BIOSAFETY TECHNIQUES

Biosafety is discipline that addresses the safe handling and containment of infectious
microorganisms to protect workers, the public, agriculture, and the environment from exposure
to biological agents or materials that may cause disease or other detrimental effects in humans,
plants or animals. It is the prevention of loss of biological integrity, focusing both on ecology and
human health. These prevention mechanisms include conduction of regular reviews of the biosafety in
laboratory settings, as well as strict guidelines to follow. Many laboratories handling biohazards employ
an ongoing risk management assessment and enforcement process for biosafety. Failures to follow such
protocols can lead to increased risk of exposure to biohazards or pathogens. Human error and poor
technique contribute to unnecessary exposure and compromise the best safeguards set into place for
protection.

The concept of biocontainment is related to laboratory biosafety and pertains to microbiology

laboratories in which the physical containment of highly pathogenic organisms or agents
(bacteria, viruses and toxins) is required, usually by isolation in environmentally and biologically
secure cabinets or rooms, to prevent accidental infection of workers or release into the
surrounding community during scientific research.

Containment types

Primary containment is the first container in direct contact with biohazardous material as well as
protection of personnel and the immediate laboratory environment from exposure to infectious
agents. Primary containment requires using proper storage containers, good microbiological
techniques, and the use of appropriate safety equipment such as biological safety cabinets.

Secondary containment is the protection of the environment external to the laboratory from
exposure to infectious materials and is provided by a combination of facility design and
operational practices.

Biological safety cabinets (BSC), are fairly common devices designed to provide effective
primary biocontainment in laboratories working with highly infectious agents. Three general
levels and types have been devised (Class I, Class II, and Class III).

Biosafety suites are suites of laboratory rooms which are essentially equivalent to large Class III
cabinets in which positive pressure personnel suits ("space suits") serve as the "outside"
environment for workers. Examples include the biosafety suites at USAMRIID at Fort Detrick,
Maryland, USA and the Maximum Containment Facility (MCF) of the CDC in Atlanta, Georgia,
USA.

Agricultural containment: The term “biocontainment” is used differently in facilities for the
study of human pathogens versus those used for the study of agricultural pathogens. In
agricultural facilities, the definition for “biocontainment” resembles that for “biosafety,” i.e., the
safety practices and procedures used to prevent unintended infection of plants or animals or the
release of high-consequence pathogenic agents into the environment (air, soil, or water). In the
agricultural setting, worker protection and public health are always considerations; however,

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emphasis is placed on reducing the risk that agents under study could escape into the
environment.

Biosafety levels

A biosafety level (BSL) is a set of biocontainment precautions required to isolate dangerous

biological agents in an enclosed laboratory facility. They are individual safeguards designed to
protect laboratory personnel, as well as the surrounding environment and community.

These levels, which are ranked from one to four, are selected based on the agents or organisms
that are being researched or worked on in any given laboratory setting. The levels of containment
range from the lowest biosafety level 1 (BSL-1) to the highest at level 4 (BSL-4). At the lowest
level of biosafety, precautions may consist of regular hand-washing and minimal protective
equipment. At higher biosafety levels, precautions may include airflow systems, multiple
containment rooms, sealed containers, positive pressure personnel suits, established protocols for
all procedures, extensive personnel training, and high levels of security to control access to the
facility.

The Centers for Disease Control and Prevention (CDC) sets BSL lab levels as a way of
exhibiting specific controls for the containment of microbes and biological agents. Each BSL lab
level builds upon on the previous level—thereby creating layer upon layer of constraints and
barriers. These lab levels are determined by the following:

 Risks related to containment

 Severity of infection
 Transmissibility
 Nature of the work conducted
 Origin of the microbe
 Agent in question
 Route of exposure

The reason biosafety levels are so important is because they dictate the type of work practices
that are allowed to take place in a lab setting. They also heavily influence the overall design of
the facility in question, as well as the type of specialized safety equipment used within it. The
following are explanations of each biosafety level.

BSL–1

As the lowest of the four, biosafety level 1 applies to laboratory settings in which personnel work
with low-risk microbes that pose little to no threat of infection in healthy adults. This level of
biosafety is appropriate for work with several kinds of microorganisms including non-pathogenic
Escherichia coli, Bacillus subtilis, Saccharomyces cerevisiae and other organisms not suspected
to contribute to human disease.

