RECOGNIZINGTHEBIOSAFETYLEVELS 
BiosafetyLevels 
AUTHORIZED PERSONNEL ONLY 
Prepared by : 
DebbraMarcel, Monoclonal Antibody Section, 
Veterinary Research Institute (VRI), Ipoh, Perak, Malaysia
OBJECTIVE 
Define the major characteristics of each stage in BiosafetyLevels. 
Relate the Risk Groups with the BiosafetyLevels. 
Prepared by : 
DebbraMarcel, Monoclonal Antibody Section, 
Veterinary Research Institute (VRI) 
Prepared by: 
MONOCLONAL ANTIBODY SECTION
WHYDOWENEEDTOHAVEKNOWLEDGEABOUTTHEBIOSAFETY LEVELS? 
Most of us are always dealing with a potentially harmful microbe. 
Precautions must be taken in the laboratory to make sure ourselves and others are not infected. 
Ask questions for better understanding… 
Where in the lab would you complete your work? 
What protective equipment and practices would you use? 
How would you contain the microbe to limit contamination or accidental infection? 
Prepared by: 
MONOCLONAL ANTIBODY SECTION
DEFINITION 
What is Biosafety? -the application of safety precautions that reduce a laboratorian’srisk of exposure to a potentially infectious microbe and limit contamination of the work environment and, ultimately, the community. 
What are BiosafetyLevels (BSLs)? -the level of safety from exposure to infectious agents; depends on work practices and safety equipment and facilities 
Prepared by: 
MONOCLONAL ANTIBODY SECTION
BIOSAFETY LEVEL SPECIFIC CONTAINMENT CONTROLS 
Each level are required for the following: 
1. Laboratory practices (esp. GLP) 
2. Safety equipment (e.gPPE) 
3. Facility construction (BSC) 
Prepared by: 
MONOCLONAL ANTIBODY SECTION
4LEVELS OF BIOSAFETY 
Low risk microbes 
High risk microbesEVERY LEVEL MUST FOLLOW THE STANDARD MICROBIOLOGICAL PRACTICES. 
Prepared by: 
MONOCLONAL ANTIBODY SECTION
STANDARDMICROBIOLOGICALPRACTICES 
1.Not eating, drinking, or applying cosmetics in the lab 
2.Washing hands after working with infectious materials and before leaving the lab 
3.Routinely decontaminating work surface 
Prepared by: 
MONOCLONAL ANTIBODY SECTION
LEVELS OF CONTAINMENT ? 
The primary risks: - 
1.Infectivity 
2.Severity of disease 
3.Transmissibility 
4.The nature of the work conducted 
5.Origin of the microbe/agent in question 
6.The root of exposure 
Prepared by: 
MONOCLONAL ANTIBODY SECTION
BSL-1 (BIOSAFETYLEVEL1) 
•Well-characterized agents with minimal potential hazard 
•Laboratories not necessarilyseparated from the general building 
•Work typically conducted on open bench tops 
•Specific training in the procedures conducted in the laboratory and must be supervised by a scientist with training in microbiology or a related science. 
•e.gnon-pathogenicE. Coli 
Prepared by: 
MONOCLONAL ANTIBODY SECTION
BSL-1 (BIOSAFETYLEVEL1) 
1. Laboratory practices 
Standard microbiological practices 
Work can be performed on an open lab bench or table. 
2.Safety equipment 
Personnel protection equipment (e.glab coats, gloves, eye protection) are worn as needed. 
3.Facility construction 
Not required special design on containment facility 
Laboratory doors to separate the working space with the rest of the facility. 
A sink for hand washing 
AB 
Prepared by: 
MONOCLONAL ANTIBODY SECTION
BSL-2 (BIOSAFETYLEVEL2) 
builds upon bsl-1. 
the microbes pose moderate hazardsto laboratoriansand the environment 
laboratory personnel have specific training in handling pathogenic agents 
access to the laboratory is restricted when work is being conducted 
personnel are supervised by scientists competent in handling infectious agents and associated procedures 
microbes are typically indigeneousand associated with diseases of varying severity (e.gStaphylococcus aureus) 
Prepared by: 
MONOCLONAL ANTIBODY SECTION
BSL-2 (BIOSAFETYLEVEL2) 
BSL-2 laboratories containment requirements: 
1.Laboratory practices 
Access to the laboratory is restricted when work is being conducted. 
2.Safety equipment 
Appropriate PPE is worn ( lab coats, gloves, eye protection, face shields, etc.) 
All procedures that can cause infection from aerosols performed in a biosafetycabinet (BSC) 
Autoclave/method of decontamination is available for proper disposals. 
3.Facility construction 
The laboratory has self-closing doors. 
A sink and eyewash are readily available. 
BA 
Prepared by: 
MONOCLONAL ANTIBODY SECTION
BIOHAZARD !!! 
•Mandatory Warning Sign 
•Designate BiosafetyLevel 
•Special Entry Procedures 
–Immunizations 
–PPE 
•Contact Information 
Prepared by: 
MONOCLONAL ANTIBODY SECTION
BSL-3 (BIOSAFETYLEVEL3) 
builds upon BSL-2. 
clinical, diagnostic, teaching, research, or production facilities. 
Work indigenous or exotic agents (serious or potentially lethal disease) ~ inhalation route exposure. 
Laboratory personnel MUST receive specific trainingin handling pathogenic & potentially lethal agents. 
Must be supervised by competent scientists. 
BSL-2 + procedures involving the manipulation of infectious materials must be conducted within BSCs. 
