyl alcohol is generally more effective against vegetative bacteria; ethyl alcohol is a more
virucidal agent.
Alcohols are also very flammable, so precautions should be taken to prevent exposure to spark or
flame sources.
7.2.1.3 Aldehydes (Formaldehyde, Glutaraldehyde)

Aldehydes are commonly-used fixative agents in laboratories; however, during the process of
fixation, materials are decontaminated. Formaldehyde for use as a decontaminant is usually
marketed as a solution of about 37% concentration referred to as formalin, or as a solid
polymerized compound called paraformaldehyde. Glutaraldehyde is commonly used in
concentrations of 2-4%.
Formaldehyde at a concentration of 5% active ingredient is an effective liquid decontaminant. A

glutaraldehyde-based commercial disinfectant (Cidex®) is used in some hospital settings.
Aldehyde disinfectants lose considerable activity at refrigeration temperatures, and the pungent,
irritating odors and health risks make formaldehyde solutions difficult to use in the laboratory.
The use of Cidex® has been restricted in the U.K. because of the carcinogenic effects. All
materials disinfected with aldehydes must be disposed as chemical wastes to adhere to EPA
standards.
Formaldehyde vapor generated from solution is an effective space decontaminant for buildings or
rooms, but in the vapor state in the presence of water tends to polymerize on surfaces to form
paraformaldehyde, which is persistent and unpleasant. Heating paraformaldehyde to depolymerize
it can liberate formaldehyde gas. In the absence of high moisture content in the air, formaldehyde
released in the gaseous state forms less polymerized residues on surfaces and less time is required
to clear treated areas of fumes than is the case in the vapor state.
7.2.1.4 Phenols
Phenol itself is not often used as a decontaminant. The odor is somewhat unpleasant and a sticky,
gummy residue remains on treated surfaces. This is especially true during steam sterilization.
Although phenol itself may not be in widespread use, phenol homologs and phenolic compounds
are basic to a number of popular decontaminants, such as the original Lysol® and Amphyl®.

7-5
Phenolic compounds are effective decontaminants against some viruses, fungi, and vegetative
bacteria, including rickettsiae. Phenolics are not effective in ordinary use against bacterial spores.
7.2.1.5 Quaternary Ammonium Compounds (a.k.a. “Quats”)

After 40 years of testing and use, there is still considerable controversy about the efficacy of the
Quats as decontaminants. These cationic detergents are strongly surface-active and are effective
against lipid-containing viruses and often vegetative gram positive bacterial however, they have
variable activities against gram negative bacteria and fungi, and are not very effective against non-
lipid enveloped viruses. The Quats will attach to protein so that dilute solutions will quickly loose
effectiveness in the presence of proteins. Quats tend to clump microorganisms and are neutralized
by anionic detergents such as soap. They have the advantages of being nontoxic, odorless, stable,
non-staining, non-corrosive to metals, and inexpensive. Many common household and laboratory
disinfectants are Quats (e.g., Roccal®, Germex®, Coverage Plus®).
7.2.1.6 Iodine
The characteristics of chlorine and iodine are similar. One of the most popular groups of
decontaminants for laboratory use are the iodophors, including Wescodyne®, Betadyne® and
Providone®.
The range of dilution of Wescodyne® recommended by the manufacturer is 1 oz. in 5 gal. of

water (25 ppm available iodine) to 3 oz. in 5 gal. of water (75 ppm available iodine). The small
amount of free iodine available in this range can rapidly be taken up by extraneous protein that
may be present. Clean surfaces or clear water can be effectively treated with 75-ppm available
iodine, but difficulties may be experienced if any appreciable amount of protein is present. For
iodophors such as Wescodyne®, it is critical that the manufacturer’s written instructions are
followed. Higher concentrations of iodophores are actually less effective, as the iodine is bound to
itself or the carrier molecule. For washing the hands or for use as a sporicide, it is recommended
that Wescodyne® be diluted 1 to 10 in 50% ethyl alcohol (a reasonably good decontaminant
itself.) This will give 1,600 ppm of available iodine, at which concentration relatively rapid
inactivation of any and all microorganisms will occur.
7.2.1.7 Peroxygens
Peroxide-based disinfection has become more popular in recent years with the increasing
popularity of vapor-phase hydrogen peroxide generators, which are an effected area
decontamination method. Liquid peroxygen disinfectants, such as Virkon-S® are also available as
surface decontamination methods. Peroxygens have broad-spectrum disinfectant properties and are
generally effective against vegetative bacteria, viruses and some spores. Peroxygens have variable
efficacy in the presence of organic matter. However, peroxygens can be incompatible with some
materials (such as Aluminum, Copper, Zinc, Brass, Natural rubber and some plastics), which
should be considered when selecting disinfectants.
7.2.2 Selecting Chemical Disinfectants

No single chemical disinfectant or method will be effective or practical for all situations in which
decontamination is required. Selection of chemical disinfectants and procedures must be preceded by
practical consideration of the purposes for the decontamination and the interacting factors that will
ultimately determine how that purpose is to be achieved. Selection of any given procedure will be
influenced by the information derived from answers to the following questions:
What is the target organism(s)?

What disinfectants, in what form, are known to, or can be expected to, inactivate the target
organism(s)?
What degree of inactivation is required?

7-6
In what menstruum is the organism suspended (i.e. simple or complex, on solid or porous surface,
and/or airborne)?
What is the highest concentration of organisms anticipated to be encountered?
Can the disinfectant, either as a liquid, vapor, or gas, be expected to contact the organism and can
effective duration of contact be maintained?
What restrictions apply with respect to compatibility of materials?
What is the stability of the disinfectant in use concentrations, and does the anticipated use
situation require immediate availability of the disinfectant or will sufficient time be available for
preparation of the working concentration shortly before its anticipated use?
Will conditions permit safe use of some disinfectants (e.g., is ventilation sufficient to safely use
aldehydes)?
The primary target of decontamination in the laboratory is the organism(s) under investigation. Laboratory
preparations or cultures usually have titers in excess of those normally observed in nature. Inactivation of
these materials presents other problems since agar, proteinaceous nutrients, and cellular materials can
effectively retard or chemically bind the active moieties of chemical disinfectants. Such interference with
the desired action of disinfectants may require higher concentrations and longer contact times than those
shown to be effective in the test tube. Similarly, a major portion of the contact time required to achieve a
given level of agent inactivation may be expended in inactivating a relatively small number of the more
resistant members of the population. The current state of the art provides little information with which to
predict the probable virulence of these more resistant cells. These problems are, however, common to all
potentially pathogenic agents and must always be considered in selecting disinfectants and procedures for
their use.
Organisms exhibit a range of resistance to chemical disinfectants. In terms of practical decontamination,

most vegetative bacteria, fungi, and lipid-containing viruses are relatively susceptible to chemical
disinfection. The non-lipid-containing viruses and bacteria with a waxy coating, such as tubercule bacillus,
occupy a mid-range of resistance. Spore forms and unconventional (slow) viruses are the most resistant.
A disinfectant selected on the basis of its effectiveness against organisms on any range of the resistance
scale generally will be effective against organisms lower on the scale. Therefore, if disinfectants that
effectively control spore forms are selected for routine laboratory decontamination, it can be assumed that
any other organism generated by laboratory operations, even in higher concentrations, would also be
inactivated.
Pertinent characteristics and potential applications for several categories of chemical disinfectants most
likely to be used in the biological laboratory are summarized in the table on the following pages. Practical
concentrations and contact times that may differ markedly from the recommendations of manufacturers of
proprietary products are suggested. It has been assumed that microorganisms will be afforded a high
degree of potential protection by organic menstruums. It has not been assumed that a sterile state will result
from application of the indicated concentrations and contact times. It should be emphasized that these data
are only indicative of efficacy under artificial test conditions. Individual investigators should conclusively
determine the efficacy of any of the disinfectants. It is readily evident that each of the disinfectants has a
range of advantages and disadvantages as well as a range of potential for inactivation of a diverse
microflora. Equally evident is the need for compromise as an alternative to maintaining a veritable “drug
store” of disinfectants.
7.2.3 Characteristics of Some Liquid Disinfectants Table

The following tables, prepared by the Iowa State University Center for Food Security and Public Health are
provided for reference on properties of various chemical disinfectants.

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The Antimicrobial Spectrum of Disinfectants
Chemical Disinfectants
Note: Removal of organic material must always precede the use of any disinfectant.
Acids Alcohols Aldehydes Alkalis Biguanides Halogens Oxidizing Agents Phenolic Quaternary
(hydrochloric (ethyl alcohol, (formaldehyde, (sodium or (chlorhexidine®, hypochlorite iodine (hydrogen peroxide, Compounds Ammonium
acid, isopropyl alcohol) paraformaldehyde, ammonium Nolvasan®, peroxyacetic acid, (Lysol®,Osyl®, Compounds
acetic acid, gluteraldehyde) hydroxide, Chlorhex®, Trifectant®, Virkon-S®, Amphyl®, (Roccal®, Zepharin®,
citric acid) sodium Virosan®, Oxy-Sept 333®) TekTrol®, DiQuat®, Parvosol®,
carbonate) Hibistat®) Pheno-Tek II®) D-256®)
most susceptible
mycoplasmas
gram-positive bacteria
gram-negative bacteria
pseudomonads
susceptibility of microorganisms
rickettsiae
to chemical disinfectants
a
enveloped viruses
chlamydiae
non-enveloped viruses
fungal spores
picornaviruses (i.e. FMD)
parvoviruses
acid-fast bacteria
b
bacterial spores
coccidia c d
prions
highly effective a–varies with composition

most resistant
LEGEND
b–peracetic acid, a strong

effective no activity oxidizing agent, is sporicidal
c–ammonium hydroxide
limited activity information not available d–some have activity against coccidia
Adapted from Linton AH, Hugo WB, Russell AD. Disinfection in Veterinary and Farm Practice. 1987. Blackwell Scientific Publications; Oxford, England.
Disclaimer: Use of trade names does not in any way signify endorsement of a particular product. For additional product names, please consult the most recent Compendium of Veterinary Products.
Characteristics of Selected Disinfectants
Halogens: Quaternary
Disinfectant Halogens: Oxidizing
Alcohols Aldehydes Biguanides Iodine Phenols Ammonium
Category Hypochlorites Agents
Compounds Compounds (QAC)
Ethyl alcohol Formaldehyde Chlorhexidine Bleach Betadyne® Hydrogen peroxide One-Stroke Environ® Roccal�
Isopropyl alcohol Paraformaldehyde Nolvasan® Chlorox® Providone® Peroxyacetic acid Pheno-Tek II® DiQuat�
Sample Glutaraldehyde Chlorhex® Trifectant® Parvosol�
Trade Names Tek-Trol®
Virosan® Virkon S® Pine-Sol, Lysol Zepharin�
Oxy-Sept 333® D-256�
•Precipitates •Denatures proteins •Alters membrane •Denatures proteins •Denatures proteins • Denature proteins • Alters cell wall • Binds phospholipids
Mechanism proteins •Alkylates permeability and lipids permeability of cell membrane
of Action •Denatures lipids nucleic acids • Denatures proteins • Denatures proteins
•Fast acting •Broad spectrum •Broad spectrum •Broad spectrum •Stable in storage • Broad spectrum • Good efficacy with • Stable in storage
•Leaves no residue •Short contact time •Relatively safe organic material • Non-irritating to skin
•Inexpensive • Non-corrosive • Effective at high
Advantages
• Stable in storage temperatures and
• Effective over high pH (9-10)
large pH range
•Rapid evaporation •Carcinogenic •Only functions in •Inactivated •Stains clothes or • Damaging to • Toxic to animals • NOT effective for
•Flammable •Irritation limited pH range by sunlight, treated surfaces some metals • Can cause skin and FMD or Johne’s
to mucous (5–7) some metals •Inactivated by eye irritation
membranes and •Toxic to fish •Requires organic debris and • NOT effective
Disadvantages
tissues (environmental frequent application QACs for FMD
•Only use in well concern) •Corrodes metals •Requires frequent
ventilated areas •Irritating to mucous application
membranes, skin •Corrosive
Never mix with acids;
Toxic to animals,
Precautions Flammable Carcinogenic will release toxic
especially cats
chlorine gas
Vegetative Effective Effective Effective Effective Effective Effective Effective
YES—Gram Positive
Bacteria Limited—Gram Negative
Mycobacteria Effective Effective Variable Effective Limited Effective Variable Variable
Enveloped Effective Effective Limited Effective Effective Effective Effective Variable

Viruses
Non-enveloped Variable Effective Limited Effective Limited Effective Variable Not Effective
Viruses
Spores Not Effective Effective Not Effective Variable Limited Variable Not Effective Not Effective
Fungi Effective Effective Limited Effective Effective Variable Variable Variable
Efficacy with Reduced Reduced ? Rapidly reduced Rapidly reduced Variable Effective Inactivated
Organic Matter
Efficacy with ? Reduced ? Effective ? ? Effective Inactivated
Hard Water
Efficacy
with Soap/ ? Reduced Inactivated Inactivated Effective ? Effective Inactivated
Detergents
? Information not documented
DISCLAIMER: Use of trade names does not in any way signify endorsement of a particular product.
For additional product names, please consult the most recent Compendium of Veterinary Products.
7.3 DEACTIVATION OF BIOLOGICAL TOXINS
The following information is compiled directly from the CDC/NIH BMBL: Appendix I
(http://www.cdc.gov/od/ohs/biosfty/bmbl5/BMBL_5th_Edition.pdf) and the Georgia State University
Biosafety Manual: Further reference material can be found at the URL, above.
Toxin stability varies considerably outside of physiological conditions depending upon the temperature,
pH, ionic strength, availability of co-factors and other characteristics of the surrounding matrix. Literature
values for dry heat inactivation of toxins can be misleading due to variations in experimental conditions,
matrix composition, and experimental criteria for assessing toxin activity. Moreover, inactivation is not
always a linear function of heating time, and some protein toxins possess a capacity to re-fold, and partially
reverse inactivation caused by heating. In addition, the conditions for denaturizing toxins in aqueous
solutions are not necessarily applicable for inactivating dry, powdered toxin preparations.
General guidelines for laboratory decontamination of selected toxins are summarized in Tables 7.3A, B and
C, but inactivation procedures should not be assumed to be 100% effective without validation using
specific toxin bioassays. Many toxins are susceptible to inactivation with dilute sodium hydroxide (NaOH)
at concentrations of 0.1-0.25N, and/or sodium hypochlorite (NaOCl) bleach solutions at concentrations of
0.1-0.5% (w/v). Use freshly prepared bleach solutions for decontamination; undiluted, commercially
available bleach solutions typically contain 3-6% (w/v) NaOCl. Special care should be taken while
deactivating acute biological toxins to protect the handler, but also to ensure thorough decontamination.
See Tables 7.3A, B and C for further guidance on disinfectant procedures.
To chemically decontaminate toxins, perform all operations in a fume hood or biosafety cabinet with the
sash at the lowest reasonable sash height for safe and effective work. Place plastic-backed absorbent paper
(e.g. Benchkote or diaper) on the work surface of the fume hood or BSC to protect the surfaces. Wear
long-sleeved protective clothing (lab coat, gown), gloves and eye protection while decontaminating toxins.
Place toxin into solution in a non-glass primary container, which can be placed in a secondary container,
such as a beaker or rack. Slowly dispense equal volume of the concentrations of NaOCl and/or NaOH or
similar disinfectant to deactivate the toxin. Do not replace the cap on the primary container and allow for a
minimum of 30 minutes exposure time or as recommended in the tables below.
Depending upon the toxin, contaminated materials and toxin waste solutions can be inactivated by
incineration or extensive autoclaving, or by soaking in suitable decontamination solutions (See Table
7.3B). Autoclaving should not be used for destruction of any low molecular weight toxins (e.g.,
mycotoxins, marine and reptile venoms). To autoclave, the cap on the primary container should be
loosened in a fume hood or BSC to allow for steam penetration, then placed in secondary containers before
autoclaving at 121°C for 1 hour on liquid cycle (slow exhaust). Allow time for the materials to cool before
handling and dispose materials as of as toxic waste.
Contaminated or potentially contaminated protective clothing and equipment should be decontaminated

using suitable chemical methods or autoclaving before removal from the laboratory for disposal, cleaning
or repair. If decontamination is impracticable, materials should be disposed of as toxic waste.

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Table 7.2A
PHYSICAL INACTIVATION OF SELECTED TOXINS
(Reproduced from Appendix I, CDC/NIH BMBL http://www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm )
TOXIN STEAM DRY HEAT FREEZE-THAW GAMMA-

AUTOCLAVE (10 MIN) IRRADIATION
Botulinium Yesa > 100°Cb Noc Incompleted
neurotoxin (BoNT)
Staphylococcal Yese > 100°C; refoldsf Nog Incomplete
Enterotoxin
Ricin Yesi > 100°Ci Noj Incompletek
Microcystin Nol > 260°Cm Non ND
Saxitoxin Nol > 260°Cm Non ND
Palytoxin Nol > 260°Cm Non ND
Tetrodotoxin Nol > 260°Cm Non ND
T-2 mycotoxin Nol > 815°Cm Non ND
Brevetoxin (PbTx-2) Nol > 815°Cm Non ND
Table 7.2 A Notes:

ND indicates “not determined” from available decontamination literature
a
Steam autoclaving should be at ≥ 121°C for 1 h. For volumes larger than 1 liter, especially those containing Clostridium botulinum
spores, autoclave at ≥ 121°C for 2 h to ensure that sufficient heat has penetrated to kill all spores.
b
Exposure to 100°C for 10 min. inactivates BoNT. Heat denaturation of BoNT as a function of time is biphasic with most of the
activity destroyed relatively rapidly, but with some residual toxin (e.g., 1-5%) inactivated much more slowly.
c
Measured using BoNT serotype A at -20°C in food matrices at pH 4.1-6.2 over a period of 180 days.
d
Measured using BoNT serotype A and B with gamma irradiation from a 60Co source.
e
Protracted steam autoclaving, similar to that described for BoNT, followed by incineration is recommended for disposal of SE-
contaminated materials.
f
Inactivation may not be complete depending upon the extent of toxin re-folding after denaturation. Biological activate of SE can be
retained despite heat and pressure treatment routinely used in canned food product processing.
g
SE toxins are resistant to degradation from freezing, chilling or storage at ambient temperature. Active SEB in the freeze-dried state
can be stored for years.
i
Dry heat of ≥100°C for 60 min in an ashing oven or steam autoclave treatment at >121°C for 1 h reduced the activity of pure ricin by
>99%. Heat inactivation of impure toxin preparations (e.g., crude ricin plant extracts) may vary. Heat-denatured ricin can undergo
limited refolding (<1%) to yield active toxin.
j
Ricin holotoxin is not inactivated significantly by freezing, chilling or storage at ambient temperature. In the liquid state with a
preservative (sodium azide), ricin can be stored at 4°C for years with little loss in potency.
k
Irradiation causes a dose-dependent loss of activity for aqueous solutions of ricin, but complete inactivation is difficult to achieve; 75
MRad reduced activity 90%, but complete inactivation was not achieved even at 100MRad. Gamma irradiation from a laboratory
60
Co source can be used to partically inactivate aqueous solutions of ricin, but dried ricin powders are significantly resistant to
inactivation by this method.
l
Autoclaving with 17 lb pressure (121-132°C) for 30 min failed to inactivate LMW toxins. All burnable waste from LMW toxins
should be incinerated at temperatures in excess of 815°C (1,500°F).
m
Toxin solutions were dried at 150°C in a crucible, placed in an ashing oven at various temperatures for either 10 or 30 min,
reconstituted and tested for concentration and/or activity; tabulated values are temperatures exceeding those required to achieve 99%
toxin inactivation.
n
LMW toxins are generally very resistant to temperature fluctuations and can be stored in the freeze-dried state for years and retain
toxicity.
Please see: CDC/NIH BMBL, Appendix I (http://www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm) for references and further
information.

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Table 7.2B
CHEMICAL INACTIVATION OF SELECTED TOXINS
(Reproduced from Appendix I, CDC/NIH BMBL http://www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm )
TOXIN NaOCl (30 NaOH (30 NaOCl + NaOH OZONE

MIN) MIN) (30 MIN) TREATMENT
Botulinium >0.1%a >0.25 N ND Yesb
neurotoxin
Staphylococcal >0.5%c >0.25 N ND ND
Enterotoxin
Ricin >1.0%d ND >0.1% + 0.25 Ne ND
Microcystin ≥0.1%e ND 0.25% + 0.25 Ne ND
Saxitoxin ≥0.1%e ND 0.25% + 0.25 Ne ND
Palytoxin ≥0.5%e ND 0.25% + 0.25 Ne ND
Tetrodotoxin ≥0.5%e ND 0.25% + 0.25 Ne ND
T-2 mycotoxin ≥2.5%f ND 0.25% + 0.25 Ne ND
Brevetoxin (PbTx-2) ≥2.5%e,f ND 0.25% + 0.25 Ne ND
Table 7.2 b Notes:

ND indicates “not determined” from available decontamination literature
a
Solutions of NaOCl (≥0.1%) or NaOH (>0.25 N) for 30 min inactivate BoNT and are recommended for decontaminating work
surfaces and spills of C. botulinum or BoNT. Chlorine at a concentration of 0.3-0.5 mg/L as a solution of hypochlorite rapidly
inactivates BoNT (serotypes B or E tested) in water. Chlorine dioxide inactivates BoNT, but chloramines are less effective.
b
Ozone (>2 mg/L) or powdered activated charcoal treatment also completely inactivate BoNT (serotypes A, B tested) in water under
defined condition.
c
SEB is inactivated with 0.5% hypochlorite for 10-15 min.
d
Ricin is inactivated by a 30 min exposure to concentrations of NaOCl ranging from 0.1-2.5%, or by a mixture of 0.25% NaOCl plus
0.25 N NaOH. In general, solutions of 1.0% NaOCl are effective for decontamination of ricin from laboratory surfaces, equipment,
animal cages, or small spills.
e
The minimal effective concentration of NaOCl was dependent on toxin and contact time; all LMW toxins tested were inactivated at
least 99% by treatment with 2.5% NaOCl, or with a combination of 0.25% NaOCl and 0.25N NaOH.
f
For T-2 mycotoxin and brevetoxin, liquid samples, accidental spills, and nonburnable waste should be soaked in 2.5% NaOCl with
0.25% N NaOH for 4 h. Cages and bedding from animals exposed to T-2 mycotoxin or brevetoxin should be treated with 0.25% N
NaOCl and 0.025 N NaOH for 4 h. Exposure for 30 min to 1.0% NaOCl is an effective procedure for the laboratory (working
solutions, equipment, animal cages, working area and spills) for the inactivation of saxitoxin or tetrodotoxin.
Alternate methods of chemical decontamination:

1 N sulfuric or hydrochloric acid did not inactivate T-2 mycotoxin and only partially inactivated microcystin-LR, saxitoxin, and
brevetoxin (PbTx-2). Tetrodotoxin and palytoxin were inactivated by hydrochloric acid, but only at relatively high molar
concentrations. T2 was not inactivated by exposure to 18% formaldehyde plus methanol (16 h), 90% Freon-114+ 10% acetic acid,
calcium hypochlorite, sodium bisulfate, or mild oxidizing. Hydrogen peroxide was ineffective in inactivating T-2 mycotoxin. This
agent did cause some inactivation of saxitoxin and tetrodotoxin, but required 16 h contact time in the presence of ultraviolet light.
Please see: CDC/NIH BMBL, Appendix I (http://www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm) for references and further
information.

7-12
Table 7.3C. Inactivation Procedures for Select Agent Toxins
7.4 DEACTIVATION OF PRIONS

Work with neural tissues presents the possible remote risk for the presence of prions. Prions are
characterized by resistance to conventional inactivation procedures including irradiation, boiling, dry heat,
and chemicals (formalin, betapropiolactone, alcohols). While prion infectivity in purified samples is
diminished by prolonged digestion with proteases, results from boiling in sodium dodecyl sulfate and urea
are variable. Likewise, denaturing organic solvents such as phenol or chaotropic reagents such as guanidine
isothiocyanate have also resulted in greatly reduced but not complete inactivation. The use of conventional
autoclaves as the sole treatment has not resulted in complete inactivation of prions.
Formalin-fixed and paraffin-embedded tissues, especially of the brain, remain infectious. Some
investigators recommend that formalin-fixed tissues from suspected cases of prion disease be immersed for
30 min in 96% formic acid or phenol before histopathologic processing, but such treatment may severely
distort the microscopic neuropathology.
The safest and most unambiguous method for ensuring that there is no risk of residual infectivity on
contaminated instruments and other materials is to discard and destroy them by incineration. Current
recommendations for inactivation of prions on instruments and other materials are based on the use of
sodium hypochlorite, NaOH, Environ LpH and the moist heat of autoclaving with combinations of heat and
chemical being most effective (See Table 7.4). Unfortunately, these solutions are corrosive and require
suitable personal protective equipment and proper secondary containment. These strong corrosive solutions
require careful disposal in accordance with local regulations.
Precautions in using NaOH or sodium hypochlorite solutions in autoclaves NaOH spills or gas may damage
the autoclave if proper containers are not used. The use of containers with a rim and lid designed for
condensation to collect and drip back into the pan is recommended. Persons who use this procedure should
be cautious in handling hot NaOH solution (post-autoclave) and in avoiding potential exposure to gaseous
NaOH, exercise caution during all sterilization steps, and allow the autoclave, instruments, and solutions to

7-13
cool down before removal. Immersion in sodium hypochlorite bleach can cause severe damage to some
instruments.
Further information about biosafety measures for handling prions and the risks of prion diseases can be
found in Section VIII-H of the CDC/NIH BMBL:
http://www.cdc.gov/od/ohs/biosfty/bmbl5/sections/SectionVIIIH-PrionDiseases.pdf .
Table 7.4
PRION INACTIVATION METHODS FOR REUSABLE INSTRUMENTS AND SURFACES
1. Immerse in 1 N NaOH, and heat in a gravity displacement autoclave at 121°C for 30 minutes. Clean
and sterilize by conventional means.
2. Immerse in 1 N NaOH or sodium hypochlorite (20,000 ppm) for 1 hour. Transfer into water and
autoclave (gravity displacement) at 121°C for 1 hour. Clean and sterilize by conventional means.
3. Immerse in 1N NaOH or sodium hypochlorite (20,000 ppm) for 1 hour. Rinse instruments with water,
transfer to open pan and autoclave at 121°C (gravity displacement) or 134°C (porous load) for 1 hour.
Clean and sterilize by conventional means.
4. Surfaces or heat-sensitive instruments can be treated with 2N NaOH or sodium hypochlorite (20,000
ppm) for 1 hour. Ensure surfaces remain wet for entire time period, then rinse well with water. Before
chemical treatment, it is strongly recommended that gross contamination of surfaces be reduced because
the presence of excess organic material will reduce the strength of either NaOH or sodium hypochlorite
solutions.
5. Environ LpH (EPA Reg. No. 1043-118) may be used on washable, hard, non-porous surfaces (such as
floors, tables, equipment, and counters), items (such as non-disposable instruments, sharps, and sharp
containers), and/or laboratory waste solutions (such as formalin or other liquids). This product is
currently being used under FIFRA Section 18 exemptions in a number of States. Users should consult
with the State environmental protection office prior to use.

