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Safety in

Laboratory
and Hospital
dr. Tri Wibawa, PhD
Dept of Microbiology
FM UGM
Laboratory and Hospital Hazards

The 4 types of workplace hazards are:

• CHEMICAL HAZARDS
• PHYSICAL HAZARDS
• BIOLOGICAL HAZARDS
• ERGONOMIC HAZARDS
Physical Hazards
• Include:
– Equipment
– Machinery
– Electricity
– Fire
– Noise
– Heat
– Cold
Ergonomic Hazards
Ergonomics means adapting the workplace to
the worker, instead of trying to adapt the
worker to the workplace.
If the work station, tools and equipment,
physical environment including lighting,
temperature and noise are not properly
designed, undue stresses and strains can be
placed on the musculoskeletal system.
What is a Biohazard?
A potential hazard to humans, animals or the
environment caused by a biological organism,
or by material produced by such an organism

Examples;
• Viruses, bacteria, fungi, and parasites and their toxins.
• Blood and body fluids, as well as tissues from humans and
animals.
• Transformed cell lines and certain types of nucleic acids .

5
Why are we concerned about
biohazardous materials?
• Potential for acquiring a laboratory-associated
infection (LAI)
• Contamination of the environment
• Contamination of research
• Public perception*
Laboratory Associated Infections

Infection Source Susceptible Host


• Cultures and stocks • Immune system
• Research animals • Vaccination status
• Specimens • Age
• Items contaminated with
above

Route of Transmission
• Percutaneous inoculation
• Inhalation of aerosols
• Contact of mucous membranes
• Ingestion
LAI’s
• Only 20% causative or defined event
• 80% of which are caused by human error
• 20% are caused by equipment failure

• Top 4 accidents resulting in infection


1. Spillages & splashes
2. Needle and syringe
3. Sharp object, broken glass
4. Bite or scratch from animals or ectoparasites

http://www.weizmann.ac.il/safety/bio2.html
LAI’s
WHO WHAT WHERE WHEN HOW
Researcher SARS Taiwan December 2003

Microbiologist West Nile Virus United States August 2002 Laceration

Laboratory Meningococcal United States 2000 Aerosol?


Worker Disease
Laboratory Vaccinia virus Europe 2002 Contact
Worker
Bloodborne Pathogens (BBP)
• Sources
– Blood
– Semen
– Vaginal Secretions
– Body Fluids
• Cerebrospinal
• Amniotic
• Synovial
– Tissue Cultures
– Organ Cultures
– Infected Experimental Animals
Risk of Exposure

• Pathogen involved
• Type of body fluid
• Route of exposure
• Duration of exposure
• Volume of blood involved in exposure
• Concentration of pathogens at time of
exposure
• PPE worn
Specific Examples of
Bloodborne Pathogens

Hepatitis B

Hepatitis C

HIV
Issues to Consider
• Symptoms
• Mode of transmission
• Incubation period
• Survival outside host
• Communicability
• Immunization
• Prophylaxis / Treatment
If An Exposure Occurs
(or the possibility of exposure)

1. Initiate first aid


2. Notify your supervisor / designated
person
3. Report to hospital emergency
department or University’s Health
Services
Universal Precautions
• Minimum standard of practice for preventing the
transmission of BBP
Includes: - Education
- Hand washing
- Wearing protective barriers
- Use safe work practices

If samples cannot be guaranteed non-


infective …… treat as infectious!
Classification of Biohazards
• Conventional Agents
• Unconventional Agents
• Recombinant DNA
• Tissue Culture
• Animal Work
• Anatomical Specimens
Classification of Biohazards
As the level  so does:
BSL 4
 The risk of the organism to humans,
animals, plants and/or the environment
BSL 3
 The procedural and facility requirements

 The level of containment required


BSL 2
 The degree of protection for personnel,
the environment and the community.
BSL 1
Conventional Agents
Risk Individual Community Containment Examples
Group Risk Risk

Low Low Level 1 E.coli, bacillus subtilis


1
Unlikely to cause disease in
healthy workers or animals

Moderate Limited Level 2 e.coli 0157:H7, several


2 streptococcus & salmonella species,
Rarely cause serious human adenoviruses, Hepatitis A, B & C,
or animal disease Influenza, Measles

