Risk Group Classification of

Microorganisms and Bio-safety Levels

AHMAD NAEEM SAJED

Lesson Two

Bio-safety and Bio-Risk Management Training

course
BIO-HAZARDOUS AGENTS
CLASSIFICATION
BACTERIA:

This single celled organism lack a nucleus and is

classified in three phenotypes:
 Gram-positive (cocci or bacilli)
 Gram-negative (cocci or bacilli)
 Mycoplasmas (those that lack a cell wall)
Note: When a bacterium causes harm and disease in human
and/or animal hosts, they are referred to as bacterial
pathogens.
Examples: Escherichia coli, Mycobacterium tuberculosis,
Staphylococcus aureus, and Bacillus anthracis.
VIRUSES:
Viruses are infectious entities, smallest of the
replicating organisms have no metabolism of their
own and are obligate parasites that depend on their
host’s machinery for replication.
Classification:
 DNA viruses
 RNA viruses
Note: When a bacterium causes harm and disease in human
and/or animal hosts, they are referred to as bacterial
pathogens.
Examples: Influenza virus, Human Immunodeficiency Virus (HIV),
Rabies virus, and Ebola virus.
FUNGI:
Fungi are heterotrophic eukaryotic microorganisms are
larger in size and have organelles, including a nucleus,
vacuoles, and mitochondria.
Classification:
 Yeasts
 Molds
Note: These are two main categories known to have species that
cause disease in human and/or animal hosts. Molds have been
known to release spores and some toxins, thus increasing the
risk to using certain strains.
Examples: Aspergillus nigher, Candida albicans, and Histoplasma
capsulatum.
PARASITES:
Protozoa and helminthes that live on or within a larger
host organism at the host’s expense are considered
parasites.
Classification:
 Ecto-parasites live on the external surface or within the skin of
their host and cause infestation.
 Endo-parasites live within the tissues or cells, but cause
infections.
Examples:
Protozoa: Plasodium falciparum, Leishmania donovani, and
Giardialamblia.
Pathogenic helminthes: include: Trichinella spiralis (nematode),
Enterobius vermicularis (pinworm), and Hymenolepis nana
(tapeworm).
ZOONOTIC PATHOGENS:
Zoonotic pathogen refers to pathogens naturally
transmitted between animals and humans.
Pathways: Zoonoses can invade humans are via
breathing air, drinking water, eating contaminated
food, or hand to mouth contact.

Examples: Salmonellosis, Plague, Rabies, Toxoplasmosis, and

Prions.
TOXINS:
Toxins are poisonous substances that are produced by
bacteria, animals or plants. They are usually active at
very low concentration and vary in size. They range
from small molecules to larger molecules such as,
peptides or proteins.
Classification:
 Endo-toxins are structural molecules that are embedded in
the outer layer of the cell wall of certain Gram negative
bacteria.
 Exo-toxins often heat labile proteins and polypeptides that are
produced and secreted or released by a variety of species.
Examples: venom and tetrodotoxin
HUMAN BLOOD AND TISSUE:

Human blood and tissues is with what they may contain.

There are numerous blood borne pathogens such as
HIV, hepatitis B & C, and TB.
-Infected health-care worker can touch a patient and directly
transmit a large number of microorganisms to the new host.

-The infected patient touches—and contaminates—an object, an

instrument, or a surface. Subsequent contact between that item
and another patient is likely to contaminate the second individual
who may then develop an infection.
BIOSAFETY LEVELS
 BIO-SAFETY LEVEL 1 (BSL-1)
 Agents not known to cause disease in healthy adults
– Some organisms may cause disease in immuno-
compromised individuals
 Agents include Bacillus subtilis, Naegleria gruberi,
infectious canine hepatitis virus, non-pathogenic E. coli
species
 Standard practices required:
– frequent hand washing
– no smoking, eating, drinking, storage of food in
Ambulance;
– decontamination of work surfaces after every use of
Ambulance;
BIO-SAFETY LEVEL 1 (BSL-1)
• Standard practices (continued):
– decontamination of Ambulance Equipment;
– "sharps" precautions, including special containers
for disposing of needles and other sharp objects;
– maintenance of insect/rodent control program;
– use of personal protective equipment (Gown, latex
gloves, eye protection or face shields)
• Open bench top sink for hand washing
BIO-SAFETY 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
 Primary hazards:
– accidental needle sticks
– exposure to eyes and nose (mucous membranes)
– ingestion of infectious materials
 BIO-SAFETY LEVEL 2 (BSL-2)
 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 instruments
 Standard practices include BSL-1 plus:
– surveillance of EMS personnel with appropriate
immunizations offered;
BIO-SAFETY LEVEL 2 (BSL-2)
Standard practices include BSL-1 plus:
– Bio-safety manual with definitions of needed waste
decontamination or medical surveillance policies;
– supervisory staff who have experience working
with infectious agents and specific training for EMS
personnel in handling these agents
 Personal protective equipment: , gloves, face
protection as needed
 Protective clothing removed when personnel leave
contaminated area
 Secondary barriers: BSL-1 barriers plus autoclave for
bleeding kit and Surgical kit
BIO-SAFETY LEVEL 3 (BSL-3)
 Agents with potential for respiratory transmission,
may cause serious and potentially lethal infection
 Agents include Mycobacterium tuberculosis, St. Louis
encephalitis virus, Francisella tularensis, Coxiella burnetii
 Primary hazards: needle sticks, ingestion, exposure to
infectious aerosols
 Standard practices include BSL-2 plus:
– decontamination
– Similar to BSL-2 personal protective equipment
– Respiratory equipment if risk of infection through
inhalation
BIO-SAFETY 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 Congo-Crimean hemorrhagic fever,
Lassa fever
• 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
BIO-SAFETY LEVEL 4 (BSL-4)
• For example
– Workers studying infected monkeys from Uganda
– First documented naturally-occurring human case
occurred in 1975
• Personnel must receive specialized training to enter the
scene for handling extremely dangerous infectious
agents, containment equipment.
• Immuno-compromised persons are never allowed to
expose to these agents
BIO-SAFETY LEVEL 4 (BSL-4)
• Standard practices include BSL-3 plus:
– changing clothing before entering and exiting
contaminated area (showering upon exiting
recommended);
– decontaminating all material exiting facility
– All physical barriers at BSL-3
 REFERENCES
1. 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.
2. Clemson University Environmental Health and Safety. Biological Safety
[online training].http://ehs.clemson.edu/training/biosafety/index.html.
Accessed February 6, 2008.
3. 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.htmlAccessed February 6, 2008.
4. 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.
Thank you!