7-14
8. BIOLOGICAL WASTE MANAGEMENT
8.1 PURPOSE OF BIOLOGICAL WASTE MANAGEMENT PROGRAM

This section is intended to guide MCG personnel in the safe and legal way to dispose of biological waste.
Our program is designed to protect the people who handle, transport and dispose of your waste; protect the
Environment; protect the public perception of the university and minimize MCG’s regulatory liability.
Some people believe they can save money by working around this program. These attempts are counter
productive. They may place other people and the University at risk. The costs associated with one injury, or
violation fines can easily exceed annual operational costs. We would much rather hear and consider your
suggestions for program improvement than have you implement unauthorized procedures.
The biological waste management program does not supersede the requirements for radioactive
and/or hazardous chemical waste programs. Radioactive or hazardous chemical wastes shall be
disposed of through the radioactive waste stream or the hazardous chemical waste stream respectively. In
fact, in mixed waste situations (biological/chemical or biological/radiological), the waste disposal
requirements of the chemical or radiological waste disposal procedures will take precedence over the
biological, particularly, since biological wastes are more capable of being decontaminated/deactivated prior
to placing the waste in the chemical or radiological waste streams.
The Division of Environmental Health and Safety is continually working behind the scenes to improve this
program and control its cost. Direct any questions or suggestions to the Environmental Health and Safety
Office at x1-2663. Call if you have questions about unusual situations or anything not covered in this
guide.
8.2 DEFINITION OF BIOMEDICAL WASTE

“Biological waste”, “biomedical waste” and “biohazardous wastes” are terms commonly applied
interchangeably on MCG’s campus. These materials must be excluded from the general waste stream
unless appropriately decontaminated or deactivated as per the Georgia Environmental Protection Division
(EPD) as defined in the Georgia EPD biomedical waste regulations Chapter 391-3-4-.15
http://rules.sos.state.ga.us/docs/391/3/4/15.pdf. These include:
1. Waste Cultures and Stocks of Microorganisms or Etiologic Agents Including:

a) Cultures and stocks of infectious agents or microorganisms from facilities assigned
to Biosafety Levels 1 through 3 (BL1, BL2, BL3).
b) Cultures of specimens from medical and pathological laboratories.
c) Disposable containers, materials, and supplies that may have been contaminated
during the manipulation and transportation of microbial cultures and stocks. This
includes culture dishes and devices used to transfer, inoculate, and mix cultures.
d) Wastes from the production and handling of biological materials (including all tissue
culture materials.)
e) Live and attenuated vaccines.
2. Human Pathological Wastes

Pathological waste consists of all recognizable human tissues and body parts (except
teeth) which are removed during surgery, obstetrical procedures, autopsy, and laboratory
procedures. This also includes blood and blood products, exudates secretions,

8-1
suctionings, and other dialysate; cerebrospinal, synovial, pleural, peritoneal, and
pericardial fluids and other bodily fluids; and their respective containers.
3. Waste Human Blood and Blood Products and Their Containers Including:
a) Waste human blood and blood products (e.g. blood plasma, platelets, red or white
corpuscles, and other derived licensed products such as interferon, etc.)
b) Items contaminated with human blood or blood products.
c) Items caked with dried human blood or blood products.
d) Intravenous bags.
4. Used Sharps Waste

This category includes used hypodermic needles, syringes (with or without the attached
needles), pasteur pipettes, disposable plastic pipettes, scalpel blades, razor blades, blood
vials, test tubes, needles with attached tubing, broken plastic culture dishes, unbroken
glass culture dishes, and other types of broken and unbroken glassware that was in
contact with biological material including microscope slides and coverslips.
5. Unused Sharps Waste

Unused hypodermic needles, suture needles, syringes, and scalpel blades. This includes
those which have never been in contact with any biological materials.
6. Waste Animal Carcasses, Body Parts, and Bedding

All animal carcasses, bedding and body parts are to be disposed through the Division of
Laboratory Animal Services (LAS). As per LAS policies any animal bedding, caging or
carcasses which have been purposely infected or known to have been exposed to Risk
Group 2 agents shall be autoclaved prior to disposal in Biowaste containers.
All animal carcass and body parts will be stored in the appropriate DLAS freezers and
placed in the Stericycle® waste carts for transport to Stericycle® and incineration.
7. Isolation Wastes
Isolation wastes are defined as biological wastes and discarded materials contaminated
with blood, excretion, exudates, or secretions from humans or animals isolated due to
infection with Class 4 microbial agents. If a human or animal is known to be infected
with a Risk Group 3 or 4 agent, contact the Biological Safety Officer (x1-2663)
immediately.
8. Chemotherapy waste
Any disposable material which has come in contact with cytotoxic/antineoplastic agents
(agents toxic to cells) and/or antineoplastic agents (agents that inhibit or prevent the
growth and spread of tumors or malignant cells) during the preparation, handling, and
administration of such agents. Such waste includes, but is not limited to, masks, gloves,
gowns, empty IV tubing bags and vials, and other contaminated materials. Liquid waste
containers must first be classified as empty which means a quantity that it is not subject
to other federal or state waste management regulations prior to being handled as
biomedical waste (typically less than 3%).
9. Discarded medical equipment and parts

This excludes expendable supplies and materials, which have not been decontaminated,
and that were in contact with infectious agents.
In general, biological waste at MCG can be divided into five categories:

1. Solid biomedical waste (including sharps)
2. Pathological waste*
3. Liquid biomedical waste

8-2
4. Chemotherapy agent wastes*
5. Mixed biological waste (bio/rad or bio/chem.) *
*
These wastes must be incinerated and may not be placed in biohazardous waste cardboard
cartons unless a special incineration sticker has been applied. For proper disposal information,
contact the Biosafety Office or the EHOS office at x1-2663.
8.3 SOLID BIOMEDICAL WASTE

Following collection from the laboratories and clinics, all solid biological waste generated by MCG is
congregated for transport in a truck for eventual transportation by a professional biomedical waste handling
company (as of June 2008, that contractor is Stericycle®). Stericycle® will transport the waste containers
to their facilities for decontamination and disposal. Except for the material in the animal waste carts, those
demarcated as chemotherapy waste, and mixed waste (e.g., radiologically contaminated biohazardous waste
decayed to background levels), most of the biological waste at MCG is autoclaved and land-filled by
Stericycle®.
Even though the waste is transported away from MCG for decontamination and disposal, MCG is still
liable for the Environmental repercussions of inappropriate transport or disposal of any waste generated at
MCG; therefore it is crucial that all personnel who may handle biologicals understand the appropriate waste
disposal procedures:
Liquids or soggy materials should NOT be placed inside the Stericycle red-bag lined boxes.
Although the red bags are of high grade, there is still a potential for these biohazardous wastes
to leak from the bags before or during transport. This not only exposes MCG and/or Stericycle
staff to this material, but transport of uncontained biohazardous waste constitutes a violations
of the U.S. Department of Transportation regulations and can result in steep fines.
No chemicals or radiological materials should be placed in the general biomedical waste

stream due to the potential risk posed to the health and safety of both MCG and Stericycle®
handlers, but also to the risks of contaminating the landfill into which this waste is placed.
Biohazardous waste, by definition, is hazardous material and must remain secured at all times.
Biohazard waste boxes or containers should not be left in unsecured areas (e.g., in hallways or
on loading docks) where non-trained personnel or personnel with unknown health status may
encounter them. They should be removed from the laboratories and clinics and directly
transferred and secured in the Stericycle® truck. Placing this waste in secondary accumulation
areas (e.g., closets within buildings) is not permitted. Care should be taken to not allow the
boxes to become wet or damaged or exposed to vermin.
Never remove a leaking biohazard waste box from a laboratory. Notify the laboratory staff
and/or the Biosafety Office to enable the staff to safely re-package the wastes prior to removal.
If leakage is noticed after removal from the laboratory, the Environmental Services staff should
contact the Environmental Health and Safety Office (x1-2663) immediately, and provide
relevant information to identify the laboratory of the waste’s origin.
Because it is significantly more costly to dispose of biomedical waste than regular trash, items
which are not biological waste or potentially contaminated with biological materials should
NOT be placed in the biohazardous waste containers. This includes pipette wrappers,
notebook papers, paper towels, which have not been in contact with any biological material.
However, if one is unsure whether a particular item is contaminated with biological material,
one should default to the biohazardous waste container. Since food items are not permitted in
the laboratory, soda cans, candy wrappers, etc. should never be found in biohazard waste
containers.

8-3
Biohazardous waste bags/boxes/sharps containers should NOT be used for alternate purposes
other than for collection of biohazardous wastes. These are provided by the biohazardous
waste contractor and should be used expressly for collection of biohazardous wastes.
8.3.1 Proper Solid Biohazardous Waste Disposal Procedures

The MCG Environmental Services (a.k.a. housekeeping) staff is responsible for providing
the appropriate biohazardous waste containers and removing these items from the
laboratory and clinics. To request additional biohazardous waste containers, or to request
waste pick-ups, please contact x1-2434.
Biowaste boxes, red bags, sealing tape and animal carts are delivered in each arriving
empty truck from Stericycle®. Environmental Services staff is responsible for removing
these materials from the truck prior to loading any biohazard waste containers into the
trailer for disposal. Supplies should be stored in the Environmental Services supply areas
throughout campus.
The MCG Environmental Services Office will provide and handle the four different types
of authorized biohazardous waste containers, shown below. Alternate waste containers
need to be provided by the laboratory and placed into one of the authorized Stericycle®
waste containers by the laboratory staff for removal by the Environmental Services staff.
The biohazard “red bag” box

No liquids or soggy materials should be placed
in these containers
No loose sharp items should be placed in these
containers which would puncture the red bags
These containers have a 50 lb. maximum
weight limit.
Any material of higher hazard (≥BSL-2) should
be decontaminated prior to placement into
these containers to prevent inadvertent
contamination of those who may close the bag
as per IBC approval agreements.
The box should be closed and removed before
the contents leak or spill over the edge. Do not
overfill boxes.
When preparing these boxes, Environmental Services Staff should use at least three (3) strips of packing
tape to secure the bottoms of the boxes, each overlapping the edges by at least six (6) inches on either side.
One strip of tape should be placed across the seam of the box; the other two strips should be placed
perpendicular to the first strip of tape to secure it. The box is turned upright, and the top flaps bent
backwards. Each box should be lined with one red biohazard plastic bag, folded over the box flaps, as
shown above.
After filling, while wearing appropriate PPE, the laboratory staff should carefully gather the edges of the
bag together to avoid production of aerosols, and grasp the bag approximately 10 inches below the edges
and twist the bag closed. The twisted bag should be secured with several pieces of packing tape.
After the bags are securely closed, before removal from the laboratory, while wearing PPE (gloves, eye
protection, coat/gown) the Environmental Services staff should tape the top box flaps closed with three

8-4
strips of tape, overlapping each edge by at least six (6) inches (similarly to the way the bottom of the box
was closed). They should place a Stericycle® label (white labels designate the box to be autoclaved and
landfilled; yellow labels designate the box for incineration). The label should include the date the waste
was removed from the laboratory and the building and room number where the waste originated. The
boxes should be removed by Environmental Services staff and placed on the Stericycle® truck for transport
to their facilities for decontamination and disposal.
Special dollies are made to facilitate transport of vendor biowaste cartons. Ordering information (as of
March 2008) is:
Stericycle: Phone number: 866-783-7422-4-2-1
Catalog number 1818V2 (lid and dolly)
Cost: $91.30
Please note: the purchaser must provide departmental payment information at
the time of order. These purchases will not be invoiced to MCG with biowaste
manifests.
Should laboratory staff notice waste that requires disposal, please contact Environmental Services (x1-
2434).
The Sharps Containers:

There are three varieties of sharps containers available upon request through MCG Environmental Services
(x1-2434). These are shown below:
The Standard/Small Sharps Container The Large Sharps Container

with Slotted Lid
Each of these containers all comply with OSHA BBP standard

requirements dictated for sharps containers. These are:
closable, puncture-resistant, leakproof on the sides and bottom, labeled
and/or color-coded. In addition, to comply with these standards these
containers need to be maintained properly within the laboratory. They
should be:
Easily accessible and located close to work areas where
sharp materials are used (place containers near sharps; do
not walk across rooms handling sharps to dispose into the
containers).
The containers must be maintained upright.
These containers need to be replaced routinely. Do not
allow these containers to become overfull. As a rule, no The 7.5 gallon Biohazard Waste
sharps container should be allowed to become >2/3 full. Bucket
Many sharps containers have handy “Full” arrow marks on
the containers to remind users to replace the containers
once they have reached ~2/3 full.

8-5
Sharp items which should be placed in these containers include anything which is capable of puncturing the
biohazard waste bags, including hypodermic needles, syringes (with or without the attached
needles), pasteur pipettes, disposable plastic pipettes, scalpel blades, razor blades, blood vials, test tubes,
needles with attached tubing, broken plastic culture dishes, unbroken glass culture dishes, and other types
of broken and unbroken glassware, including microscope slides and coverslips, that was in contact with
biological material.
In addition, hypodermic needles, suture needles, syringes, and scalpel blades which have never been in
contact with biological materials should also be placed in these sharps wastes containers, since they pose a
risk of injury to any staff member who may come in contact with these. In addition, some of these items
(e.g., syringes without needles) may raise a public perception issue should they be placed in the general
waste stream. Therefore, these should all be disposed in the sharps biomedical waste containers.
Because these containers have leak-proof sides and bottoms, soggy items and small amounts of liquids
(e.g., a few ml of blood remaining in a tube) can be disposed in these containers. However, larger volumes
of liquids should be handled as liquid waste (described in Section 8.5, below).
Environmental Services is responsible for closing and removing these containers from the laboratories.
These are typically placed UPRIGHT in the red bag/boxes prior to sealing the box as described above.
8.3.2 Non-contaminated glass

Unbroken or non-broken glass which has not been contaminated with any hazardous materials should be
collected in solid-sided 5-gallon white plastic buckets with fitted lids or closed heavy duty cardboard
cartons before disposal in the general waste stream.
8.3.3 Improper Solid Biohazardous Waste Disposal

Environmental Services Staff members are to handle only the four authorized waste containers above.
They are not to:
Lift any biohazard (red) bag from any waste receptacle (lifting only increases the Environmental
Services staff members’ potential exposure risk to biohazardous aerosols which may be generated
while lifting or materials which may leak from the bag should the bag puncture).
Combine the contents of multiple waste containers in order to compile one full container for
disposal (combining only increases the risk of exposure to biohazardous aerosols, but also risks
contaminating floors and surrounding areas).
Leave any waste container unsecured or unprotected outside of the laboratory/clinic.
Leave the Stericycle® truck unsecured.
Accumulate wastes in any secondary waste collection sites (e.g., closets). The waste should be
removed from the laboratory or clinic and taken to the Stericycle® truck.
Please note: like all sharps containers, the 7.5 gallon biohazard
buckets are not intended to be re-usable waste receptacles. Bag
liners are not to be used in conjunction with these buckets, and
they should always be used with their fitted plastic lids.

8-6
Laboratory staff may choose to use alternate intermediary biohazard
waste containers provided that these containers are:
Clearly demarked as biohazardous waste with color-coded
labels (e.g., by using red bags or biohazard stickers).
The waste in these containers are removed as promptly after
use and not allowed to linger in these intermediary waste
containers.
Capable of being decontaminated and are decontaminated
often.
No sharps are disposed in these intermediary containers (sharps must be directly placed into
authorized sharps containers).
The waste is transferred to the authorized waste receptacle by the laboratory staff (not the
Environmental Services staff).
A word of caution about the use of lifting biohazard bags:

The act of lifting a bag containing biohazardous materials can
increase ones risks of exposure to biohazardous materials. First, the
act of lifting may generate aerosols; therefore staff should be fully
protected with PPE (at least lab coat, gloves and eye protection), and
the bag should be tightly closed before lifting. In addition, any
pointed items that may penetrate the bag (e.g., pipettes, micropipette
tips) should not be placed in bags that are to be lifted, since they may
also increase the exposure risks if the bag were to be punctured
during lifting. Care should be taken by any staff member lifting a
bag. These should be placed in the red bag/box for eventual
disposal.
8.4 PATHOLOGICAL WASTES

Any item which is identifiable as a human or animal body part or an animal carcass needs to be disposed
pathological waste. These items must be appropriately labeled for incineration. At MCG, the majority of
the pathological waste is generated through animal waste disposal procedures. Therefore, it is critical that
all animal carcasses be packaged and labeled according to LAS/IACUC compliant procedures and placed
in one of the necropsy freezers (e.g., in CB-1344, CA-1105, CA-1126, CA-1135, BG-1135A, CL-1119 or
CL-1133) or in the cold storage room (CB-3100). Contact LAS (x1-3421) for further information about
labeling procedures. These wastes will be placed in the animal waste carts and placed on the Stericycle®
trailer just prior to departure. Contact the Division of Environmental Health and Safety (x1-2663) if your
have any other questions related to pathological waste disposal.
8.5 LIQUID BIOLOGICAL WASTES

Liquids which are derived from biological organisms (e.g., blood) or have been exposed to biological
materials (e.g., tissue culture media, cell extracts) must be decontaminated prior to disposal. Liquid waste
decontamination and disposal methods must be documented in each laboratory’s or clinic’s Standard
Operating Procedures (SOPs).
The sanitary sewer was designed for the disposal of certain liquid wastes. Use of the sanitary sewer
reduces the chance for leaks or spills during transport and reduces disposal costs. Whenever possible,
decontamination of liquids by autoclaving or use of chemicals which can be disposed in the sewer system
(namely, bleach), is highly recommended (See Section 7, Decontamination). Remember to rinse sink with
copious water after disposal of decontaminated/deactivated biological materials.
Other chemical disinfectant methods may require subsequent disposal of the wastes through the chemical
waste stream. Similarly, if the biological wastes are contaminated with chemicals or radiological materials,

8-7
disposal procedures will have to be modified to comply with disposal requirements for these materials (see
Section 8.7, Mixed Waste).
Caution must be paid to disposal of any material which may clog or clog sewer disposal pipes. This would
include large amounts of blood or agar. Disinfection of large amounts of blood may be accomplished by
treatment with Isolyzer Plus®. This chlorine-based granular product solidifies and disinfects the blood or
body fluids so they may be placed in general (clear), not biowaste (red) waste bags. Agar should be
allowed to solidify prior to disposal in the regular waste stream (if not used in conjunction with biological
materials), or placed in solid-sided waste containers, such as the 7.5 gallon biohazard waste bucket (if
potentially contaminated with biological materials).
8.6 CHEMOTHERAPY WASTES

Chemotherapy agents need to be segregated from the general biohazardous waste, as per Georgia EPD
Chapter 391-3-4-.15 http://rules.sos.state.ga.us/docs/391/3/4/15.pdf. While these agents are often used in
clinical therapy, many of these agents may also be used for in vitro and in vivo animal research applications
(e.g., Actinomycin D, Mitomycin-C, Bleomycin, GM-CSF, Interleukin-2, INF-α, Gleevec) and must be
disposed as chemotherapy waste. A list of chemotherapy agents can be found at:
http://www.chemocare.com/bio/
According to GA EPD, chemotherapy wastes include:

“Any disposable material which has come in contact with cytotoxic/antineoplastic agents (agents
toxic to cells) and/or antineoplastic agents (agents that inhibit or prevent the growth and spread of
tumors or malignant cells) during the preparation, handling, and administration of such agents.
Such waste includes, but is not limited to, masks, gloves, gowns, empty IV tubing bags and vials,
and other contaminated materials. The above waste must first be classified as empty which means
such quantity that it is not subject to other federal or state waste management regulations prior to
being handled as biomedical waste.”
(Note: "Empty" is generally defined as containing less than 3% by weight of the total capacity of the
container).
Stock solutions of these chemicals and items that are heavily contaminated are disposed of through the
Chemical Hazardous Waste Program. Call the Chemical Safety Office (x1-2663) for guidelines concerning
the disposal of chemical hazardous waste.
8.7 MIXED WASTE

“Mixed waste” are those wastes which are contaminated with more than one type of hazardous material.
For the purposes of this Guide, focus will be on those materials which are contaminated with biologicals
and either chemical or radiological materials.
8.7.1 Biological/Chemical Mixed Waste

Many chemicals serve to disinfect biological materials through lysis, dehydration or protein-crosslinking.
Common chemical fixatives (formalin, glutaraldehyde) may serve to decontaminate a biological material,
leaving only the chemical waste disposal issues to address. Whenever possible, in biological/chemical
mixed waste situations, efforts should be made to determine whether the chemicals have adequately
decontaminated the biologicals. If they have not, seek assistance from the Biological and Chemical Safety
Offices (x1-2663) to determine whether a chemically-compatible decontamination method can be devised
which would then allow these items to be disposed as chemical waste. These should be documented in
your laboratory SOPs prior to initiation of any experiment which would produce the mixed waste.
Items contaminated with ethidium bromide, diaminobenzidine (DAB), phorbol, or phenol-chloroform

mixtures should not be mixed with other medical waste. These are already chemically decontaminated and

8-8
do not autoclave or bleach. These items should be segregated into their own containers and disposed as
chemical waste.
Do not dispose of biological items which contain mercury in the biowaste. Biological items which may
contain mercury (e.g., extracted teeth which contain mercury amalgams fillings) should first be
decontaminated using a broad-spectrum chemical disinfectant (never autoclave mercury!) and disposed as
mercury waste through the EHOS office (x1-2663).
8.7.2 Biological/Radioactive Mixed Waste

All Radioactive wastes are required to be disposed through the Radiation Safety Office (x1-9826). Any
biological materials that can be decontaminated with bleach, first, should be decontaminated and the pH of
the resultant waste adjusted by the addition of non-hazardous buffering agents (sodium bicarbonate, Tris)
prior to disposal as radioactive waste.
Radioactive sharps waste should be disposed of in sharps containers to which Radioactive warning labels
have been clearly affixed. These should be disposed through the Radiation Safety Office.
Animal carcasses, tissue/parts, and excreta containing/contaminated with radioactive materials shall be
handled and collected by those with proper radioactive material training and exposure badging (contact the
Radiation Safety Office x1-9826 for further information).
8.8 REQUISITIONS FOR NEW OR ADDITIONAL BIOHAZARDOUS

WASTE CONTAINERS
Extra Stericycle box-bag units or authorized sharps containers can be obtained from Environmental
Services at no cost to your laboratory (x1-2434 to make these requests). Any alternate required
intermediary waste containers (small biohazard bags, small sharps containers) must be provided by the
laboratory director.
If an unusual situation arises which requires disposal of large amounts (>15 gallons) of soggy materials or
liquid materials, please contact the Environmental Health and Safety Office (x1-2663) for assistance.
8.9 REQUISITION FOR BIOHAZARDOUS WASTE PICK-UP

The Environmental Services staff (a.k.a. Housekeeping) is responsible for removal of any full authorized
waste containers. If laboratory staff members notice containers that are full and require removal, please
contact the Environmental Services Office at x1-2434.

8-9
9. PLACARD SIGNS AND LABELS
Signs and labels which identify laboratories and equipment in which biological materials may be
encountered are requirements of:
• OSHA Bloodborne Pathogen standards (29 CFR 1910.1030)
http://www.osha.gov/pls/oshaweb/owadisp.show_document?p_table=STANDARDS&p_id=10051
• The State of Georgia’s Bloodborne Pathogen Act (O.C.G.A. § 31-12-13 http://www.lexis-
nexis.com/hottopics/gacode/)
• The CDC Biosafety in Microbiological and Biomedical Laboratories (BMBL)
http://www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm
• The NIH Guidelines for Research with Recombinant DNA Molecules (NIH Guidelines)
According to the CDC BMBL and the NIH Guidelines BSL-1 and BSL-2 standards, the laboratory
supervisor is expected to limit access to the laboratory and establish policies and procedures whereby only
persons who have been advised of the potential hazard and meet any specific entry requirements (e.g.,
immunization) may enter the laboratory or animal rooms. The Principal Investigator has the final
responsibility for assessing each circumstance and determining who may enter or work in the laboratory.
Therefore, the purpose of the door placard is to advise all persons entering the laboratory of the potential
hazards the specific entry/exit requirements that must be met as per the laboratory’s policies and
procedures.
9.1 DOOR PLACARDS

Entryways into any laboratory or clinic area in which
biological materials are handled must be labeled with a
biohazard door placard (see Figure 9.1). These signs are
available through the Biosafety Office (x1-2663 or
BIOSAFETY@mcg.edu). Biohazard door placards must be
fluorescent orange-red (or predominantly so) with lettering or
symbols in a contrasting color and include the following
information:
1. The universal biohazard symbol with the legend

“BIOHAZARD”.
2. The building and room number.
3. The name of the responsible investigator, laboratory
director, or clinic director. This person must have
knowledge of the types of biological material, the
hazards they pose, and the appropriate laboratory
incident emergency response standard operating
procedures.
4. The name of an alternate contact person who may be
contacted in the case of an emergency.
5. The daytime phone numbers and emergency after-
hours contact numbers for the responsible
investigator, laboratory or clinic direction and those
of the alternate contact person.
6. The Biosafety Containment Level (BSL) of the
room. (See Section 4 for more information). Figure 9.1 Example of required Biohazard Door Placard
7. A description of the biological hazards within the
laboratory.
8. The appropriate PPE that must be worn by all personnel within the laboratory.
9-1
9. Any additional entry requirements or special provisions for entry as established by the PI in the
laboratory SOPs (e.g. required vaccination, respirators, authorizations, etc.) as described in NIH
Guidelines and/or the CDC BMBL.
9.2 LABELS AND COLOR-CODING

Inside any facility in which biological materials are handled, warning labels shall be affixed to containers
of medical waste, refrigerators, freezers, incubators, and centrifuges which may contain biohazardous
material for any period of time without supervision of laboratory personnel. Labels of various sizes are
available in the Biosafety Office upon request (x1-2663 or BIOSAFETY@mcg.edu). Other equipment
such as waterbaths, sonicators, and biological safety cabinets do not require a permanent biohazard label if
decontaminated after each use. In these situations, a biohazard labels should be temporarily posted on the
equipment while in use with human blood, other potentially infectious materials, or an infectious agent.
Warning labels shall also be affixed to other containers used to store or transport
biological materials. This includes temporary bench-top waste containers (e.g.,
beakers used to collect pipette tips) or transport containers. Labels required must be
fluorescent orange-red (or predominantly so) with lettering or symbols in a
contrasting color, have the international biohazard symbol and bear the legend
"Biohazard". Labels shall be affixed as close as feasible to the container by string,
wire, adhesive, or any other method that prevents their loss or unintentional
removal.
The use of warning labels may be waived if: (1) waste is placed in red bags or red containers; (2) containers
of blood, blood components, or blood products are labeled as to their contents and have been released for
transfusion or other clinical use within MCGHI facilities as per MCGHI policies; or (3) individual primary
containers of biological materials are placed inside a labeled secondary container during storage, transport,
shipment or disposal.
9.3 LABELING EQUIPMENT SENT OUT FOR REPAIR OR DISPOSAL

Contaminated and potentially contaminated equipment sent out for repair or disposal must be
decontaminated as thoroughly as possible as per the laboratory standard operating procedures (SOPs).
Affix a tag to the equipment indicating when the equipment was decontaminated, what disinfectant was
used, and the name of the person who performed the decontamination.
Thorough decontamination of highly

technical or sensitive equipment or
equipment with limited access to
contaminated areas may not be possible.
Decontaminate the equipment to the best
degree possible (flushing lines or wiping
down the exterior) and affix a label to the
equipment before sending it out for repair.
The label must indicate what portions of
the equipment may remain contaminated,
with which agent(s) and include the
biohazard symbol as well as the legend
"Biohazard". The label must convey this
information to all affected workers
(service representatives, manufacturer,
etc.). A sample of this tag appears at left
and in Appendix J.

9-2
10. SHIPPING/EXTRAMURAL TRANSPORT
The following Section is an adapted version of materials originally developed by Andy
Glode and David R. Gillum at the University of New Hampshire. The MCG Biosafety
Office wishes to thank Mr. Glode and Mr. Gillum for their work in producing the original
document and their generosity in sharing their document with MCG.
This Section includes information about how to properly classify, package, mark and label your biological
materials for shipment or extramural transport. This Section also describes the training requirements
necessary to ship biological materials and dry ice. Requirements for intramural transport were previously
discused in Section 4.1.5. Information on the regulations and procedures for transport/shipping of live
animals can be found through the Division of Laboratory Animal Services.
Shipped/transported biological specimens, infectious agents and other biological materials are regulated by
governmental and non-governmental, consensus development organizations. Penalties for non-compliance
with the rules are significant and could result in the following fines:
• Up to $250,000 and up to a year jail sentence for individuals.

• Up to $500,000 per incident for organizations.
Several agencies regulate the shipment and transport of biological materials including:
• International Air Transport Association (IATA).

• US Department of Transportation (DOT).
• US Public Health Service (PHS).
• Occupational Health and Safety Administration (OSHA).
• United States Postal Service (USPS).
Infectious substances and other dangerous goods must always be transported according to the appropriate
regulations. Carrying dangerous goods by hand, for example in a vial in your pocket or in luggage, is
strictly prohibited. IATA and DOT regulations cover your checked luggage, materials you carry on, or
materials you carry in your pockets when you board an airplane. Persons who violate regulations are
subject to fines and criminal prosecution.
IATA regulations are commonly encountered since they regulate materials transported by air and are
generally the most restrictive. For these reasons, this guide pays special attention to IATA protocols;
however the DOT standards often reflect those of IATA and also pertain to ground transportation of your
materials.
10.1 TRAINING REQUIREMENTS

Federal rules require that anyone wishing to ship biological materials or dry ice must first have shipping
training. If you intend to package biological materials or dry ice for shipment, you must complete a special
training module, and provide documentation for training to the MCG Biosafety Office. MCG must be able
to document training certification for anyone shipping dangerous goods during regulatory audits. Training
consists of:
1. Read this section of the Biosafety Guide. This document will provide familiarity with the general
provisions relating to the regulations and will direct you to obtain more detailed training in the
requirements applicable to shipping biological materials and/or dry ice.

10-1
2. Have a current Biosafety training and bloodborne pathogen training. This training ensures that
you are familiar with hazards presented by infectious materials, proper handling and emergency
response procedures.
3. Complete an IATA-compliant shipping training module. This must provide you with a
certificate of completion and a copy of this certificate must be submitted to the Biosafety Office.
This training is required once every two years. Currently, the Biosafety Office maintains several
copies of a commercial training module developed by the company Saf-T-Pak; however, the
Biosafety Office may accept alternate acceptable forms of training, as long as the content of the
class can be documented and complies with the DOT/IATA requirements.
4. Develop a shipping/transport Standard Operating Procedure & submit to the Biosafety Office.
The IBC will ask that you document your intent to ship or transport biological materials, your
safety and security plans as required by law. Documenting the steps that you intend to follow for
packaging, labeling and transport will enable the IBC to perform a risk assessment of your
proposed operations. See template (Figure 10.1, below)
Shipping regulations change frequently so it is necessary to repeat training certification every two years.

10-2
Figure 10.1 Sample SOP template to Ship/Transport Hazardous Materials
After reading Section 10 of the MCG Biosafety Guide on Shipping/Extramural Transport, and completion of Saf-T-Pak or equivalent
IATA/DOT compliant training, fill out this form to qualify to ship dangerous materials at MCG. This SOP will be reviewed by
Biosafety Office and IBC and upon successful completion that will demonstrate knowledge of applicable regulations you will be
certified to ship those materials designated on this form.
1. What biological or other regulated material(s) might you ship/transport? List all hazardous materials that you intend to
ship/transport. Also, list the mode of transport (shipment, vehicular ground transport, courier) and the names of any
couriers/shipping services you may use.
2. What packaging will you use to ship your material(s)? Include company name and product number for chosen packaging for
each material you intend to ship.
3. Check those that should appear on your package:

Class 6.2 label
Class 9 label
UN3373 label
Cargo Aircraft label
Dry ice, UN1845, net weight _____ kg
Infectious substance, affecting humans, UN2814, net quantity _____
Infectious substance, affecting animals, UN2900, net quantity _____
Name, Address and Phone Number of Shipper
Name and Address of Consignee
Person Responsible: [name, contact phone number as listed, below]
List name of 24/7 emergency contact: _________________________________

List phone number of 24/7 emergency contact: __________________________
Overpack
“Exempt Human Specimen,” or “Exempt Animal Specimen.”
Genetically modified microorganisms, UN3245, net quantity _____
Diagnostic Specimens
4. Do any of your materials require permits for transport (e.g., etiological agent may require CDC or USDA/APHIS permits? If so,
please list these and attach a copy of these permits.
5. Are any of your shipments/transports international? Indicate whether these require importation or exportation permits,
explanation letters or special courier or special transport instructions? If so, please indicate these materials and requirements for
each. Attach a copy of these permits/letters.
6. Fill out and attach a Shipper’s Declaration for Dangerous Goods (if your shipments require one) or a completed waybill.. An
example of each material you intend to ship must be included in the “Nature and Quantity of Dangerous Goods” section.
7. Location of shipping records for laboratory/Person responsible for maintaining records (Records must be maintained at least 2
years).
8. List of authorized shippers in the laboratory, their signature and date of expiration of training (please attach copies of certificate
of completion of IATA/DOT compliant training class). Training is required once/2 years. By signing, each of the authorized
shippers below acknowledge that they understand the hazards associated with the materials noted above and they understand the
shipping requirements for those materials, as outlined in their training.
Authorized Shipper (print or type) Signature Date of expiration of Shipping certificate

10-3
10.2 SHIPPING OVERVIEW
Follow these steps when shipping biological materials and dry ice.
1. Classify your materials for shipment/transport. See Section 10.3.

