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• 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
• 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
From-
•Bacteria
•Viruses
•Fungi
•Parasites
•Prions
•Recombinant DNA
Source-
•Various specimens
•Human blood
•Unfixed tissue
•Human cell lines
To-
•Lab personnel
•Community
Infections of special
concern
•Tuberculosis
•Hepatitis B
•HIV
•Enteric infections
Routes of infections
•Inoculation
•Ingestion
•Inhalation
• 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.
• 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.
• 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.
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.
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.
• NOT permitted in
laboratories:
Eating
Drinking
Storing food and drink
Smoking
Handling contact lenses
Pipetting by mouth
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
• - Primary barriers
• - Secondary barriers
• Primary barriers: Physical barriers or personal
protective equipments for lab worker
• Gloves, masks, Goggles, aprons, suits, special
breathing apparatuses
• 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
• Tuberculosis
• Rubella
• Hepatitis B
• Typhoid
• Cholera
• Anthrax
• Rabies
• Poliomyelitis
• Diphtheria
• 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
• 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
• 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
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,
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
• BSL1 - microorganisms
that don’t consistently
cause disease in healthy
adults
• E. coli , polyoma virus
• Basic laboratory
• Standard
Microbiological
Practices
• 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;
• 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
• 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
• 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
• 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
• 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
• 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
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
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.
• 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.
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
• 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
• 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.
• 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.
• 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
• 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
• 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
• 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)
• 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
• 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
• 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
• 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
• 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)
• 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
• 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
• Four standard biosafety levels are also
described for activities involving infectious
disease work with commonly used
experimental animals.
• 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.

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Biosafety in the laboratories

  • 1.
  • 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
  • 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.
  • 9.
  • 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
  • 29.
  • 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
  • 31.
  • 32.
  • 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)
  • 49.
  • 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.