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This laboratory setting typically consists of research taking place on benches without the use of
special contaminant equipment. A BSL-1 lab, which is not required to be isolated from
surrounding facilities, houses activities that require only standard microbial practices, such as:

 Mechanical pipetting only (no mouth pipetting allowed)

 Safe sharps handling
 Avoidance of splashes or aerosols
 Daily decontamination of all work surfaces when work is complete
 Hand washing
 Prohibition of food, drink and smoking materials in lab setting
 Personal protective equipment, such as; eye protection, gloves and a lab coat or gown
 Biohazard signs

BSL-1 labs also require immediate decontamination after spills. Infection materials are also
decontaminated prior to disposal, generally through the use of an autoclave. BSL-1 laboratories
must have a door which can be locked to limit access to the lab, however it is not necessary for
BSL-1 labs to be isolated from the general building.

BSL–2

This biosafety level covers laboratories that work with agents associated with human diseases
(i.e. pathogenic or infections organisms) that pose a moderate health hazard. Examples of agents
typically worked with in a BSL-2 include Hepatitis A, B, and C viruses, human
immunodeficiency virus (HIV), pathogenic Escherichia coli, Staphylococcus aureus,
Salmonella, Plasmodium falciparum, and Toxoplasma gondii.

BSL-2 laboratories maintain the same standard microbial practices as BSL-1 labs, but also
includes enhanced measures due to the potential risk of the mentioned microbes. Personnel
working in BSL-2 labs are expected to take even greater care to prevent injuries such as cuts and
other pricks of the skin, as well as ingestion and mucous membrane exposures.

In addition to BSL 1 expectation, the following practices are required in a BSL 2 lab setting:

 Appropriate personal protective equipment (PPE) must be worn, including lab coats and
gloves. Eye protection and face shields can also be worn, as needed.
 All procedures that can cause infection from aerosols or splashes are performed within a
biological safety cabinet (BSC).
 An autoclave or an alternative method of decontamination is available for proper
disposals.
 The laboratory has self-closing, lockable doors.
 A sink and eyewash station should be readily available.
 Biohazard warning signs
 Laboratory personnel have specific training in handling pathogenic agents and are
directed by scientists with advanced training.
 Access to the laboratory is limited when work is being conducted.
 Extreme precautions are taken with contaminated sharp items.

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Access to a BSL-2 lab is far more restrictive than a BSL-1 lab. Outside personnel, or those with
an increased risk of contamination, are often restricted from entering when work is being
conducted.

BSL-3

Again building upon the two prior biosafety levels, a BSL-3 laboratory typically includes work
on microbes that are either indigenous or exotic, and can cause serious or potentially lethal
disease through inhalation. Biosafety level 3 is commonly used for research and diagnostic work
involving various microbes which can be transmitted by aerosols and/or cause severe disease.
These include Francisella tularensis, Mycobacterium tuberculosis, Chlamydia psittaci,
Venezuelan equine encephalitis virus, Eastern equine encephalitis virus, SARS coronavirus,
Coxiella burnetii, Rift Valley fever virus, Rickettsia rickettsii, several species of Brucella,
chikungunya, yellow fever virus, and West Nile virus.

The microbes are so serious that the work is often strictly controlled and registered with the
appropriate government agencies. Laboratory personnel are also under medical surveillance and
could receive immunizations for microbes they work with. This type of work can be done in
clinical, diagnostic, teaching, research, or production facilities.

Common requirements in a BSL-3 laboratory include:

 Standard personal protective equipment must be worn, and respirators might be required
This cannot be worn outside of the laboratory and must be discarded or decontaminated
after each use.
 Solid-front wraparound gowns, scrub suits or coveralls are often required
 All work with microbes must be performed within an appropriate BSC
 Access hands-free sink and eyewash are available near the exit
 Sustained directional airflow to draw air into the laboratory from clean areas towards
potentially contaminated areas (Exhaust air cannot be re-circulated)
 A self closing set of locking doors with access away from general building corridors
 A laboratory-specific biosafety manual must be drafted which details how the laboratory
will operate in compliance with all safety requirements.

Access to a BSL-3 laboratory is restricted and controlled at all times.

In addition, the facility which houses the BSL-3 laboratory must have certain features to ensure
appropriate containment. The entrance to the laboratory must be separated from areas of the
building with unrestricted traffic flow. Additionally, the laboratory must be behind two sets of
self-closing doors (to reduce the risk of aerosols escaping).The construction of the laboratory is
such that it can be easily cleaned. Carpets are not permitted, and any seams in the floors, walls,
and ceilings are sealed to allow for easy cleaning and decontamination. Additionally, windows
must be sealed, and a ventilation system installed which forces air to flow from the "clean" areas

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of the lab to the areas where infectious agents are handled. Air from the laboratory must be
filtered before it can be recirculated.

