Safe Handling of Biological Agents

in the Laboratory
Four risk or hazard groups are often used to categorize biological agents based on several
key factors associated with their inherent characteristics. These are:

• ability to cause disease

• severity of disease
• likelihood of spreading to the wider community
• availability of prophylaxis or treatment
• These groups are then often associated with an equivalent biosafety level (BSL; also
known as containment level).
• Each BSL (ranging from levels 1 to 4) has a specific set of facility and operational
requirements that must be complied with before work with the associated biological
agents can be started.
• Most routine microbiological analytical work is undertaken in laboratories operated at
BSL-2. Although biological agents handled at BSL-2 can cause disease, these typically
present a low-to-moderate risk to employees and are unlikely to spread to the wider
community because effective treatment or prophylaxis is available. 1,2 
• Examples of these include common bacteria such as Staphylococcus
aureus, Mycobacterium fortuitum, and Acinetobacter baumannii as well as viruses such as
respiratory syncytial virus and norovirus. Fungal isolates such as Aspergillus
fumigatus and Microsporum species also fall into this category. Many other examples exist
and are listed by most national regulatory bodies as well as by international agencies
RISK ASSESSMENT
• The aim of a risk assessment is to decide whether appropriate control measures
are in place to eliminate or effectively control risks, or to determine whether
more are required. The first step in assessing risk from biological agents in the
laboratory is to identify and characterize the hazard(s) (ie, sources of harm), the
nature of that harm, who might be affected, and how they may be harmed (ie,
what they are doing and what could happen to cause harm).
Therefore, it is necessary to identify
• What biological agents will be used
• The activities or procedures to be undertaken, including whether sharps are used,
and the potential for causing an exposure, for example, to generate aerosols or
splash
• The potential health consequences to both laboratory personnel and others who
could be exposed
Characterizing Biohazards
When making a biological risk assessment, the hazards relate directly to the biological agents to be
handled. The risk associated with a biological agent will be dependent on several key factors
including
• the likelihood of infection if exposed
• severity of disease (morbidity or mortality)
• infectious dose
• route(s) of transmission (natural versus laboratory; see in the following text)
• communicability/R0 (basic reproduction rate—used to measure the transmission potential of a
disease)
• epidemiology (exotic or endemic)
• effective prophylaxis and treatments (availability, practical, appropriate)
• susceptibility of local population (naive)
Characterizing Biohazards
Other factors that should be considered can include
• environmental robustness (persistence outside of host/culture conditions) of
the biological agent
• natural host range (ie, coming across agents in laboratory not normally
encountered), such as rabies being exotic in United Kingdom but endemic in
mainland Europe and multi-drug-resistant tuberculosis (MDR TB) considered a
greater risk pathology in laboratories at an inner city compared to a rural
location
• zoonosis
• history of laboratory-acquired infection
Characterizing Biohazards
• susceptibility of staff working in the laboratory
(immunocompromised)
• symptoms (possibility of asymptomatic disease)
• shedding (likelihood and type)
• official control (is the agent on a national notifiable disease
database, subject to human, plant, animal, or other regulations)
• the concentration and volume of the biological agent to be
handled
Percutaneous
Within the laboratory, routes of percutaneous transmission can
include
• sharps injury such as needles, broken glass, scalpel blades
• splashing of the mucous membranes of the eye, nose, or
mouth
• contamination of unprotected skin lesions, for example,
eczema
• animal bites or scratches
• presence of a vector, for example, biting or piercing insects
Contact

• Direct contact with microbiologically contaminated objects or

surfaces (fomites) can readily result in
• ingestion of microorganisms, typically after pick up and hand-
to-mouth transfer
• their contact with mucus membranes
• contact with the skin (commonly through broken skin but can
occur via contact with intact skin)
Airborne

• The airborne route of exposure is commonly implicated in

occupational exposures to all types of hazards including
chemicals, nonbiological particulates, and bioaerosols. It is
also an exposure that can be readily controlled in the
laboratory setting using good laboratory techniques and
effective engineering solutions, especially ventilation controls.
Failure to do this can result in inhalation of infectious
aerosols, conjunctival exposure, inhalation of infectious
aerosols followed by swallowing/ingestion (eg, with norovirus),
and droplet exposure.
Infectious Dose and Environmental
Robustness
• The ability of a pathogen to cause infection requires its contact with susceptible
host receptors (a combination of routes of transmission and exposure, as
mentioned earlier) in a sufficient quantity, that is, at an infectious dose.
• Data on infectious dose for the wide range of human pathogens are limited.
However, some notable examples illustrate the importance of taking this into
consideration. 
• Escherichia coli is an ubiquitous bacterium. Most E coli strains can cause minor
infection via the oral route (hand-to-mouth transfer) but require a high dose to do
so; therefore, it is safe to work with them in a general-purpose microbiology
laboratory (BSL-2)
• using routine laboratory precautions. Disabled strains are commonly used tools in
laboratory work, especially in genetic and molecular biology modifications and
because of this attenuation, they are of minimal hazard (equivalent of BSL-1).
EXAMPLES OF LABORATORY ACTIVITIES
AND INHERENT RISK
Estimating the Risks

• To estimate the risks associated with the handling of

microorganisms, the following need to be considered:
consequence of release, consequence of exposure, likelihood
of exposure, what could go wrong, and how likely it would be
to go wrong.
Consequence of Release
• Considerations in determining the consequences if the
biological agent was released can include
• impact/consequence
• environmental survival
• natural distribution (of the agent in the country)
• novelty (new or emerging biological agent, eg, pandemic
influenza)
• country preparedness (public health context)
• epidemiology (endemic, exotic)
Consequence of Exposure
• The main exposure risks associated with working in this
environment are therefore well identified, with some of these
reports extending back many years. Some key examples are
presented in the following text and provide an international
insight into LAIs that help to put in perspective the biological
agents implicated in these exposures and how they occurred.
Most are based on human error, rather than failure of
engineering controls.