2. Package, mark, and label your material(s) appropriately. See Section 10.4.
3. Fill out the Shipper’s Declaration for Dangerous Goods form. See Section 10.5.
4. If you are shipping any Federally regulated materials, including Select Agents, special regulations
may apply and/or permits may be required. Consult Section 10.6.
5. If you plan on importing or exporting biological materials, permits may be required. Consult
Section 10.7.
10.3 SHIPMENT TYPE

For shipment purposes, biological material will fit into one of the following categories:
• Unregulated biological material;

• Category A infectious substances;
• Category B infectious substances;
• Patient specimens;
• Genetically modified organisms and microorganisms; or
• Regulated Medical/Clinical Waste
Read each material section carefully to determine how to classify a material. If you are shipping a
biological material that cannot cause disease, infectious substance regulations do not apply, unless sent by
mail (see Section 10.9). Refer to the classification guide to assist with classification of materials (Figure
10.3). Note: All specimens or packaging containing dry ice or liquid nitrogen must be shipped properly
(see Other Packaging Requirements, Section 10.4.2). All samples preserved with flammable or corrosive
materials, such as ethanol or formalin, must receive consultation and pre-authorization from the MCG
Chemical Safety Office (x1-2663) and be shipped appropriately (also please see Section 10.4.2). Materials
which are both biological and radioactive require consultation with and pre-authorization from the
Radiation Safety Office (x1-9826).
10.3.1 Unregulated Biological Material

The materials listed below are technically not subject to IATA or DOT infectious substance shipping
regulations. However, guidance for shipping these materials (e.g., patient specimens, biological products)
are found in subsequent sections and these may require a permit for shipment abroad. Please check with the
Biosafety Office if you have any questions about these materials. All shipments of blood and blood
products must be labeled with a biohazard symbol.
• Substances which do not contain infectious substances or which are unlikely to cause
disease in humans or animals;
• Non-infectious biological materials from humans, animals or plants. Examples include
non-infectious cells, tissue cultures, blood or plasma from individuals not suspected of
having an infectious disease, DNA, RNA, or other genetic elements;
• Substances containing microorganisms, which are non-pathogenic to humans or animals;
• Substances that have been neutralized or inactivated such that they no longer pose a
health risk;
• Environmental samples which are not considered to pose a significant risk of infection;
• Dried blood spots*;
• Fecal occult blood screening tests*;
• An infectious substance, other than a Category A infectious substance (See Section
10.3.2.1), contained in a patient sample being transported for research, diagnosis,

10-4
investigational activities, or disease treatment and prevention, or a biological product,
when such materials are being transported by a private or contract carrier in a motor
vehicle used exclusively to transport such materials;
• Blood or blood components which have been collected for the purpose of transfusion or
the preparation of blood products to be used for transfusion or transplantation*;
• Tissues or organs intended for use in transplantation*;
• A material with a low probability of containing an infectious disease or where the
concentration of the infectious substance is at a level naturally occurring in the
environment so it cannot cause disease when exposure to it occurs. Examples of these
materials include foodstuffs and environmental samples (such as water or a sample of
dust or mold); or
• A biological product, including an experimental or investigational product or component
of a product, subject to federal approval, permit, review or licensing requirements such as
those required by the Food and Drug Administration (see, e.g.,
http://www.fda.gov/importeddrugs/ or http://www.fda.gov/CBER/gdlns/exprtcert.htm ),
or the US Department of Agriculture* (see, e.g.,
http://www.aphis.usda.gov/import_export/index.shtml and
http://www.aphis.usda.gov/vs/ncie/fac_imp.html for guidance).
* When mailing these items with the USPS, follow packaging guidelines for non-
regulated items. See Section 10.9.

10-5
Figure 10.3 The Classification Guide
Is your sample expected to

YES contain any human, animal or
NO
plant pathogens?
Is your sample: Is your sample a direct patient

• Dried blood spots? specimen and packaged as an exempt
NO
• Fecal occult blood screening tests? human or animal specimen?
YES • Blood, blood components, organs or tissues
for purposes of transfusion or transplantation
or for the preparation of products for the YES
purpos of transfusion or transplantation?
NO
Is your sample a
genetically
modified
NO
organism or
micro-organism?
Is your sample on the indicative
examples of Category A Infectious
YES Substances (See Table 10.3.2.1)
OR
Is your sample capable of causing YES
permanent disability, life threatening or
fatal disease?
Does your sample affect
humans only or humans
& animals? NO NO
Does your sample YES Is your sample

YES affect animals only? Medical waste?
YES NO
UN 2814 UN 2900
Category A Category A UN 3373
UN 3291 UN 3245
Infectious Infectious Biological Patient specimens
Biomedical Genetically modified
substance, substance, substance, (PI 650
Not waste, n.o.s. organisms and micro- Not
affecting afecting Category B recommended)
regulated (PI 650) organisms regulated
humans animals (PI 650) See Section
See Section (PI 913)
(PI 602) (PI 602) See Section 10.3.3
10.3.6 See Section 10.3.5
See Section See Section 10.3.2.2
10.3.2.1 10.3.2.1
Note: This flow chart only refers to IATA classificaztion of biological materials and does not cover other hazards &/or considerations for:
•Special permits for transfer (See Section 10.6)
•Dry ice (See Section 10.4.2.2)
•Liquid Nitrogen (See Section 10.4.2.3)
•Radiological hazards (Refer to Radiation Safety Office)
•Chemical hazards (Refer to Chemical Safety Office)

10-6
10.3.2 Infectious Substances
Infectious substances are materials known to be, or are reasonably suspected to contain, an animal or
human pathogen. A pathogen is a virus, microorganism (including bacteria, plasmids, or other genetic
elements), proteinaceous infectious particle (prion) or recombinant microorganism (hybrid or mutant) that
is known or reasonably expected to cause disease in humans or animals. Microorganisms that are unlikely
to cause human or animal disease are not subject to biological shipping regulations.
10.3.2.1 Category A Infectious Substances

Category A infectious substances are capable of causing permanent disability, life threatening or
fatal disease in humans or animals when exposure to them occurs. Category A infectious
substances are shipped as infectious substances, affecting humans (UN2814), or infectious
substances affecting animals (UN2900). Indicative examples of Category A infectious substances
are listed in Table 10.3.2.1.
10.3.2.1.1 Packaging
The triple packaging concept (explained in Section 10.4) applies to Category A infectious
substances. Purchase packaging compliant with IATA Packing Instruction 602 as detailed
in the IATA Dangerous Goods Regulations (DGR), which is available in the Biosafety
Office. See Table 10.3.2.1.1 for a list of packaging suppliers. Make sure to specify if you
are shipping a refrigerated sample (ice packs or dry ice). The maximum quantity of
infectious substance that can be shipped by air in one package is 4 L or 4 kg. The
maximum quantity that may be shipped via passenger aircraft is 50 mL or 50 g.

10-7
Table 10.3.2.1. Indicative Examples of Category A Infectious Substances
UN # and Proper
Shipping Name Microorganism
- Bacillus anthracis cultures - Japanese Encephalitis virus cultures
- Brucella abortus cultures - Junin virus
Infectious substance affecting
- Brucella melitensis cultures - Kyasanur Forest disease virus

- Brucella suis cultures - Lassa virus
- Burkholderia mallei - Pseudomonas mallei - Glanders cultures - Machupo virus
- Burkholderia pseudomallei - Pseudomonas pseudomallei cultures - Marburg virus
- Chlamydia psittaci - avian strains cultures - Monkeypox virus
- Clostridium botulinum cultures - Mycobacterium tuberculosis cultures
- Coccidioides immitis cultures - Nipah virus
- Coxiella burnetii cultures - Omsk hemorrhagic fever virus
UN 2814
humans
- Crimean-Congo hemorrhagic fever virus - Poliovirus cultures

- Dengue virus cultures - Rabies virus cultures
- Eastern equine encephalitis virus cultures - Rickettsia prowazekii cultures
- Escherichia coli, verotoxigenic cultures - Rickettsia rickettsia cultures
- Ebola virus - Rift Valley fever virus
- Flexal virus - Russian spring-summer encephalitis virus cultures
- Francisella tularensis cultures - Sabia virus
- Guanarito virus - Shigella dysenteriae type 1 cultures
- Hantaan virus - Tick-borne encephalitis virus cultures
- Hantavirus causing hemorrhagic fever with renal syndrome - Variola virus
- Hendra virus - Venezuelan equine encephalitis virus
- Hepatitis B virus cultures - West Nile virus cultures
- Herpes B virus cultures - Yellow fever virus cultures
- Human immunodeficiency virus cultures - Yersinia pestis cultures
- Highly pathogenic avian influenza virus cultures
- African swine fever virus cultures
substance affecting
- Avian paramyxovirus Type 1 – Velogenic Newcastle disease virus cultures

- Classical swine fever virus cultures
- Foot and mouth disease virus cultures
- Lumpy skin disease virus cultures
Infectious
UN 2900
- Mycoplasma mycoides - Contagious bovine pleuropneumonia cultures

animals
- Peste des petits ruminants virus cultures

- Rinderpest virus cultures
- Sheep pox virus cultures
- Goatpox virus cultures
- Swine vesicular disease virus cultures
- Vesicular stomatitis virus cultures
* This list is not exhaustive. New or emerging pathogens not on the list may meet the criteria to be included in Category A.

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Table 10.3.2.1.1. Manufacturers of Shipping Containers for Infectious Substances
and Dry Ice
Air Sea Atlanta All-Pak, Inc. CARGOpak Corporation

1234 Logan Circle Corporate One West 3215-A Wellington Court
Atlanta GA 30318 1195 Washington Pike Raleigh, NC 27615
Phone: 404-351-8600 Bridgeville, PA 15017 Phone: 800-266-0652
http://www.airseaatlanta.com Phone: 800-245-2283 http://www.cargopak.com
http://www.all-pak.com
DG Supplies, Inc. EXAKT Technologies, Inc. HAZMATPAC, Inc

5 Boxal Drive 7416 N Broadway Ext., Suite E 5301 Polk St., Bldg 18
Cranbury, NJ 08512 Oklahoma City, OK 73116 Houston, TX 77023
Phone: 800-347-7879 Phone: 800-923-9123 Phone: 800-347-7879
http://www.dgsupplies.com http://www.exaktpak.com http://www.hazmatpac.com
Inmark, Inc. JIT Certified, Inc. Polyfoam Packers Corporation

220 Fisk Drive S.W. 1740 Fenpark Drive 2320 S. Foster Avenue
Atlanta, GA 30336-0309 Fenton, MO 63026 Wheeling, IL 60090
Phone: 800-646-6275 Phone: 800-962-8636 Phone: 888-765-9362
http://www.inmarkinc.com http://www.jitcertifed.com http://www.polyfoam.com
SAF-T-PAK, Inc. Source Packaging of New England, Therapak Corporation

10807 - 182 Street Edmonton, Alberta, Inc. 1440 Arrow Highway, Unit A
Canada, T5S 1J5 405 Kilvert St. Irwindale, California 91706
Phone: 800-814-7484 Warwick, RI 02886 Phone: 888-505-7377
http://www.saftpak.com Phone: 800-200-0366 http://www.therapak.com
http://www.sourcepak.com

10-9
10.3.2.1.2 Labeling
The outer container of a Category A infectious substance shipment must display the
following information:
Sender and recipient’s full name and address;

Infectious substance label (see below);
“UN2814, Infectious substance, affecting humans” and net quantity or
“UN2900, Infectious substance, affecting animals” and net quantity;
The text “Person responsible: [name, phone number]” You must provide a
24 hr/day, 7 days/week contact for a person who knows what material has
been shipped and emergency response information. This should be
documented in your shipping/transport SOP;
Class 9 label (see below), including UN1845 and net weight, if packaged
with dry ice; and
Cargo Aircraft Label (see below), when shipping over 50 mL or 50 g.
Infectious substance
Label Class 9 Label Cargo Aircraft Label
DANGER
10.3.2.2 Category B Infectious Substances

Category B infectious substances are materials that are infectious, but do not meet the standard for
inclusion in Category A. Category B infectious substances are assigned to UN3373.
10.3.2.2.1 Packaging
The basic triple packaging concept applies to Category B infectious substances. Purchase
packaging that complies with IATA Packing Instruction 650. See Table 10.3.2.1.1 for a
list of some packaging suppliers. Be sure to specify if the shipment is a refrigerated
sample (e.g., ice packs or dry ice) or will be sent at ambient
(room) temperature.
For Category B infectious substances, the maximum quantity of

liquid per primary receptacle is 1 liter and outer packaging
must not contain more than 4 L or 4 kg.
10.3.2.2.2 Labeling
The outer container of a Category B infectious substance
shipment must display the following information:
UN3373 Label

10-10
The sender and recipient’s full name and address;
The words “Biological Substance, Category B”;
UN3373 label (see right);
The text “Person responsible: [name, phone number]” You must provide a
24 hr/day, 7 days/week contact for a person who knows what material has
been shipped and emergency response information. This should be
documented in your shipping/transport SOP; and
Class 9 label (see Section 10.2.2.1.2), if packaged with dry ice.
10.3.3 Patient Specimens

Patient specimens that have a minimal likelihood of containing pathogens are exempt from many shipping
requirements. Professional judgment is used to determine if a specimen contains pathogens and should be
based on the patient’s medical history, symptoms, local conditions and individual circumstances.
If there is more than a “minimal likelihood” that a patient specimen contains pathogens, it must be shipped
as a Category A infectious substance (UN2814 or UN2900) or a Category B infectious substance
(UN3373).
Patient specimens unlikely to contain pathogens must be prepared for shipment as follows:
10.3.3.1 Packaging
• Leak-proof primary container;
• Leak-proof secondary packaging;
• Fragile primary containers must be wrapped or separated to prevent breakage;
• Absorbent material must be placed between the primary and secondary containers to
absorb entire contents so that no liquid release will reach the outer packaging; and
• Outer packaging must be durable enough for its intended use with at least one side 100
mm x 100 mm or larger.
10.3.3.2 Labeling
The outer package must be marked with “Exempt human specimen,” or “Exempt animal
specimen.”
10.3.3.3 Dried blood

Special guidance has been provided by the CDC for shipment of dried blood spot specimens
which vary from those above. In particular, these should not be packaged in airtight, leak-proof
plastic bags because the lack of air exchange in the inner environment of a sealed plastic bag
causes heat buildup and moisture accumulation that can damage the dried blood spot test
substances. In addition, various chemicals that can adversely affect the test substances in the dried
blood spots could leach from these plastics and thus cause incorrect analytical test results. See:
http://www.cdc.gov/od/ohs/biosfty/driblood.htm for further information.
10.3.4 Biological Products

Biological products are derived from living organisms and manufactured for use in the prevention,
diagnosis, treatment or cure of diseases in humans or animals and are certified by the USDA, FDA or other
national authority.
Examples of biological products include certain viruses, therapeutic serums, toxins, antitoxins, vaccines,
blood, and blood products. Materials which contain incidental blood products, such as fetal calf serum,
may also be regulated, particularly internationally. For further information and guidance for shipping
biological materials, see:

10-11
Food and Drug Administration Guidelines
Importation: http://www.fda.gov/ora/import/
Exportation: http://www.fda.gov/CBER/gdlns/exprtcert.htm
US Department of Agriculture
Importation/exportation URL: http://www.aphis.usda.gov/import_export/index.shtml
Materials which do not require permits: http://www.aphis.usda.gov/vs/ncie/fac_imp.html
Biological products transported for final packaging, distribution, or use by medical professionals are not
subject to biological shipping regulations (although international shipments may have importation or
exportation standards, See Section 10.7). Biological products that do not meet these criteria must be
assigned to UN2814, UN2900 or UN3373, as appropriate.
10.3.5 Genetically Modified Organisms or Microorganisms

Genetically modified organisms (GMO) or microorganisms (GMMO) are organisms and microorganisms
in which genetic material has been purposely altered through genetic engineering in a way that does not
occur naturally. GMOs and GMMOs that are not infectious but that can alter animals, plants or
microorganisms in a way that is not normally the result of natural reproduction are considered a
miscellaneous hazard (Class 9) and are assigned to UN3245. GMOs and GMMOs that are infectious must
be assigned to UN2814, UN2900 or UN3373.
10.3.5.1 Packaging
These materials are packed for shipment in the same way as Category A infectious substances,
except there are no testing requirements for the packaging; this packaging variation is IATA
Packing Instruction 913. Packages designed for Packing Instruction 913 may not be available from
most vendors. In this case, use packages compliant with Packing Instruction 602.
The maximum allowable quantity per primary receptacle is 100 mL or 100 g. There is no
maximum net quantity per package.
10.3.5.2 Labeling
The outer container of a GMO or GMMO assigned to UN3245 must display the following
information:
The sender and recipient’s full name and address;

Class 9 label (See Section 10.3.2.1.2); and
Genetically modified microorganisms, UN3245, and net quantity.
10.3.6 Regulated Medical Waste

Regulated Medical Waste (also sometimes referred to as Regulated Biomedical Waste or Clinical Waste) is
defined differently by many state and federal agencies. Under DOT rules, regulated medical waste (RMW)
is a waste or reusable material suspected or known to contain an infectious substance, and is generated in
the diagnosis, treatment, immunization, or biomedical research of humans or animals. Regulated Medical
Waste (RMW) is assigned UN 3291 and is generally packaged consistently with IATA packing instruction
650 (See Section 10.3.2.2, Category B Infectious Substances for further information about packing
instruction 650), although some special exceptions exist in the DOT regulations (49 CFR 173.137 (c) and
(d) and 179.197).
MCG currently packages all RMW in boxes and carts provided by Stericycle® (See Section 8, Waste
Management); which comply with the regulations for packaging and labeling of RMW. Typically, these
are transported from MCG campus by Stericycle® personnel who have to adhere to the requirements for

10-12
transport of these materials, and are generally not transported by most MCG personnel in vehicles on public
roads. However, special considerations should be made should occasion arise that any regulated medical
wastes are to be transported, offered for transport or shipped if these wastes are to be:
Transported via vehicles traveling on public thoroughfares (e.g., to MCG or to the

Stericycle® truck from remote campus locations)
Any material transported under the Regulated Medical Wastes must be in a vehicle
dedicated for RMW transport if it may contain agents of Risk Group 2 or higher.
Transported outside of campus in containers other than the large Stericycle® boxes or carts
(e.g., in sharps containers), which may not have the required markings. These should be
appropriately packaged and labeled prior to transport.
Please contact the Biosafety Office (x1-2663, BIOSAFETY@mcg.edu) to declare your intention to
transport RMW and for further guidance prior to transport.
10.4 PACKAGING BIOLOGICAL MATERIALS

Potentially hazardous biological materials must be packaged to withstand leakage of contents, shocks,
temperature, pressure changes and other conditions that can occur during ordinary handling in
transportation. Packaging your material(s) appropriately is accomplished by purchasing certified
packaging. Refer to Table 10.3.2.1.1 for vendors that can supply certified packaging for biological
materials. When ordering, specify what type of material(s) you will be shipping: Category A infectious
substances, Category B infectious substances, etc. Different categories have slightly different packaging
needs, but all follow the basic triple packaging requirements described below.
10.4.1 Triple Packaging

Biological materials must be packaged according to the triple packaging principle. The three elements of
triple packaging include: primary receptacle, leak-proof secondary container, and durable outer container.
Infectious substances in Category A and B, patient specimens and genetically modified microorganisms
must be packaged in this way, with slight variations. An example of triple packaging is illustrated in Figure
10.4.1.
The primary container holds the biological material; it must be leak-proof. It must be labeled with the
name of the contents. A leak-proof seal, such as a heat seal, skirted stopper or metal crimp, is required. If
the container has a threaded lid, it must be secured with waterproof tape (e.g., Parafilm, etc.). Petri plates
cannot be used as primary receptacles. Lyophilized substances can only be shipped in flame sealed glass
ampoules or rubber stopped glass vials with metal seals. Packaging purchased for shipping infectious
substances usually does not include the primary container.
The secondary container holds one or more primary containers, and must also be leak-proof. Secondary
containers for all Category A and liquid Category B infectious substances must meet specific pressure test
standards when shipping liquids. Containers purchased from commercial vendors are designed to meet the
necessary standards. If you are shipping any liquid, there must be enough absorbent material in the
secondary container to absorb all of the liquid in the primary receptacle(s). If multiple primary containers
are used, they must be wrapped to prevent contact between them so they do not break during transport.
The outer container must be rigid and have one side that is at least 100 mm X 100 mm, in order for
required markings and labels to fit. The outer package must be of adequate strength for its capacity, mass,
and intended use. An itemized list of package contents must be included between the outer and secondary
container. The outer package should be marked to identify hazardous contents, including the proper
shipping name, UN number and net quantity for each substance, if required.

10-13
Figure 10.4.1
Source: Transporting Infectious Substances Safely, US DOT Document PHH50-0079-0706.
10.4.2 Other Packaging Requirements
10.4.2.1 Overpacks
An overpack can be used to combine several triple packages into one large package. This may be
done to save on shipping charges when shipping multiple samples. Each triple package inside the
overpack must be properly marked and labeled. The outside of the overpack must bear the same
markings and labels as the triple packages within including hazard labels and proper shipping
names. The outer container of the overpack must also be marked with the word, “Overpack.”
10.4.2.2 Dry Ice
10.4.2.2.1 Hazard Identification

Dry ice is classified by DOT and IATA as a “miscellaneous” hazard, class 9. Dry ice is
considered hazardous during transportation for three reasons:
Explosion hazard: Dry ice releases a large volume of carbon dioxide gas as it
sublimates. If packaged in a container which does not allow for release of the gas, it may
explode, causing potential injury and/or property damage.
Suffocation hazard: a large volume of carbon dioxide gas emitted in a confined space
may create an oxygen-deficient atmosphere.
Contact hazard: Dry ice is a cryogenic material that causes severe frostbite upon contact
with skin..
Packaging dry ice properly will minimize the risk to personnel transporting the material.
The explosion hazard will be eliminated with a package designed to vent gaseous carbon

10-14
dioxide. Suffocation and contact hazards will be greatly reduced by labeling the package
correctly, so those who come in contact with it will be aware of the contents.
10.4.2.2.2 Packaging Dry Ice

There are five basic requirements for all shipments of dry ice:
a. Gas venting: packages must allow for release of carbon dioxide gas. Dry ice must
never be sealed in a container with an airtight seal such as a jar with a threaded lid or
a plastic cooler. When transporting in a vehicle, the box should not be placed inside
the passenger compartment to prevent carbon dioxide accumulation within the
vehicle.
b. Package integrity: a package containing dry ice must be of adequate strength for
intended use. It must be strong enough to withstand the loading and unloading
normally encountered in transport. It must also be constructed and closed in order to
prevent any loss of contents that might be caused by vibration of by changes in
temperature, humidity, or altitude.
c. Package materials: do not use plastics that can be rendered brittle or permeable by
the temperature of dry ice. This problem can be avoided by using commercially
available packages intended to contain dry ice (see Table 10.3.2.1.1).
d. Waybill: the waybill (also referred to as the airbill) must include the statement “Dry
ice, 9, UN1845, [number of packages] X [net weight in kilograms]” FedEx has a
check box on their waybill to satisfy this requirement (see Figure 10.4.2.2.2A).
Airborne Express requires a slightly different format (see Figure 10.4.2.2.2B). Check
with your courier to make sure you have made the proper notation on their
paperwork.
e. Labeling: the outermost container must be labeled with a hazard class 9 label,
UN1845m and net weight of dry ice in kilograms. See Figure 10.4.2.2.2C, below.
This must be a specific size (5” x 5”). A full-scale printable version is included in
Appendix K.
Figure 10.4.2.2.2A. FedEx Waybill which properly documents 1 box containing 6 kg of dry ice.

10-15
Figure 10.4.2.2.2B. AirBorne Express waybill which properly documents 1 box containing 5 kg
of dry ice.
If shipping biological materials with your dry ice, you

must comply with the requirements for both shipping of
biological materials and dry ice.
When shipping biological materials and dry ice

together:
a. Dry ice must be placed outside the secondary
packaging (See Figure 10.4.2.2D)
b. Secure your samples in such a way that when the
dry ice sublimates, they will not move freely inside
the insulated box. This can be accomplished by
wedging your samples in place with cardboard or
Styrofoam. Fragile containers such as glass tubes
or vials should be wrapped in cushioning material. Figure 10.2.2.2C Dry ice label (not to size)
c. A Shipper’s Declaration for Dangerous Goods is
not required for shipments in which dry ice is the only hazardous material; however,
dry ice is included on declarations for shipments that include other hazardous
materials such as infectious substances.
Other issues that you should consider when shipping with dry ice:
a. Refer to your package manufacturer’s recommendations. Make arrangements with
your cosignee to make sure your package will be received on its intended delivery
date. Take into account local holidays or closings that might delay package receipt.
b. Minimize the volume of air to which the dry ice is exposed in order to slow the rate
of sublimation. If there is any air space after you fill the package with dry ice, fill it
with packing peanuts or other material to reduce the volume of air space.
c. Shipments are generally recommended to contain 5-10 pounds (2.27-4.54 kg) of dry
ice per 24 hours.

10-16
d. Dry ice shipments can be made with FedEx and DHL. UPS and the U.S. Postal
Service have extremely restrictive policies concerning shipments of hazardous
materials. Do not ship dry ice with UPS or with the U.S. Postal Service.
Material packaged in
primary and
secondary packages
10.4.2.3 Liquid Nitrogen (as if for ambient
shipping– See
Biological materials can be shipped refrigerated with liquid Figure 10.4.1)
nitrogen in cryogenic dry shippers, which are insulated packages

containing refrigerated liquid nitrogen fully absorbed in a porous
DRY ICE
material. The dry shippers, themselves, do not contain hazardous
materials, do no allow for the build-up of pressure within the
container and will not permit the release of any refrigerated Cooler (with lid
liquid nitrogen regardless of the dry shipper’s orientation. which allows for
gas venting)
Properly used, the dry shippers, alone, do not contain free liquid
nitrogen, and are not subject to hazardous material regulations
by the DOT or IATA. Dry shippers are capable of maintaining
cryogenic temperatures normally associated with liquid nitrogen Outer box
for approximately 24 hours (depending on the manufacturer)

without risk of liquid nitrogen spilling. However, be aware,
improperly filled dry shippers present a risk of liquid nitrogen
leakage and are subject to regulation should spillage occur.
Follow the manufacturer’s instructions for filling the dry

shipper. Some general practices when filling the dry shipper
are: Figure 10.2.2.2D Packaging for Dry Ice
a. Wear insulated gloves made for handling liquid
nitrogen and a face shield.
b. Add the liquid nitrogen slowly since a significant volume of nitrogen gas will form as the
cold liquid contacts the warm surfaces.
c. When the liquid level reaches the neck of the dry shipper, stop filling. Replace the cap
and set the dry shipper aside for the period specified by the manufacturer to allow the
liquid nitrogen to saturate the absorbent.
d. Repeat steps a-c until the liquid level no longer drops on standing. Special packing
regulations apply to shipments containing nitrogen. Contact the Biosafety Office at x1-
2663 or BIOSAFETY@mcg.edu if you need to ship materials with liquid nitrogen.
Some manufacturers have empty and full weights for their dry shippers. Dry shippers that will not
achieve their full weight may indicate a problem with the absorbent’s ability to hold the nitrogen.
This may prevent the dry shipper from maintaining the proper cryogenic temperatures during
shipment and may damage your samples. Contact the manufacturer to determine if the dry shipper
is safe to use.
When you are ready to ship, follow the following steps:

a. Remove all free liquid nitrogen from the dry shipper before transport.
b. Wear insulated gloves and a face shield when emptying the dry shipper.
c. Do not pour liquid nitrogen on to the floor since it could splash on your shoes or legs and
cause severe burns. It is recommended to pour the excess liquid nitrogen back into a
large liquid nitrogen dewar.
d. Hold the dry shipper upside down until the liquid stops flowing.
e. Stand the dry shipper upright for the period specified by the manufacturer.
f. Repeat steps a-e, above as many times as necessary to remove remaining liquid nitrogen.
g. Place your canes of material into the dry shipper and replace the cap.
h. Place the dry shipper into the case supplied by the manufacturer.

10-17
Make sure that the materials you are transporting are not hazardous chemicals such as samples
frozen in propane, ethane, halocarbon or other hazardous gas. If you are shipping biological
materials, your specimens will till have to comply with the IATA/DOT standards for biologicals,
as described above.
Special packing regulations apply to shipments containing liquid nitrogen. Contact the Biosafety
Office at x1-2663 or BIOSAFETY@mcg.edu if you need ship or transport materials with liquid
nitrogen.
10.4.2.4 Samples Preserved in Fixative

Special consultation with both the Biosafety and Chemical Safety Offices are required before
shipping materials which may be preserved in chemical, such as formalin or ethanol. However,
below are reference guidelines for shipping materials preserved in aqueous solutions of formalin
or ethanol. Packages prepared according to these guidelines must not contain any materials other
than those described (i.e. containers holding formalin- or ethanol-preserved specimens and related
absorbent or packaging materials). laboratory or sampling equipment, unrelated documents, or
other goods must be packaged and shipped in separate boxes.
10.4.2.4.1 In Aqueous Formaldehyde Solutions (Formalin) <25%

Aqueous formaldehyde solutions of less than 25% are considered hazardous materials
when shipped by air. This is because formalin can cause eye, skin, and respiratory tract
irritation. Formaldehyde is regulated by OSHA as a carcinogen. Additionally, exposure
to formaldehyde solutions may cause an allergic respiratory reaction. Be sure to review
the MSDS before handling or shipping any hazardous material.
Proper packaging shipments of formalin will minimize the chance of leakage during
transportation. Properly labeling and documenting these shipments will communicate the
hazard to transport workers who may be exposed to the formaldehyde in the event of a
leak.
Formaldehyde solutions are assigned to hazard class 9, packing group II. As such, each
inner packaging may not contain more than 30 ml. Each outer package may contain not
more than 500 ml. At this amount, these are considered “excepted quantities”.
Packaging for excepted quantities must have three basic components:

a. Inner (primary) packaging, such as a vial, tube, jar, etc. Do not completely
fill inner packagings; allow 10% head-space for liquid expansion. Liquids
must not completely fill inner packagings at a temperature of 55°C (130°F).
Closures of inner packagings must be held securely in place with tape, wire,
metal crimps, or other positive means.
b. Intermediate (secondary) packaging, such as a ziplock or other plastic bag.
Use good quality bags that are well sealed. Intermediate packaging must
contain enough absorbent material to absorb all contents and must not react
with formaldehyde. Use two plastic bags: put the absorbent and the inner
container(s) in the first bag and seal it well with tape. Then seal this bag in
another bag for added protection.
c. Outer packaging, such as a cardboard box. Formaldehyde solutions may not
be shipped in envelopes, Tyvek sleeves, or other non-rigid mailers. The
dimensions of the outer box must be at least 100 m (~4 inches) on two sides.

