Biosafety

in
Microbiology laboratory

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Fundamental objective biosafety program
 - the containment of potentially harmful biological agents.
 “Containment”- safe methods, facilities and equipment for
managing infectious materials in the laboratory
environment where they are being handled or maintained.
 The purpose of containment-
to reduce or eliminate exposure of laboratory workers,
other persons, and the outside environment to potentially
hazardous agents.
 The use of vaccines may provide an increased level of
personal protection.
 Why is Biosafety Important?
 Laboratories should recognize hazards of
processing infectious agents

 Guidelines should be developed to protect workers

in microbiological and medical labs through
i. engineering controls
ii. management policies
iii. standard work practices.
 BIOSAFETY: Preventing lab-acquired
infections
• It is a system for the safe
handling of toxic and
dangerous biological and
chemical substances
• Bacteria
• Viruses
• Fungi
• Human blood, unfixed
tissue
• Human cell lines
• Recombinant DNA

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 Standard Microbiological Practices

• NOT permitted in
laboratories:
 Eating
 Drinking
 Smoking
 Handling contact lenses
 Pipetting by mouth
 Storing food and drink

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Standard Microbiological Practices
• NEVER
– recap, bend, or
break needles
– discard needles or
sharps into
biological waste
bags
– discard needles
into regular trash

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 Components of Safety in All labs

 Safe handling, storage and disposal of

-Specimens
-Chemicals
-Instruments
-Radio active components
 Fire safety
 Electrical safety

 Disaster management in areas risk prone for natural

calamities like Earthquakes, Hurricanes or snowstorms
Lab Safety Symbols

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 Biosafety Levels
• Precautions so people researching to 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

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 Laboratories divided on basis of nature of
microbes

• 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

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 Barriers - Primary barriers
• Primary barriers:
physical barriers or
personal protective
equipment
between lab
worker and
pathogen
– Gloves, masks,
special breathing
apparatuses
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Barriers Secondary barriers:

• Secondary barriers:
structural aspects of the
laboratory that make
working environment
safer against infection
– Sinks for handwashing,
special containment
areas, special air
ventilation patterns

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Biosafety Cabinets

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Biosafety Cabinets

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Biosafety Cabinets

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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, non-
pathogenic E. coli species

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 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;

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 Biosafety Level 2 (BSL-2)
• Agents associated with human
disease
– Generally required for
any human-derived
blood, bodily fluids,
tissues in which
infectious agent may be
unknown
– Agents include
measles virus,
Salmonella species,
pathogenic
Toxoplasma,
Clostridium botulinum,
hepatitis B virus

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 Biosafety Level 2 (BSL-2)
• Primary hazards:
– accidental needle sticks
– exposure to eyes and nose (mucous membranes)
– ingestion of infectious materials
• Agents do not cause lethal infections, are not transmissible via
airborne route
– (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

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 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

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 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
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 Levels of Containment
• BL3 - microorganisms that
cause serious disease,
transmitted by inhalation
– M. tuberculosis, yellow
fever virus, hantavirus, Y.
pestis (plague)
– Containment lab:
double door entry;
directional airflow; all
work in biosafety cabinet

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 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
()
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 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

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 Biosafety Level 4
• microorganisms that
cause lethal disease, with
no known treatment or
vaccine
Ebola virus, Marburg
virus
Maximum
containment lab;
positive pressure
ventilated suits
(moon suits)

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 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)

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Risk groups, biosafety levels, practices and
equipment
BSL Laboratory type Laboratory Safety equipment
practices
1 Basic teaching, Good microbiological None
research techniques Open bench work
2 Primary health Good microbiological Open bench PLUS
services; diagnostic techniques, biological safety cabinet for potential
services, research protective clothing, aerosols
biohazard sign

3 Special diagnostic As BSL 2 PLUS Biological safety cabinet and/or other

services, research special clothing, primary devices for all activities
controlled access,
directional airflow

4 Dangerous As BSL 3 PLUS Class III biological safety cabinet,

pathogen units airlock entry, shower positive pressure suits, double ended
exit, special waste autoclave (through the wall), filtered air
 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

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