Editor's Notes

  1. Biosafety is the section of Environmental Safety that addresses working safely in the laboratory with microorganisms and rDNA (organisms whose genetic material has been altered). Its goal is to help prevent lab-acquired infections while allowing researchers to work with organisms of various levels of hazard. This is done by advising researchers of the containment practices that are appropriate for the organisms they are handling, based on the level of risk presented by the organism.
  2. Laboratories have long recognized the hazards of processing infectious agents. In response to these hazards, guidelines have been developed to protect workers in microbiological and medical labs through a combination of safeguards including engineering controls, management policies and work practices. (1)  
  3. These are the basic practices when working with any microorganisms, and are the foundation for BL1 containment. They are designed to prevent transmission by contact, ingestion, and puncture. These practices are appropriate for ALL laboratories that work with radioisotopes and chemicals as well as microorganisms.
  4. In any laboratory, precautions must be taken so that the people researching or trying to identify organisms do not become infected themselves. According to the Centers for Disease Control and Prevention (CDC), scientists and lab technicians have to be very aware of microorganisms; while handling or testing clinical specimens, they could accidentally infect themselves or their coworkers. Because of this danger, labs must adhere to very specific safety regulations to work with organisms that pose a threat to human health. (1)  
  5. There are 4 levels of biological containment that allow laboratory researchers to work safely with infectious agents of various levels of risk. They are designed to prevent lab-associated infections. BL1 is the lowest level of containment. At UM there are many BL1 labs, most of which are molecular biology labs that use non-pathogenic strains of E. coli for cloning. .
  6. Only standard practices are required for laboratory work at this level. Standard practices include: frequent hand washing, especially after removing gloves and before leaving the laboratory; a door that can be kept closed when working; limits on access to the lab space when working; no smoking, eating, drinking, or storage of food in the laboratory; care to minimize splashes and actions that may create aerosols (tiny droplets); decontamination of work surfaces after every use and after any spills;
  7. 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 an insect and rodent control program; and use of personal protective equipment (such as lab coats, latex gloves, and eye protection or face shields as needed depending on the type of work being done). An open bench top sink is required for hand washing. No other major facility structures are required for BSL-1 labs.  
  8. In working with BSL-2 agents, the primary hazards to personnel are accidental needle sticks, potential infection through exposure to the eyes and nose (mucous membranes), and ingestion of infectious materials. BSL-2 agents do not cause lethal infections and are not transmissible via the airborne route. This means that they do not cause infection if tiny droplets of the material become airborne (i.e., aerosolized) and are inhaled, which might occur if the material were spattered. In addition, agents studied in a BSL-2 lab are pathogens for which immunization or antibiotic treatment is available. However, extreme care should still be taken with needles and sharp lab instruments when they are contaminated with agents.
  9. To reduce accidental infection, special procedures for BSL-2 labs include all standard practices for BSL-1 labs, plus a few others: special policies and procedures to restrict access to the lab when work is being conducted; biohazard warning signs posted outside the lab (see Figure 3); surveillance of laboratory personnel with appropriate immunizations offered; a biosafety manual that includes definitions of any needed waste decontamination or medical surveillance policies specific to the activities and agents in that lab; and supervisory staff who have experience in working with infectious agents and specific training for laboratory personnel in handling these agents. Some primary barriers in BSL-2 labs are biosafety cabinets or other approved containment devices. These areas minimize potential contamination while working with an agent, particularly if there may be splashes or aerosolization of infectious materials. Personal protective equipment includes lab coats, gloves, and face protection as needed when working with infectious agents. Protective clothing must be removed when personnel leave the laboratory area. Cabinets should be thoroughly decontaminated daily and if radioactive materials are used, monitored for radiation as a method of personal protection. Secondary barriers include all BSL-1 barriers, plus an autoclave (sterilization machine) for lab glassware.
  10. Some primary barriers in BSL-2 labs are biosafety cabinets or other approved containment devices. These areas minimize potential contamination while working with an agent, particularly if there may be splashes or aerosolization of infectious materials. Personal protective equipment includes lab coats, gloves, and face protection as needed when working with infectious agents. Protective clothing must be removed when personnel leave the laboratory area. Cabinets should be thoroughly decontaminated daily and if radioactive materials are used, monitored for radiation as a method of personal protection. Secondary barriers include all BSL-1 barriers, plus an autoclave (sterilization machine) for lab glassware.
  11. BL3 containment is used for work with infectious agents that are transmitted by inhalation. We have 2 BL3 labs at UM, but neither is currently in use.
  12. Established lab practices that must be in place in BSL-3 laboratories include all BSL-2 practices, 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 and decontaminating all lab clothing before laundering; and institutional policies regarding specimen (serum) collection and storage from lab workers to establish exposure to infectious agents.  
  13. Primary and secondary protective barriers in the BSL-3 lab emphasize protecting lab personnel, as well as personnel in nearby lab areas, the community, and the environment from exposure to potentially infectious aerosols. Primary barriers are similar to BSL-2 personal protective equipment, but may also include respiratory equipment if there is a risk of infection through inhalation. Secondary barriers at BSL-3 labs include all BSL-2 barriers, plus a few more sophisticated barriers. Corridors must be separated from direct access to the laboratory. Access must be through self-closing double doors. Air handling systems must be designed to ensure negative air flow, so that air around doors and windows flows into the laboratory rather than out of the laboratory. Air pumped into the laboratory is not re-circulated in the building. This measure is to prevent infectious aerosols from being carried outside the lab through the air.
  14. There are only a very few BL4 labs in the country: CDC in Atlanta; NIH and Ft. Detrick in MD. BL4 is used for work with the most hazardous microorganisms. Hot Zone described BL4 containment.
  15. Dangerous and exotic agents that pose high risk of life-threatening disease and aerosol-transmitted infections are found in BSL-4 labs. Related agents with an unknown risk of transmission are also studied in these labs. These agents pose a high risk of life-threatening disease, can be transmitted via the aerosol (respiratory) route, and have no available vaccine or therapy. All BSL-4 agents are viruses. Examples are Marburg virus, Ebola virus (see Figure 5), and viruses that cause Congo-Crimean hemorrhagic fever and Lassa fever.  
  16. Laboratory work with materials potentially infected with these agents, such as diagnostic samples or naturally and experimentally infected animals, pose a high risk of exposure and infection to laboratory personnel, and possibly even to the community and the environment. Personnel working with BSL-4 agents may be at risk of respiratory exposure to infectious aerosols, mucous membrane exposure to infectious droplets, and accidental needle sticks with needles or other sharp objects contaminated with infectious material. For example, in the late 1960s, there were 25 laboratory-acquired Marburg infections, including 5 deaths. These workers had been studying infected monkeys from Uganda. The first documented naturally-occurring human case occurred in 1975. (1)  
  17. Because of these risks, all lab personnel must receive specialized training in handling extremely dangerous infectious agents and in containment equipment and functions. Furthermore, access to the lab is restricted. Immunocompromised persons (including children and pregnant women) are never allowed to enter the lab. Laboratory practices for the BSL-4 include all BSL-3 practices, plus: strictly controlled access to the laboratory; changing clothing before entering and exiting the lab (showering upon exiting the lab is recommended); and decontaminating all material exiting the facility.  
  18. Primary barriers include conducting procedures in the biosafety cabinets used at the other biosafety levels in combination with a full-body, air-supplied, positive pressure personnel suit. Thus BSL-4 lab workers do not enter the lab unless they are wearing a “space suit” (see Figure 6). Secondary barriers in BSL-4 facilities include all the physical barriers at BSL-3 labs, plus: an isolated zone or a separate building; dedicated supply and exhaust, vacuum, and decontamination systems; and a recommended absence of windows—any windows must be sealed and resistant to breakage.  
  19. BSL-1 laboratories are fairly ubiquitous. Many are located in high schools, community colleges, and municipal drinking water treatment facilities. (1) BSL-2 facilities may be found in local health departments, universities, state laboratories, private laboratories (e.g., hospitals or health care systems), and industrial laboratories (e.g., clinical diagnostic companies). Most facilities that conduct infectious disease research have BSL-3 laboratories. They may be located in state health departments, universities, private companies, industry, and the federal government (e.g., the National Institutes of Health [NIH] and the Centers for Disease Control and Prevention [CDC]). In 2007, there were only 15 BSL-4 facilities in the United States: 9 federal facilities (including the CDC in Atlanta, GA, and the National Institutes of Health in Bethesda, MD), 4 university facilities (including Georgia State University in Atlanta, GA, and the University of Texas Medical Branch in Galveston, TX), 1 state facility, and 1 private facility. Renovations are underway at several of these labs, and new BSL-4 facilities are being proposed at additional sites. (4) Â