2. • Biosafety is the prevention of large-scale loss of
biological integrity, focusing both on ecology and
human health.
• It is a system for the safe handling of toxic and
dangerous biological and chemical substances
• In Medicine-
• It refers to the levels of lab containment
protocols, measured as Bio Safety Level (BSL)
1, 2, 3, 4 in rising order of danger

3. • 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

4. From-
•Bacteria
•Viruses
•Fungi
•Parasites
•Prions
•Recombinant DNA
Source-
•Various specimens
•Human blood
•Unfixed tissue
•Human cell lines
To-
•Lab personnel
•Community

5. Infections of special
concern
•Tuberculosis
•Hepatitis B
•HIV
•Enteric infections
Routes of infections
•Inoculation
•Ingestion
•Inhalation

6. • 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- 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.

7. • Laboratories should recognize hazards of
processing infectious agents
• Guidelines should be developed to protect workers
in microbiological and medical labs through
engineering controls, management policies and
standard work practices.

8. • It is issued under the Environmental (Protection)
Act of 1986
• Rules for manufacture, use/import/export and
storage of hazardous microorganisms/genetically
engineered organisms or Cells, 1989, Dec 5
• There is a new law—the Biotechnology Authority
of India (BRAI)—under consideration of the
Central government.

10. 1. Consider all the specimens potentially infectious for HIV
and other blood borne infections
2. All specimens should be placed in a leak-proof
impervious container for transport
3. Use gloves while handling all samples, especially when
there is contact with body fluids, non-intact skin or
mucous membrane.
4. If there is likelihood of spattering, use face mask with
glasses and gowns. Wrap around gowns should be
preferred. These should not be used outside the lab.
5. Cover cuts or abrasions present over skin with
waterproof bandage.

11. 6. Decontaminate the laboratory work surfaces immediately
in case of spillage of blood or any other body fluids
7. Follow ‘no needle recapping’ strategy
8. All sharps should be collected and disposed away
properly.
9. Never pipette by mouth. Use mechanical pipetting
devices.
10. There should always be a system working efficiently for
management of hospital generated waste.
11. It is advisable for the laboratory personnel to be
vaccinated against Hepatitis-B
12. Facilities should be available easily for post exposure
prophylaxis in case of exposure to HIV & HBV.

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

13. Do’s
•controlled access to
the laboratory
•Frequent hand wash
•Mechanical pipetting
•Appropriate waste
management &
Sterilization and
disinfection measures
•Training to the workers
Don’ts
•Eating, drinking,
smoking, handling
contact lenses,
•storing food
•Mouth pipetting

14. • - Primary barriers
• - Secondary barriers
• Primary barriers: Physical barriers or personal
protective equipments for lab worker
• Gloves, masks, Goggles, aprons, suits, special
breathing apparatuses

15. • structural aspects of the
laboratory that make
working environment
safer against infection
• Sinks for hand washing
• Special containment areas
• Special air ventilation
patterns
• Sterilization equipments

16. • Tuberculosis
• Rubella
• Hepatitis B
• Typhoid
• Cholera
• Anthrax
• Rabies
• Poliomyelitis
• Diphtheria

17. • Precautions to be taken by people researching or
trying to identify organisms
• Labs must adhere to these specific safety regulations
• A biosafety level is the level of the bio containment
precautions, required to undertake while handling
dangerous biological agents in an enclosed facility

18. • These are the guidelines developed to protect
workers in microbiological and medical labs
through a combination of safeguards which
include
-engineering controls
-management policies
-standard work practices.
• These BSL help you to understand why every lab
cannot perform tests for every organism

19. • Labs are divided into 4 biosafety levels; in which
protective practices increase with each level
• 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

20. BSL Laboratory type Laboratory practices Safety equipment
1 Basic teaching, research Good microbiological
techniques
None
Open bench work
2 Primary health
services; diagnostic
services, research
Good microbiological
techniques,
protective clothing,
biohazard sign
Open bench PLUS
biological safety cabinet for potential
aerosols
3 Special diagnostic
services, research
As BSL 2 PLUS
special clothing,
controlled access,
directional airflow
Biological safety cabinet and/or other
primary devices for all activities
4 Dangerous
pathogen units
As BSL 3 PLUS
airlock entry, shower exit,
Class III biological safety cabinet, positive
pressure suits,

