Biosafety

 Biosafety is the prevention of large-scale loss of biological

integrity, focusing both on ecology and human health.
 The maintenance of safe conditions in biological research to
prevent harm to workers, non-laboratory organisms, or the
environment.

A fundamental objective of any biosafety program is the containment of

potentially harmful biological agents.

The term “containment” is used in describing safe methods, facilities and

equipment for managing infectious materials in the laboratory environment
where they are being handled or maintained.
 Technical Definitions
• Biohazard: An agent of biological origin that has the capacity
to produce deleterious effects on humans, i.e.
microorganisms, toxins and allergens derived from those
organisms; and allergens and toxins derived from higher
plants and animals.

• Biosafety: The containment principles, technologies and

practices that are implemented to prevent the accidental
exposure to pathogens and toxins, or their accidental release.

• Biosecurity: Control of accidental and deliberate release of

biohazardous material
Why we need Biosafety
Some unfortunate examples of disease outbreaks in research labs:
1950‐1976:
•A survey of 5000 labs showed 3921 cases of disease outbreaks
•Most commonly reported were: Hepatitis, tuberculosis, typhoid, brucellosis,
rabbit fever
2003-2014:
•SARS infects researchers in a lab in Singapore
•A US and a russian scientist are infected by Ebola. One survives, one dies.
•Polio virus escapes from two idian labs
•Scientists from Boston University contract rabbit fever (a serious bacterial
disease)
•Anthrax exposure in a Huston lab due to aerosols leaked inside an unshielded
Centrifuge.
 Biosafety in Various Disciplines

Biosafety is related to several fields

• ECOLOGY
• AGRICULTURE
• MEDICINE
• CHEMISTRY
• BIOLOGY
 Biosafety in Academic Research
Research Universities:
• Promoting safe laboratory
practices,
• Procedures; proper use of
containment equipment and
facilities;
• Provides advice on laboratory
design and risk assesment of
experiments involving infectious
agents, rDNA in-vitro and in-
vivo.
 Biohazard Symbol

• In 1966, Charles
Baldwin at National
Cancer Institute at
NIH.
 Biosafety Issues

• Laboratory Safety
• Bloodborne pathogens (BBP)
• Recombinant DNA (rDNA)
• Biological waste disposal
• Infectious substance and
diagnostic specimen shipping
 Biosafety Issues

• Respiratory Protection
• Bioterrorism and Select agents
• outdoor and indoor air quality
• Occupational safety and health in the use
of research animals
• Biohazards used in animal models
 Biohazardous Materials
• Viruses
• Bacteria
• Fungi
• Rickettsiae
• Prions
• Recombinant DNA
 Types of pathogens
Bacteria
– Size: 0.3 to 2 μm
– Single‐celled organisms
– Various morphologies
– Examples: Salmonella spp., E.
coli, Vibrio spp. (Cholera),
Mycobacterium (Tuberculosis)
 Virus

– Size: 18‐200 nm
– Basic structure: capsid (protein) +
nucleic acid
– Obligate parasites
– Enveloped vs. non‐enveloped
– Examples: Hepatitis, polio, HIV
 Protozoa
– Size: 5‐10 μm
– Single‐celled eukaryotes
– Numerous morphologies
– Examples: Cryptosporidium spp., Plasmodium
spp. (Malaria), Giardia spp.
 Helminths
• – Size: 20‐100 μm
• – Multi‐cellular eukaryotes
• – For transmission mainly concerned with
eggs
 Biohazardous Materials
• Transgenic Plants, Animals and Insects
 Biohazardous Materials

• Human and Primate Cells, Tissues, and

Body Fluids
• Brain Tissue from crazy Patients
• Viral Vectors
– Replication deficient viruses
 Biosafety containment levels
• Biosafety levels
– Level 1& 2: basic laboratories
– Level 3: containment laboratories
– Level 4 : high containment laboratories
• Each level associated with appropriate
– Equipment, practices, work procedures
• Diagnostic and health-care laboratories must
be biosafety level 2 or above
 Risk group classification

Risk Individual risk Community risk

Group
1 no, low no, low

2 moderate low

3 high low

4 high high
 Risk Group 1
• Unlikely to cause animal or human disease
• Non pathogenic agent
 Risk Group 2
Pathogenic for humans
Unlikely a serious hazard
Treatment and preventive
measures available
Limited risk of spread of
infection

CDC, Yersinia pestis laboratory

 Risk Group 3

Pathogenic, cause serious disease

Effective treatment and
preventive measures usually
available
Little person-to-person spread

Laboratory in Lyon France

 Risk Group 4
Lethal, pathogenic agent
Readily transmittable
– direct, indirect
Effective treatment and
preventive measures not
usually available

National Institute for Infectious

Diseases, Rome, Italy
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
 Disinfection