10-18
Labels and Marks on the outer packaging must include the following:
a. Dangerous Goods in Excepted Quantities Label, See Figure 10.4.2.4A. This
label must be filled out with the signature, title, name and address of the
shipper and the date. It must be affixed to the outer container on a vertical
side. For formaldehyde solutions of less than 25%, check the box for Class 9
material and enter “UN 3334” as the applicable UN number. The printable
label may be found in Appendix K and must be printed in color. Its overall
dimensions must be at least 100 mm x 100 mm (~4 in. x 4 in).
b. Name and Address: The outer container must display the name and address of
the shipper and consignee.
Many printer inks run when exposed to small amounts of water, such as rain or snow.
Therefore, it may be necessary to fully cover each label you have affixed to the box with
clear plastic tape. Also, when re-using shipping boxes, completely obliterate all
unnecessary labels and marks.
Figure 10.4.2.4A. Dangerous Goods in

Excepted Quantities Label (not to scale)
Package Tests must be performed and documented to ensure package compliance. A

representative example of packaging used for excepted quantities of formaldehyde
solutions must pass a drop test and compressive load test without any breakage or leakage
of any inner packaging and without any significant reduction in package effectiveness.
Perform the following tests on representative example of your packaging and keep a
record of the results.
Drop Test. Drop a representative package from a height of 1.8 m (5.9 feet)
directly on to a solid unyielding surface:
• One drop flat on bottom;
• One drop flat on top
• One drop flat on the long side;
• One drop flat on the short side; and
• One drop on a corner at the junction of three intersecting edges.
Compressive Load Test. Apply a force to the top surface of a representative

package for a duration of 24 hours, equivalent to the total weight of identical
packages if stacked to a height of 3 meters.

10-19
Proper documentation is required for all shipments of hazardous materials. Incorrect
documentation is the most common cause of package refusal. If using documentation for
couriers other than FedEx and DHL, contact EHS for assistance.
For domestic shipments with FedEx Express, fill out the standard US waybill. Fill out the form
completely and be sure to include the following information:
In section 6, Special Handling, check the box “Yes, Shipper’s Declaration not required.”
On the top of the form above the FedEx tracking number, include the statement, “Dangerous
Goods in Excepted Quantities”. See example in Figure 10.4.2.4B.
For DHL shipments, under the “Nature and Quantity of Goods” box on the air waybill, include the
words “Dangerous Goods in Excepted Quantities”.
Figure 10.4.2.4B Example of FedEx waybill for excepted quantity shipment
10.4.2.4.2 In Aqueous Ethanol Solutions of 55-100%

Ethanol solutions of 55-100% are considered hazardous materials when shipped by air.
This is because ethanol is a flammable liquid (NFPA rating = 3), and its vapor can travel
a considerable distance to an ignition source and “flash back”. Contact of ethanol with
strong oxidizers, peroxides, strong alkalis, and strong acids may cause fires and
explosions. Be sure to review the manufacturer MSDS before handling or shipping any
hazardous material.

10-20
Ethanol solutions are assigned to hazard class 3, packing group II. As such, each inner
packaging may not contain more than 30 ml. Each outer package may contain no more
than 500 ml.
Packaging, labeling requirements, package tests and documentation are similar to those
indicated above for formaldehyde solutions (See Section 10.4.2.4.1) with the exception
that for ethanol solutions of 55-100%, the Dangerous Goods in Excepted Quantities
Label (as seen in Figure 10.4.2.4A for formaldehyde solutions) should have the Class 3
material box checked and “UN 1170” as the applicable UN number.
10.5 SHIPPER’S DECLARATION FOR DANGEROUS GOODS

A Shipper’s Declaration for Dangerous Goods must be completed when shipping a Category A infectious
substance assigned to UN2814 or UN2900or a GMO or GMMO assigned to UN3245. A declaration is not
required for shipments in which dry ice is the only hazardous material. A declaration is not required for
shipments of Category B infectious substances assigned to UN3373. Improperly completed declarations are
the most common cause of package refusal.
Refer to the Shipper’s Declaration for Dangerous Goods (Figure 10.5) for an explanation of each section:
A. Shipper: Enter your full name, address and telephone number.

B. Consignee: Enter full name and address of recipient. When shipping infectious substances,
include the text, “Person responsible:” [then you must provide a 24 hr/day, 7 days/week
contact for a person who knows what material has been shipped and emergency response
information. This should be documented in your shipping/transport SOP].
C. Transport Details: Indicate here if your shipment is restricted to cargo aircraft only (if it is
more than 50 ml or 50 g of an infectious substance). Airport of departure and airport of
destination will be filled out by the carrier, leave blank.
D. Shipment Type: Cross out “radioactive” to indicate you are shipping a non-radioactive
substance.
E. UN or ID Number: Enter appropriate UN number as found in Table 10.5.
F. Proper Shipping Name: Enter the proper shipping name exactly as it appears in Table 10.5.
G. Class or Division: Enter appropriate hazard class as found in Table 10.5.
H. Packing Group: For dry ice, enter “III” in this column. Biological materials are not assigned
packing groups.
I. Quantity and Type of Packaging: Enter the net quantity for each material here. Use only
metric units. At the bottom of this column, indicate the number and type of packages used
(usually, “All packed in one fibreboard box.”) If using an overpack, indicate here with
“Overpack Used.”
J. Packing Instructions: Enter appropriate packing instruction number. Refer to Table 10.5.
K. Authorization: Leave this column blank.
L. Additional Handling Instructions: You must provide an emergency contact name and
phone number which can be reached 24 hours per day, 7 days per week to provide emergency
response information should a question develop about your package during transport. The
statement “Emergency Contact: [fill in contact name; phone number]” must be provided.
M. This section is self-explanatory. Sign and date each copy of your Shipper’s Declaration.
A blank Shipper’s Declaration for Dangerous Goods is available in Adobe PDF format at
http://www.unh.edu/ehs/shipping. Please note the following:
• Declarations must be typewritten or computer-generated; handwritten declarations will not be

accepted.
• Declarations must be printed in color to display the red-striped border.
• Always print at least four copies: provide three to the carrier and keep one for your records.

10-21
• Remember to sign and date each copy.
• Regulations require that you must retain your copy for 2 years.
A completed sample declaration can be found in Figure 10.5B. Contact the Biosafety Office x1-2663 or
BIOSAFETY@mcg.edu with any questions regarding the Shipper’s Declaration.
Table 10.5. Summary of Shipping Information.
Max. Net
Max. qty. Max. Net
Hazar Packing Packing qty./pkg.
Proper Shipping UN per qty./pkg.
Shipment Type d Group Instructi for
Name Number primary for Cargo
Class (PG) on (PI) Passenger
receptacle Aircraft
Aircraft
Category A
infectious
Infectious
substance, Liquids: 4 L 50 ml or 50 4 L or 4
substance, affecting UN2814 6.2 - 602
affecting humans Solids: 4 kg g kg
humans
and possibly
animals
Category A
infectious
Infectious
substance, Liquids: 4 L 50 ml or 50 4 L or 4
substance, affecting UN2900 6.2 - 602
affecting only Solids: 4 kg g kg
animals
animals (not
humans)
Category B Biological
Liquids: 1 L 4 L or 4
infectious substance, Category UN3373 6.2 - 650 4 L or 4 kg
Solids: 4 kg kg
substance B
Dry Ice
or
Dry Ice UN1845 9 III 904 N/A 200 kg 200 kg
Carbon Dioxide,
solid
Non-infectious,
transducing Genetically
genetically modified UN3245 9 - 913 No limit No limit No limit
modified organism microorganisms
or microorganism
Clinical waste,
Biomedical waste Regulated medical
UN3291 6.2 II 650 No limit No limit No limit
Regulated medical waste, n.o.s.
waste
Dangerous
Aqueous Goods in
formaldehyde Aviation regulated Excepted
UN3334 9 II 30 ml - 500 ml
solutions of less liquid, n.o.s. Quantities
instructions
than 25% (IATA 2.7)
Dangerous
Aqueous ethanol Goods in
solutions (55- Ethanol solution UN1170 3 II Excepted 30 ml - 500 ml
100%) Quantities
(IATA 2.7)

10-22
Figure 10.5A. Shippers Declaration

10-23
Figure 10.5B. Example of Completed Shippers Declaration
Medical College of GA
1120 15th St., CB-4100
Augusta, GA 30912
Ben Thompson (706)555-1212
Ben Thompson (706)555-1212
Augusta, GA, March 30, 2008

10-24
10.6 REGULATED AGENTS WHICH MAY REQUIRE SPECIAL PERMITS
FOR TRANSFER
10.6.1 CDC/USDA Select Agents and Toxins

The U.S. Department of Health and Human Services has developed a list of biological agents (see Section 2.6.2.2)
that have the potential to pose a severe threat to public health. Special regulations apply to the use and transfer of
these materials, including registration with the MCG Institutional Biosafety Committee and the Centers for Disease
Control and Prevention (CDC) or United States Department of Agriculture (USDA). If you are planning to, or
currently possess, use or transfer any of the select agents and have not registered, contact the Biosafety Officer at
x1-2663 (BIOSAFETY@mcg.edu ). Specific shipping restrictions apply to these agents which are not discussed in
this document.
10.6.2 Agricultural Pests, Pathogens and Biological Agents

The USDA/APHIS requires permits to transfer any plant or agricultural animal pest or pathogens. In addition,
permits may be required to export or import the agents shown below. Further information can be obtained from the
URL and phone number shown below:
APHIS Agricultural Permits

http://aphisweb.aphis.usda.gov/ppq/permits/
htt p://www.aphis.usda.gov/import_export/index.shtml
http
Material which do not require permits instruction: http://www.aphis.usda.gov/vs/ncie/fac_imp.html
Telephone: 1-877-770-5990
EXPORT/IMPORT
• Arthropods (insects and mites) • Livestock
• Arthropods inhabiting dung or of medical/veterinary significance • Moths
• Bees and bee related articles • Noxious weeds
• Biological materials containing animal material • Nursery stocks (including seeds)
• Butterflies • Parasitic plants
• Cell cultures of bovine or other livestock origins • Plant pathogens
• Cut flowers • Predators and parasitoids of arthropods
• Earthworms • Prohibited material for research purposes
• Endangered species • Rice and rice related articles
• Endangered species of wild fauna and flora • Seeds
• Entomopathogens • Snails and slugs
• Farm animals • Soil
• Foreign cotton and covers • Sugarcane products and by-products (including parts of the
• Fruits and vegetables sugarcane plant)
• High consequence livestock pathogens and toxins • Tissue culture materials of bovine or other livestock origins
• Indian corn or maize, broomcorn and related plants • Weed biocontrol
• Infectious agents of livestock • Wildlife
• Khapra beetle products • Wood products
• Live arthropods for display or educational purpose

10-25
10.6.3 Agents of Vectors of Human Disease
CDC permits are required when shipping any infectious agent known or suspected to cause disease in humans,
unsterilized specimens of human or animal tissues (including blood and other fluids), or biological vectors of
infectious animals, bats, insects, arthropods and snails. This includes the materials in the table below; further
information can be obtained at the URL and phone numbers below.
CDC Permit to Import or Transport Agents or Vectors of Human Disease

[http://www.cdc.gov/od/ohs/biosfty/imprtper.htm]]
Telephone: 1-404-498-2260
INFECTIOUS SUBSTANCES
• It is impractical to list all of the several hundred species of infectious substances. In general, an import permit is needed for any infectious
substance known or suspected to cause disease in man.
BIOLOGICAL MATERIALS
• Unsterilized specimens of human and animal tissues (such as blood, body discharges, fluids, excretions or similar material) containing an
infectious agent requires a permit in order to be imported.
VECTORS
• Animals: Any animal known or suspected of being infected with an organism capable of causing disease transmissible to man may require a
CDC permit. Importation of live turtles of less than 4 inches in shell length and all nonhuman primates requires an importation permit issued
by the Division of Quarantine.
• Bats: All live bats require an import permit from the CDC and the U.S. Department of Interior, Fish and Wildlife Services.
• Insects or Arthropods: All live fleas, flies, lice, mites, mosquitoes, or ticks require a CDC import permit, regardless of infection status.
Permits are required for adult forms, as well as eggs, larvae, pupae, and nymph stages. Any other living insect or arthropod, known or
suspected of being infected with any disease transmissible to man requires a CDC import permit.
• Snails: Any snail species capable of transmitting a human pathogen require a permit from the Centers for Disease Control.
10.6.4 Department of Commerce- Bureau of Industry and Security (BIS) Regulated

Agents
A permit may be required from the Commerce Department, when exporting infectious agents of human, plant, and
animal diseases, including genetic material, and products which might be used for culture of large amounts of agents
(Commerce Control List Supplement No. 1 to Part 774 Category 1, pages 54 - 59). In fact, in some instances,
permits may be required to domestically ship commerce-controlled materials to certain foreign nationals or U.S.
nationals who may fall on U.S. Governmental prohibition lists. In fact, these same individuals may even require a
permit in order to access to these materials or their information while at MCG (known as “deemed exports”). See
the list below for the biological agents which fall on the Commerce Control List. For further information on export
compliance, view the URL below and contact the MCG Legal Office for further guidance.

10-26
Commerce Department – Bureau of Industry and Security (BIS)
[http://www.bis.doc.gov/index.htm]]
HUMAN PATHOGENS and TOXINS
Bacteria Viruses
• Bacillus anthracis • Chikungunya virus
• Brucella abortus • Congo-Crimean haemorrhagic fever virus
• Brucella melitensis • Dengue fever virus
• Brucella suis • Eastern equine encephalitis virus
• Burkholderia mallei (Pseudomonas mallei) • Ebola virus
• Burkholderia pseudomallei (Pseudomonas pseudomallei) • Hantaan virus
• Chlamydia psittaci • Hendra virus (Equine morbillivirus)
• Clostridium botulinum • Japanese encephalitis virus
• Clostridium perfringens, epsilon toxin producing types • Junin virus
• Enterohaemorrhagic Escherichia coli, serotype O157 and other verotoxin • Kyasanur Forest virus
producing serotypes • Lassa fever virus
• Francisella tularensis • Louping ill virus
• Salmonella typhi • Lymphocytic choriomeningitis virus
• Shigella dysenteriae • Machupo virus
• Vibrio cholerae • Marburg virus
• Yersinia pestis • Monkey pox virus
• Murray Valley encephalitis virus
• Nipah Virus
• Omsk haemorrhagic fever virus
Toxins
• Oropouche virus
• Abrin
• Powassan virus
• Aflatoxins
• Pulmonary and renal syndrome-haemorrhagic fever viruses (Seoul,
• Botulinum toxins
• Cholera toxin Dobrava, Puumala, Sin Nombre)
• Rabies virus cultures
• Clostridium perfringens toxins
• Rift Valley fever virus cultures
• Conotoxin
• Rocio virus
• Diacetoxyscirpenol toxin
• South American haemorrhagic fever virus (Sabia, Flexal, Guanarito)
• HT-2 toxin
• St. Louis encephalitis virus
• Microcystin (Cyanginosin)
• Tick-borne encephalitis virus (Russian Spring-Summer encephalitis virus)
• Modeccin toxin
• Variola virus
• Ricin
• Venezuelan equine encephalitis virus cultures
• Saxitoxin
• Western equine encephalitis virus
• Shiga toxin
• White pox
• Staphylococcal aureus toxins
• Yellow fever virus
• T-2 toxin
• Tetrodotoxin
• Verotoxin
• Volkensin toxin
• Viscum Album Lectin 1 (Viscumin)
Rickettsiae
• Bartonella quintana (Rochalimea quintana, Rickettsia quintana)
• Coxiella burnetii
• Rickettsia prowasecki
• Rickettsia rickettsii
ANIMAL PATHOGENS and TOXINS
Bacteria
Mycoplasma mycoides
Viruses
• African horse sickness virus • Newcastle disease virus
• African swine fever virus • Peste des petits ruminants virus
• Avian influenza virus (certain highly pathogenic strains – see the Export • Porcine enterovirus type 9 (swine vesicular disease virus)
Administration Regulations for more information) • Porcine herpes virus (Aujeszky’s disease)
• Bluetongue virus • Rinderpest virus
• Foot and mouth disease virus • Sheep pox virus
• Goat pox virus • Swine fever virus (Hog cholera virus)
• Lumpy skin disease virus • Teschen disease virus
• Lassa virus • Vesicular stomatitis virus

10-27
Commerce Department – Bureau of Industry and Security (BIS) - CONTINUED
[http://www.bis.doc.gov/index.htm]]
GENETIC ELEMENTS/GENETICALLY MODIFIED ORGANISMS
• Genetic elements that contain nucleic acid sequences associated with the • Genetically modified organisms that contain nucleic acid sequences
pathogenicity of controlled microorganisms. associated with the pathogenicity of controlled microorganisms.
• Genetic elements that contain nucleic acid sequences coding for any • Genetically modified organisms that contain nucleic acid sequences coding
controlled “toxins” or “sub-units of toxins.” for any controlled “toxins” or “sub-units of toxins.”
• Technical Note: Genetic elements include, inter alia, chromosomes,
genomes, plasmids, transposons, and vectors, whether genetically modified
or unmodified.
PLANT PATHOGENS
Bacteria Fungi
• Xanthomonas albilineans • Colletotrichum coffeanum var. virulans (Colletotrichum kahawae)
• Xanthomonas campestris pv. citri including strains referred to as Xanthomonas • Cochliobolus miyabeanus (Helminthosporium oryzae)
campestris pv. citri types A,B,C,D,E or otherwise classified as Xanthomonas • Magnaporthe grisea (pyricularia grisea/pyricularia oryzae)
citri, Xanthomonas campestris pv. aurantifolia or Xanthomonas campestris pv. • Microcyclus ulei (Dothidella ulei)
Citrumelo. • Puccinia graminis (Puccinia graminis f. sp. tritici)
• Puccinia striiformis (Puccinia glumarum)
10.6.5 FDA Import Permits

All food (except most meat and poultry), drugs, biologics, cosmetics, medical devices, and electronic products that
emit radiation require a permit or registration before importation into the United States. See:
http://www.fda.gov/ora/import/ for more information about FDA import permits.
Export permits requirement information can be found at: http://www.fda.gov/CBER/gdlns/exprtcert.htm
10.6.6 Fish and Wildlife Service Permits

A permit may be required for transporting fish, wildlife, endangered species, or materials found in the list below.
Fish and Wildlife Service Permit Station

[http://international.fws.gov/permits/permits.html]]
Telephone: 1-800-770-0150
EXPORT
• African elephant ivory • Ginseng
• Animals • Marine mammals
• Artificially propagated plants • Museum specimens
• Asian elephant ivory • Personal pet
• Biological samples • Plants
• Captive-born export • Raptors
• Circuses/traveling animal exhibitions • Trophies by taxidermist
• Goldenseal • Wildlife
IMPORT
• African elephant • Marine mammals
• African elephant ivory • Museum specimens
• African leopard • Personal pet
• Argali • Plants
• Asian elephant ivory • Polar bears
• Biological samples • Scientific and zoological breeding or display
• Birds • Sport hunted trophy
• Bontebok • White rhinoceros
• Circuses/traveling animal exhibitions • Wildlife

10-28
10.7 INTERNATIONAL SHIPMENTS
Shipping and receiving animals and animal-derived materials, infectious or biohazardous agents, biological toxins,
and genetically modified organisms may require the approval of federal agencies, both domestic and foreign.
Regulations that govern the transfer of biological materials help to minimize or eliminate the possible threats to
public health and agriculture. In addition, the Departments of Commerce, Treasury and State regulate exportation
based on special considerations on the material’s economic impact, commercial value, ecological impact and/or
military dual-use.
Some countries, couriers and airlines restrict the importation or transportation of some hazardous materials (e.g., dry
ice). It is advisable for the shipper to determine these restrictions prior to shipment/transport. Contact the Biosafety
Office for assistance x1-2663 or BIOSAFETY@mcg.edu.
Packages shipped internationally generally require increased preparation time due to the additional paperwork
required for such packages. An import/export permit may be required when shipping biological materials
internationally (See Section 10.6). Check the following U.S. governmental agencies for permits and additional
information.
10.7.1 Exporting from the United States

Depending on the nature of the shipment, a U.S. export permit may be required when sending your package.
Additionally, an import permit may be required in the country where the package is being shipped. If your shipment
requires an export permit, it must be completed and approved by the appropriate government agency prior to
shipment. Typically, a copy of the import permit of the country of destination is included in the shipping
documentation and should be obtained from the consignee prior to shipment. For more information on whether your
shipment requires an export permit, please contact the MCG Biosafety Office x1-2663 or BIOSAFETY@mcg.edu .
Note: Packages may be opened and inspected when leaving the United States or at any time by any inspection
service provided by other countries. In order to assure that your package is safely delivered to its intended
destination, always consider the following:
1. If necessary, obtain an export permit from the appropriate governmental organization prior to
shipment.
2. Package and label the material according to the guidelines listed in this manual.
3. Include a courtesy letter with the shipment describing the contents in detail including information
about whether the material is infectious. Copies of importation paperwork from the consignee should
be included, if required.
10.7.2 Importing into the United States

All shipments entering the United States are processed by the U.S. Bureau of Customs and Border Protection. An
import permit may be required to deliver the package even if a permit is not required by the originating country.
Check with the appropriate governmental organization prior to shipment of the material.
Note: Packages may be opened and inspected upon entry into the United States. In order to assure that your package
is safely delivered to its intended destination, always consider the following:
If necessary, obtain an import permit from the appropriate governmental organization prior to shipment.
1. Package and label the material according to the guidelines listed in this manual.
2. Consider including a courtesy letter with the shipment.

10-29
The importer is legally responsible for assuring that foreign personnel package, label, and ship the infectious
materials according to USPHS and IATA regulations. Shipping labels containing the universal biohazard symbol,
the address of the importer, the permit number, and the expiration date are also issued to the importer with the
permit. The importer must send the labels and one or more copies of the permit to the shipper. The permit and
labels inform the U.S. Customs and Border Protection and U.S. Division of Quarantine personnel of the package
contents.
10.8 SHIPPING COMPANY RESTRICTIONS

Some shipping companies may have requirements that are more restrictive than those discussed in this document.
Consider the following information before planning a shipment.
10.8.1 DHL
DHL will accept shipments made according to IATA or DOT regulations. Shipments made according to instructions
in this manual will be acceptable to DHL.
10.8.2 FedEx
FedEx Express and FedEx Ground will accept shipments prepared according to instructions in this manual. FedEx
will not accept any material considered to be in Risk Group 4. A Risk Group 4 pathogen is one that usually causes
serious human or animal disease and that can be readily transmitted from one individual to another, directly or
indirectly, and for which effective treatments and preventive measures are not usually available.
10.8.3 United Parcel Service (UPS)

UPS will not accept shipments of Category A materials. UPS will accept shipments of UN3373 and exempt patient
specimens.
10.8.4 United States Postal Service (USPS)

The USPS has highly restrictive regulations concerning the shipment of hazardous materials by mail. Category A
materials may not be mailed with the USPS. USPS will accept shipments of UN3373 and exempt patient specimens.
For more information, refer to Section 10.9.
10.9 UNITED STATES POSTAL SERVICE MAILINGS

The United States Postal Service (USPS) does not allow Category A infectious substances to be mailed. Follow the
procedures below when mailing Category B substances, exempt patient specimens and non-regulated items.
10.9.1 Mailing Category B Substances

Follow packaging and labeling requirements listed in Section 10.3.2.2 and note the following variations:
• Shipments of both liquid and solid substances must be packaged in a pressure tested primary
or secondary container; and
• Category B substances may be mailed as First-Class, Priority, or Express mail.

10-30
10.9.2 Mailing Exempt Human and Animal Specimens
Follow packaging and labeling requirements listed in Section 10.3.3 and note the following variations:
• Inner containers and the total volume per package are limited to 500 mL or 500 g;
• Outer packaging must me rigid; and
• Exempt specimens must be mailed as First-Class, Priority, Express, or Package Services mail.
10.9.3 Mailing Non-Regulated Materials

According to USPS regulations, specific packing instructions apply when mailing non-regulated materials.
The following are examples of non-regulated biological materials:
• Biological products not containing Category A or Category B substances;

• Blood or blood products collected for transfusion or preparation of blood products;
• Tissues or organs intended for transplantation;
• Dried blood spots; and
• Dried specimens for fecal occult blood detection.
Quantity limits and form of substance (liquid or solid) determine the packaging requirements for non-
regulated materials. Refer to the appropriate category below to determine how to package you material.
10.9.3.1 Non-Regulated Liquid Substance, Not Exceeding 50 ml

Primary container and total package contents may not exceed 50 ml. Primary receptacle must be
leak-proof and properly sealed. Include cushioning and enough absorbent to absorb entire contents
of liquid. Enclose the primary container(s) in a leak-proof secondary container (e.g. plastic bag).
Label primary or secondary container with a biohazard symbol. No other labeling is required.
Secondary container may serve as the outer container.
10.9.3.2 Non-Regulated Liquid Substance, Exceeding 50 ml

Primary container must not exceed 50 ml; total package may not exceed 500 ml. Package in triple
packaging. Include cushioning and enough absorbent to absorb entire contents of liquid. Label
primary or secondary container with a biohazard symbol. No other labeling is required.
10.9.3.3 Non-Regulated Dry Substance

Primary container must be sift-proof and must be enclosed in a sift-proof secondary container.
Label primary or secondary container with a biohazard symbol. No other labeling is required.
Secondary container may serve as the outer container.
10.10 TRANSPORT AS AIRLINE BAGGAGE

Hazardous materials should never be carried in the passenger compartment of an airplane. Do not even attempt to
carry these aboard an airline. Although occasionally, limited non-infectious biological materials and small amounts
of dry ice may be transported as checked baggage on some passenger flights, these must not only comply with
DOT/IATA and other standards as described above, but must adhere to each airlines policies and prerogative to
carry such materials. Some airlines may refuse to carry these. Because of this, the practice is highly discouraged
and it is recommended that MCG personnel make arrangements to ship their materials prior to departure rather than
attempt to check their baggage. However, in the exceptional situations where shipment is not possible, those
wishing to transport non-infectious biological materials and/or dry ice as checked baggage must contact the

10-31
airline(s) well ahead of time to confirm that the airline’s policies will permit this and any special
limitations/instructions they may have for preparing such packages. All packages must comply with the IATA/DOT
packing, marking and labeling standards, and comply with any special permit standards as described in earlier
sections. Those who intend on transporting such materials in this manner must have documented shipping training
and declare their intentions to carry these materials on a passenger airline on their IBC-approved Biosafety Protocols
and SOPs and fully disclose the nature of these materials to airline and TSA personnel.
10.11 TRANSPORT IN GROUND VEHICLES

USDOT regulations do not apply to private or contract motor carriers used exclusively to transport biological
materials, diagnostic specimens or biological products; however, other standards (e.g., permits) may still be
required. Medical or clinical equipment and laboratory products may be transported aboard the same vehicle
provided they are properly packaged and secured against exposure or contamination. Note, in order for the vehicle to
be “dedicated,” the vehicle can not be utilized for other purposes at the same time (e.g., patient or passenger
transport, food transport). Although no specific packing instructions are required by law, packages should be
prepared in accordance with the packaging guidelines outlined in Section 10.4 and the carrier’s specifications, if
applicable.
All those transporting materials must be licensed drivers and comply with all applicable driving laws and accepted
safety standards (e.g., seat belt use).
Intention to transport biological materials or dry ice via vehicles by MCG personnel should be fully disclosed in
each IBC-approved Biosafety Protocol and SOPs prior to transport. These SOPs should include a description of the
steps which should be taken if one is involved in a motor vehicle accident en route. These include:
• Call for emergency assistance, if needed
• Let all emergency response teams know that you are transporting potential biohazards
• Notify your supervisor to contact the shipper and recipient of the sample status
• Arrange for alternate transportation if you are not able to get to your destination
10.11.1 Automobiles
Special considerations should be made for the locations and security of the package in a passenger vehicle. During
transport, the vehicle must be dedicated to the purpose. Biological materials should not be transported in any area
where food/beverages are transported and these areas should be fully decontaminated prior to transport of
food/beverages. Packages with dry ice or liquid nitrogen should never be transported in the passenger compartment
of the vehicle due to the suffocation hazards (see Section 10.4.2.2). Materials should be secured from theft. Any
material transported in an open truck must be secured to prevent loss via jostling of the vehicle during transport.
Transport via privately-owned vehicle is discouraged as many private insurance companies do not cover this
activity. Check with your insurance company to verify the terms of your policy prior to transport.
10.11.2 Public Modes of Transportation (e.g., Shuttles busses)

Transport of biological materials or other hazardous materials is not permitted in passenger compartments of
vehicles used for public transportation, such as shuttle busses.
10.11.3 Courier Services

Several commercial courier services are available to transport patient (diagnostic) specimens or biological products.
Although USDOT regulations do not apply to private or contract motor carriers used exclusively to transport patient
(diagnostic) specimens or biological products, and therefore no specific packing instructions are required by law,

10-32
those wishing to offer their packages of biological materials to a commercial carrier for transport must prepare
shipments in accordance to the courier’s standards and must fully declare the nature of the materials which are being
offered for transport. Often, these will be similar to that described in Section 10.4.