High Low Level 3 Bacillus anthracis, mycobacterium


3 tuberculosis, Hantaan virus, HIV,
Yellow fever virus
May cause serious disease

High High Level 4 Lassa virus, Ebola virus, Marburg


4 Likely to cause very serious
virus
disease
Unconventional Pathogens
• Transmissible Spongiform Encephalopathy
(TSE) prion diseases; lethal transmissible
neurodegenerative conditions
• Creutzfeld-Jakob disease, Variant C-J Disease, Mad
Cow Disease, Scrapie, Chronic Wasting Disease.

• Resistant to destruction by procedures that


normally inactivate viruses.
Recombinant DNA
• Genetic Engineering = in
vitro incorporation of
genetic material from one
cell into another

• BSL was assign according


to it components
Tissue Culture
• Have the potential to contain pathogenic organisms
• In general;

Human & non-human primate, Level 2


and mycoplasma-containing cell
lines
Others Level 1

A detailed risk assessment should be undertaken


when using a new cell line.
Animal Work
• Animals can harbour infectious organisms
(naturally or introduced)
• Level dependent on type of work being conducted.
• Special Animal Care training is required for all
personnel working with animals.
• All work involving animal use must receive prior
approval from the Animal Care Committee
• Potential Hazards:
– Zoonosis
– Allergen
– Scratch/bite
Anatomical Specimens
• All specimens should be considered
infectious due to potential presence
of infectious agents
• Important to consider the type of
specimen
• blood, organs, tissues
• Spinal sample, brain tissue
• From infectious patient
• In general Level 2 but it depends on
the nature of the work.
Infection/Biohazard Control
Engineering Controls

Administrative Controls

Practices and Procedures

Personal Protective Equipment


Practices and Procedures
• General Safety Guidelines
• Good Microbiological Practice
• Handwashing
• Specific Procedures
• Centrifuges
• Needles & Syringes and other sharps
• Pipettes
• Blenders, Grinders, Sonicators & Lyophilizers
• Inoculation Loops
• Cryostats
General Laboratory Safety
Guidelines
• Mostly common sense, but you must understand the
hazards you face in the laboratory and be adequately
trained to deal with them.
• Basic must knows for all labs.
• Protect yourself, others and the environment and
indirectly the product.

b i o s a f e t y
Good Microbiological Practice
(GMP)

• Basic code of practice that should be applied to


all types of work involving microorganisms
• Objectives
• prevent contamination of laboratory workers and the
environment
• prevent contamination of the experiment/samples
• Application of aseptic technique, minimization
of aerosols, contamination control, personal
protection, emergency response
Handwashing
• One of the single effective means of preventing
infections
• IF done properly and frequently
• When to wash?
• Before starting any manipulations
• Before leaving the lab
• When hands are obviously soiled
• Before and after completing any task in a BSC
• Every time gloves are removed
• Before contact with one’s face or mouth
• At the end of the day
Pipettes
• Mouth pipetting is prohibited.
• All biohazardous materials should be pipetted in
BSC’s.
• Never force fluids out, use ‘to deliver’ pipettes.
• To avoid splashes, allow discharge to run down
dispense the receiving container wall.
• Never mix material by suction and expulsion.
• Reusable pipettes should be placed horizontally in a
disinfectant filled pan.
Inoculation Loops
• Sterilization in an open flame may create aerosols
which may contain viable microorganisms.
• Shorter handles minimize vibrations.
• Disposable plastic loops are good alternatives.
Personal Protective Equipment
• PPE can become an
important line of
defence (last line of
defense).
• Responsibility of
both the user and the
supervisor to ensure
that PPE is worn
PPE
• Criteria for consideration
• Routes of exposure that need to be blocked
• Degree of protection offered
• Ease of use

• Only effective if correctly selected, fitted, used and


cared for, and the individual is trained
• Ensure PPE is removed before leaving the lab.
PPE
Footwear
• Closed toed shoes should always be worn. Booties
are worn in some higher containment labs and
animal facilities.