BSL-4

Biosafety level 4 (BSL-4) is the highest level of biosafety precautions, and is appropriate for
work with agents that could easily be aerosol-transmitted within the laboratory and cause severe
to fatal disease in humans for which there are no available vaccines or treatments. BSL-4 labs are
rare. However some do exist in a small number of places in the US and around the world. As the
highest level of biological safety, a BSL-4 lab consists of work with highly dangerous and exotic
microbes. Infections caused by these types of microbes are frequently fatal, and come without
treatment or vaccines. Two examples of such microbes include Ebola and Marburg viruses.

In addition to BSL-3 considerations, BSL-4 laboratories have the following containment

requirements:

 Personnel are required to change clothing before entering, shower upon exiting
 Decontamination of all materials before exiting
 Personnel must wear appropriate personal protective equipment from prior BSL levels, as
well as a full body, air-supplied, positive pressure suit
 A Class III biological safety cabinet

A BSL-4 laboratory is extremely isolated—often located in a separate building or in an isolated

and restricted zone of the building. The laboratory also features a dedicated supply and exhaust
air, as well as vacuum lines and decontamination systems. BSL-4 laboratories are generally set
up to be either cabinet laboratories or protective-suit laboratories. In cabinet laboratories, all
work must be done within a class III biosafety cabinet. Materials leaving the cabinet must be
decontaminated by passing through an autoclave or a tank of disinfectant. The cabinets
themselves are required to have seamless edges to allow for easy cleaning. Additionally the
cabinet and all materials within must be free of sharp edges in order to reduce the risk of damage
to the gloves. In a protective-suit laboratory, all work must be done in a class II biosafety cabinet
by personnel wearing a positive pressure suit. In order to exit the BSL-4 laboratory, personnel
must pass through a chemical shower for decontamination, then a room for removing the
positive-pressure suit, followed by a personal shower. Entry into the BSL-4 laboratory is
restricted to trained and authorized individuals, and all persons entering and exiting the
laboratory must be recorded.

As with BSL-3 laboratories, BSL-4 laboratories must be separated from areas that receive
unrestricted traffic. Additionally airflow is tightly controlled to ensure that air always flows from
"clean" areas of the lab to areas where work with infectious agents is being performed. The
entrance to the BSL-4 lab must also employ airlocks to minimize the possibility that aerosols
from the lab could be removed from the lab. All laboratory waste, including filtered air, water,
and trash must also be decontaminated before it can leave the facility.Biosafety-level-4
laboratories are used for diagnostic work and research on easily transmitted pathogens which can
cause fatal disease. These include a number of viruses known to cause viral hemorrhagic fever
such as Marburg virus, Ebola virus, Lassa virus, Crimean-Congo hemorrhagic fever. Other

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pathogens handled at BSL-4 include Hendra virus, Nipah virus, and some Flaviviruses.
Additionally, poorly characterized pathogens which appear closely related to dangerous
pathogens are often handled at this level until sufficient data are obtained either to confirm
continued work at this level, or to work with them at a lower level. This level is also used for
work with Variola virus, the causative agent of smallpox, though this work can only be done at
the World Health Organization approved facilities at the Centers for Disease Control and
Prevention in Atlanta, U.S.A., as well as the State Research Center of Virology and
Biotechnology in Koltsovo, Russia.

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Basic Biological Safety – Principles and Practices

Practices and Procedures – The Second Element of BL Selection

The nature of the agent is the starting point for BL selection. Procedures to be carried out could
be considered the final selection criteria. There are a number of procedures that can increase the
risk for personnel as well as environmental contamination potential. These include:

 Work with large quantities of samples

 Work with an agent involving live animals
 Work with concentrated cultures
 Work with large single volumes (i.e. greater than 1 liter) of biologically active materials.

Under these conditions, an increase of the biosafety level assignment or enhancement of

practices and techniques may be warranted. These decisions should be made by the primary
investigator before work involving infectious agents is undertaken. This process is part of the
Biological Risk Assessment.

Biosafety Level 1 – Good Microbiological Practices

All lab operations involving viable biological agents must be conducted at BL1. Most campus
laboratories are BL1.

Facilities features are minimal at this level but do include:

 Doors for access control

 A sink for hand washing
 Bench tops that are impervious so they can be disinfected
 Study lab furniture
 Spaces accessible for cleaning
 Fly screens if the lab has window that open

The core of BL1 and the foundation of all biosafety levels are Standard Microbiological
Practices. The following are the standard microbiological practices for BSL.