10-33
Appendix A: Biosafety Protocol Application Forms
Avialable in fillable *.pdf format at: http://www.mcg.edu/research/ibc/apps.htm

Medical College of Georgia
Biosafety Protocol (BSP) Application for Research Involving
Biological Materials and/or Recombinant DNA
INSTRUCTIONS
Approval of the Institutional Biosafety Committee (IBC) is required for all research involving biological material or recombinant
DNA before implementation or research or transfer of material to MCG. Please complete this Biosafety Protocol (BSP)
application form, in addition to the forms and training described below and submit to the Biosafety Officer, Environmental
Health & Safety, CI-1002 or kraemers@mcg.edu. Sample applications can be viewed at:
http:///www.mcg.edu/research/ibc/examples
When your BSP authorization is approved, you will receive an authorization memo from the IBC containing an IBC
Authorization Number (a Principal Investigator (PI)-specific identifier) and a Biosafety Protocol (BSP) approval number, and the
title of your BSP. This information is required on MCG Extra and Intramural Grant and Contract Routing forms. Your approval
is protocol-specific. In other words:
Your authorization is granted ONLY for those biological agents and recombinant DNA and techniques
listed in your approved BSP. You are approved to have or work with these agents only in those
locations indicated in your BSP, and only by the personnel listed on this application, and under the
constraints indicated in your approval agreement.
If your research involves the future addition, deletion or changes to your biological agents, recombinant DNA, personnel or
experimental protocols, you will need to amend your application. This can be done either by simply updating the appropriate
form or schedule and re-submitting those pages to the Biosafety Office or you may fully detail the amendment in an email to the
Biosafety Office. Any amendments which may alter the risk assessment of your protocol will be submitted for approval to the
IBC; others may be handled administratively as per IBC policies. If you have any questions, please contact the Biosafety Officer
at (706) 721-2663 or kraemers@mcg.edu.
The IBC requires the Biosafety Officer conduct a yearly review of your protocol and perform a laboratory safety assessment in
order to maintain your IBC authorization. At this time, you will also be asked to re-affirm the status of your BSP and your
SOPs will be reviewed. If you have any questions please contact the Biosafety Officer at (706) 721-2663.
In addition to completing the Biosafety Protocol (BSP) application primary form and appropriate
associated schedules, you will need to submit the following additional forms and complete the
following training requirements:
REQUIRED ADDITIONAL FORMS:
1. Your Laboratory’s Standard Operating Procedures (SOPs).

These are a list of rules and procexpected to be followed by those working in your laboratory, and therefore
should address the specific hazards and risks in your laboratory. These should be easily available for
reference in the laboratories and PIs should review this material with all research personnel prior to initiation
of research. Principal Investigators bear responsibility for ensuring that all research personnel understand
and comply with these rules at all times.
Templates for developing some common SOPs are available online. Principal Investigators (PIs) are highly
encouraged to work together with the Biosafety Office to modify these forms as needed to reflect any specific
procedures which may be employed in the laboratory.
SOP templates: http://www.mcg.edu/research/ibc/pdf/BSLSOP.pdf
2. A Self-Audit Form for each of your laboratory rooms.

Self-audits for BSL-1 and BSL-2 laboratories are designed to allow principal investigators to ensure the
laboratory complies with the appropriate biosafety guidelines issued by the Centers of Disease Control
(CDC) and the National Institutes of Health (NIH) prior to an audit from the Biosafety Office. This form
should be completed and answered accurately to reflect the situation in the laboratory.
BSL-1 Self Audit Form: http://www.mcg.edu/research/ibc/pdf/BSL1Audit.pdf
BSL-2 Self Audit Form: http://www.mcg.edu/research/ibc/pdf/BSL2Audit.pdf
i
3. Certificates for Dangerous Goods Transport Training (if applicable)
If you intend to ship or transport to/from campus any biological material, including diagnostic specimens,
cells, tissues, infectious substances, genetically modified organisms, toxins of biological origin or anything
on dry ice or in liquid nitrogen, you will need to include a copy of the certificate of completion of the
Shipping of Dangerous Goods (Saf-T-Pak) training course for the person(s) assuming the responsibility of
shipping/transport. Copies of the Saf-T-Pak training CD are available for loan from the Biosafety Office (x1-
2663). See “transportation of biological specimens and infectious material”, Schedule L.
TRAINING REQUIREMENTS:
The IBC requires ALL staff who will physically handle biological agents or recombinant DNA molecules and are conceivably at
risk from research procedures involving the use of these biological materials and those who are responsible for supervising those
who handle this material (i.e. Principal Investigators) to document appropriate Biosafety training and Right to Know (RTK)
training. The following training is required for all PIs and any personnel listed on this protocol. Collaborators at other institutions
are exempt unless they are conducting research in MCG laboratories. This application WILL NOT be processed without
documentation of completion of training by all personnel.
Biosafety Training:
All new MCG employees are required to take an Initial Laboratory/Chemical/Biosafety Training Course. This course
is offered didactically once per month. If you and/or personnel on the protocol need to register for this class, please
contact Ken Erondu (kerondu@mcg.edu or at (706)721-2663).
Any current MCG employees are required to take annual refresher Biosafety training. This training can be satisfied in
two manners:
A didactic refresher training course is available for departments, centers, or large laboratory groups (to
schedule a training session, please contact the Biosafety Office).
An online refresher training module is available at: "LIVE" Tegrity Training
Georgia Board of Reagents online Right-to-Know (RTK) Training:
The Georgia Board of Reagents (BOR) requires all laboratory personnel to take three primary RTK training modules:
Basic Awareness RTK (http://www.usg.edu/ehs/training/rtkbasic) - required upon initial hire
Chemical Hazard RTK (http://www.usg.edu/ehs/training/chemical)- required annually
Hazardous Waste Awareness RTK Training (http://www.usg.edu/ehs/training/hazwaste)- required
annually
The BOR also requires an additional RTK training module to be completed by those researchers working with fresh
human or non-human primate material.
Blood-Borne Pathogen RTK (http://www.usg.edu/ehs/training/pathogens/)-required annually.
Specialized Training Requirements:
Additional training requirements may be added for particular personnel or protocols at the discretion of the IBC, based
on their risk assessment of each protocol. PIs will be notified if additional training requirements have been placed upon
their personnel, and arrangements will be made for this training through the Biosafety Office.
Laboratory personnel who will be performing work either with Select Agents or under BSL-3 containment conditions
are required to have annual BSL-3 and/or Select-Agent-specific training. Additional training requirements may also be
placed upon these personnel, at the discretion of the IBC. Contact the Biosafety Officer (x1-2663) for further
information or assistance.
Any PI who desires additional application-specific safety training for proper handling of any biological material for
themselves or their laboratory personnel can request assistance from the Biosafety Office (x1-2663). All efforts will be
made to accommodate or facilitate these requests.
ii
SPECIAL NOTE FOR ANY PROTOCOL INVOLVING ANIMALS AND/OR HUMAN PATIENTS OR THEIR
SPECIMENS:
If your research involves the use of animals, human patients, or primary specimens from them, you will also need to submit
applications for approval to the Institutional Animal Care and Use Committee (IACUC) or an MCG-approved Institutional
Review Board (IRB) [either the Human Use Assurance Committee (HAC) or Chesapeake Research Review Inc. (CRRI)] in
addition to your submission to the IBC. You will need these committees’ approval in addition to IBC approval before
beginning your research. Please be aware that each compliance committee has separate applications and approval processes,
and the review criteria for IBC, IACUC, and IRBs differ.
IACUC applications and instructions can be accessed at: http://www.mcg.edu/research/animal/index.html . Please contact Jenny
Whitlock in the Laboratory Animal Services office at jwhitlock@mcg.edu or x1-0198 for further assistance.
HAC applications and instructions can be accessed at: http://www.mcg.edu/Research/OHRP/hac/ . Please contact the HAC
Administrative Office at hac@mcg.edu or one of the following extensions: x1-3110, 1-1482 or 1-8397 for further assistance.
CCRI applications and instructions can be accessed at: http://www.mcg.edu/research/ohrp/CRRI.htm . Please contact Angela
Randazzo in the Office of Human Research Protection at arandazzo@mcg.edu or x1-9346 for further assistance.
BIOHAZARDOUS MATERIAL AND COMPLIANCE WORKSHEET

This worksheet is intended as a guide to help you assess which, if any, additional supplemental schedules you may need to complete in addition
to this Biosafety Primary form in order to provide the IBC with a complete and accurate Biosafety Protocol application. This form does not
have to be submitted as part of the application.
Does the research in this application involve:
Yes No The use of non-exempt recombinant DNA? If “yes”, please complete & attach Biosafety Schedule A
For further explanation of Recombinant DNA classifications, please see page 2 of the
BSP Primary Form (Section 3, Part A) or NIH Guidelines for Recombinant DNA
Research1 (Sections III-A through F).
Yes No Mammalian cells, tissues, fluids, organs or cultures requiring >BSL-2 If “yes”, please complete & attach Biosafety Schedule B
containment2?
This includes: human or primate blood, tissues or fluids; cell, tissue or organ cultures
of human or non-human primate origin, (e.g. COS cells, HeLa cells, etc.) or animal
cells, tissues or organs which have been potentially infected with Risk Group >2
agents which were not documented in Schedule A.
Yes No Any large-scale cultures (> 10 liters at any one time)? If “yes”, please complete & attach Biosafety Schedule C
Yes No The use of potentially infectious microbial agents If “yes”, please complete & attach Biosafety Schedule D
Consider all microbial agents >Risk Group 22 or requiring BSL2 containment2
(viruses, bacteria, yeast, fungi, parasites, prions) or materials which have been
exposed to potentially infectious agents. Note: this excludes any material previously
described in either Schedule A or B (recombinant viral vectors or materials exposed to
recombinant viral vectors or mammalian tissues/cultures/fluids which might be
potentially contaminated with microbial.agents, respectively).
Yes No The use of biological materials in association with live animals (other than If “yes”, please complete & attach Biosafety Schedule E
human or non-human primates)?
This would include, e.g., delivery of recombinant DNA or genetically engineered cells,
tissues, or organs into animals (including creation of transgenic or knock-out/in
animals), injection of toxins of biological origin with LD50<100 μg/kg into animals,
transplantation of cells, tissues or organs into animals, or injection of microbial
agents into animals.
Yes No Human gene transfer/therapy2? If “yes”, please complete & attach Biosafety Schedule F
Yes No The use of CDC/USDA regulated Select Agents/Toxins? (see: If “yes”, please complete & attach Biosafety Schedule G
http://www.cdc.gov/od/sap/docs/salist.pdf)
Yes No An application for sponsored funding (e.g. a grant or contract) or additional If “yes”, please complete & attach Biosafety Schedule O
applications for IRB approval?
Yes No Shipping, extramural transport, or importation or exportation of any If “yes”, please make sure a current SOP which is related to
biological material (including diagnostic/clinical specimens, cell cultures, shipping of any material in this protocol and copies of
tissues, genetically modified organisms and/or infectious agents, toxins of current Saf T Pak training certificates for all designated
biological origin) or items on dry ice or liquid nitrogen4? shippers are on file with the Biosafety Office.
Yes No The use of toxins of biological origin with an LD50 <100 μg/kg body weight If “yes”, please make sure a current SOP related to
or poisonous, toxic or venomous plants, animals or insects? handling, storing and possessing any toxins of biological
origin associated with this project is on file in the Biosafety
Office.
1
NIH Guidelines for Recombinant DNA Research: http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Guidelines_Apr_02.htm
2
American Biosafety Association Risk Group Guide: http://www.absa.org/XriskgroupsX/index.html
Public Health Agency of Canada MSDSs: http://www.phac-aspc.gc.ca/msds-ftss/
NIH Guidelines for Recombinant DNA Research: http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Guidelines_Apr_02.htm
CDC’s Biosafety in Microbiological and Biomedical Laboratories (BMBL): http://www.cdc.gov/od/ohs/biosfty/bmbl4/bmbl4toc.htm
3
For definition, please see: Section III-C of NIH Guidelines for Recombinant DNA Research: http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Guidelines_Apr_02.htm
4
Shipping, transport, importation and exportation of biological agents, dry ice and liquid nitrogen must comply with multiple International and Federal (including DOT, USDA, CDC, DOC) laws
and guidelines for Transport of Dangerous Goods. High civil and criminal liability and fines may be associated with non-compliance with these laws.
iii
Application Date: _______________ Principal Investigator: ___________________________________________
For EH&S office use only:

BSL-1 rDNA-exempt Human source IRB app MCGHI or
mat. VA
BSL-2 rDNA-non- Microbial Animals Potential
exempt Dual-use
BSL-3 Infectious agents Biotoxins DSPA Shipping
Select Agent Hazardous cells Med Surv. Req Rad/Chem Import/Export
Medical College of Georgia Biosafety Primary Form

SECTION 1: GENERAL INFORMATION
Principal Investigator: Academic Title:
Office Phone Number: Department:
Email Address: Fax number:
Campus Address: Emergency contact phone number:
BSP Title:
DESCRIPTION OF RESEARCH: Provide a short summary of the research you will be conducting, focusing on the biological agents
you propose to use and how you intend to use them. If describing recombinant DNA, generally describe vectors, inserts and uses. See
instructions for further details.
(Please do not copy/paste your grant abstract or Specific Aims; merely provide the IBC with a description of the overall aims of your research, which biological
agents or recombinant DNA this involves and how these will be manipulated to accomplish the research aims).
SECTION 2: LOCATIONS
• Please list ALL locations in which a biological agent or recombinant DNA may be found at any moment in time including culture rooms, laboratory,
microbial growth chamber areas, benchtop laboratories, storage areas, clinical areas, animal facilities, core facilities, etc. See instructions for further
details.
• The proposed containment Biosafety Level (BSL) of these rooms. The proposed BSL should reflect the risk group of your agents, your practices and
facilities. Note: the BSL of any room is dictated by the highest BSL containment required by any agent utilized in that room. Please see instructions for
further details.
Lab Room List Biological Agents BSL of
Number (e.g. Recombinant DNA; Animals or animal cell lines, tissues, fluids or organs; Human or Non-Human Primate cell lines, tissue, fluids or organs, room
(Bldg./room) Potentially infectious or infected material, Toxins of biological origin, Microbial pathogens)
EMERGENCY CONTACT INFORMATION: Placards designating emergency contact information are required on doors of
biological laboratories which list emergency contact information. Contacts must have knowledge of the biological hazards in the laboratory. Please provide the
names, office phone numbers, and emergency (after hours, cell, pager) of a primary and secondary contact person which should be listed on your door placards
(typically the PI and a laboratory manager):
Contact name Office Phone Emergency Phone
ver. 120606 BSP- Primary p. 1

SECTION 3: PROTOCOL DETAILS

PART A
Does your research involve recombinant DNA (rDNA)? Yes No
If “yes”, please complete the chart below. If “no”, please skip to Part B, below.
Please check below which column(s) your rDNA experiments fall, based on categorizations as defined by the NIH Guidelines for
Recombinant DNA Research Section III-A through F1:
“Non-exempt” Recombinant DNA1 “Exempt”1

Experiments which must be registered and Experiments that require registration Recombinant DNA experiments that do not require
approved prior to initiation: simultaneous with initiation: IBC approval:
1. Deliberate transfer of a drug trait to a 1. Experiments using as vectors < 2/3 of the genome 1. rDNA containing less than 1/2 of an eukaryotic
microorganism not known to acquire it naturally of a eukaryotic virus, demonstrated to be free of viral genome propagated in cell culture (with the
(if it could compromise the use of the drug to helper virus or complementing helper virus exception of expression of DNA from Risk
control disease agents in humans, animals or components. Group4 2, 3, 4 or restricted agents5);
agriculture)2. (Note: this would likely exclude
most subcloning procedures using antibiotic
selectable markers in E. coli K12 derivatives) 2. Transgenic or knockout rodent experiments for 2. rDNA work involving E. coli K12 derivatives, S.
which BSL-14 containment is appropriate (NOTE: cerevisiae, and B. subtilis host-vector systems
2. Cloning of DNA encoding toxic molecules lethal the purchase of transgenic rodents for BSL-1 (with the exception of expression of DNA from
to vertebrates at an LD50 of <100 µg/kg body experiments falls into the “exempt” category; Risk Group 2, 3, 44 or restricted agents).
weight2,3. however, breeding of these animals with different
strains of mice is considered non-exempt rDNA
3. Human gene transfer/therapy experiments2; experiments).
4. Cloning using human or animal pathogens as 3. All experiments not specified in this chart.
host-vector systems.
5. Cloning of DNA from all Risk Group4 2, 3, 4 or

restricted human or animal pathogens (including
HIV and related viruses and viruses capable of
infecting human cells).
6. Experiments using more than 2/3 of the genome

of infectious animal or plant viruses or defective
viruses grown in the presence of helper virus or
complementing helper virus components.
7. Recombinant DNA experiments involving whole

animals, including trangenic or knockout rodent
experiments requiring BSL24 containment; or
transplantation of genetically engineered cells
into animals.
8. Large scale DNA projects (>10 liter cultures at

any moment in time).
Yes No Yes No Yes No
Experiments which fall within these two categories can be initiated by the
researcher upon notification to the IBC.
1
For further explanations/details, please see: http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Guidelines_Apr_02.htm
2
Please note that these experiments may require approval of NIH Director, NIH’s Office of Biotechnology Activies (OBA) approvals or registration and/or the Recombinant DNA Advisory
Committee (RAC) review. Please contact the Biosafety Office (x1-2663) as soon as possible if your experiments fall into these categories for assistance.
3
LD50 for many toxins can be viewed on commercially available Material Safety Data Sheets (MSDSs), e.g.
http://www.sigmaaldrich.com/Area_of_Interest/The_Americas/United_States.html or contact the Biosafety Office x1-2663 if you need assistance.
4
• American Biosafety Association Risk Group Guide: http://www.absa.org/XriskgroupsX/index.html
• Public Health Agency of Canada MSDSs: http://www.phac-aspc.gc.ca/msds-ftss/
• NIH Guidelines for Recombinant DNA Research: http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Guidelines_Apr_02.htm
• CDC’s Biosafety in Microbiological and Biomedical Laboratories (BMBL): http://www.cdc.gov/od/ohs/biosfty/bmbl4/bmbl4toc.htm
PART B
Please refer to the chart in Part A, above, and “Biohazardous Material and Compliance Worksheet” on page iii (in the
instructions). You may proceed to page 4 of the BSP Primary Form to the Principal Investigator’s Statement of
Responsibility, if:
1. You indicated, above, that your research does not involve recombinant DNA or it involves only “exempt” recombinant DNA;
AND
2. Your research involves only agents which can be appropriately contained using BSL-1 practices and facilities, and your
application will not require completion of any other additional schedules as assessed on the “Biohazardous Material and
Compliance Worksheet” on p. iii (in the instructions).

SECTION 4: PERSONNEL
The following list of personnel should include all staff who will physically handle the biohazardous agents or recombinant DNA molecules and
are therefore conceivably at risk from research procedures involving the use of these biological materials, as well as those who supervise those
who handle the material. Approval is given only for the identified personnel listed below. The Biosafety Officer must be notified of any changes
in personnel. Amendments to the personnel list can be submitted as an updated version of this form or via email to the Biosafety Office
(kraemers@mcg.edu). List additional personnel on a copy of this sheet, if needed.
NAME TITLE DEPARTMENT E-MAIL PHONE
EXPERIENCE PERSONNEL HAVE WITH NON-EXEMPT rDNA AND AGENTS REQUIRING

>BSL2 CONTAINMENT LISTED ON APPLICATION:
*The IBC does not require a full resume, merely an idea of how much experience with the agents the personnel has and in what
setting(s) this experience was gained.
Name List biological agents/techniques with which this Where was this Years of Experience with
person has experience experience obtained?* agents*

PRINCIPAL INVESTIGATOR’S STATEMENT OF RESPONSIBILITY FOR

RESEARCH INVOLVING RECOMBINANT DNA AND BIOLOGICAL AGENTS
Please review each of the following terms of this agreement prior to signing this agreement, below.
I attest that the information contained in the attached application and supplements is accurate and complete.
I agree to comply with the requirements pertaining to the use, shipment and transfer of biological agents, non-
exempt recombinant DNA, and/or CDC or USDA-regulated agents.
I am familiar with and agree to abide by the provisions of the current CDC Guideline “Biosafety in Microbiological
and Biomedical Laboratories” (BMBL), the NIH Guidelines for Research with Recombinant DNA Molecules,
and Federal Regulations governing CDC/USDA regulated Select Agents as well as any other specific granting
agency instructions pertaining to the proposed project.
I will ensure that before entering my laboratory, any person is advised of the potential hazards.
I agree to accept responsibility and accountability that all laboratory personnel are familiar with and trained to
employ the proposed safety precautions, appropriate emergency procedures, and the practices and techniques
described in this BSP.
I agree to amend this protocol to include any changes in agents, personnel, locations, applications or major
equipment (e.g. biosafety cabinets, autoclaves) prior to implementation of the changes.
I agree to report any accident that results in overt exposure of personnel or the environment to biological agents or
recombinant DNA including inoculation, ingestion, mucous membrane exposure or inhalation by personnel of
any agent to the Biosafety Officer (x1-2663), followed by a written report to the IBC.
I agree to immediately report any high hazard spills (large volumes, high risk group agents) or spills in public (non-
laboratory) areas to the Biosafety Officer (x1-2663).
I agree to report any problems pertaining to operation and implementation of biological and physical containment
safety procedures or equipment or facility failures to the Biosafety Officer (x1-2663) as soon as possible,
followed by a written report to the IBC.
I agree to bring any new information bearing on the biosafety risks of any research project, such as unanticipated
toxic or oncogenic reactions, to the immediate attention of the Biosafety Officer (x1-2663), followed by a written
report to the IBC.
I agree to bring any new information bearing on the compliance of my research with NIH Guidelines, the CDC
BMBL or Federal Select Agent Laws such as technical information relating to hazards and safety procedures
and/or innovations, to the attention of the Biosafety Officer (x1-2663).
I understand that the IBC may be obligated to report any non-compliance to Biosafety guidelines or regulations to
my funding agencies (e.g. NIH) or Federal authorities.
I will not carry out the work described in the attached application until it has been approved by the Institutional
Biosafety Committee (IBC).
Protocol Principal Investigator: _____________________________ ____________

(Signature) (Date)

BIOSAFETY SCHEDULE A: NON-EXEMPT RECOMBINANT DNA
1. (a) Are you using recombinant viral vectors or other non-exempt vector backbones? Yes No
• If “no”, please proceed to Question 2 of this Schedule.
• If “yes”, please complete charts and description(s) below to describe which viral sequences have been deleted from the
wild-type vector (e.g. rendering a replication competent vector replication incompetent), and the packaging systems in
which these vectors will be used for the production of viral particles.
Vector Technical Viral (or non-exempt Regions of viral genome deleted or altered (if any) % of viral genome Is the vector
Name vector) Backbone Source to produce viral vector included in designed to be
(e.g. MLV, MSCV, HIV, (i.e. what is the basis of vector attenuation or constructs (or replication
provide map or reference competent?
Vaccinia, Adenoviral, replication incompetence, if any)
for vector construction)
AAV, Plasmids from
Risk Group 2 organisms)
Yes No
Yes No
Yes No
Yes No
Yes No
Name of Packaging Cell Source(s) of cells (i.e. Tropism Source of envelope glycoproteins
line(s) or Helper plasmids species) (i.e. what species of cells can the virus infect?) (if retro- or lentivirus)
used in co-transfect ion to
produce viral particles
(b) Please describe the insert(s) (if any) to be used in the above vectors.
Include sufficient information for a Risk Assessment by the IBC, which may include answers to the following
questions, as appropriate:
i. What is the gene source (species and/or strain)
ii. What is the nature of the insert(s) (e.g. which protein(s), siRNA are encoded by the insert)
iii. What is the anticipated effect upon expression of the insert(s) (if known)?
iv. How will these constructs be utilized (what are the target cells and/or animals for these constructs)?
v. Have these agents been previously passaged through animals or other cells or cell lines?
vi. At what point in your experiments will your agents be inactivated or lysed?
ver. 120606 BSP Schedule A, page 1

(c) Will you be assaying for the production of wild-type/helper/replication competent viral particles? Yes No
If “yes”, please describe methods and stage in your experiment at which these assays will be performed.
2. Are you using any non-exempt recombinant DNA inserts (e.g. toxins with LD50<100 µg/kg body weight, antibiotic
resistance cassettes or regions of genomes from Risk Group 2, 3 or 4 or restricted organisms)? Yes No
• If “yes”, please describe these inserts and their associated vectors. Include sufficient pertinent information for a Risk
Assessment by the IBC, which may include answers to the following questions, as appropriate:
a. What is the gene source (species and/or strain) of the insert?
b. Into which vectors are these non-exempt inserts being cloned?
c. What is the nature of the insert(s) (e.g. which protein(s), siRNA are encoded by the insert)
d. What is the anticipated effect upon expression of the insert(s)?
e. How will these constructs be utilized (what are the target cells and/or animals for these constructs)?
f. At what point in your experiments will your agents be inactivated or lysed?

3. Are you expressing any recombinant DNA in microbial pathogens (e.g. in risk group 2, 3, or 4 bacteria, yeast, fungi
or parasites or microbes requiring >BSL2 containment)? Yes No
• If “yes”, please describe the pertinent information to enable the risk assessment process, including:
a. The nature of the recombinant DNA construct (vector and insert)
b. Which microbial host cells is this recombinant DNA will be introduced.
c. What are the anticipated effects of introduction of the recombinant DNA into the host (if known)
d. At what point in your experiments will your agents be inactivated or lysed?
4. List approximate maximum volume(s) of cultures of these agent(s) generated at any one time. Note: if any cultures are
>10 liters, please include Schedule C in application.
5. Describe any procedures that will be performed with this material which may be associated with an increased
potential risk of exposure of personnel (increased risk of exposure may be associated with generation of splashes, sprays
or aerosols from centrifugation, sonication, homogenization, vortexing, FACS, use of sharps (needles, glass or syringes),
cage cleaning of infected animals, animal surgeries, etc. Management of these risks should be addressed in the PI’s
laboratory-specific SOPs)

6. Will you be introducing any recombinant DNA or genetic engineered material into animals? Yes No
If “yes”, you will also need to apply for IACUC approval in addition to IBC approval before beginning your research.
You will also need to include Biosafety Schedule E with this application.
7. Will you be introducing these biological agents into humans? Yes No

If “yes”, in addition to IBC approval, your research will require IRB approval and likely review by the Recombinant
DNA Advisory Committee of NIH before beginning your research. You will also need to include Biosafety
Schedule B with this application.
8. Will these experiments result in acquisition of new characteristics of any infectious agents, such as altered virulence or
infectivity, or changes in resistance/susceptibility to drug therapy or changes in host range? Yes No
If “yes”, please describe:

BIOSAFETY SCHEDULE B: HANDLING AND/OR CULTURE OF MAMMALIAN

CELLS, TISSUES, AND/OR ORGANS REQUIRING > BSL-2 CONTAINMENT
(This includes human and non-human primate blood, fluid or tissue specimens,
but excludes those experiments involving non-exempt recombinant DNA documented in Schedule A)
Species of Cells/Tissues/Organs/Specimens Primary (fresh) Established and High likelihood of Genetically Potentially Will these be
origin (If possible, cluster similar items together by material* characterized cell being infected or engineered tumorigenic cultured?
species and/or risks they present. For established lines (e.g. from infectious
cell lines, provide examples of cell lines within ATCC)
each grouping—comprehensive lists are not
required unless they may present additional risk
considerations.)
Human Cell lines (e.g.: 293T, MCF-7, HUVEC) x x Yes No
Human Blood, serum and buccal specimens from x x Yes No

HIV-infected patients
Yes No
Yes No
Yes No
Yes No
Yes No
Yes No
Yes No
*If you will be handling primary (fresh) human or non-human primate material, all personnel will be required to complete the
Georgia Board of Regents Blood-Borne Pathogen Right-to-Know Training module. See instructions for further details.
1. List approximate maximum volume(s) of these agents handled or cultured at any one time. Note: if any cultures are >10
liters, please include Schedule C in application.
2. Describe any procedures to be performed which may be associated with an increased potential risk of exposure of personnel
(e.g. increased risk of exposure may be associated with generation of splashes, sprays or aerosols from centrifugation,
sonication, homogenization, vortexing, FACS, use of sharps (needles or glass), cage cleaning, intranasal inoculation of
animals, etc. Management of these risks should be addressed in the PI’s laboratory-specific SOPs.)
ver. 120606 BSP Schedule B, page 1

3. Will you be introducing these biological agents into animals*? Yes No

If “yes, you will also need to apply for IACUC approval in addition to IBC approval before beginning your research.
4. Will you be introducing these biological agents into humans*? Yes No

If “yes”:
(a) Provide pertinent information to facilitate a comprehensive risk assessment by the IBC, such as:
i. The nature of the material to be introduced into the patients.
ii. Method of delivery and health status of the patients
iii. Frequency of administration
iv. The anticipated effect of the introduction of the agent upon the patient (if known)
v. The expected persistence of the agents after administration (if known)
vi. How long after administration will you obtain specimens to be analyzed? What types of specimens will be obtained?
vii. What types of analyses will be performed? (e.g behavioral analyses, in vivo instrumentation, blood tests, analyses of tissue biopsies?)
viii. What biosafety precautions will be taken to avoid inadvertent exposure to other patients, researchers, or health care providers.
* If you are introducing cells into humans, you will also need to apply to IRB approval in addition to IBC
approval before beginning your research.
(b) Indicate any safety tests or pathogen screening which will be performed on these cells/tissues prior to delivery into
humans:
5. Will any of these experiments require MCG IRB approval? Yes No

If “yes”, have you applied for and/or received IRB approval for your research project(s)? Yes No Pending
If “yes”, indicate HAC or CCRI file number(s) and project title(s) that are covered by this BSP
6. Is your research in the Augusta VA facility? Yes No

If “yes”, please answer the following questions:
Does your research involve any MCG personnel, or transfer to MCG property? Yes No
Have you applied for and/or received VA Biosafety approval for your research? Yes No Pending
If “yes”, please indicate VA safety approval #
If “pending”, please indicate date of submission:
ver. 120606 BSP Schedule B, page 2