Lab Coats/Gowns
• Long-sleeved, knee length with snaps
• Elastic cuffs *
• Back-closing gowns *
• Periodic cleaning required
PPE
Gloves
• Latex, nitrile & vinyl for work with biological agents
• Exam gloves should not be reused, change frequently.
Utility gloves can be disinfected and reused if they
show no sign of degradation.
• Consider tensile characteristics, length of cuff
• Double gloving
• ORM can provide assistance finding an alternative for
people with allergies.
• Gloves are not to be worn in public places
PPE
Eye & Face Production
• Goggles, safety glasses to protect the eyes
• Full face shield to protect facial skin.

Respirators
• Only personnel who have been fit-tested and trained
should wear respirators.
Spills
• Spill response will vary depending on:
• What was spilled?
• How much was spilled?
• Where was the spill?
• What is the potential for release to the environment?
• Spills should be cleaned up immediately (unless an
aerosol was generated), to ensure proper
decontamination.
• Ensure appropriate PPE is worn and clean-up
equipment is readily available.
Spills – General Clean-up
• Cover spill area with absorbent material
• Soak the spill area with an appropriate disinfectant (i.e. 10%
bleach)
• Pour disinfectant from the outside surface of the absorbent material
towards the inside
• Ensure any broken glass is picked up (with forceps!) and placed in
a sharps container
• Leave on for 20 to 30 minutes
• Wipe up with absorbent material
• Waste should be disposed in appropriate biohazard bags and where
possible autoclaved
Decontamination, Disinfection and
Sterilization
• Decontamination: Free of contamination, the
destruction of microorganisms to a lower level such
that it removes danger of infection to individuals.

• Sterilization: The complete destruction of all viable


microorganisms.

• Disinfection: Use of agents (physical or chemical) to


destroy harmful organisms on inanimate objects (not
necessarily all organisms)
Decontamination: Physical

• Heat:
• Autoclaving (most practical and recommended)
• Incineration (for disposal of sharps and tissues)
• Irradiation:
• UV light (wavelength of 253 nm is germicidal)
• Gamma (disrupts DNA and RNA)
• Filtration
• HEPA (biological safety cabinets, ventilation)
Hazardous Chemicals
• Can enter the body in several ways:
– Inhalation
– Ingestion
– Absorbed through the skin
Main Issues
• Access (Security) • Exceeding storage
– Drugs, select agents, limits
terrorism potential – Time
• Adequate warning • Safety
• Efficacy
– Signs
– Quantity
– Labels
• Fire Code
– Abbreviation lists – Building limits
• Incompatible • Homeland Security
– Address limits
chemicals mixing
• Building Design Limits
– Earthquakes – Green Buildings
Chemical Hazard Classes
• Corrosives
• Flammables
• Oxidizers
• Toxins
• Reactive Chemicals
Corrosives
• Acids
– Strong acids give up protons (accepts electron pairs)
• Bases (Alkalis, Caustics)
– Strong bases accept protons (donate an electron pair)
• Storage Segregation
Acid Type Examples
• Inorganic:
– hydrochloric acid • Organic:
– nitric acid – lactic acid
– phosphoric acid – acetic acid
– sulfuric acid – formic acid
– boric acid – citric acid
– hydrofluoric acid – oxalic acid
– hydrobromic acid • Oxidizing:
– nitric acid
– perchloric acid
– chromic acid
Flammables
• NFPA Class IA, IB, IC
Oxidize
4Fe + 3O2  2[Fe2O3]
Fe0  Fe+3 (O0  O-2)
1. combine with oxygen
2. dehydrogenate esp by action of oxygen
3. change (an element or ion) from a lower
to a higher positive valence : remove one
or more electrons
4. coat with oxide; make into an oxide
Oxidizing Polyatomic Ions
Chemical Group Chemical Formula
peroxides O2-2
nitrates NO3-
nitrites NO2-
perchlorates ClO4-
chlorates ClO3-
chlorites ClO2-
hypochlorites ClO-
dichromates Cr2O7-2
permanganates MnO4-
persulfates S2O8-2
Oxidizers (Class 1, 2, 3, 4)