BL1 – Standard Microbiological Practices

1. Access to the lab is limited or restricted at the discretion of the lab director when
experiments to work with cultures and specimens are in progress. This practice can
ensure that distractions are kept to a minimum.
2. Persons wash their hands after handling viable materials, after removing gloves, and
before leaving the lab. This practice is not only necessary for good hygiene; it will also
minimize any potential for contamination to remove from one lab to another.
3. Eating, drinking, smoking, handling or applying cosmetics, and storing food for human
use are not permitted in the work area. Food is stored outside of the work area in an area
designated for that purpose. All hand creams, lip balms and other lotions should not be

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 stored or applied in the open lab environment where container surfaces can become
contaminated by activities in the lab.
4. Mouth pipetting is prohibited. Use mechanical pipetting devices. Mouth pipetting
presents an opportunity for accidental ingestions and should not be performed.
5. Policies for safe handling of sharps are implemented. Safe sharps procedures are
discussed later in this module.
6. All procedures are performed carefully to minimize the creation of aerosols or splashes.
Perform your procedures in the safest, cleanest manner you can. Use only the quantity of
material you need and do dry runs of new procedures to refine and learn the process
before you begin work with viable materials, etc.
7. Work surfaces are decontaminated at least once a day and after any spill of viable
material. The basic procedures for proper cleaning and disinfection are also presented
later in this module.
8. All cultures, stocks and other regulated wastes are decontaminated by an approved
decontamination method.

Biosafety Level 1: Primary Barriers

There are no special containment devices or equipment like biosafety cabinets required at this
level. The following recommendations are made regarding personnel protective equipment:

 Lab coats are recommended to prevent contamination of street clothes.

 Fluid-resistant gloves are recommended, especially if you have broken skin or rashes on
your hands. Nitrile and/or vinyl gloves should be available for use in the lab.

Protective eyewear should be worn when conducting procedures that involve manipulation of
viable biological materials or other hazardous materials. Minimally, safety glasses should be
worn. Where procedures are likely to generate a splash, splash goggles should be worn.

Biosafety Level 2 Laboratories

Biosafety level 2 work requires attention to aerosol minimization, limiting handling of viable
biological materials, and safe sharps procedures. Therefore, facilities where BL2 work is to be
carried out must meet the following requirements (in addition to the BL1 Facility requirements):

 Doors are lockable

 Lab is easily cleaned and no carpets are rugs are permitted
 Chairs and other lab furniture used in lab are covered with non-fabric materials that can
be easily decontaminated.
 Biosafety cabinets are properly installed and located.
 An eyewash station is readily available
 Lab illumination is adequate for all activities, avoiding reflections and glare that could
impede vision.

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  While there are no specific ventilation requirements, new facilities should be designed so
that the ventilation systems provide an inward flow of air without recirculation to spaces
outside the lab.

The standard microbiological practices are the basis of biosafety containment practices at all
levels. At BL2, the following additional special practices are required.

 Regarding restricted access, persons who are at increased risk for acquiring infection or
for whom infection may have serious consequences are not permitted in the lab.
 Policies and procedures are in place to ensure that only personnel who have been advised
of the biohazard present and meet the vaccination requirements (if applicable) are
permitted in the lab.
 A biohazard sign must be posted on the entrance to the lab when etiologic agents are in
use. The sign must include the biosafety level, the agents in use, the primary
investigator’s name and contact numbers, required PPE and exit procedures. The
Biosafety program will provide signs for BL2 labs to ensure compliance with current
requirements.
 When appropriate, considering the agents handled, baseline serum samples for lab or
other at-risk personnel are collected and stored. Serum banking is not generally practiced
at UWO.
 Biosafety procedures are incorporated into standard operating procedures or in a
biosafety manual adopted or prepared specifically for the lab. Personnel are advised or
special hazards and are required to read and follow instructions on practices and
procedures.

Use of Sharps in BSL2 Labs

Adequate precautions must be always taken with any contaminated sharp items. Sharps handling
is discussed later in this module, but at BL2 some specific practices are emphasized and should
be followed. These include:

 Sharps use restricted in the lab and used only when there is no alternative.
 Glassware should be replaced with plastic-ware whenever possible.
 Needle-locking or disposable syringe-needle units are used for injection or aspiration of
infectious materials.
 Non-disposable sharps are placed in hard-walled containers for reprocessing.
 Broken glassware is not handled directly by hand. Use a broom and dustpan to pick up
broken pieces.
 Use sharp devices with safety features.