BIOSAFETY SCHEDULE C: RESEARCH INVOLVING LARGE-

SCALE (>10 LITER) CULTURES
Please be aware that special precautions may be required for large-scale cultures, particularly those involving recombinant DNA.
These can be reviewed at: http://www4.od.nih.gov/oba/rac/guidelines_02/Appendix_K.htm#_Toc7255533
Lab Room Agents to be cultured under large- In this room, the Agent is Biosafety Level of Biosafety Maximum
Number(s) scale conditions and/or stored in this Room cabinet Volume at
area available? any one time
Decontam-
Used Stored inated
1. Describe the vessels in which you intend to perform large-scale cultures. Indicate whether this is a closed system, and
whether seals are available to prevent leakage enclosed in ventilated housings. Describe whether high efficiency
particulate/HEPA filters are available and where. Describe your monitoring or sensing devices to monitor containment:
2. Describe how you intend to transfer material from your growth chambers and how this material will be decontaminated:
3. Describe how your system exhaust gases will be treated to prevent release of viable organisms:
4. How will your system be sterilized? How will this be validated and tested?
ver. 120606 BSP Schedule C, page 1

5. Describe your transport protocol for transport of agent(s) between locations (i.e. through hallways and other non-laboratory
areas).
6. Describe potential risk of infection of personnel while using the agents indicated (e.g. increased risk of exposure may be
associated with generation of splashes, sprays or aerosols from pouring, centrifugation, sonication, homogenization,
vortexing, FACS, etc. Management of these risks should be addressed in the PI’s laboratory-specific SOPs).
ver. 120606 BSP Schedule C, page 2

BIOSAFETY SCHEDULE D: RESEARCH INVOLVING POTENTIALLY

INFECTIOUS MICROBIAL AGENTS (Risk Group >2) OR SPECIMENS
EXPOSED TO THESE INFECTIOUS MICROBIAL AGENTS
(excluding any material previously described in either Schedule A or B)
Is this a Select Is agent Is agent Is agent Risk Group2

Names of Agents Agent1? infectious to infectious to replication
humans? animals? competent?
Yes No Yes No Yes No Yes No

1
for a list of Select Agents, please see: http://www.cdc.gov/od/sap/docs/salist.pdf
2
See the following links for assistance in Risk Group classification:
http://www4.od.nih.gov/oba/rac/guidelines_02/APPENDIX_B.htm
http://www.absa.org/XriskgroupsX/index.html
http://www.phac-aspc.gc.ca/msds-ftss/
1. List approximate maximum volume(s) of cultures of these agent(s) generated at any one time. Note: if any cultures are >10
liters, please include Schedule C in application.
2. Describe any procedures that will be performed with this material which may be associated with an increased potential risk of
exposure of personnel (e.g. increased risk of exposure may be associated with generation of splashes, sprays or aerosols from
centrifugation, sonication, homogenization, vortexing, FACS, use of sharps (needles or glass), cage cleaning of infected
animals, etc. Management of these risks should be addressed in the PI’s laboratory-specific SOPs)
3. Will you be introducing these biological agents into animals? Yes No

If “yes”, you will also need to apply for IACUC approval in addition to IBC approval before beginning your research.
ver. 120606 BSP Schedule D, page 1

4. Will you be introducing these biological agents into humans? Yes No

If “yes:
(a) Please provide sufficient pertinent information to facilitate the risk assessment by the IBC, such as:
i. The nature of the potentially infectious material to be introduced into the patients.
ii. Have these agents been passaged through animals or other cells or cell lines?
iii. Method of delivery and health status of the patients
iv. Frequency of administration
v. The anticipated effect of the introduction of the agent upon the patient (if known)
vi. The expected persistence of the infection after administration (if known)
vii. How long after administration will you obtain specimens to be analyzed? What types of specimens
will be obtained?
viii. What types of analyses will be performed? (e.g behavioral analyses, in vivo instrumentation, blood
tests, analyses of tissue biopsies?)
ix. At what stage of your experiments will the infectious agent(s) be inactivated or lysed?
x. What biosafety precautions will be taken to avoid inadvertent exposure to other patients,
researchers, or health care providers.
* If you are introducing agents into humans, you will also need to apply to IRB approval in addition to IBC approval
before beginning your research. You may also need to include Biosafety Schedule B with this application.
(b) Indicate any safety tests or pathogen screening which will be performed on these agents prior to delivery into
humans:
5. Will these experiments result in acquisition of new characteristics of these infectious agents, such as altered virulence or
infectivity, or changes in resistance/susceptibility to drug therapy or changes in host range? ? Yes No
If “yes”, please describe:
ver. 120606 BSP Schedule D, page 2

BIOSAFETY SCHEDULE E: USE OF ANIMALS
Please be aware that you will also require approval of the IACUC before initiation of these experiments.
1. Please indicate species of animals you propose to utilize in your research, the biological agent or rDNA you intend to
introduce into these animals, and the locations where you propose to perform these experiments with whole animals or
primary isolates of animal blood and/or tissues.
Species of Biological Agent/rDNA Containment/BSL requirements Building or Location(s) of

housing prior to during housing post- Experiments (if
Animal/Strain known/assigned)
procedures procedures procedures
2. Please provide sufficient pertinent information to facilitate the risk assessment process by the IBC, including:
i. The nature of the biological material to be introduced into the animals (cells, rDNA, genetically
engineered cells, toxins of biological origin).
ii. Method of delivery and which animals will be used.
iii. Frequency of administration
iv. The anticipated effect of the introduction of the biological agent upon the animal (if known).
v. The expected persistence of the infectious material, cells, toxins and/or heterologous gene expression
after administration (if known)
vi. At what point in your experiments will your biological agents be inactivated or lysed?
vii. How long after administration will your specimens be analyzed?
viii. What types of analyses will be performed? (e.g behavioral analyses, in vivo instrumentation, post-
necropsy analyses of tissues?)
3. Are you preparing to create a new strain of genetically engineered (transgenic or knock-out) animals? (Note: this would
include breeding of one genetically engineered animal into a different genetic background or cross-breeding two strains of
genetically engineered animals). Yes No
(a) If “yes”, please describe these.

Please list the genes that will be inserted or knocked out by stable introduction of rDNA into the germ-line (or provide
a copy of the MCG ES/Tg core application if not already received by Biosafety Office), or research plans.
ver. 120606 BSP Schedule E, page 1

4. Will the delivery of any biological material or genetic modification cause the animal to potentially shed infectious material,
toxins or potentially create a hazard to animal handlers, research staff or other animals in the facility? Yes No
If “yes”, describe any procedures that researchers or animal care givers may perform which would be associated with
an increased potential risk of exposure. (e.g. increased risk of exposure may be associated with generation of splashes,
sprays or aerosols from animal cage changing, intranasal inoculation, injection, the use of sharps or glass materials,
or necropsies):
5. Will your experiments require researchers or animal care givers to observe any special safety precautions (beyond those
required by LAS SOPs) in order to prevent potential exposures to hazards associated with your research? Yes No
If “yes”, please describe these:
6. Will you be returning animals to LAS facilities after introduction of biological material? Yes No
If “yes”, indicate location (building/rooms #) or the animals will be housed after return.
7. Do you intend to perform any safety tests or pathogen screening prior to introduction of biological agents into animals (eg.
viral assays of cells or helper viral assays or MAP testing) or monitoring for agents after introduction of the agents into
animals ? Yes No
If “yes”, please describe
ver. 120606 BSP Schedule E, page 2

BIOSAFETY SCHEDULE F: HUMAN GENE TRANSFER/THERAPY
Human gene transfer is defined in NIH Guidelines for Research with Recombinant DNA Molecules as:
Deliberate transfer of recombinant DNA, or DNA or RNA derived from recombinant DNA into one or more
human research participants (Section III-C-1). Please note that this definition would also include research
involving ex vivo transduction of cells for human application.
1. Please provide the Federal Drug Administration (FDA) Investigational New Drug (IND) number and date that the
application went into effect.
2. Does this protocol fit the following criteria for exemption from the NIH/OBA requirements for protocol submission, review
and reporting process as described below (from Appendix M-VI-A of the NIH Guidelines):
Human studies in which induction or enhancement of an immune response to a vector-encoded microbial immunogen is
the major goal, such an immune response has been demonstrated in model systems, and the persistence of the vector-
encoded immunogen is not expected. Yes No
• If “yes”, please address questions and provide material as described in Section A, below.
• If “no”, please provide material as described in Section B, below.
¾ Section A: For DNA Vaccine Human Gene Therapy Protocols which fall under the
Appendix M-VI-A exception:
a. Describe below what evidence exists to suggest that the vector-encoded immunogen expression is not
expected to persist in the patient?
b. Attach responses to Appendices M-II through M-V of NIH Guidelines, Description of the Proposal,
Informed Consent, Privacy and Confidentiality, and Special Issues. Responses to Appendices M-II
through M-V may be provided either as an appendix to the clinical protocol or incorporated in the
clinical protocol. If responses to Appendices M-II through M-V are incorporated in the clinical
protocol, each response must refer to the appropriate Appendix M-II through M-V.
¾ Section B: Human Gene Transfer protocols which do not fall under the Appendix M-
VI-A exception:
a. Attach a copy of the complete protocol submitted to NIH OBA, as described in Appendix M-I-A
b. Attach a copy of the written response from NIH OBA to the protocol submission that either:
i. Indicates the submission does not present characteristics that warrant public RAC review and
discussion; or
ii. Provides a summary of the RAC’s key comments and recommendations after public review.
3. Include a copy of your Human Gene Therapy SOP along with this Schedule.
ver. 120606 BSP Schedule F, page 1

BIOSAFETY SCHEDULE G: HHS/USDA REGULATED SELECT

AGENTS AND TOXINS
1. Do you plan to possess, use, and/or transfer with any toxins of biological origin that are on the HHS and/or USDA Select Agent
and Toxin (SAT) list as defined in 7 CFR Part 331, 9 CFR Part 121, and/or 42 CFR Part 73? Yes No
If “yes”, please:
(a) Include a copy of your laboratory’s toxin-specific SOP.
(b) Place a check in the box to the left of the toxin(s) you intend to possess, use and/or transfer, and complete the information in the table to
the right of the toxin(s).
Names of Toxin(s) Maximum Types(s) or Class of Check (9) Indicate maximum amount (in Check (9) here if
Toxin toxin (if applicable) here if toxin mg) you plan to possess in your your toxins are
amounts has been laboratory at any moment in not eligible for
permitted inactivated time exemption or
9 under SAT prior to exclusion from
exemptions receipt SAT regulations4
Abrin1 100 mg
Botulinum neurotoxins2 0.5 mg
2
Clostridium perfringens epsilon toxin 100 mg
1
Conotoxins 100 mg
Diacetoxyscirpenol (DAS) 1 1000 mg
1
Ricin 100 mg
Saxitoxin1 100 mg
2
Shigatoxin 100 mg
Shiga-like ribosome inactivating proteins 100 mg
(Verotoxins) 1
Staphylococcus enterotoxins2 100 mg
2
T-2 toxin 1000 mg
Tetrodotoxin1 100 mg
1
Regulated by HHS
2
Regulated by HHS and USDA
3
Conotoxins specifically excluded from SAT regulations are: the class of sodium channel antagonist μ-conotoxins, including GIIIA; the class of calcium channel
antagonist ω-conotoxins, including GVIA, GVII, MVIIA, MVIIC, and their analogs or synthetic derivatives; the class of NMDA-antagonist conantokins, including
con-G, con-R, con-T and their analogs or synthetic derivatives; and the putative neurotensin agonist, contulakin-G and its synthetic derivatives.
4
If the amount of toxin you plan to possess exceeds the maximum toxin amounts permitted under SAT exemptions (see above or
http://www.cdc.gov/od/sap/sap/toxinamt.htm), you will need to contact the Biosafety Office immediately to address further SAT regulation issues.
2. Do you plan to possess, use, and/or transfer with any microbial/prion agents that are on the HHS and/or USDA Select Agent and
Toxin (SAT) list as defined in 7 CFR Part 331, 9 CFR Part 121, and/or 42 CFR Part? The SAT list can be viewed at:
http://www.cdc.gov/od/sap/docs/salist.pdf Yes No
If “yes”, please:
(a) Provide the information requested below about your microbial/prion agents
(b) In addition to this Schedule, you will need to include Biosafety Schedule D with this application.
Names of Microbial Agent and Strain Source from which this agent will be If agent has been Check (9) here if you
received inactivated prior to will not be using a
receipt, please indicate strain which is
what method was used excluded from SAT
regulations5
5
Excluded strains of Select Agents can be viewed at: http://www.cdc.gov/od/sap/sap/exclusion.htm
4
If the agent you plan to possess is not excluded from SAT regulations, you will need to contact the Biosafety Office immediately to address
further SAT regulation issues.
ver. 120606 BSP Schedule G, page 1

BIOSAFETY SCHEDULE O: USE OF BIOLOGICAL AGENTS AND/OR RECOMBINANT

DNA IN APPLICATIONS FOR SPONSORED FUNDING OR IRB APPROVAL
1. In order to verify that experiments proposed in a particular sponsored project application to the funding agencies indicated below are fully detailed in the associated
Biosafety Protocol (BSP), please provide information for communication between the Biosafety Office and DSPA.
By providing this information in the chart below, the PI is verifying that the agents, experiments and personnel involved in the sponsored project application have been fully detailed in the associated BSP as indication. Random
reviews will be performed by the Biosafety Office, and grant material may be requested for review at that time.
Funding Anticipated
PI listed on Agency Date of Funding period
BSP # PI listed on BSP BSP Title Grant/Contract Grant/Contract Title Source Submission (MM/YY-MM/YY)
2. In order to verify that experiments proposed in a particular HAC or CCRI application are fully detailed in the associated Biosafety Protocol (BSP), please provide
information for communication between the Biosafety Office and OHRP.
By providing this information in the chart below, the PI is verifying that the agents, experiments and personnel involved in the IRB application have been fully detailed in the associated BSP as indication. Random reviews may be
performed by the Biosafety Office, and protocol material may be requested for review at that time.
Funding Anticipated
PI listed on IRB Agency Date of Funding period
BSP # PI listed on BSP BSP Title Application Title of Project/Protocol Source Submission (MM/YY-MM/YY)
120506 BSP Schedule O, p. 1

Appendix B Classification of Human Etiologic Agents on the Basis of Hazard
This section has been reprinted from the NIH Guidelines for Research Involving Recombinant DNA
Molecules (NIH Guidelines), October 1997.
This appendix includes those biological agents known to infect humans as well as selected animal agents
that may pose theoretical risks if inoculated into humans. Included are lists of representative genera and
species known to be pathogenic; mutated, recombined, and non-pathogenic species and strains are not
considered. Non-infectious life cycle stages of parasites are excluded.
This appendix reflects the current state of knowledge and should be considered a resource document.
Included are the more commonly encountered agents and is not meant to be all-inclusive. Information on
agent risk assessment may be found in the Agent Summary Statements of the CDC/NIH publication,
Biosafety in Microbiological and Biomedical Laboratories (see Sections V-C, V-D, V-E, and V-F,
Footnotes and References of Sections I through IV. Further guidance on agents not listed in Appendix B
may be obtained through: Centers for Disease Control and Prevention, Biosafety Branch, Atlanta, Georgia
30333, Phone: (404) 639-3883, Fax: (404) 639-2294; National Institutes of Health, Division of Safety,
Bethesda, Maryland 20892, Phone: (301) 496-1357; National Animal Disease Center, U.S. Department of
Agriculture, Ames, Iowa 50010, Phone: (515) 862-8258.
A special committee of the American Society for Microbiology will conduct an annual review of this
appendix and its recommendation for changes will be presented to the Recombinant DNA Advisory
Committee as proposed amendments to the NIH Guidelines.
Appendix B - Table 1. Basis for the Classification of Biohazardous Agents by Risk Group (RG)
Risk Group 1 (RG1) Agents that are not associated with disease in healthy adult humans
Risk Group 2 (RG2) Agents that are associated with human disease which is rarely serious and
for which preventive or therapeutic interventions are often available
Risk Group 3 (RG3) Agents that are associated with serious or lethal human disease for which
preventive or therapeutic interventions may be available (high individual
risk but low community risk)
Risk Group 4 (RG4) Agents that are likely to cause serious or lethal human disease for which
preventive or therapeutic interventions are not usually available (high
individual risk and high community risk)
Appendix B-I. Risk Group 1 (RG1) Agents

RG1 agents are not associated with disease in healthy adult humans. Examples of RG1 agents include
asporogenic Bacillus subtilis or Bacillus licheniformis (see Appendix C-IV-A, Bacillus subtilis or Bacillus
licheniformis Host-Vector Systems, Exceptions), Escherichia coli-K12 (see Appendix C-II-A, Escherichia
coli K-12 Host-Vector Systems, Exceptions), and adeno-associated virus types 1 through 4.
Those agents not listed in Risk Groups (RGs) 2, 3 and 4 are not automatically or implicitly classified in
RG1; a risk assessment must be conducted based on the known and potential properties of the agents and
their relationship to agents that are listed.
Appendix B-II. Risk Group 2 (RG2) Agents

RG2 agents are associated with human disease that is rarely serious and for which preventive or therapeutic
interventions are often available.
Appendix B-II-A. Risk Group 2 (RG2) - Bacterial Agents Including Chlamydia

♦ Acinetobacter baumannii (formerly Acinetobacter calcoaceticus)

B-1
♦ Actinobacillus
♦ Actinomyces pyogenes (formerly Corynebacterium pyogenes)
♦ Aeromonas hydrophila
♦ Amycolata autotrophica
♦ Archanobacterium haemolyticum (formerly Corynebacterium haemolyticum)
♦ Arizona hinshawii - all serotypes
♦ Bacillus anthracis
♦ Bartonella henselae, B. quintana, B. vinsonii
♦ Bordetella including B. pertussis
♦ Borrelia recurrentis, B. burgdorferi
♦ Burkholderia (formerly Pseudomonas species) except those listed in Appendix B-III-A (RG3)
♦ Campylobacter coli, C. fetus, C. jejuni
♦ Chlamydia psittaci, C. trachomatis, C. pneumoniae
♦ Clostridium botulinum, Cl. chauvoei, Cl. haemolyticum, Cl. histolyticum, Cl. novyi, Cl.
septicum, Cl.tetani
♦ Corynebacterium diphtheriae, C. pseudotuberculosis, C. renale
♦ Dermatophilus congolensis
♦ Edwardsiella tarda
♦ Erysipelothrix rhusiopathiae
♦ Escherichia coli - all enteropathogenic, enterotoxigenic, enteroinvasive and strains bearing K1
antigen, including E. coli O157:H7
♦ Haemophilus ducreyi, H. influenzae
♦ Helicobacter pylori
♦ Klebsiella - all species except K. oxytoca (RG1)
♦ Legionella including L. pneumophila
♦ Leptospira interrogans - all serotypes
♦ Listeria
♦ Moraxella
♦ Mycobacterium (except those listed in Appendix B-III-A (RG3)) including M. avium complex,
M.asiaticum, M. bovis BCG vaccine strain, M. chelonei, M. fortuitum, M. kansasii, M. leprae,
M. malmoense, M. marinum, M. paratuberculosis, M. scrofulaceum, M. simiae, M. szulgai,
M. ulcerans, M. xenopi
♦ Mycoplasma, except M. mycoides and M. agalactiae which are restricted animal pathogens
♦ Neisseria gonorrhoeae, N. meningitidis
♦ Nocardia asteroides, N. brasiliensis, N. otitidiscaviarum, N. transvalensis
♦ Rhodococcus equi
♦ Salmonella including S. arizonae, S. cholerasuis, S. enteritidis, S. gallinarum-pullorum, S.
meleagridis, S. paratyphi, A, B, C, S. typhi, S. typhimurium
♦ Shigella including S. boydii, S. dysenteriae, type 1, S. flexneri, S. sonnei
♦ Sphaerophorus necrophorus
♦ Staphylococcus aureus
♦ Streptobacillus moniliformis
♦ Streptococcus including S. pneumoniae, S. pyogenes
♦ Treponema pallidum, T. carateum
♦ Vibrio cholerae, V. parahemolyticus, V. vulnificus
♦ Yersinia enterocolitica
Appendix B-II-B. Risk Group 2 (RG2) - Fungal Agents

♦ Blastomyces dermatitidis
♦ Cladosporium bantianum, C. (Xylohypha) trichoides
♦ Cryptococcus neoformans
♦ Dactylaria galopava (Ochroconis gallopavum)
♦ Epidermophyton
♦ Exophiala (Wangiella) dermatitidis
♦ Fonsecaea pedrosoi
♦ Microsporum

B-2
♦ Paracoccidioides braziliensis
♦ Penicillium marneffei
♦ Sporothrix schenckii
♦ Trichophyton
Appendix B-II-C. Risk Group 2 (RG2) - Parasitic Agents

♦ Ancylostoma human hookworms including A. duodenale, A. ceylanicum
♦ Ascaris including Ascaris lumbricoides suum
♦ Babesia including B. divergens, B. microti
♦ Brugia filaria worms including B. malayi, B. timori
♦ Coccidia
♦ Cryptosporidium including C. parvum
♦ Cysticercus cellulosae (hydatid cyst, larva of T. solium)
♦ Echinococcus including E. granulosis, E. multilocularis, E. vogeli
♦ Entamoeba histolytica
♦ Enterobius
♦ Fasciola including F. gigantica, F. hepatica
♦ Giardia including G. lamblia
♦ Heterophyes
♦ Hymenolepis including H. diminuta, H. nana
♦ Isospora
♦ Leishmania including L. braziliensis, L. donovani, L. ethiopia, L. major, L. mexicana, L.
peruvania, L. tropica
♦ Loa loa filaria worms
♦ Microsporidium
♦ Naegleria fowleri
♦ Necator human hookworms including N. americanus
♦ Onchocerca filaria worms including, O. volvulus
♦ Plasmodium including simian species, P. cynomologi, P. falciparum, P. malariae, P. ovale, P.
vivax
♦ Sarcocystis including S. sui hominis
♦ Schistosoma including S. haematobium, S. intercalatum, S. japonicum, S. mansoni, S. mekongi
♦ Strongyloides including S. stercoralis
♦ Taenia solium
♦ Toxocara including T. canis
♦ Toxoplasma including T. gondii
♦ Trichinella spiralis
♦ Trypanosoma including T. brucei brucei, T. brucei gambiense, T. brucei rhodesiense, T. cruzi
♦ Wuchereria bancrofti filaria worms
Appendix B-II-D. Risk Group 2 (RG2) - Viruses

♦ Adenoviruses, human - all types
♦ Alphaviruses (Togaviruses) - Group A Arboviruses
• Eastern equine encephalomyelitis virus
• Venezuelan equine encephalomyelitis vaccine strain TC-83
• Western equine encephalomyelitis virus
♦ Arenaviruses
• Lymphocytic choriomeningitis virus (non-neurotropic strains)
• Tacaribe virus complex
• Other viruses as listed in the reference source (see Section V-C, Footnotes and
References of Sections I through IV)
♦ Bunyaviruses
• Bunyamwera virus
• Rift Valley fever virus vaccine strain MP-12

B-3
♦ Calciviruses
♦ Coronaviruses
♦ Flaviviruses (Togaviruses) - Group B Arboviruses
• Dengue virus serotypes 1, 2, 3, and 4
• Yellow fever virus vaccine strain 17D
♦ Hepatitis A, B, C, D, and E viruses
♦ Herpesviruses - except Herpesvirus simiae (Monkey B virus) (see Appendix B-IV-D, Risk
Group 4 (RG4) -Viral Agents)
• Cytomegalovirus
• Epstein Barr virus
• Herpes simplex types 1 and 2
• Herpes zoster
• Human herpesvirus types 6 and 7
♦ Orthomyxoviruses
• Influenza viruses types A, B, and C
• Other tick-borne orthomyxoviruses as listed in the reference source (see Section V-C,
Footnotes and References of Sections I through IV)
♦ Papovaviruses
• All human papilloma viruses
♦ Paramyxoviruses
• Newcastle disease virus
• Measles virus
• Mumps virus
• Parainfluenza viruses types 1, 2, 3, and 4
• Respiratory syncytial virus
♦ Parvoviruses
• Human parvovirus (B19)
♦ Picornaviruses
• Coxsackie viruses types A and B
• Echoviruses - all types
• Polioviruses - all types, wild and attenuated
• Rhinoviruses - all types
♦ Poxviruses - all types except Monkeypox virus (see Appendix B-III-D, Risk Group 3 (RG3) –
Viruses and Prions) and restricted poxviruses including Alastrim, Smallpox, and Whitepox
(see Section V-L,Footnotes and References of Sections I through IV)
♦ Reoviruses - all types including Coltivirus, human Rotavirus, and Orbivirus (Colorado tick fever
virus)
♦ Rhabdoviruses
• Rabies virus - all strains
• Vesicular stomatitis virus - laboratory adapted strains including VSV-Indiana, San Juan,
and Glasgow
♦ Togaviruses (see Alphaviruses and Flaviviruses)
• Rubivirus (rubella)
Appendix B-III. Risk Group 3 (RG3) Agents

RG3 agents are associated with serious or lethal human disease for which preventive or therapeutic
interventions may be available.
Appendix B-III-A. Risk Group 3 (RG3) - Bacterial Agents Including Rickettsia

♦ Bartonella

B-4
♦ Brucella including B. abortus, B. canis, B. suis
♦ Burkholderia (Pseudomonas) mallei, B. pseudomallei
♦ Coxiella burnetii
♦ Francisella tularensis
♦ Mycobacterium bovis (except BCG strain, see Appendix B-II-A, Risk Group 2 (RG2) - Bacterial
Agents Including Chlamydia), M. tuberculosis
♦ Pasteurella multocida type B -"buffalo" and other virulent strains
♦ Rickettsia akari, R. australis, R. canada, R. conorii, R. prowazekii, R. rickettsii, R, siberica, R.
tsutsugamushi, R. typhi (R. mooseri)
♦ Yersinia pestis
Appendix B-III-B. Risk Group 3 (RG3) - Fungal Agents

♦ Coccidioides immitis (sporulating cultures; contaminated soil)
♦ Histoplasma capsulatum, H. capsulatum var. duboisii
Appendix B-III-C. Risk Group 3 (RG3) - Parasitic Agents

None
Appendix B-III-D. Risk Group 3 (RG3) - Viruses and Prions

♦ Alphaviruses (Togaviruses) - Group A Arboviruses
• Semliki Forest virus
• St. Louis encephalitis virus
• Venezuelan equine encephalomyelitis virus (except the vaccine strain TC-83, see
Appendix BII-D (RG2))
♦ Arenaviruses
• Flexal
• Lymphocytic choriomeningitis virus (LCM) (neurotropic strains)
♦ Bunyaviruses
• Hantaviruses including Hantaan virus
• Rift Valley fever virus
♦ Flaviviruses (Togaviruses) - Group B Arboviruses
• Japanese encephalitis virus
• Yellow fever virus
♦ Poxviruses
• Monkeypox virus
♦ Prions
• Transmissible spongioform encephalopathies (TME) agents (Creutzfeldt-Jacob disease
and kuru agents)(see Section V-C, Footnotes and References of Sections I through IV, for
containment instruction)
♦ Retroviruses
• Human immunodeficiency virus (HIV) types 1 and 2
• Human T cell lymphotropic virus (HTLV) types 1 and 2
• Simian immunodeficiency virus (SIV)
♦ Rhabdoviruses
• Vesicular stomatitis virus
Appendix B-IV. Risk Group 4 (RG4) Agents

RG4 agents are likely to cause serious or lethal human disease for which preventive or therapeutic
interventions are not usually available.
Appendix B-IV-A. Risk Group 4 (RG4) - Bacterial Agents

B-5
None
Appendix B-IV-B. Risk Group 4 (RG4) - Fungal Agents

None
Appendix B-IV-C. Risk Group 4 (RG4) - Parasitic Agents

None
Appendix B-IV-D. Risk Group 4 (RG4) - Viral Agents

♦ Arenaviruses
• Guanarito virus
• Lassa virus
• Junin virus
• Machupo virus
• Sabia
♦ Bunyaviruses (Nairovirus)
• Crimean-Congo hemorrhagic fever virus
♦ Filoviruses
• Ebola virus
• Marburg virus
♦ Flaviruses (Togaviruses) - Group B Arboviruses
• Tick-borne encephalitis virus complex including Absetterov, Central European
encephalitis, Hanzalova, Hypr, Kumlinge, Kyasanur Forest disease, Omsk hemorrhagic
fever, and Russian spring-summer encephalitis viruses
♦ Herpesviruses (alpha)
• Herpesvirus simiae (Herpes B or Monkey B virus)
♦ Paramyxoviruses
• Equine morbillivirus
♦ Hemorrhagic fever agents and viruses as yet undefined
Appendix B-V. Animal Viral Etiologic Agents in Common Use

The following list of animal etiologic agents is appended to the list of human etiologic agents. None of
these agents is associated with disease in healthy adult humans; they are commonly used in laboratory
experimental work.
A containment level appropriate for RG1 human agents is recommended for their use. For agents that are
infectious to human cells, e.g., amphotropic and xenotropic strains of murine leukemia virus, a containment
level appropriate for RG2 human agents is recommended.
♦ Baculoviruses
♦ Herpesviruses
• Herpesvirus ateles
• Herpesvirus saimiri
• Marek's disease virus
• Murine cytomegalovirus
• Papovaviruses
• Bovine papilloma virus
• Polyoma virus
• Shope papilloma virus
• Simian virus 40 (SV40)
♦ Retroviruses
• Avian leukosis virus
• Avian sarcoma virus
• Bovine leukemia virus

B-6
• Feline leukemia virus
• Feline sarcoma virus
• Gibbon leukemia virus
• Mason-Pfizer monkey virus
• Mouse mammary tumor virus
• Murine leukemia virus
• Murine sarcoma virus
• Rat leukemia virus
Appendix B-V-1. Murine Retroviral Vectors

Murine retroviral vectors to be used for human transfer experiments (less than 10 liters) that contain less
than 50% of their respective parental viral genome and that have been demonstrated to be free of detectable
replication competent retrovirus can be maintained, handled, and administered, under BL1 containment.