Disinfection requirements depend on the

experimental work and nature of the agents
being handled
Contact time and concentration for disinfectants
are specific for each material and manufacturer
Efficacy is a function of surface, contact time,
product, dilution
 Using detergents
• Use licensed detergents/disinfectants
• Follow manufacturers’ recommendations
• Wear adequate personal protective equipment
• Perform hand hygiene
• Avoid difficult to clean surfaces/ materials
(carpet, wood)
• Apply disinfectant to a large area - wiping,
soaking
• Avoid aerosolizing specimens while handling
Disinfection with household bleach
Work areas
-disinfect with 0.5% bleach after
every procedure

Contaminated supplies
-soak with 0.5 % household
bleach for 30 minutes
-wash in soapy water
-sterilize as necessary

Photo: WHO
 Activity spectrum of
select
detergents and disinfectants
Bact Bact Myco
Spores Yeast Virus Prions
eria+ eria- B
Alcohol 70° ++ ++ ++ 0 + + 0
Aldehydes +++ +++ ++ + +++ ++ 0
Ammonium IV +++ + 0 0 + + 0
Anilides + 0 0 0 0 0 0
Chlorhexidine +++ ++ 0 0 + + 0
Cl compounds +++ +++ ++ ++ ++ ++ + (a)
Iodine (+ der.) +++ +++ ++ ++ ++ ++ 0
Hg compounds ++ ++ 0 0 + 0 ou + 0
Phenols : Variable activity depending on components (b)
Hexachlorophene +++ + 0 0 + 0 0
(a) Bleach (6%) during 60 min at 20°C ; (b) discussion on efficacy of phenol on prions
 Personal Protection: Gloves
• Whenever possible, suitable gloves should be
worn when handling biohazardous materials
– not a substitute for regular and proper hand
hygiene
Personal Protection: Hand hygiene

Alcohol based hand rubs are gold standard in health care

settings (if hands not visibly soiled)
Must complement with hand washing with normal soap

Photos: WHO
 Category A
Bacillus anthracis (cultures only)
Brucella abortus (cultures only)
Brucella melitensis (cultures only)
Brucella suis (cultures only)
Burkholderia mallei [Pseudomonas mallei –
Glanders](cultures only)
Burkholderia pseudomallei [Pseudomonas
pseudomallei] (cultures only)
Chlamydia psittaci [avian strains] (cultures
only)
Clostridium botulinum (cultures only)
Coccidioides immitis (cultures only)
Coxiella burnetii (cultures only)
Crimean-Congo hemorrhagic fever virus
Dengue virus (cultures only)
Eastern equine encephalitis virus (cultures only)
Escherichia coli, verotoxigenic (cultures only)
Ebola virus
Flexal virus
Francisella tularensis (cultures only)
 Category A (cont’d)
Guanarito virus Junin virus
Hantaan virus Kyasanur Forest disease virus
Hantaviruses causing haemorrhagic fever Lassa virus
with renal syndrome
Machupo virus
Hendra virus
Hepatitis B virus (cultures only) Marburg virus
Herpes B virus (cultures only) Monkeypox virus
Human immunodeficiency virus (cultures Mycobacterium tuberculosis (cultures
only) only)
Highly pathogenic avian influenza virus Nipah virus
(cultures only) Francisella tularensis (cultures only)
Japanese Encephalitis virus (cultures only)
 Category A (cont’d)
Omsk hemorrhagic fever virus
Poliovirus (cultures only)
Rabies virus (cultures only)
Rickettsia prowasekii (cultures only)
Rickettsia rickettsii (cultures only)
Rift Valley fever virus (cultures only)
Russian spring-summer encephalitis virus
(cultures only)
Sabia virus
Shigella dysenteriae type 1 (cultures only)
Tick-borne encephalitis virus (cultures
only)
Variola virus
Venezuelan equine encephalitis virus
(cultures only)
West Nile virus (cultures only)
Yellow fever virus (cultures only)
Yersinia pestis (cultures only)
 Category A (cont’d)
•Mycoplasma mycoides [Contagious
•African swine fever virus (cultures only)
bovine pleuropneumonia] (cultures only)
•Avian paramyxovirus Type 1 [Velogenic
•Peste des petits ruminants virus (cultures
Newcastle disease virus] (cultures only)
only)
•Classical swine fever virus (cultures only)
•Rinderpest virus (cultures only)
•Foot and mouth disease virus (cultures
•Sheep-pox virus (cultures only)
only)
•Goatpox virus (cultures only)
•Lumpy skin disease virus (cultures only)
•Swine vesicular disease virus (cultures
only)
•Vesicular stomatitis virus (cultures only)
 Sample collection and shipping

Developed by the Department of Epidemic and

Pandemic Alert and Response of the World Health
Organization with assistance from:

European Program for Intervention

Epidemiology Training

Canadian Field Epidemiology Program

Thailand Ministry of Health

Institut Pasteur

Laboratory Training for FieldEEpidemiologists

P I D E M I C A L E R T A N D R E S P O N S E