“Safer Sharps”

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A “safer sharp” is one that has a safety mechanism built into the design that allows the use to
enclose or retract the sharp end without recapping or otherwise manipulating the device.
Examples include:

 Disposable scalpels with shields

 Needles that are equipped with a shield
 Blood collection systems that retract the needle with a spring

History shows that the bulk of sharps accidents occur from recapping or other actions involved
with the disposal process. The design of a safer sharp is to minimize that exposure risk. OSHA
regulations now require that “safer sharps” be used in human health care and clinical lab settings.

BLS 2 – Standard Microbiological Practices

 Cultures, tissues, specimens of body fluids, or potentially infectious wastes are placed in
a container with a cover that prevents leakage during collection, handling, processing,
storage transport or shipping.
 Lab equipment and work surfaces should be decontaminated with an effective
disinfectant on a routine basis, at the conclusion of procedures with potentially infectious
materials, and whenever a spill or splash occurs. Contaminated equipment must be
decontaminated before it is sent for repair or maintenance.
 Spills and accidents that result in exposure to potentially infectious materials (including
viable clinical specimens or cell materials) must be immediately reported to the primary
investigator. Proper incident response and reporting is critical to ensure that you receive
the best possible care in the event that the exposure results in infection.
 Animals not involved in the work being performed are not permitted in the lab. The lab is
no place for pets.

Avoid Aerosols

Doing aerosol-generating activities with biohazardous materials is a major concern. It is

important to note that pathogens not normally transmitted by the aerosol route can be an aerosol
hazard when you perform procedures that generate aerosols.

Some common activities that generate aerosols include:

 Vortexing
 Popping tube caps
 Pipetting, pouring
 Homogenization
 Loading and injecting syringes
 Centrifugation

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  Changing bedding of infected animals
 Blending

If possible do aerosol-generating activities in containment such as a Biosafety Cabinet (BSC).

When aerosol-generating activities need to be done outside containment, use appropriate safety
practices.

BLS 2 Primary Barriers

 Using a Biological Safety Cabinet (BSC) to Minimize Your Exposure Risk

A biological safety cabinet should be used whenever:

 Conducting procedures with a potential for creating aerosols or splashes

 Using high concentrations of infectious agents
 Using large volumes of infectious agents

A biosafety cabinet is equipped with high efficiency particulate air (HEPA) filters that are
capable of capturing particulates. This feature protects not only the user but the materials inside
the cabinet as well. However, this is only possible when the cabinet is used properly.

 Proper Use of A Biological Safety Cabinet

o Turn it on – check the gauge
o Maintain a constant air curtain
o Establish a “Clean to Dirty” work pattern
o Avoid clutter; keep grille area clear
o Disinfect working surface and interior
o Ensure that BSC is certified annually
 Biosafety Cabinets – No Substitute for a Chemical Fume Hood

Biosafety cabinets are designed to capture particulates ONLY. They do not capture vapors.
Additionally, whether a BSC is exhausted to the outdoors or to the room, most BSCs will
recirculate the air within the cabinet.

Hazardous chemicals, especially volatiles, should not be used in a BSC. The cabinet will simply
recirculate and concentrate these vapors and increase your chemical exposure and the risk for an
explosion.

 Flames in the BSC: Not Recommended

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 When used in conjunction with flammables (i.e., ethanol) in the BSC, flames are a significant
fire hazard and must not be used. Flames are also disruptive to the airflow, which is critical for
the proper function of the BSC, and they can damage the HEPA filters.

Alternatives to flame sterilization would include the use of pre-sterilized equipment and
materials or flameless incinerating equipment.

Ultraviolet (UV) Lights: A Possible Burn Hazard

Some biosafety cabinets are equipped with UV lights. These lights (usually tubes that are
purplish-black in color) are intended to be germicical but cannot be used as a stand-alone means
of disinfection under most circumstances. They are a much greater burn/exposure hazard to
personnel working in the immediate environment than an effective means of disinfection.

If your BSC is equipped with a UV light, know where the “ON/OFF” switch is and how it
functions. Do not work in the BSC with the UV light on and do not work in close proximity (in
the same culture room) to a BSC where the light is on.

BLS 2 – Personnel Protective Equipment (PPE)

At biosafety level 2, the biological materials that you work with are assumed to be a biological
exposure risk through:

 Accidental ingestion
 Contact with the mucous membranes (primarily through splashes to the eyes nose, or
mouth)
 Entry through broken skin (either through cuts with contaminated objects, contact with
broken skin and possibly through prolonged contact with skin that appears to be
undamaged)

Therefore, you must use PPE that is designed to effectively block the route of exposure that your
procedures are likely to create. PPE selection will vary depending on the nature of the work that
you will be doing and the materials encountered. Even so, we will review the principles of PPE
selection in this section.

