Laboratory Biosafety

Levels

Goals

Define barriers and procedures used by

laboratories to protect workers and others
from infection
Describe the four biosafety levels and the
protective measures used by each type of
laboratory when handling infectious
materials
Provide examples of the types of biological
agents handled in each type of laboratory
Describe typical places where each type of
laboratory can be found in the US

Why is Biosafety
Important?

Laboratorians recognize hazards of

processing infectious agents
Guidelines developed to protect
workers in microbiological and
medical labs through engineering
controls, management policies,
work practices

Biosafety Levels

Precautions so people researching or

trying to identify organisms do not
become infected
While handling or testing clinical
specimens, workers could accidentally
infect themselves or coworkers
Labs must adhere to very specific safety
regulations to work with organisms that
pose a threat to human health

Biosafety Levels

Regulations outline precautions, special

practices, decontamination procedures
Labs divided into 4 biosafety levels;
protective practices increase with each

Biosafety Level 1 labs - work with least

dangerous agents, require fewest precautions
Biosafety Level 4 labs - have strictest
methods because dealing with agents that
are most dangerous to human health

About this Information

Information summarized here should

not be used to establish laboratory
safety protocols
Complete information and
recommendations can be found in
Biosafety in Microbiological and
Biomedical Laboratories5th Edition at
http://www.cdc.gov/od/ohs/biosfty/bmbl
4/ bmbl4s6.htm

Barriers

Primary barriers: physical barriers or

personal protective equipment between
lab worker and pathogen

Gloves, masks, special breathing apparatuses

Secondary barriers: structural aspects of

the laboratory that make working
environment safer against infection

Sinks for handwashing, special containment

areas, special air ventilation patterns

Universal Precautions

Universal precautions developed to protect health

professionals

Most often apply in a clinical setting

May also be important for field epidemiology practices
during an outbreak investigation (e.g., collecting lab
specimens)

Include hand hygiene, gloves, gown, masks, eye

protection, face shields, safe injection practices
Require that all equipment or contaminated items
are handled to prevent transmission of infectious
agents
Special circumstances may require additional
precautions

Protective clothing, special site decontamination

Biosafety Level 1 (BSL-1)

Agents not known to cause disease

in healthy adults

Some organisms may cause disease

in immunocompromised individuals

Agents include Bacillus subtilis,

Naegleria gruberi, infectious
canine hepatitis virus, nonpathogenic E. coli species

(transmission electron micrograph of E. coli)

Biosafety Level 1 (BSL-1)

Standard practices required:

frequent handwashing
door that can be kept closed when working;
limits on access to the lab space when working;
no smoking, eating, drinking, storage of food in
laboratory;
care to minimize splashes and actions that may
create aerosols (tiny droplets);
decontamination of work surfaces after every
use after any spills;

(continued on next slide)

Biosafety Level 1 (BSL-1)

Standard practices (continued):

decontamination of laboratory wastes;

use of mechanical pipettes only (no mouth
pipetting);
"sharps" precautions, including special
containers for disposing of needles and other
sharp objects;
maintenance of insect/rodent control program;
use of personal protective equipment (lab coats,
latex gloves, eye protection or face shields)

Open bench top sink for hand washing

Biosafety Level 2 (BSL-2)

Agents associated with human

disease

Generally required for any humanderived blood, bodily fluids, tissues in

which
infectious
agent
may be
Agents
include
measles
virus,
unknown
Salmonella species, pathogenic
Toxoplasma, Clostridium
botulinum, hepatitis B virus
(transmission electron micrograph of hepatitis B virus)

Biosafety Level 2 (BSL-2)

Primary hazards:

Agentsdo not cause lethal infections, are not

transmissible via airborne route

accidental needle sticks

exposure to eyes and nose (mucous membranes)
ingestion of infectious materials

(do not cause infection if tiny droplets become

airborne and are inhaled, which might occur if the
material were spattered)

Agents are pathogens for which immunization

or antibiotic treatment is available
Extreme care should be taken with
contaminated needles and sharp lab
instruments

Biosafety Level 2 (BSL-2)

Standard practices include BSL-1 plus:

policies to restrict access to lab;

biohazard warning signs posted outside lab;
surveillance of laboratory personnel with
appropriate immunizations offered;
biosafety manual with definitions of needed
waste decontamination or medical surveillance
policies;
supervisory staff who have experience working
with infectious agents and specific training for
laboratory personnel in handling these agents

Biosafety Level 2 (BSL-2)

Primary barriers: biosafety cabinets or

other approved containment devices
Personal protective equipment: lab
coats, gloves, face protection as needed
Protective clothing removed when
personnel leave laboratory area
Cabinets thoroughly decontaminated
daily and monitored for radiation for
personal protection
Secondary barriers: BSL-1 barriers plus
autoclave for glassware

Biosafety Level 2 (BSL-2)

Example of
biosafety sign
posted outside
lab working with
infectious agents
Labs biosafety level
Infectious agents
under study
Contact information
for responsible
person and 2
emergency contacts

Biosafety Level 3 (BSL-3)

Agents with potential for respiratory

transmission, may cause serious
and potentially lethal infection

May be studied at BSL-2 for diagnosis

Agents include Mycobacterium

tuberculosis, St. Louis
encephalitis virus, Francisella
tularensis, Coxiella burnetii