Least  Most Reactive


Class 4
 An oxidizing material that can undergo an explosive reaction when
catalyzed or exposed to heat, shock or friction
Class 3
 An oxidizing material that will cause a severe increase in the burning
rate of combustible material which it contacts or will undergo
vigorous self-sustained decomposition when catalyzed or heat
Class 2
 An oxidizing material that will moderately increase the burning rate
or which may cause spontaneous ignition of combustible material
which it contacts
Class 1
 An oxidizing material whose primary hazard is that it may increase the
burning rate of combustible material with which it comes in contact
Oxidizer examples (Classified by NFPA)
• Class 4
– ammonium perchlorate NH4+

– ammonium permanganate

– guanidine nitrate H2O2


– hydrogen peroxide (>91% conc.)

– perchloric acid (>72.5%)


KO2

– potassium superoxide
Oxidizer Examples (continued)
• Class 3
– ammonium dichromate
– potassium chlorate
– hydrogen peroxide (52-91% conc.) H2O2
– potassium dichloroisocyanurate K+
– calcium hypochlorite (>50% wgt.)
– sodium chlorate
– perchloric acid (60-72.5% conc.)
– sodium chlorite (>40% wgt.)
– potassium bromate
– sodium dichloro-s-triazinetrione
Example
• Linseed oil (flax seed oil)

• Class 2 oxidizer  spontaneous ignition of


combustible material which it contacts
Toxins
• Acute & Chronic Poisons; Highly Toxic
• Carcinogens
– Select Carcinogens – Occupational (31 substances -
www.dir.ca.gov/Title8/sb7g16a110.html)
– Prop 65 (845 items - www.oehha.org/prop65/prop65_list/Newlist.html)
– National Toxicology Program, Report on Carcinogens (245 -
http://ntp.niehs.nih.gov/go/roc)
– International Agency for Research on Cancer
(http://monographs.iarc.fr/ENG/Classification/index.php )
• Group 1: Carcinogenic to humans (108 agents)
• Group 2A: Probably carcinogenic to humans (63 agents)

• Reproductive & Developmental Toxins


– www.cdc.gov/niosh/topics/repro

• Neurotoxins, organ specific toxins, Irritants


Reactive Chemicals
Purchase small amounts, use up rapidly

• Water Reactive
– Store in an isolated area within the lab, in a water-
tight cabinet or secondary container
• Pyrophoric
– Store in air-tight containers without oxygen or
moisture (sure seal bottles, glove boxes)
– Separate from flammables, but in flame resistant
container when practical
• Self Decomposing
– Check frequently
Water Reactive Substances
Dangerous When Wet
Reactions
Reactions results
• Alkali metals (Na, K, Li)
• flammable gas release
+ water  detonation,
• strong oxidizing gas release heat, formation of
• toxic gas release hydroxide, hydrogen gas
• metal oxide fume release • Alkaline earths (Mg, Be,
• corrosive acids formation Ca, Ba) + water 
detonation, hydrogen gas
Dangerous When Wet Reactions (continued)

• Hydrides (LiH, LiAlH4)


+ water  hydrogen,
• Metallic (inorganic, such
caustic solution
as potassium) peroxides +
• Carbides of Al, Ca, Mg + water  oxygen gas
water  acetylene and heat
liberated
• Chlorides of group III
• Phosphides + water  metals, transition
phosphine gas metals, non-metals (Al,
• Nitrides + water  Ti, S) + water 
ammonia, caustic solution hydrogen chloride gas
Pyrophoric
• Can spontaneously ignite in air
• Derived from Greek for “fire-bearing”
• Includes organometallic reagents:
– Alkyllithiums
– Alkylzincs
– alkylmagnesiums (Grignards)
– and some finely divided metal powders
• Specific ex.
– Diborane
– Diethylzinc
– tert-butyllithium
– diphosphine
Decomposition by Fission & Fusion

+ O2

NO2 + CO2 +H2O + …


Before Leaving the Lab
• Turn off
 Gas
 Water
 Power lines
 Other non necessary
equipment
• Clean your work area
• Return ALL chemicals to
storage
• Lower fume hood sash
• Wash your hands

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