PPE: Body Protection at BLS 2

In lab applications and animal applications where anticipated contamination is minimal (i.e.
work with diagnostic specimens, small rodents or birds), a lab coat is the minimum body
protection that must be worn. In some research applications, disposable fluid-resistant coveralls
may be warranted based on the risk assessment.

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PPE: Eye and Face Protection at BLS 2

Eye protection requirements at BLS 2 are essentially the same as for BLS 1. Safety glasses are
required at a minimum for all lab applications where work with chemical or biological materials
is underway. Where procedures are likely to create a splash or spray of biological materials,
goggles must be worn unless this will hinder your ability to safely execute the procedure.

Face shields should be considered to further protect against contamination of your face that can
lead to accidental ingestion or mucous membrane exposure. However, a face shield is not a
substitute for eye protection and must be worn in conjunction with eye protection.

PPE: Gloves at BLS 2

Glove use at BLS 2 is required whenever work with viable biological material is underway.
Gloves must be changed when contaminated or when the integrity of the glove is compromised.
Dispose of gloves at BLS 2 as biohazardous waste. Disposable gloves can degrade quickly. If
your gloves are "sticky" or are discolored in the cuffs or fingertips, degradation has probably
already started. Always inspect your gloves thoroughly before you begin working to minimize
your exposure risk.

Biosafety Level 3: Facility Features

Biosafety Level 3 facilities and procedures are typically required for diagnostic or research work
with agents that may cause serious or potentially lethal disease as a result of exposure by the
inhalation route. With this said, the safety features of most common university labs will not meet
BL3 containment requirements. In addition the facility features required for BLS2, lab
environments operating at BLS 3 must have the following features:

 Access to the lab is restricted. The lab is separated from areas that are open to
unrestricted flow within the building.
 Passage through a series of two self-closing doors into lab from the access corridor is
standard. (This creates a buffer zone that prevents air from drifting out of the lab directly
into the corridor).
 A ducted exhaust air ventilation system is provided that draws air from the "cleanest"
area to the "dirtiest" area. Exhaust air is not recirculated to other parts of the building, and
must be dispersed away from occupied areas and air intakes.

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BIOSAFETY COMMITEE

Typically, institutions that experiment with or create potentially harmful biological material will
have a committee or board of supervisors that is in charge of the institution's biosafety. They
create and monitor the biosafety standards that must be met by labs in order to prevent the
accidental release of potentially destructive biological material. Biosafety is related to several
fields:

 In ecology (referring to imported life forms from beyond ecoregion borders),

 In agriculture (reducing the risk of alien viral or transgenic genes, genetic engineering or
prions such as BSE/"MadCow", reducing the risk of food bacterial contamination)
 In medicine (referring to organs or tissues from biological origin, or genetic therapy
products, virus; levels of lab containment protocols measured as 1, 2, 3, 4 in rising order
of danger),
 In chemistry (i.e., nitrates in water, PCB levels affecting fertility)
 In exobiology (i.e., NASA's policy for containing alien microbes that may exist on space
samples. See planetary protection and interplanetary contamination), and
 In synthetic biology (referring to the risks associated with this type of lab practice)

The international Cartagena Protocol on Biosafety deals primarily with the agricultural definition
but many advocacy groups seek to expand it to include post-genetic threats: new molecules,
artificial life forms, and even robots which may compete directly in the natural food chain.

In synthetic biology

High-security facilities are necessary when working with synthetic biology as there are
possibilities of bioterrorism acts or release of harmful chemicals and or organisms into the
environment. A complete understanding of experimental risks associated with synthetic biology
is helping to enforce the knowledge and effectiveness of biosafety. With the potential future
creation of man-made unicellular organisms, some are beginning to consider the effect that these
organisms will have on biomass already present. Scientists estimate that within the next few
decades, organism design will be sophisticated enough to accomplish tasks such as creating
biofuels and lowering the levels of harmful substances in the atmosphere. Scientist that favor the
development of synthetic biology claim that the use of biosafety mechanisms such as suicide
genes and nutrient dependencies will ensure the organisms cannot survive outside of the lab
setting in which they were originally created. Organizations like the ETC Group argue that
regulations should control the creation of organisms that could potentially harm existing life.
They also argue that the development of these organisms will simply shift the consumption of
petroleum to the utilization of biomass in order to create energy. These organisms can harm
existing life by affecting the prey/predator food chain, reproduction between species, as well as
competition against other species (species at risk, or act as an invasive species). Synthetic
vaccines are now being produced in the lab. These have caused a lot of excitement in the
pharmaceutical industry as they will be cheaper to produce, allow quicker production, as well
enhance the knowledge of virology and immunology.