B-7
Appendix C Biosafety Levels
Only work at biosafety levels 1,2 and 3 is allowed at MCG. There are no biosafety level 4 facilities for
biosafety level 4 work allowed at MCG.
The CDC and NIH have established biosafety guidelines that are found in the CDC/NIH publication
Biosafety in Microbiological and Biomedical Laboratories (BMBL). The following is reprinted from that
publication. For additional information please contact the Biosafety Office at x1-2663 or
BIOSAFETY@mcg.edu.
PRINCIPLES OF BIOSAFETY
The term "containment" is used in describing safe methods for managing infectious agents in the laboratory
environment where they are being handled or maintained. The purpose of containment is to reduce or
eliminate exposure of laboratory workers, other persons, and the outside environment to potentially
hazardous agents.
Primary containment, the protection of personnel and the immediate laboratory environment from exposure
to infectious agents, is provided by both good microbiological technique and the use of appropriate safety
equipment. The use of vaccines may provide an increased level of personal protection. Secondary
containment, the protection of the environment external to the laboratory from exposure to infectious
materials, is provided by a combination of facility design and operational practices. Therefore, the three
elements of containment include laboratory practice and technique, safety equipment, and facility design.
The risk assessment of the work to be done with a specific agent will determine the appropriate
combination of these elements.
LABORATORY PRACTICE AND TECHNIQUE

The most important element of containment is strict adherence to standard microbiological practices and
techniques. Persons working with infectious agents or potentially infected materials must be aware of
potential hazards, and must be trained and proficient in the practices and techniques required for handling
such material safely. The director or person in charge of the laboratory is responsible for providing or
arranging for appropriate training of personnel.
Each laboratory should develop or adopt a biosafety or operations manual that identifies the hazards that
will or may be encountered and that specifies practices and procedures designed to minimize or eliminate
risks. Personnel should be advised of special hazards and should be required to read and to follow the
required practices and procedures. A scientist trained and knowledgeable in appropriate laboratory
techniques, safety procedures, and hazards associated with handling infectious agents must direct
laboratory activities.
When standard laboratory practices are not sufficient to control the hazard associated with a particular
agent or laboratory procedure, additional measures may be needed. The laboratory director is responsible
for selecting additional safety practices, which must be in keeping with the hazard associated with the agent
or procedure.
Laboratory personnel, safety practices, and techniques must be supplemented by appropriate facility design
and engineering features, safety equipment, and management practices.
SAFETY EQUIPMENT (PRIMARY BARRIERS)

Safety equipment includes biological safety cabinets (BSCs), enclosed containers, and other engineering
controls designed to remove or minimize exposures to hazardous biological materials. The biological safety
cabinet (BSC) is the principal device used to provide containment of infectious splashes or aerosols
generated by many microbiological procedures.

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Three types of biological safety cabinets (Class I, II, III) used in microbiological laboratories are described
and illustrated in Appendix A of the CDC/NIH publication. Class I and Class II biological safety cabinets
are primary barriers that offer significant levels of protection to laboratory personnel and the environment
when used with good microbiological techniques. The Class II biological safety cabinet also provides
protection from external contamination of the materials (e.g., cell cultures, microbiological stocks) being
manipulated inside the cabinet. The gas-tight Class III biological safety cabinet provides the highest
attainable level of protection to personnel and the environment. An example of another primary barrier is
the safety centrifuge cup; an enclosed container designed to prevent aerosols from being released during
centrifugation. To minimize this hazard, containment controls such as BSCs or centrifuge cups must be
used for handling infectious agents that can be transmitted through the aerosol route of exposure.
Safety equipment also may include items for personal protection such as gloves, coats, gowns, shoe covers,
boots, bouffant head covers, respirators, face shields, safety glasses, or goggles. Personal protective
equipment is often used in combination with biological safety cabinets and other devices that contain the
agents, animals, or materials being worked with. In some situations in which it is impractical to work in
biological safety cabinets, personal protective equipment may form the primary barrier between personnel
and the infectious materials. Examples include certain animal studies, animal necropsy, agent production
activities, and activities relating to maintenance, service, or support of the laboratory facility.
FACILITY DESIGN (SECONDARY BARRIERS)

The design of the facility is important in providing a barrier to protect persons working inside and outside
of the laboratory within the facility, and to protect persons or animals in the community from infectious
agents that may be accidentally released from the laboratory. Laboratory management is responsible for
providing facilities commensurate with the laboratory's function and the recommended biosafety level for
the agents being manipulated.
The recommended secondary barrier(s) will depend on the risk of transmission of specific agents. For
example, the exposure risks for most laboratory work in Biosafety Level 1 and 2 facilities will be direct
contact with the agents, or inadvertent contact exposures through contaminated work environments.
Secondary barriers in these laboratories may include separation of the laboratory work area from public
access, availability of a decontamination facility (e.g., autoclave), and hand-washing facilities.
As the risk for aerosol transmission increases, higher levels of primary containment and multiple secondary
barriers may become necessary to prevent infectious agents from escaping into the environment. Such
design features could include specialized ventilation systems to assure directional air flow, air treatment
systems to decontaminate or remove agents from exhaust air, controlled access zones, airlocks as
laboratory entrances, or separate buildings or modules for isolation of the laboratory. Design engineers for
laboratories may refer to specific ventilation recommendations as found in the Applications Handbook for
Heating, Ventilation, and Air-Conditioning (HVAC) published by the American Society of Heating,
Refrigerating, and Air-Conditioning Engineers (ASHRAE).
BIOSAFETY LEVELS
Four biosafety levels (BSLs) are described which consist of combinations of laboratory practices and
techniques, safety equipment, and laboratory facilities. Each combination is specifically appropriate for the
operations performed, the documented or suspected routes of transmission of the infectious agents, and for
the laboratory function or activity.
BIOSAFETY LEVEL 1 practices, safety equipment, and facilities are appropriate for undergraduate and
secondary educational training and teaching laboratories, and for other facilities in which work is done with
defined and characterized strains of viable microorganisms not known to cause disease in healthy adult
humans. Bacillus subtilis, Naegleria gruberi, and infectious canine hepatitis virus are representative of
those microorganisms meeting these criteria. Many agents not ordinarily associated with disease processes
in humans are, however, opportunistic pathogens and may cause infection in the young, the aged, and

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immunodeficient or immunosuppressed individuals. Vaccine strains which have undergone multiple in vivo
passages should not be considered avirulent simply because they are vaccine strains.
Biosafety Level 1 represents a basic level of containment that relies on standard microbiological practices
with no special primary or secondary barriers recommended, other than a sink for hand-washing.
BIOSAFETY LEVEL 2 practices, equipment, and facilities are applicable to clinical, diagnostic, teaching
and other facilities in which work is done with the broad spectrum of indigenous moderate-risk agents
present in the community and associated with human disease of varying severity. With good
microbiological techniques, these agents can be used safely in activities conducted on the open bench,
provided the potential for producing splashes or aerosols is low. Hepatitis B virus, the salmonellae, and
Toxoplasma spp. are representative of microorganisms assigned to this containment level. Biosafety Level
2 is appropriate when work is done with any human-derived blood, body fluids, or tissues where the
presence of an infectious agent may be unknown. (Laboratory personnel working with human-derived
materials should refer to the Bloodborne Pathogen Standard for specific, required precautions).
Primary hazards to personnel working with these agents relate to accidental percutaneous or mucous
membrane exposures, or ingestion of infectious materials. Extreme precaution with contaminated needles
or sharp instruments must be emphasized. Even though organisms routinely manipulated at BSL2 are not
known to be transmissible by the aerosol route, procedures with aerosol or high splash potential that may
increase the risk of such personnel exposure must be conducted in primary containment equipment, or
devices such as a BSC or safety centrifuge cups. Other primary barriers should be used as appropriate such
as splash shields, face protection, gowns, and gloves.
Secondary barriers such as hand-washing and waste decontamination facilities must be available to reduce
potential environmental contamination.
BIOSAFETY LEVEL 3 practices, safety equipment, and facilities are applicable to clinical, diagnostic,
teaching, research, or production facilities in which work is done with indigenous or exotic agents with a
potential for respiratory transmission, and which may cause serious and potentially lethal infection.
Mycobacterium tuberculosis, St. Louis encephalitis virus, and Coxiella burnetii are representative of
microorganisms assigned to this level. Primary hazards to personnel working with these agents relate to
autoinoculation, ingestion, and exposure to infectious aerosols.
At Biosafety Level 3, more emphasis is placed on primary and secondary barriers to protect personnel in
contiguous areas, the community, and the environment from exposure to potentially infectious aerosols. For
example, all laboratory manipulations should be performed in a BSC or other enclosed equipment, such as
a gas-tight aerosol generation chamber. Secondary barriers for this level include controlled access to the
laboratory and a specialized ventilation system that minimizes the release of infectious aerosols from the
laboratory.
ANIMAL FACILITIES
Four biosafety levels are also described for activities involving infectious disease work with experimental
mammals. These four combinations of practices, safety equipment, and facilities are designated Animal
Biosafety Levels 1, 2, 3, and 4, and provide increasing levels of protection to personnel and the
environment.
CLINICAL LABORATORIES
Clinical laboratories, especially those in health care facilities, receive clinical specimens with requests for a
variety of diagnostic and clinical support services. Typically, the infectious nature of clinical material is
unknown, and specimens are often submitted with a broad request for microbiological examination for
multiple agents (e.g., sputa submitted for "routine," acid-fast, and fungal cultures). It is the responsibility of
the laboratory director to establish standard procedures in the laboratory which realistically address the
issue of the infective hazard of clinical specimens. Except in extraordinary circumstances (e.g., suspected

C-3
hemorrhagic fever), the initial processing of clinical specimens and identification of isolates can be done
safely at Biosafety Level 2, the recommended level for work with bloodborne pathogens such as hepatitis B
virus and HIV. The containment elements described in Biosafety Level 2 are consistent with the
Occupational Exposure to Bloodborne Pathogens Standard from the Occupational Safety and Health
Administration (OSHA), that requires the use of specific precautions with all clinical specimens of blood or
other potentially infectious material (Universal Precautions). Additionally, other recommendations specific
for clinical laboratories may be obtained from the National Committee for Clinical Laboratory Standards.
Biosafety Level 2 recommendations and OSHA requirements focus on the prevention of percutaneous and
mucous membrane exposures to clinical material. Primary barriers such as biological safety cabinets should
be used when performing procedures that might cause splashing, spraying, or splattering of droplets.
Biological safety cabinets should also be used for the initial processing of clinical specimens when the
nature of the test requested or other information is suggestive that an agent readily transmissible by
infectious aerosols is likely to be present (e.g., M. tuberculosis), or when the use of a biological safety
cabinet (Class II) is indicated to protect the integrity of the specimen. The segregation of clinical laboratory
functions and limiting or restricting access to such areas is the responsibility of the laboratory director. It is
also the director's responsibility to establish standard, written procedures that address the potential hazards
and the required precautions to be implemented.

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Appendix D BL2+ Work Practices
Biosafety Level 2 Plus (BL2+) is the designation utilized for those biohazard experiments that require
practices that are more stringent than standard BL2 procedures. Generally, BL3 practices are mandated in a
space designed for BL2 work. It is preferred that the BL2 laboratory be self-contained; that is, all
equipment required for the experiment should be located within the lab. A sign is posted on the door while
BL2+ work is in progress, and access is restricted to those involved in the experiment. When work is
completed, and equipment has been decontaminated, the sign is removed and the lab returns to standard
BL2 or BL1 usage.
BL3 practices require that all work be conducted under physical containment. Therefore, all manipulations
with BL2+ material are conducted within a Class II biological safety cabinet and secondary containment is
utilized for centrifugation and other potential aerosol generating procedures. The following notes further
describe the requirements for work at BL2+.
Personal Protective Equipment (PPE)

♦ Dedicate PPE for the experiment. PPE worn for BL2+ work should not be worn in other areas.
Remove before leaving the laboratory.
♦ Wear a lab coat or solid-front gown, preferably with a knit or grip cuff.
♦ Double glove for all work within the biological safety cabinet (BSC). Remove the outer pair
before exiting the BSC, and don a new pair each time you reenter the BSC.
♦ Ensure that your gloves extend over the sleeve of your lab coat. An opening at the wrist will
allow aerosols generated within the BSC to contaminate your wrist and forearm, extending
hand-washing to your elbow.
♦ Sleeve covers can be worn to ensure coverage of the wrist and will also minimize contamination
of the sleeves of your lab coat.
♦ Face Protection (mask and eyewear, can also be worn and will protect mucous membranes from
exposure in the event a spill outside the BSC during transfer of material to and from the
incubator. It will also help to prevent you from touching your eyes, nose and mouth when
working within the BSC.
♦ Remove PPE before leaving the laboratory. Placing a coat hook within the BL2+ area will
facilitate this requirement. Remove your outer gloves first, then your lab coat or gown,
followed by the inner gloves. Take your face protection off last. Don’t touch your face with
gloved hands. Remove gloves and other clothing aseptically, from the inside out, and avoid
touching the contaminated outer side of the glove.
♦ Decontaminate reusable PPE as soon as feasible after it has been contaminated. Small areas can
be spot treated with a suitable disinfectant, such as 1-10% household bleach. Lab coats can
also be autoclaved, or sent to a laundry facility equipped to handle biohazardous PPE.
Disposable PPE can be placed within a biohazard bag, treated and discarded as biomedical
waste.
♦ Wash your hands with soap and water after removing PPE and before leaving the laboratory.
Work Practices In the Biological Safety Cabinet (BSC)

♦ Perform all work within a BSC. This includes discarding waste within the BSC. Moving your
hands in and out of the BSC will disrupt the protective air curtain at the front access opening.
♦ Place all items required for the experiment within the BSC before starting work.
♦ Wipe items down with disinfectant prior to placement within the BSC.
♦ Segregate clean areas from contaminated areas within the BSC (by at least 12-14”).
♦ Keep the front and rear grilles clear when working within the BSC. Avoid blocking the rear
grille. Don’t store items on top of the BSC. Remind fellow researchers to minimize traffic and
work behind the operator, as this may interfere with cabinet airflow. Depending on the
location of the BSC within the room, opening and closing the room door can significantly
interfere with BSC airlfow.

D-1
♦ Avoid the use of a flame within the BSC. In addition to presenting a fire hazard, an open flame
can disrupt airflow and possibly damage the paper filter located above the work surface. If the
use of flame is absolutely necessary, use a burner with a pilot light that provides a flame only
when depressed and releases after contact. Never leave an open flame (burner or pilot light)
unattended in your BSC.
♦ Store tissue culture flasks in the incubator within small secondary trays to help minimize
contamination. Trays will also facilitate transfer to and from the BSC.
♦ Keep your hands away from your face (face protection helps to minimize the potential for this
route of exposure).
♦ Avoid the use of glass Pasteur pipettes or needles and syringes. Substitute plastic for glass
whenever feasible. Alternatives to glass Pasteur pipettes include: plastic pipettes, plastic
transfer pipettes, plastic gel loading pipette tips and pipette tip extendors, aspirators, and
flexible plastic aspiration pipettes. Some researchers will either score and break the end off of
a 1 ml or 5 ml plastic pipette or remove the wool plug and use for aspirating cultures.
♦ If the use of sharps cannot be avoided, maintain a sharps container in the immediate vicinity of
use. Discard intact needles and syringes immediately after use. Use a one-handed disposal
method (keep a hand behind your back or by your side, don’t place on or near the opening of
the sharps container). Never recap, bend, break or otherwise manipulate sharps by hand. If
you must remove the needle from the syringe, use the small opening on the top of the
needlebox for this purpose. Forceps, tweezers, or small pliers may also be utilized.
♦ Protect the house vacuum system or pump from contamination by installing a trap and filter
system. Use a primary collection flask containing disinfectant, followed by an overflow flask,
which leads through a HEPA or hydrophobic filter.
♦ Collect all waste within the BSC. Smaller biohazard waste bags may be utilized along with
beakers or shallow trays containing disinfectant for the collection and disinfection of pipettes
and other contaminated items. Waste can also be collected within the BSC in the following
manner.
• Horizontal collection: Horizontal trays containing disinfectant allow total immersion of
pipettes.
• Vertical collection: Beakers containing disinfectant can be used if disinfectant is drawn
up inside the pipette and allowed to run down the interior wall upon disposal into the
beaker.
• Bags: Bags have the potential for creating aerosols when moved. At BL2+, seal autoclave
bags within the cabinet and place within a second bag. Carefully add water to the primary
bag before sealing (25 ml for smaller bags, 200 ml for larger bags). The addition of water
will help to generate steam within the bag during the autoclave cycle.
♦ Wipe items down with disinfectant prior to removal from the BSC.
♦ Wipe down BSC with disinfectant after use (work surface, grilles, sides, back and inside front
view screen).
♦ Decontaminate liquid waste with household bleach diluted 10% against the volume of the waste.
Allow at least a 30 minute contact time for full decontamination.
♦ Transport waste to autoclave in a leakproof container.
Centrifugation
♦ Use sealed rotors or safety buckets as secondary containment for centrifugation.
♦ Load and unload the rotor or safety buckets within the BSC.
♦ Don’t overfill primary containers, limit to < ¾ full. Wipe exterior of tube with disinfectant
before loading.
♦ Seal rotor or bucket and wipe down with disinfectant, remove outer gloves, and transport to the
centrifuge.
♦ Post a sign on centrifuge that includes the biohazard symbol, name of the agent with Biosafety
Level, and your name.
♦ Wait 2-5 minutes after the run to allow aerosols to settle in the event of a spill. Transport sealed
rotor or safety bucket to cabinet to complete your experiment. Don new pair of outer gloves.

D-2
♦ Decontaminate the rotor or safety bucket by spraying with 70% ethanol and allowing to air dry.
Wipe the throw line within the centrifuge with disinfectant and remove your biohazard sign.
In the event of a spill during centrifugation, follow the spill response procedures outlined in
the the laboratory SOPs.
♦ Avoid the use of microfuge, which is difficult to contain. If you cannot avoid using a microfuge,
use a model that has built in secondary containment (a sealed rotor) along with microfuge
tubes equipped with a O-ring seal. You can also operate your microfuge in the rear of your
BSC (don’t perform any work within the BSC while the microfuge is in operation, and wait 2-
5 minutes after the run before opening the microfuge).
Labels
♦ Post a biohazard sign at the entry to the BL2+ laboratory.
♦ Ensure that any specific entry requirements (vaccination), the name of the agent, the Biosafety
Level, and the name of an emergency contact person ARE posted on the door placards.
♦ Place the BL2 wall notice (not a door sign) inside your laboratory to remind researchers of the
core safety practices.
♦ Label equipment housing the agent (incubators, freezers) with the universal biohazard symbol
and agent name.
Transport of Biohazards on Campus (between labs or buildings):

♦ Must have two leakproof containers, including the following:
• a sealed primary container
• a sealed secondary container
• absorbent (paper towels) between the primary and secondary containers suitable for the
volume transported
• a biohazard sticker on the outside of the secondary container with agent name
• lab address and phone number on the outside of the secondary container
♦ Utilize plastic containers whenever feasible. Avoid glass.
♦ Sealed plastic (not glass) primary vials can be transported within sealed, labeled plastic bags.
♦ If glass primary containers must be used, place containers within a sealed rigid plastic container
with absorbent and padding to cushion vials during transport.
♦ Decontaminated the outside of the primary container before placing into the secondary
container.
♦ Decontaminate the secondary container before leaving the laboratory
Hand-washing
♦ Wash hands whenever PPE is removed and before leaving the laboratory.
♦ Wash with soap and warm water for at least 15 seconds. Since the contact time of most soaps is
quite extensive for actual decontamination, mechanical friction from scrubbing and water
dilution are essential for complete cleaning.
♦ No glove is 100% leakproof.
♦ Never wet or handwash your gloves with water or disinfectant, as this will encourage wicking
and increase permeability of the protective barrier.
Spills and Exposure Incidents

♦ All researchers must be familiar with the applicable exposure response procedures before
initiating their experiments.
♦ Review the attached Biosafety Spill and Incident Response SOPs before starting work.

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Appendix E: Standard Operating Procedure Development
Template
Avialable in MS Word format at: http://www.mcg.edu/research/ibc/apps.htm
This template should be considered a starting point to initiate the SOP development process. It covers only
the basic standard issues addressed in most biological laboratory settings; however, this should be edited
and/or modify to address the risks within each laboratory or clinic.
Special considerations to document practices such as extramural transport, possession, use or handling of
biological toxins or any specific issues relevant to the laboratory should be added.

E-1
LABORATORY
STANDARD OPERATING PROCEDURE
DEVELOPMENT GUIDANCE TEMPLATE
Please edit and complete as necessary to address Biosafety Risks within your
laboratory
Date:
Principal Investigator:
Department:
Room(s) + BSL of each room:
Phone:
______________________________________________________________________________
1. Access to the laboratory is limited to staff, or other persons with permission of the Principal Investigator,
when work with BSL-2 pathogens is being conducted. Access is limited according to attached procedures.
2. All laboratory personnel must be screened by Employee Health before working with potentially infectious
materials, including fresh, unfixed, human or non-human primate specimens, uncharacterized cell lines, etc.
Any available vaccinations which would reduce the risks associated with exposure to any of the agents in the
protocol must be offered to all personnel or a signed waiver must be obtained by the PI.
3. Persons who have increased risk of infection, or for whom infection may have serious consequences, must
not be allowed to enter laboratory when work with infectious agents is in progress without permission of the
Principal Investigator.
4. All persons entering the laboratory must be advised of potential hazards. Laboratory workers must be trained
and made aware of the hazards and appropriate safety precautions before working with any of the biological
agents.
5. Spills and accidents that result in overt exposures to infectious materials must be immediately reported to the
Principal Investigator and appropriate medical evaluation must be provided.
6. Laboratory staff must not eat, drink, smoke, handle contact lenses, chew gum, apply cosmetics in laboratory.
7. Food or drink for human consumption or utensils or cups must stored outside laboratory work area in
refrigerators designated for that purpose only.
8. Laboratory staff and all other persons working with infectious substances must wear gloves. Contaminated
gloves must be changed IMMEDIATELY. Under NO CIRCUMSTANCES will gloves be reused.
9. Laboratory staff must wash hands after handling infectious materials, after removing gloves, and before
leaving the laboratory.
10. Face protection (goggles, mask, face shield, or other splatter guard) must be used for all procedures when
such procedures could produce splashes or sprays of infectious or other hazardous materials or when
microorganisms are manipulated outside the biological safety cabinet. See attached procedures for more
detailed PPE requirements.
11. Protective clothing must be removed and left in laboratory before going to non-laboratory areas (cafeteria,
library, administrative areas).
12. Protective clothing must be either disposed of in laboratory or laundered by institution. (NEVER taken
home!)
13. Only mechanical pipetting devices must be used in the laboratory.

E-2
14. All sharps must be placed in red (approved) sharps containers.
15. Sharps containers must be turned in when ¾ full.
16. All procedures must be performed to minimize creation of splashes or aerosols according to attached
procedures.
17. A Biosafety Cabinet must be used for all procedures with potential for creating infectious aerosols or
splashes, or whenever handling high concentrations of infectious materials.
18. Biological Safety Cabinet must have a current (annual) certification.
19. Biological Safety Cabinet must not be used until it is recertified after relocation, any movement, or
dislocation.
20. Centrifuges used for high concentrations or large volumes of infectious agents must have sealed rotor heads
or centrifuge safety cups. Safety cups must be opened only in a Biosafety cabinet.
21. All infectious animal carcasses must be disposed of by placing in a plastic bag and placing in infected animal
carcass freezer or according to attached procedures.
22. All non-liquid contaminated cultures, stocks, plasticware and other biologically contaminated waste must be
disinfected and placed in biomedical waste containers for disposal according to attached procedures.
23. All infectious liquids must be decontaminated or disinfected prior to being poured into drain according to
attached procedures.
24. All work surfaces must be decontaminated at the completion of work, at the end of the day, or after any spill
or splash of viable material according to attached procedures.
25. Laboratory equipment, surfaces and other contaminated materials must be decontaminated with an effective
disinfectant on a routine basis, after work with infectious materials is finished, and especially after overt
spills, splashes, or other contamination according to attached procedures. Equipment must be
decontaminated before removal from the laboratory (for repair maintenance or other purposes).
Decontamination protocol is attached.
26. Laboratory doors must be locked when the laboratory is not occupied.
27. Spaces between benches, cabinets, and equipment must be kept accessible for cleaning.
28. Eyewash must be flushed at least monthly and kept clear of obstructions.
29. All personnel must know the location of the nearest safety shower and must ensure that area under the shower
is kept free of obstructions.
30. Any personnel with overt exposure will be taken to Employee Health and/or the ER for immediate (<1 hour)
post-exposure medical intervention and monitoring.
31. Any overt exposures of personnel to biological agents will be reported to the PI and the BSO as soon as
possible.

E-3
SOP ADDENUM- DETAILED LABORATORY-SPECIFIC PROTOCOLS
1. Storage and access control of cells or cultures include the following. Consider storage in all locations, including freezers,
cryotanks, cold rooms.
Yes No Agents are stored inside well-labeled tubes/vials.

Yes No Agents are stored inside well-labeled storage boxes.
Yes No Biohazard labels are clearly affixed on any storage device clearly identifying the hazards inside
Yes No Storage refrigerators/freezers/cryotanks are in rooms whose access is limited to authorized personnel.
Yes No Refrigerators are kept locked and accessible only to authorized personnel
Yes No Freezers are kept locked and accessible only to authorized personnel
Yes No Cryotanks are kept locked and accessed only by authorized personnel
Yes No Lockboxes are utilized and accessible only to authorized personnel
Yes No Policies and procedures are in place whereby only persons who have been advised of the potential hazard
and meet any specific entry requirements (e.g., immunization) are authorized to enter the laboratory or
have access to agents.
Yes No Others (please specify)
2. Procedures employed which may result in the generation of aerosols, splashes and/or sprays of biological material and safety
precautions that should be followed by personnel performing these procedures are as follows:
Centrifugation
Sonication
Vortexing
Homogenization
Opening Vacuum vials
Syringe filtration
Placing biological materials under pressure (through small bore hoses or needles)
Animal cage changing
Intranasal inoculation
Necropsy of infected animals
Fluorescence activated cell sorting
Fluorescence activated cell analysis
Other (please specify):
Safety precautions (e.g. use of Biosafety Cabinets, shields, respirators, containment devices):
3. Transport procedures for transport of agent(s)* between locations (i.e. through hallways and other non-laboratory areas). Note:
For BSL2 agents, consider transporting agents inside a sealed, leakproof primary container inside a well-labeled sealed,
leakproof, durable secondary container.
*If transporting animals, please describe procedure.
*If transporting any biological agent off-campus, please refer to Biosafety Schedule L for transport guidelines.