21. BSL The Micro organisms Remarks
1 canine hepatitis virus, non-pathogenic Escherichia coli, other
non-infectious bacteria, B.subtilis
Minimal protection required
2 C. difficile, most Chlamydiae, hepatitis A, B, and C virus,
HIV, orthomyxoviruses (other than smallpox), influenza A,
Lyme disease, Salmonella, mumps, measles, scrapie, MRSA,
and VRSA, B. anthracis
Can cause only mild disease to
humans, or are difficult to contract
via aerosol in a lab setting
3 Yersinia pestis, Francisella tularensis, Leishmania donovani,
Mycobacterium tuberculosis, Chlamydia psittaci, West Nile
virus, Venezuelan equine encephalitis virus, Eastern equine
encephalitis virus, SARS coronavirus, Coxiella burnetii, Rift
Valley fever virus, Rickettsia rickettsii, several species of
Brucella, rabies virus, and yellow fever virus
Can cause serious or potentially
lethal disease after inhalation in
humans but for which treatments
DOES exist
4 Bolivian and Argentine hemorrhagic fevers, Marburg virus,
Ebola virus, Lassa virus, Crimean-Congo hemorrhagic fever,
and various other hemorrhagic diseases
Dangerous and exotic agents that
pose a high individual risk of
aerosol-transmitted laboratory
infections, these cause severe to
fatal disease in humans for which
vaccines or other treatments are
NOT available

22. • BSL1 - microorganisms
that don’t consistently
cause disease in healthy
adults
• E. coli , polyoma virus
• Basic laboratory
• Standard
Microbiological
Practices

23. • Standard practices required:
• frequent hand washing
• 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;

24. • 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

25. • 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

26. • 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

27. • 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 of working with
infectious agents and specific training for laboratory
personnel in handling these agents

28. • 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

30. • Also called as biological safety cabinet or
microbiological safety cabinet
• It is an enclosed, ventilated laboratory workspace for
safely working with materials contaminated with (or
potentially contaminated with) pathogens requiring a
defined biosafety level
• BSCs first became commercially available in 1950

33. CATEGORYCATEGORY TYPE OF WASTETYPE OF WASTE TREATMENT &TREATMENT &
DISPOSALDISPOSAL
Category 1Category 1 Human anatomical wastesHuman anatomical wastes Incineration/ deep burialIncineration/ deep burial
Category 2Category 2 Animal wastesAnimal wastes Incineration/ deep burialIncineration/ deep burial
Category 3Category 3 Microbiology & biotechnologyMicrobiology & biotechnology
waste,waste, Liquid wastes, wasteLiquid wastes, waste
from Laboratory, bloodfrom Laboratory, blood
banks, hospitals, house etc.banks, hospitals, house etc.
Local autoclaving/Local autoclaving/
microwaving/microwaving/
incineration/incineration/
Disinfection byDisinfection by
chemicalschemicals
Category 4Category 4 Waste sharps like needles,Waste sharps like needles,
syringes, scalpels, blades, glasssyringes, scalpels, blades, glass
etcetc
DisinfectionDisinfection
(Chemical/autoclaving/(Chemical/autoclaving/
micro waving &micro waving &
mutilation/shredding)mutilation/shredding)
Category 5Category 5 Discarded Medicines & cytotoxicDiscarded Medicines & cytotoxic
drugsdrugs
Incineration/ destructionIncineration/ destruction
& disposal in land fills& disposal in land fills

34. CATEGORYCATEGORY TYPE OF WASTETYPE OF WASTE TREATMENT &TREATMENT &
DISPOSALDISPOSAL
Category 6Category 6 Soiled wastesSoiled wastes
Items contaminated withItems contaminated with
blood, body fluids includingblood, body fluids including
cotton, dressings etccotton, dressings etc
Incineration,Incineration,
autoclaving,autoclaving,
microwavingmicrowaving
Category 7Category 7 Solid wastes like catheters, IVSolid wastes like catheters, IV
sets etc.sets etc.
Disinfection byDisinfection by
chemicalchemical
treatment/autoclavingtreatment/autoclaving
/micro waving and/micro waving and
mutilation &mutilation &
shreddingshredding
Category 8Category 8 Chemical wastesChemical wastes Chemical treatment &Chemical treatment &
discharge into drainsdischarge into drains
for liquid and securedfor liquid and secured
land fills for solids.land fills for solids.

35. • And these 8 categories are again sorted according to
special colour coded receptacles for waste of those
categories.
• The waste is collected in these colour coded containers
and processed further accordingly.
• The following table depicts this categorization.