(F. tularensis under direct fluorescent antibody stain)

Biosafety Level 3 (BSL-3)

Primary hazards: needle sticks, ingestion,

exposure to infectious aerosols
For example:

Public health surveillance for West Nile virus

includes testing birds
In August 2002, state laboratory worker cut
finger while dissecting bird; 4 days later, had
symptoms of fever, myalgia, recurring sweats,
hot flashes
Worker and bird both diagnosed with West Nile
2 other lab-acquired cases in 2002

Biosafety Level 3 (BSL-3)

Tularemia common source of

laboratory-acquired infection

infections occur while handling infected

animals or experimenting with cultures
Laboratory-acquired infections known
to occur but not reportable before
9/11/2001
Tularemia now classified as potential
biological weapon

Biosafety Level 3 (BSL-3)

Standard practices include BSL-2 plus:

strictly controlled access to the lab;
specific training for lab personnel in
handling potentially lethal agents;
decontaminating all waste;
changing contaminated protective lab
clothing, decontaminating lab clothing
before laundering;
institutional policies regarding
specimen collection and storage from
workers to establish exposure

Biosafety Level 3 (BSL-3)

Primary barriers:

Similar to BSL-2 personal protective equipment

Respiratory equipment if risk of infection
through inhalation

Secondary barriers:

All BSL-2 barriers

Corridors separated from direct access to lab
Access through self-closing double doors
Air handling systems to ensure negative air
flow (air flows into the lab)
Air pumped into lab not re-circulated in
building

Biosafety Level 4 (BSL-4)

Dangerous and exotic agents with high risk of

life-threatening disease, aerosol-transmitted
Related agents with unknown risk of
transmission

Agents (all viruses) include

Marburg virus, Ebola virus,
viruses that cause CongoCrimean hemorrhagic fever,
Lassa fever

(transmission electron micrograph of Ebola virus)

Biosafety Level 4 (BSL-4)

Primary hazards:

respiratory exposure to infectious aerosols

mucous membrane exposure to infectious
droplets
accidental sticks with needles or other sharp
objects contaminated with infectious material

For example

In late 1960s, 25 laboratory-acquired Marburg

infections, including 5 deaths
Workers studying infected monkeys from
Uganda
First documented naturally-occurring human
case occurred in 1975

Biosafety Level 4 (BSL-4)

Personnel must receive specialized

training in handling extremely dangerous
infectious agents, containment equipment
and functions
Access to lab is restricted:
immunocompromised persons are never
allowed to enter the lab
Standard practices include BSL-3 plus:

strictly controlled access to the laboratory;

changing clothing before entering and exiting
lab (showering upon exiting recommended);
decontaminating all material exiting facility

Biosafety Level 4 (BSL-4)

Primary barriers:

Biosafety cabinets used at other biosafety

levels
Full-body, air-supplied, positive pressure
personnel suit

Secondary barriers:

All physical barriers at BSL-3

isolated zone or a separate building;
dedicated supply and exhaust, vacuum,
decontamination systems;
a recommended absence of windows
(or sealed and resistant to breakage)

Laboratory Locations

BSL-1: high schools, community colleges, municipal

drinking water treatment facilities
BSL-2: local health departments, universities, state
laboratories, private laboratories (hospitals, health
care systems), industrial laboratories (clinical
diagnostic companies)
BSL-3: state health departments, universities,
private companies, industry, federal government
(NIH, CDC)
BSL-4: only 15 facilities in the US

9 federal (CDC, NIH), 4 university (Georgia State University,

University of Texas Medical Branch), 1 state, 1 private
Renovations underway at several labs, new facilities
proposed at additional sites

Summary

Laboratorians have long recognized hazards of

processing infectious agents
Biosafety guidelines developed to protect
workers in microbiological and medical labs
through a combination of safeguards including
engineering controls, management policies and
work practices.
Issue described differences between biosafety
levels
Help you understand process labs may have to
undertake to identify microorganism, why every
lab cannot test for every organism

References
1.

2.

3.

4.

US Department of Health and Human Services, Centers for Disease

Control and Prevention and National Institutes of Health. Biosafety in
Microbiological and Biomedical Laboratories. 5th ed. Washington,
DC: US Government Printing Office; 2007.
http://www.cdc.gov/od/ohs/ biosfty/bmbl5/bmbl5toc.htm. Accessed
February 6, 2008.
Clemson University Environmental Health and Safety. Biological
Safety [online training].
http://ehs.clemson.edu/training/biosafety/index.html. Accessed
February 6, 2008.
Centers for Disease Control and Prevention. Laboratory-Acquired
West Nile Virus Infections United States, 2002. MMWR Morb Mort
Wkly Rep. 2002; 51:1133-1135.
http://www.cdc.gov/mmwr/preview/ mmwrhtml/mm5150a2.htm.
Accessed February 6, 2008.
United States Government Accountability Office. High-Containment
Biosafety Laboratories: Preliminary Observations on the Oversight of
the Proliferation of BSL-3 and BSL-4 Laboratories in the United
States. Publication GAO-08-108T.
http://www.gao.gov/docsearch/ abstract.php?rptno=GAO-08-108T.
Published October 4, 2007.