E-4
4. Disinfection/decontamination and disposal of viable agents or recombinant DNA or items which have come in contact with these
agents. See: http://www.mcg.edu/research/ibc/disinfectants for guidance in selecting appropriate disinfectants for the agents in this
protocol.
LIQUID WASTES (check all that apply):
Disinfection/Decontamination Method Treatment Final Disposal (after disinfection indicated in left column))
time (eg. 30 Check all that apply. If “other”, please specify below
min)
None necessary N/A In sharps container As chemical waste Other
Autoclaving Into Sewer In sharps container As chemical waste Other
Chemical Methods:
Bleach (freshly diluted to final 10% (v/v)) Into Sewer In sharps container As chemical waste Other
Other chlorine products (Clidox®) As chemical waste Other
Iodine/iodophors (eg. Wescodyne®) As chemical waste Other
Alcohols (e.g. final 70% (v/v) EtOH or IPA) As chemical waste Other
Phenolic agents (e.g. Biozide®, Vesphene®) As chemical waste Other
Quaternary Ammonium Agents (e.g. Roccal®, As chemical waste Other
Coverage Plus®, Cavicide®, Lysol®)
Aldehydes (e.g. 2-4% glutaraldehyde, 4% As chemical waste Other
formaldehyde, Cidex®)
Peroxygens (e.g. Virkon®) As chemical waste Other
Other, please specify:
SOLID WASTES (Plasticware, glassware, tubes, tissues) (check all that apply):
Disinfection/Decontamination Method Treatment Final Disposal (after disinfection indicated in left column)
min) All biological waste must be disposed as Biohazardous Waste
None required N/A In Biohaz. box In biohaz. sharps container Other
Autoclaving In Biohaz. box In biohaz. sharps container Other
Chemical Methods
Bleach (freshly diluted to final 10% (v/v)) In Biohaz. box In biohaz. sharps container Other
Aldehydes (e.g. fixation with 2-4% In Biohaz. box In biohaz. sharps container Other
glutaraldehyde, 4% formaldehyde)
Peroxygens (e.g. Virkon®) In Biohaz. box In biohaz. sharps container Other
ANIMAL TISSUES/CARCASS WASTE (check all that apply):
Disinfection/Decontamination Method Treatment Final Disposal (after disinfection indicated in left column)
min)
Autoclaving prior to removal from laboratory In LAS animal disposal freezer In Biohaz. waste box Other
None, transported to LAS as per protocol below N/A In LAS animal disposal freezer In Biohaz. waste box Other

E-5
5. Work surfaces, instruments, equipment will be decontaminated using the following method(s). Consider both benchtop
surfaces and stainless steel surfaces (such as those in biosafety cabinets), as well as in equipment (such as centrifuges) and
instruments (check all that apply):
Check all that apply

Disinfection/Decontamination Method Contact time Frequency Benchtops Stainless Equipment Instruments
(e.g. 30’) (e.g. daily and after Surfaces
spills)
Autoclaving
Chemical Methods:
Bleach (freshly diluted to final 10%
(v/v))
Bleach (freshly diluted to final 10%
(v/v)), + rinse with 70% Alcohol
Other chlorine products (Clidox®)
Iodine/iodophors (eg. Wescodyne®)
Alcohols (e.g. final 70% (v/v) EtOH or
IPA)
Phenolic agents (e.g. Biozide®,
Vesphene®)
Quaternary Ammonium Agents (e.g.
Roccal®, Coverage Plus®,
Cavicide®, Lysol®)
Aldehydes (e.g. 2-4% glutaraldehyde, 4%
Peroxygens (e.g. Virkon®)
6. Spills involving biological material will be handled in your laboratory using the following procedures (check all that apply):
(a) Describe which measures will be employed to handle spills in your laboratory:
Large spills or high-hazard spills will be reported to PI and BSO immediately for assistance
Spills in public (non-laboratory) areas will be reported to PI and BSO immediately
Spills involving high hazard agent(s) (e.g. Select Agents) will be reported to the PI and BSO immediately
Any injured individuals will be removed from areas and medical attention and follow-up will be provided
All personnel in area will be advised of hazard and told to avoid the area
Spill will be covered with copious absorbent material (like paper towels)
The spill will be decontaminated with an appropriate disinfectant for the agent(s) spilled for an appropriate contact
period (see below)
After decontamination, absorbent material will be disposed as biohazardous waste
After decontamination, the surface will be cleaned to remove residual disinfectant
Spills in equipment (e.g. Biosafety cabinets or centrifuges) will be disinfected according to written procedures on the
MCG Emergency Response Flip charts which have been posted in my laboratory

E-6
(b) Spills will be disinfected using the following disinfectant and contact time:
Disinfection/Decontamination Method Contact time

(e.g. 30’)
Autoclaving
Chemical Methods:
Bleach (freshly diluted to final 10% (v/v))
Bleach (freshly diluted to final 10% (v/v)),
+ rinse with 70% Alcohol
Other chlorine products (Clidox®)
Iodine/iodophors (eg. Wescodyne®)
Alcohols (e.g. final 70% (v/v) EtOH or
IPA)
Phenolic agents (e.g. Biozide®,
Vesphene®)
Quaternary Ammonium Agents (e.g.
Roccal®, Coverage Plus®, Cavicide®,
Lysol®)
Aldehydes (e.g. 2-4% glutaraldehyde, 4%
Peroxygens (e.g. Virkon®)
7. Protective equipment which will be utilized to minimize exposure of personnel to agents while in the laboratory:
(check all that apply):

Gloves (nitrile or latex) Cuffed wrists on gowns Safety glasses Surgical mask
Double Gloves (nitrile or Sleeve protectors Goggles N95 Respirator
latex)
Lab coat Scrubs Face shield PAPR Respirator
Tyvek coveralls Shoe covers/booties Plexiglass shield Aprons
Rear-closing gowns Biosafety Cabinet Safety caps/Sealed rotors Rubber boots
on Centrifuge
Other (please specify)
8. Protective equipment which will be utilized to minimize exposure of personnel to agents while working with animals (if
applicable):
Agents will be delivered into animals using which BSL containment practices and standards: BSL-1 BSL-2 BSL-3
Please indicate Personal Protective Equipment (PPE) and other safety equipment to be utilized while delivering agents into animals
(check all that apply):
Gloves (nitrile or latex) Cuffed wrists on gowns Safety glasses Surgical mask
Double Gloves (nitrile or Sleeve protectors Goggles N95 Respirator
latex)
Lab coat Scrubs Face shield PAPR Respirator
Tyvek coveralls Shoe covers/booties Plexiglass shield Aprons
Rear-closing gowns Biosafety Cabinet Rubber boots
Other (please specify): ______________________________________________________________________________
After delivery of agents into animals:

After delivery of agents, animals will be housed in (check all that apply):
Separate cages from other animals Well-labeled cages indicating possible hazards to animal staff Cages with filter-top lids
Microisolator cages Conventional/ABSL-1 facilities ABSL-2 facilities ABSL-3 facilities
Live animals will not be returned to LAS facilities after delivery Other (please specify): _________________________________
How long after delivery of agents will animals be analyzed and/or sacrificed (indicate time range)? _________________________________

E-7
Appendix F Human Gene Transfer Clinical Trials
Proposed clinical trials involving human gene transfer require registration and approval from both campus
and federal agencies before initiation. NIH defines human gene transfer as the “the deliberate transfer of
recombinant DNA, or DNA or RNA derived from recombinant DNA, into human subjects.” The MCG
Institutional Biological Safety Committee requirements for human gene therapy protocols are detailed
below. Federal requirements (NIH and FDA) for these experiments are described in significant detail in
Appendix M of the NIH Guidelines for Research Involving Recombinant DNA, May, 1999, and in the
Code of Federal Regulations, 21 CFR, Part 312 (FDA Points to Consider).
MCG/MCGHI Standards which must be met:

MCG Institutional Review Board (IRB) x1-1478 (through the Office of Human Research Protection)
MCG Institutional Biological Safety Committee (IBC) x1-2663 (through Biosafety Representatives)
MCGHI (or other hospital/clinic) Epidemiology/Infection Control Office
Federal Registration and Approval:

NIH Office of Biotechnology Activities (301) 496-9838
http://www4.od.nih.gov/oba/apndxm.htm Appendix M (Human Gene Therapy)
FDA Center for Biologics Evaluation and Research

http://www.fda.gov/cber/ind/ind.htm 21 CFR Part 312
Note: Approval from the MCG IRB and IBC are required prior to submission to the the FDA. In
accordance with the update in the NIH Guidelines for Research Involving Recombinant DNA, Appendix M,
MCG’s IRB and IBC cannot approve a HGT protocol until it has been reviewed by the NIH Recombinant
DNA Advisory Committee. Upon notification from the NIH Office of Biotechnology Activities that the
RAC has completed its review, the IRB and IBC will be eligible to complete their respective reviews and
approve if warranted.
Appendix M-1 of the NIH Guidelines includes the submission requirements and addresses for both the NIH
Office of Biotechnology Activities and FDA Center for Biologics Evaluation and Research. A copy of the
21 CFR, Part 312 detailing the FDA IND Content and Format requirements can be downloaded directly
from the FDA web address listed above.
Application for Human Gene Transfer Clinical Trials at MCG

To initiate the review of a proposed human gene transfer clinical trial, please submit a description of your
protocol in the format described in Appendix M of the attached NIH Guidelines for Research involving
recombinant DNA, May, 1999. To obtain a copy of the NIH Guidelines, access the NIH OBA web site or
contact the Biosafety Office at x1-2663 or BIOSAFETY@mcg.edu. Send a copy of your completed
Appendix M to:
The MCG Institutional Biological Safety Committee

C/O Biosafety Office
Division of Environmental Health & Safety
1405 Goss Lane, CI-1006
Augusta, GA 30912-7521
Contact person: Biosafety Officer, 706-721-2663
Only complete protocols will be sent to Committee members for review. Specifically, we’ll need:
Scientific abstract
Non-technical abstract
Your responses to NIH Guidelines Appendix M-II through M-V

F-1
Your response to Adverse Event reporting requirements detailed in Appendix M-VII
A copy of your clinical protocol (your IND Submission)
A copy of the Investigator’s Brochure
A copy of the Informed Consent Document
Curricula vitae (2 pages) for each key professional in biographical sketch format
The proposed location for vector production and description of the Good Manufacturing
or Good Clinical Practices that will be utilized to prepare the vector
A copy of the Certificate of Analysis (CoA) for sterility for each lot of vector made at
MCG or sent to the University for this experiment
Additional responsibilities of the Principal Investigator conducting a rDNA experiment are detailed in
Section IV-B-7, Roles and Responsibilities of the NIH Guidelines. The full set of PI responsibilities can be
accessed at http://www4.od.nih.gov/oba/sect4.htm
Adverse Events
All adverse events must be reported in an annual data summary that is prepared for the Yale IRB and the
IBC, the FDA, the NIH Office of Biotechnology Activities, and your sponsor. Any Serious Adverse
Events (SAE’s) must be reported by telephone within 24 hours followed by a written report within 10
days. This report must be on file with the MCG IRB and IBC, the NIH OBA, the FDA, and NIH Office for
Protection from Research Risks if applicable within 15 days. Please note that SAE’s must be reported
whether related to the protocol or not. SAE’s shall not be designated as confidential, either in whole or
in part, and the SAE reports shall be stripped of patient identifiers, such as name, address, contact
information, social security numbers, and date of birth. If the SAE occurs after the trial and deemed related
to the HGT trial, it must be reported within 15 days of the date of determination.
The NIH OBA reporting form can be downloaded from their website at http://www.nih.gov/od/oba.
MCG Institutional Review Board (IRB)--Human Assurance Committee (HAC) or Chesapeake

Research Review, Inc (CCRI)
An MCG-authorized IRB must approve all experiments involving human subjects prior to initiation. Please
contact the Office of Human Research Protection (OHRP) office at x1-1478 for information on their
requirements.

F-2
Appendix G Sources of Contamination
If contamination is experienced in the laboratory, the following items may be sources of the contamination.
For additional assistance please contact the Biosafety Office at x1-2663 or BIOSAFETY@mcg.edu.
Personal items, such as coats, hats, storm boots or overshoes, umbrellas, purses, etc., do
not belong in the laboratory. These articles should be stored elsewhere.
Nonspecific contamination by environmental organisms from humans, animals,
equipment, containers for specimens or supplies, and outside air is a complication that
may affect or invalidate the results of an experiment. Human sources of this type of
contamination are evaluated as follows:
Sneezing, coughing and talking. Sneezing, variously reported to generate as many as
32,000 or 1,000,000 droplets below 100 microns in diameter; coughing, which produces
fewer and larger droplets; and talking, which has been reported to average only 250
droplets when speaking 100 words, show great differences between persons in regard to
the number of microorganisms aerosolized. As a general rule, it may be said that these
actions by normal healthy persons may play a less important role in transmission of
airborne infection to humans or experimental materials than does liberation of
microorganisms from human skin.
Dispersal of bacteria from human skin. There is a tremendous variation in the number of
bacteria shed from the skin by a clothed subject. For instance, in one study, the number
varied from 6,000 to 60,000 per minute. These bacteria were released on skin scales of a
size that could penetrate the coarse fabric used for the laboratory and surgical clothing in
the test. Dispersal of skin bacteria was several times greater from the area below the waist
than from upper parts of the body. Effective reduction is accomplished by use of closely-
woven or impervious clothing fitted tightly at the neck, wrists, and ankles to prevent the
clothing from acting as bellows that disperses air carrying skin scales laden with bacteria.
Such clothing sometimes is too warm to work in. The purpose of this summary is to alert
laboratory personnel to the existence of this source of contamination.
Prolific dispersal of bacteria occurs from infected abrasions, small pustules, boils, and
skin disease. Washing of lesions with germicidal soap will greatly decrease the number of
organisms on the skin and dispersal into the air. Healthy nasal carriers who generate
aerosolized staphylococci usually can be identified by the presence of heavy
contamination of their fingers, face, and hair. This point may be useful in investigating
the source of staphylococcal contamination of cell lines.
Footwear. In moderate and high-risk situations, shoes reserved only for laboratory use
have been recommended as a precaution against transporting spilled infectious agents
outside the laboratory. In experiments during which reduction of potential contamination
of experimental materials is important, laboratory-only shoes can also reduce the
microbial load brought into the laboratory each day by street shoes. Shoes are efficient
transporters. In one study, there were 4 to 850 times as many bacteria per square
centimeter on the laboratory footwear as on the floor itself.
Personal Work Practices

Food, candy, gum, and beverages for human consumption will be stored and consumed
only outside the laboratory.
Smoking is not permitted in the laboratory, MCG buildings or on MCG campus.
Shaving and brushing of teeth are not permitted in the laboratory. Razors, toothbrushes,
toiletry supplies and cosmetics are permissible only in clean areas, and should never be
used until after showering or thorough washing of the face and hands.
A beard may be undesirable in the laboratory in the presence of actual or potential
airborne contamination, because it retains particulate contamination more persistently
than clean-shaven skin. A clean-shaven face is essential to the adequate facial fit of a
facemask or respirator when the work requires respiratory protection.
Develop the habit of keeping hands away from mouth, nose, eyes, face and hair. This
may prevent self-inoculation.

G-1
For product protection, person with long hair should wear a suitable hair net or head
cover that can be decontaminated. This has long been a requirement in hospital operating
rooms and in facilities where biological pharmaceutical products are manufactured. A
head cover also will protect the hair from fluids, splashes, from swinging into Bunsen
burner flames and Petri dishes, as well as reduce facial contamination caused by habitual
repetitive manual adjustment of the hair.
Long flowing hair and loose flapping clothing are dangerous in the presence of open
flame or moving machinery. Rings and wristwatches also are a mechanical hazard during
operation of some types of machines.
Contact lenses do not provide eye protection. The capillary space between the contact
lenses and the cornea may trap any material present on the surface of the eye. Caustic
chemicals trapped in this space cannot be washed off the surface of the cornea. If the
material in the eye is painful or the contact lens is displaced, muscle spasms will make it
very difficult, if not impossible, to remove the lens. For this reason, contact lenses must
not be worn by persons exposed to caustic chemicals unless safety glasses with side
shields, goggles or full face shield are worn to provide full protection.
Plants, cut flowers, an aquarium, and pets of any kind are undesirable sources of yeast,
molds and other potential microbial contaminants of biological experimental materials.
Books and journals returnable to the institutional library should be used only in the clean
areas as much as possible.
When change rooms with showers are provided, the employer should furnish skin lotion.
When employees are subject to potential occupational infection, the shower and/ or
face/handwashing facilities should be provided with germicidal soap.
Personal cloth handkerchiefs should not be used in the laboratory. Disposable cleansing
tissues should be available for use instead.
Hand washing for personal protection:
• This should be done promptly after removing protective gloves. Tests show it is not
unusual for microbial or chemical contamination to be present despite use of gloves,
due to unrecognized small holes, abrasions, tears, or entry at the wrist.
• Throughout the day, at intervals dictated by the nature of the work, the hands should
be washed. Presence of a wristwatch discourages adequate washing of the wrist.
• Hands should be washed after removing soiled protective clothing, before leaving
the laboratory area, before eating and smoking. The provision of hand cream by the
employer encourages this practice.
• A disinfectant wash or dip may be desirable in some cases, but its use must not be
carried to the point of causing roughening, desiccation or sensitization of the skin.
Anyone with a fresh or healing cut, abrasion, or skin lesion should not work with
infectious materials unless the injured area is completely protected, such as with
waterproof bandages and double gloving.
Persons vaccinated for smallpox may shed vaccinia virus during the phase of cutaneous
reaction. Therefore, vaccination requires permission of the appropriate supervisor,
because two weeks absence may be necessary before returning to work with normal cell
cultures or with susceptible animals, such as the normal mouse colony.
Use of surgeon’s mask of gauze or filter paper is of little value for personal respiratory
protection. It is designed to prevent escape of droplets from the nose or mouth. If use of
biohazards demands respiratory protection, contact the Biosafety Office(x1-2663 or
BIOSAFETY@mcg.edu) for assistance.

G-2
Appendix J Biosafety Notice for Decontamination of Equipment
BIOSAFETY NOTICE
The exterior and interior of this piece of equipment were
decontaminated and are free of any Biological Hazards. This
notice does not apply to radiation or chemical hazards (if any).
This equipment is released for (please circle one):
Service/Repair Relocation Discard/Surplus
Decontamination performed by:
Chemcial or disinfection used:
Date of decontamination:
Location of equipment:
Laboratory/clinic telephone number:
Note: The following areas indicated below on this equipment may still be contaminated with biological
hazards and a biohazard warning label has been attached near the contaminated area(s):

J-1
Appendix K Shipping Labels
All of the shipping labels on the following pages should be printed clearly in full size on white paper
(except the Cargo Aircraft Only label, which should be printed on orange paper background). Any color in
the labels is required.

K-1
DANGEROUS GOODS IN EXCEPTED QUANTITIES
This package contains dangerous goods in excepted small quantities and is

in all respects in compliance with the applicable international and national
government regulations and the IATA Dangerous Goods Regulations
Signature of Shipper
Title Date
Name and address of Shipper
This package contains substance(s) in Class(es)

(check applicable box(es))
Class: 2 3 4 5 6 8 9
and the applicable UN Numbers are:

Appendix L Map of MCG Campus

L-1
BUILDING KEY
AA Kelly Building - Administration
AB Robert B. Greenblatt, M.D. Library
AC Protein Chemistry Lab
AD School of Dentistry
AE Pavilion I
AF Pavilion II
CHAFEE AG Environmental Services
FH AH Academic Computingh & Edu Dev
FF AVEN AI Research Support Building I
UE
OAK AVE. AJ Student Health Building
AK Classroom Services/Electronic Maintenance
FE
CENTRAL AVE
AL Medical Technology/Medical Records Offices
AR Telemedicine
AVENUE
GEORGIA BIO TECH FG
PAINE COLLEGE WAR PARK AS Banking Building
VETERANS AT Vending Building
VERD
TEXACO
FM AUGUSTA
NURSING BA Hospital & Clinics - Talmadge Building
.
POPE
BRACE INC.
HOME BB Hospital & Clinics - East Wing
ERY S
FT BLISSON BC Auditoria Center

FC
T.
INN INDEPENDANT BF Murphey Building

PAINE COLLEGE LIFE BG Dugas Building
15th STREET FI BH Central Energy Plant
BO BI Hospital & Clinics - Sydenstricker Building
WARREN HITES DR. BJ Chemical Storage Shed
DG BK Harper Street Deck
BN BL Laundry/Warehouse
DB BN MCG Hospital & Clinics Deck
AD PARKING BO Parking Information Center
DA
DECK BP Ambulatory/Specialized Care Center
DH BT Childrens Medical Center
BU Seating Gazebo
DF DE DD DC SPELLMAN AVENUE CK
CA Research Center Laboratory Building
BI
GOSS LANE AT BA
AS
GILBERT MANOR CP
FIRST UNION
CJ AE NATIONAL
PARKING BB
BU VETERANS BANK
DECK ADMINISTRATION
BP HOSPITAL
BC BL
LANEY - WALKER BOULEVARD

CA
AF
BH
McDONALD'S
CUSHING LANE
HB
CMC HT
HS
HARPER STREET
BT
BF BJ
CB AA HC
Legend
BG
PARKING WILENSKY
DECK
OLD BAILIE STREET S. OC

Campus Building
LANE
AB NATIONS
CL BANK
MCGHI Building CM
CH
CS BK
OLD BAILIE STREET N.
AC EA PARKING
NEW BAILIE STREET
CT AR DECK CB Sanders R & E Building
CH Physical Therapy Building
WAY
North CI
OSLER
AG EB HF CI Research Support Building II
PARKING CJ Pavilion III
DECK AH HP CK Student Study Center
South SouthTrust Bank
WALTON
CL Hamilton Wing of R & E Building
CN AI CM South Energy Plant
HH HM
HN CN Cancer Research Center Building
HK CP Goss Lane Deck
East AJ
AK EC
MULLINS
LAB CS Material Safety Storage Facility
CT Dent Boulevard Deck
AL DA Student Center
West ST. SEBASTIAN WAY
DB Residence IV
DC MCG Village Apartments A
EF
DD MCG Village Apartments B
DE MCG Village Apartments C
DF MCG Village Apartments D
EG
DG MCG Village Laundry
DH MCG Village Security Station
EA Telemedicine Center, Admin.
EB Jennings Building
RB EC Health Sciences Building
AL
0 1 2 3 4 500 FEET RA RA SP
IT EF Walter L. Shepeard Building
EG School of Nursing
DE
HO
FC Sports Medicine
NT
TY
FE Employee Assistance
SI
RC FF Sickle Cell
ER
BO
IV
FG Psychiatry Department
UN
RD UL FH Dermatology Offices
EV FI MCG Alumni Center
FM Residence VI
SCALE IN HUNDREDS OF FEET. AR FT Biological Safety
D HB Faculty Office Building
HC Parking Deck
HF Pediatric Building
. HH Community Medicine Offices
ST HK GA Radiation Therapy Center
LLY
HO HM Outpatient Hemodialysis/Respiratory
HN Respiratory Therapy II
CAMPUS PLAN
ST
HP Physical Plant Shop

EIN
HS MCG Annex
RE NO
ER
HT Annex Building II
RT OC Child Care Center
AV
PREPARED BY
H RA Physical Plant
E.
DIVISION OF FACILITIES PLANNING RB Physical Plant Grounds Maintenance

CLAY ST. RC Physical Plant, Grounds Storage
RD Physical Plant
RE Lansing B. Lee Sr. Warehouse
ACKNOWLEDGMENTS
This Biosafety Guide was compiled based on material from multiple resources, which not have been
appropriately attributed in the body of the Guide. Therefore, the MCG Division of EH&S wishes to
acknowledge these valuable resources, and thank those who developed them, but in particular:
• Yale University Biosafety Manual

http://www.yale.edu/oehs/Documents/Bio/Biosafety%20Manual.pdf
• Andy Glode and David Gillum, Universtity of New Hampshire

UNH Shipping of Biological Materials Manual (2006)
http://www.unh.edu/ehs/pdf/UNH-Shipping-Biological-Materials.pdf
UNH Guide to Shipping with Dry Ice (2007)
http://www.unh.edu/ehs/pdf/Guide-to-Shipping-with-Dry-Ice.pdf
• Stanford University Lab Animal Safety Data Sheets

http://www.stanford.edu/dept/EHS/prod/asds/
• Iowa State University Center for Food Security and Public Health
Disinfectant Tables
http://www.cfsph.iastate.edu/BRM/resources/Disinfectants/AntimicrobialSpectr
umDisinfectants1207.pdf
http://www.cfsph.iastate.edu/BRM/resources/Disinfectants/CharacteristicsSelect
edDisinfectants.pdf
• University of Virginia Policies

Policy on the Use of Human Cell Lines for Laboratory Personnel
https://vprgsecure.web.virginia.edu/bio/uva_human_cell_line_policy.cfm
Vaccinia Virus Use & Immunization Policy for Laboratory Personnel

https://vprgsecure.web.virginia.edu/bio/vaccinia.cfm
• Georgia State University Biosafety Manual

http://www.ovpr.gsu.edu/biosafety/files/GSU%20Biosafety%20Manual_April04.pdf
• The American Biological Safety Association (ABSA) Principles and

Practices Training Materials
Material in this Guide has also been adapted from information derived from the
following resources:
Governmental Resources:
• The CDC/NIH Biosafety in Microbiological and Biomedical Laboratories (BMBL), 5th
edition (2007) http://www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm
• The NIH Guidelines for Research with Recombinant DNA Materials (2002)
http://www4.od.nih.gov/oba/rac/guidelines/guidelines.html
• The NIH Stem Cell Information Resource
http://stemcells.nih.gov/

Acknowledgments
• OSHA Blood-borne Pathogens (29 CFR 1910.1030) Standards
http://www.osha.gov/pls/oshaweb/owadisp.show_document?p_table=STANDARDS&p_
id=10051
• OSHA Respiratory Protection (29 CFR 1910.134) Standards
http://www.osha.gov/pls/oshaweb/owadisp.show_document?p_id=12716&p_table=STA
NDARDS
• National Institutes of Occupational Safety and Health (NIOSH) Resources on Respirators
http://www.cdc.gov/niosh/npptl/topics/respirators/
• National Institutes of Occupational Safety and Health (NIOSH) Resourceses on Latex
Allergies: http://www.cdc.gov/Niosh/latexalt.html
• Environmental Protection Agency (EPA) Mold References: http://www.epa.gov/mold/
Other MCG Compliance and Service Offices:

• Office of Human Research Protection: http://www.mcg.edu/research/ohrp/
• Division of Laboratory Animal Services: http://www.mcg.edu/research/animal/
• Institutional Animal Care and Use Committee:
http://www.mcg.edu/research/animal/IACUCHomePage.htm
• Office of Institutional Audits and Compliance: http://www.mcg.edu/audits/
• Compliance Oversight Council: http://www.mcg.edu/audits/coc.htm
• Office of Technology Transfer and Economic Development:
http://www.mcg.edu/research/techtransfer/

Acknowledgments
INJURIES/EXPOSURE INCIDENT PROCEDURES INVOLVING
BIOLOGICAL MATERIALS
REPORT EXPOSURE IMMEDIATELY; YOU MAY NEED IMMEDIATE THERAPY!
Needlesticks/puncture wounds:
Wash the affected area with antiseptic soap and warm water for 15 minutes
Mucous membrane exposure:

Flush the affected area continuously for 15 minutes using an eyewash.
Injured Employee’s/Student’s Responsibilities

1. Health and Safety
9 Remove gloves and contaminated PPE
9 Wash exposed area thoroughly (15 minutes wash/flush)
2. Reporting/Notification
9 Notify Supervisor
9 Report incident to GA State Dept. of Administrative Services (DOAS), MCG
Human Resources Benefits Section and/or Public Safety (x1-2911) as per MCG
policies for:
MCG Employees :
MCG Policy & Procedure 1.4.17:
http://www.mcg.edu/policies/1417.html
Persons who are not MCG employees (including students):
MCG Policy & Procedure 3.0.03:
http://www.mcg.edu/policies/3003.html
Supervisor’s Responsibilities
1. Health and Safety
9 Ensure that the health and safety of injured person is top priority
9 Assist/encourage injured person to obtain proper medical attention
9 All injury/exposure/cleanup laboratory SOPs have been followed to protect others
from hazard
9 (Later) evaluate incident with injured person and Biosafety Office and make
appropriate changes in laboratory SOPs, if necessary, to avoid future similar
situations
2. Reporting/Notification
9 Notify the Biosafety Office (x1-2663 or BIOSAFETY@mcg.edu) and IBC (and
NIH and/or sponsor) of incident
9 Recording incident in the Biosafety Manual
9 Initiate and guide employee in completing proper reporting/notification to DOAS
and HR
Emergency Phone Numbers
Police and Fire, on campus x1-2911
MCG Employee Health Office (FG-1174) x1-3418
Student Health Office (AF-1040): x1-3487
Georgia State Department of Administrative Services (DOAS) 1-877-656-7475
MCG Human Resources (Annex I building, HS-1111) x1-3770
EHS Emergency Numbers:
8:00 a.m. to 5:00 p.m. Monday-Friday: x1-2663
Other hours: x1-2911 (for on-call service)

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Medical College of Georgia

Biological Safety Guide

1 Introduction ......................................................................................... 1.1

1.2 Responsibilities .................................................................................................................................1-2

2 Biosafety Requirements ...................................................................... 2-1

2.3 Laboratory Establishment, Close-outs and Moves ........................................................................2-8

2.4 Laboratory Record-keeping ............................................................................................................2-11

2.5 Paperwork Flow ..............................................................................................................................2-11

Medical College of Georgia Biosafety Guide-June 2008

2.6.4 Animals and the Use of Biological Materials in Animals ...........................................2-33

2.7 Experiments Prohibited at MCG ....................................................................................................2-34

3 Risk Assessment .................................................................................. 3-1

3.2 Considerations in a Risk Assessement ..............................................................................................3-3

Medical College of Georgia Biosafety Guide-June 2008

4.2 Laboratory Equipment ......................................................................................................................4-21

4.3 Laboratory Facilities .........................................................................................................................4-40

Medical College of Georgia Biosafety Guide-June 2008

5 Medical Surveillance Program ........................................................... 5-1

5.2 Immunizations ..................................................................................................................................5-3

5.3 Respiratory Protection Program .......................................................................................................5-7

5.4 Medical Restrictions.........................................................................................................................5-7

5.5 Minors in Laboratories, Clinics or Other Hazardous Areas .............................................................5-12

6 Accidents, Exposures, Spill Response ................................................ 6-1

6.2 Reporting Incident.............................................................................................................................6-1

6.3 Medical Assistance............................................................................................................................6-2

6.4 Spill Clean-up Procedures................................................................................................................6-3

6.5 Investigation of Laboratory Accidents .............................................................................................6-7

7 Decontamination of Biologicals .......................................................... 7-1

Medical College of Georgia Biosafety Guide-June 2008

7.2 Characteristics of Chemical Decontaminants....................................................................................7-4

7.3 Deactivation of Biological Toxins....................................................................................................7-10

7.4 Deactivation of Prions......................................................................................................................7-13

8 Waste Management ............................................................................ 8-1

9 Placard Signs and Labels ................................................................... 9-1

9.2 Labels and Color-Coding ..................................................................................................................9-2

9.3 Labeling Equipment Sent Out for Repair or Disposal.......................................................................9-2

10 Shipping .............................................................................................. 10-1

10.2 Shipping Overview..........................................................................................................................10-4

10.3 Shipment Type ...............................................................................................................................10-4

Medical College of Georgia Biosafety Guide-June 2008

10.4 Packaging Biological Material .......................................................................................................10-13

10.5 Shipper’s Declaration for Dangerous Goods..................................................................................10-21

10.7 International Shipments................................................................................................................10-29

10.8 Shipping Company Restrictions ...................................................................................................10-30

10.9 United States Postal Service Mailings..........................................................................................10-30

10.10 Transport as Checked Airline Baggage .........................................................................................10-31

10.11 Transport in Ground Vehicles ......................................................................................................10-32

Medical College of Georgia Biosafety Guide-June 2008

Medical College of Georgia Biosafety Guide-June 2008

AAV Adeno-associated Virus

Medical College of Georgia Biosafety Guide-June 2008

Medical College of Georgia Biosafety Guide-June 2008

 To protect against exposures of personnel (MCG employees and students, community

Medical College of Georgia Biosafety Guide-June 2008

Office of Environmental Health and Safety (EHS) Contacts (CI-1001)

Office of Environmental Health and Safety Main Office (CI-1001) x1-2663

LAS Main Office (CB-1901) x1-3421

Facility Management/Research Support & Compliance Office Telephone

Facilities Management x1-2434

Medical College of Georgia Biosafety Guide- June 2008

1.2 ROLES AND RESPONSIBILITIES

1.2.2 Principal Investigators, Clinical Directors and/or Instructional Course

Medical College of Georgia Biosafety Guide- June 2008

 Registering research work involving biological materials, particularly non-exempt recombinant

To protect against exposures of personnel (MCG employees and students, community

Registering research work involving biological materials, particularly non-exempt recombinant

Maintaining a liaison with the Biosafety Office. This includes:

If inexperienced in handling human pathogens, tissue culture, recombinant DNA and/or

Performing responsibilities assigned to them by the Principal Investigator, Clinical Director or

Periodic review of rDNA experiments documented in the Biosafety Protocols to ensure

Investigation of laboratory incidents, accidents, exposures, potential exposures, illnesses which

Coordinating/consulting with other institutional safety and compliance offices to maintain

Providing advice on laboratory security

Recommend to MCG Administration appropriate sanctions for noncompliance with Biological

monitor its progress in meeting those objectives and goals, and

Developing mechanisms for ensuring adherence to Biological Safety policies.

The biological agents to be transferred to MCG