36. Colour CodingColour Coding Type of container Type of container
to be usedto be used
Waste Category Waste Category
NumberNumber
TreatmentTreatment
YellowYellow Non Chlorinated Non Chlorinated
plastic bagsplastic bags
Category 1,2,5,6Category 1,2,5,6 IncinerationIncineration
RedRed Non Chlorinated Non Chlorinated
plastic plastic
bags/puncture proof bags/puncture proof
container for sharpscontainer for sharps
Category 3,4,7Category 3,4,7 Disinfection, Disinfection,
autoclave, autoclave,
microwave, microwave,
mutilation & mutilation &
shredding, shredding,
landfillinglandfilling
BlueBlue Non Chlorinated Non Chlorinated
plastic bags plastic bags
containercontainer
Category 8Category 8
BlackBlack Non Chlorinated Non Chlorinated
plastic bagsplastic bags
Municipal WasteMunicipal Waste

37. • BSL3 - 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

38. • Care of patients with tuberculosis starts with a quality assured
diagnosis, obtained by growing and identifying Myco bacte rium
tube rculo sis from clinical specimens and conducting DST of the
organism to confirm or exclude resistance. Uptake of TBdiagnostic
technologies requires appropriate laboratory infrastructure and
adequate policy reformat country level to enable theireffective use
in TBscreening and diagnostic algorithms
• Laboratory infrastructure, appropriate biosafety measures and
maintenance Equipment validation and maintenance Specimen
transport and referral mechanisms Management of laboratory
commodities and supplies Laboratory information and data
management systems Laboratory quality management system are a
priority.

39. • With growing incidences
of MDR-TB and XMDR-
TB it is highly essential all
Microbiology laboratories
must install Grade 3
Biosafety cabinets to
prevent exposure to
Infection. If necessary
precaution's are not taken
a fraction of Medical and
Technical personal will be
infected with grave
consequences.

40. • Pathogenic agents are
grouped in Risk group
3 which can cause
serious diseases
• Effective treatment
and preventive
measures are usually
available
• Little person-to-person
spread

41. • 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 prevent exposure

42. • Primary barriers:
• Similar to BSL-2 personal protective equipment
• Respiratory equipment
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

43. • BSL4 - microorganisms
that cause lethal disease,
with no known treatment
or vaccine
Ebola virus, Marburg
virus
Maximum
containment lab;
positive pressure
ventilated suits (moon
suits)

44. • Pathogenic agents are
grouped in Risk group
4 which is lethal
• Readily transmittable-
direct or indirect
• Effective treatment
and preventive
measures are not
usually available

45. • Dangerous and exotic agents with high risk of
life-threatening disease and are aerosol-
transmittion
• Related agents with unknown risk of
transmission
• Agents (all viruses) include Marburg virus,
Ebola virus, viruses that cause Congo-
Crimean hemorrhagic fever, Lassa fever

46. • 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
• Affected workers studying infected monkeys from Uganda

47. • 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:
• More strictly controlled access to the laboratory;
• Changing clothing before entering and exiting lab
(showering upon exiting recommended);
• Decontaminating all material exiting facility

48. • 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)

50. • 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

51. • All India Institute of Medical Sciences, New Delhi
• Microbial Containment Complex, Pune
• Centre for Cellular and Molecular Biology,
Hyderabad
• High Security Animal Disease Laboratory
(HSADL), Bhopal

52. • Four standard biosafety levels are also
described for activities involving infectious
disease work with commonly used
experimental animals.

53. • Acharya, D.B. and Singh, Thebook of hospital wastemanagement 2000
• World Health Organization. Laboratory biosafety manual. 3rd ed. Geneva;
2004
• http://www.who.int/injection_safety/toolbox/docs/AM_HCW_Safety.
pdf
• WHO guidelineson laboratory training for Field Epidemiologists
• "Biohazard Symbol History".theoriginal on July 16, 2011.
• Biosafety in Microbiological and Biomedical Laboratories, 5th Edition,
U.S. Department of Health and Human Services, Public Health
ServiceCentersfor DiseaseControl and Prevention National Institutesof
Health, HHSPublication No. (CDC) 21-1112Revised December 2009
• UNC School of Public Health Laboratory Safety Levels
• U.S. Centers for Disease Control and Prevention; U.S. National
Institutes of Health (2000). Prim ary Co ntainm e nt fo r Bio haz ards:
Se le ctio n, Installatio n and Use o f Bio lo g icalSafe ty Cabine ts . U.S.
Centers for Disease Control and Prevention.