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In medicine, healthcare settings and laboratories

Biosafety, in medicine and health care settings, specifically refers to proper handling of organs or
tissues from biological origin, or genetic therapy products, viruses with respect to the
environment, to ensure the safety of health care workers, researchers, lab staff, patients, and the
general public. Laboratories are assigned a biosafety level numbered 1 through 4 based on their
potential biohazard risk level. The employing authority, through the laboratory director, is
responsible for ensuring that there is adequate surveillance of the health of laboratory personnel.
The objective of such surveillance is to monitor for occupationally acquired diseases. The World
Health Organization attributes human error and poor technique as the primary cause of
mishandling of biohazardous materials.

Biosafety is becoming also a global concern and require multilevel resources and international
collaboration to monitor, prevent and correct accidents from unintended and malicious release
and also to prevent that bioterrorists get their hands-on biologics sample to create biologic
weapons of mass destruction. Even people outside of the health sector needs to be involved as in
the case of the Ebola outbreak the impact that it had on businesses and travel required that
private sectors, international banks together pledged more than $2 billion to combat the
epidemic.The bureau of international Security and nonproliferation (ISN) is responsible for
managing a broad range of U.S. nonproliferation policies, programs, agreements, and initiatives,
and biological weapon is one their concerns Biosafety has its risks and benefits. All stakeholders
must try to find a balance between cost-effectiveness of safety measures and use evidence-based
safety practices and recommendations, measure the outcomes and consistently reevaluate the
potential benefits that biosafety represents for human health. Biosafety level designations are
based on a composite of the design features, construction, containment facilities, equipment,
practices and operational procedures required for working with agents from the various risk
groups.

Classification of biohazardous materials is subjective and the risk assessment is determined by

the individuals most familiar with the specific characteristics of the organism. There are several
factors taken into account when assessing an organism and the classification process.

 Risk Group 1: (no or low individual and community risk). A microorganism that is
unlikely to cause human or animal disease.
 Risk Group 2 : (moderate individual risk, low community risk) A pathogen that can cause
human or animal disease but is unlikely to be a serious hazard to laboratory workers, the
community, livestock or the environment. Laboratory exposures may cause serious
infection, but effective treatment and preventive measures are available and the risk of
spread of infection is limited.
 Risk Group 3 : (high individual risk, low community risk) A pathogen that usually causes
serious human or animal disease but does not ordinarily spread from one infected
individual to another. Effective treatment and preventive measures are available.
 Risk Group 4 : (high individual and community risk) A pathogen that usually causes
serious human or animal disease and that can be readily transmitted from one individual
to another, directly or indirectly. Effective treatment and preventive measures are not
usually available.

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World Health Organization Biosafety Laboratory Guidelines

Investigations have shown that there are hundreds of unreported biosafety accidents, with
laboratories self-policing the handling of biohazardous materials and lack of reporting. Poor
record keeping, improper disposal, and mishandling biohazardous materials result in increased
risks of biochemical contamination for both the public and environment.

Along with the precautions taken during the handling process of biohazardous materials, the
World Health Organization recommends: Staff training should always include information on
safe methods for highly hazardous procedures that are commonly encountered by all laboratory
personnel and which involve:

1. Inhalation risks (i.e. aerosol production) when using loops, streaking agar plates,
2. pipetting, making smears, opening cultures, taking blood/serum samples, centrifuging,
etc.
3. Ingestion risks when handling specimens, smears and cultures
4. Risks of percutaneous exposures when using syringes and needles
5. Bites and scratches when handling animals
6. Handling of blood and other potentially hazardous pathological materials
7. Decontamination and disposal of infectious material.

Agency guidance

Many government agencies have made guidelines and recommendations in an effort to increase
biosafety measures across laboratories in the United States. Agencies involved in producing
policies surrounding biosafety within a hospital, pharmacy or clinical research laboratory
include: the CDC, FDA, USDA, DHHS, DoT, EPA and potentially other local organizations
including public health departments. The federal government does set some standards and
recommendations for States to meet their standards, most of which fall under the Occupational
Health and Safety Act of 1970. But currently, there is no single federal regulating agency
directly responsible for ensuring the safety of biohazardous handling, storage, identification,
clean-up and disposal. In addition to the CDC, the Environmental Protection Agency has some of
the most accessible information on ecological impacts of biohazards, how to handle spills,
reporting guidelines and proper disposal of agents dangerous to the environment. Many of these
agencies have their own manuals and guidance documents relating to training and certain aspects
of biosafety directly tied to their agency's scope, including transportation, storage and handling
of blood borne pathogens. (OSHA, IATA). The American Biological Safety Association (ABSA)
has a list of such agencies and links to their websites, along with links to publications and
guidance documents to assist in risk assessment, lab design and adherence to laboratory exposure
control plans. Many of these agencies were members of the 2009 Task Force on BioSafety.
There was also a formation of a Blue Ribbon Study Panel on Biodefense, but this is more
concernend with national defense programs and biosecurity.

Ultimately states and local governments, as well as private industry labs, are left to make the
final determinants for their own biosafety programs, which vary widely in scope and
enforcement across the United States. Not all state programs address biosafety from all necessary

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perspectives, which should not just include personal safety, but also emphasize an full
understanding among laboratory personnel of quality control and assurance, exposure potential
impacts on the environment, and general public safety.

State occupational safety plans are often focused on transportation, disposal, and risk assessment,
allowing caveats for safety audits, but ultimately leaves the training in the hands of the employer.
22 states have approved Occupational Safety plans by OSHA that are audited annually for
effectiveness. These plans apply to private and public sector workers, and not necessarily state/
government workers, and not all specifically have a comprehensive program for all aspects of
biohazard management from start to finish. Sometimes biohazard management plans are limited
only to workers in transportation specific job titles. The enforcement and training on such
regulations can vary from lab to lab based on the State's plans for occupational health and safety.
With the exception of DoD lab personnel, CDC lab personnel, First responders, and DoT
employees, enforcement of training is inconsistent, and while training is required to be done,
specifics on the breadth and frequency of refresher training does not seem consistent from state
to state; penalties may never be assessed without larger regulating bodies being aware of non-
compliance, and enforcement is limited.

Medical waste management in the United States

Medical waste management was identified as an issue in the 1980s; with the Medical Waste
Tracking Act of 1988 becoming the new standard in biohazard waste disposal.

Although the Federal Government, EPA and DOT provide some oversight of regulated medical
waste storage, transportation, and disposal the majority of biohazard medical waste is regulated
at the state level. Each state is responsible for regulation and management of their own
bioharzardous waste with each state varying in their regulatory process. Record keeping of
biohazardous waste also varies between states.

Medical healthcare centers, hospitals veterinary clinics, clinical laboratories and other facilities
generate over one million tons of waste each year. Although the majority of this waste is as
harmless as common household waste, as much as 15 percent of this waste poses a potential
infection hazard, according to the Environmental Protection Agency (EPA). Medical waste is
required to be rendered non-infectious before it can be disposed of. There are several different
methods to treat and dispose of biohazardous waste. In the United States, the primary methods
for treatment and disposal of biohazard, medical and sharps waste may include:

 Incineration
 Microwave
 Autoclaves
 Mechanical/Chemical Disinfection
 Irradiation

Different forms of biohazardous wasted required different treatments for their proper waste
management. This is determined largely be each states regulations. Currently, there are several

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contracted companies that focus on medical, sharps and biological hazard disposal. Stericycle is
one of the leaders in medical waste and pharmaceutical disposal in the United States.

Incidents of non-compliance and reform efforts

The United States Government has made it clear that biosafety is to be taken very seriously. In
2014, incidents with Anthrax and Ebola pathogens in CDC laboratories, prompted the CDC
director Tom Frieden to issue a moratorium for research with these types of select agents. An
investigation concluded that there was a lack of adherence to safety protocols and "inadequate
safeguards" in place. This indicated a lack of proper training or reinforcement of training and
supervision on regular basis for lab personnel.

Following these incidents, the CDC established an External Laboratory Safety Workgroup
(ELSW), and suggestions have been made to reform effectiveness of the Federal Select Agent
Program. The White house issued a report on national biosafety priorities in 2015, outlining next
steps for a national biosafety and security program, and addressed biological safety needs for
health research, national defense, and public safety.

In 2016, the Association of Public Health Laboratories (APHL) had a presentation at their annual
meeting focused on improving biosafety culture. This same year, The UPMC Center for Health
Security issued a case study report including reviews of ten different nations' current biosafety
regulations, including the United States. Their goal was to "provide a foundation for identifying
national‐level biosafety norms and enable initial assessment of biosafety priorities necessary for
developing effective national biosafety regulation and oversight."

Assignment

1. How are the biological materials that you work with considered the biological exposure risk
(4marks)

2. Discuss the use of biological safety cabinet as primary biocontainment to minimize your
exposure risk (12marks)

3. Discuss the challenges that affect the implementation of biosafety measures in developing
countries (14 marks)

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