The microbes indigenous/exotic and can cause serious or potentially lethal disease through respiratory transmission. 
e.gMycobacterium tuberculosis, the bacteria that causes tuberculosis. 
Prepared by: 
MONOCLONAL ANTIBODY SECTION
BSL-3 (BIOSAFETYLEVEL3) 
1. Laboratory practices 
1.Under medical surveillance and might receive immunizations for microbes they work with. 
Access to the laboratory is restricted and controlled at all times. 
2.Safety equipment 
Appropriate PPE must be worn, and respirators might be required. 
All work with microbes must be within an appropriate BSC. 
3.Facility construction 
A hands-free sink and eyewash near the exit. 
The laboratory must have special engineering and design features directional airflow (drawing air from clean areas towards potentially contaminated areas). 
Entrance = two sets of self-closing and locking doors. A 
B 
C 
C 
Prepared by: 
MONOCLONAL ANTIBODY SECTION
BSL-4 (BIOSAFETYLEVEL4) 
builds upon BSL-3 (the highest level of biological safety) = very few. 
The microbes are dangerous and exotic, high risk of aerosol- transmitted infections (frequently fatal and without treatment or vaccines). 
Identical agents must be handled at BSL-4 until sufficient data obtained. 
All laboratory staff and supervisor must: 
1. Have specific and thorough training (handling extremely hazardous infectious agents). 
2. Understand the primary and secondary containment functions of standard and special practices, containment equipment, and laboratory design characteristics. 
3. Competent in handling agents and procedures requiring BSL-4 containment. 
e.gMarburgand Ebola viruses (high individual risk of life- threatening disease, aerosol transmission, or related agent with unknown risk of transmission). 
c 
Prepared by: 
MONOCLONAL ANTIBODY SECTION
BSL-4 (BIOSAFETYLEVEL4) 
1.Laboratory practices 
Change clothing before entering,shower upon exiting, decontaminate all materials before exiting. 2. Safety equipment 
•Must be performed within an appropriate Class III BSC, or by wearing a full body, air-supplied, positive pressuresuit. 
3.Facility construction 
The laboratory is isolated and restricted zone. 
The laboratory has dedicated supply and exhaust air, vacuum lines and decontamination systems. 
Access to the laboratory is controlled by the laboratory supervisor (Institutional Policies). 
Two types of laboratory providing absolute separation of the worker from the infectious agents = Suit Laboratory & Cabinet Laboratory 
A 
Prepared by: 
MONOCLONAL ANTIBODY SECTION
SUMMARY 
BIOSAFETY LEVEL 
LABORATORY TYPE 
LABORATORY PRACTICES 
SAFETY EQUIPMENT 
RISK GROUP 
EXAMPLE 
Basic: BSL-1 
Basic teaching, research 
GMT 
None; open bench work 
1 
canine hepatitis, non- pathogenic E. coli, some cell cultures & non- infectious bacteria 
Basic: BSL-2 
Primary health services; diagnostic services, research 
GMT plus protective clothing, biohazards sign 
Open bench plus BSC for aerosols 
2 
Staphylococcus aureus, Lyme disease, Salmonella, mumps, measles 
Containment: BSL-3 
Special diagnostic services, research 
Level 2 + special clothing, access control, directed airflow 
BSC and/or other primary devices for all activities 
3 
Mycobacterium tuberculosis, Brucellasp., rabies virus, Chikungunya 
Max. containment: BSL-4 
Dangerous pathogen units 
Level 3 + airlock entry, shower exit, special waste disposal 
Class III BSC, or positive pressure suites with class II BSCs, double ended autoclave 
4 
Marburg virus, Ebola virus, Nipahvirus 
Relation of biosafetylevels, practices and equipments to the risk group: - 
GMT= Good microbiological techniques ; BSC= Biosafetycabinet 
Prepared by: 
MONOCLONAL ANTIBODY SECTION
REFERENCES: 
CDC website: http://www.cdc.gov/ 
(e-learning, quick lessons, many interactive resources) 
Wikipedia: http://en.wikipedia.org/wiki/Biosafety_level 
Biosafetyin Microbiological and Biomedical Laboratories (BMBL) 5th Edition: e-book http://www.cdc.gov/biosafety/publications/bmbl5/index.htm 
Prepared by : 
DebbraMarcel, Monoclonal Antibody Section, 
Veterinary Research Institute (VRI)

Recognizing the Biosafety level

  • 1.
    RECOGNIZINGTHEBIOSAFETYLEVELS BiosafetyLevels AUTHORIZEDPERSONNEL ONLY Prepared by : DebbraMarcel, Monoclonal Antibody Section, Veterinary Research Institute (VRI), Ipoh, Perak, Malaysia
  • 2.
    OBJECTIVE Define themajor characteristics of each stage in BiosafetyLevels. Relate the Risk Groups with the BiosafetyLevels. Prepared by : DebbraMarcel, Monoclonal Antibody Section, Veterinary Research Institute (VRI) Prepared by: MONOCLONAL ANTIBODY SECTION
  • 3.
    WHYDOWENEEDTOHAVEKNOWLEDGEABOUTTHEBIOSAFETY LEVELS? Mostof us are always dealing with a potentially harmful microbe. Precautions must be taken in the laboratory to make sure ourselves and others are not infected. Ask questions for better understanding… Where in the lab would you complete your work? What protective equipment and practices would you use? How would you contain the microbe to limit contamination or accidental infection? Prepared by: MONOCLONAL ANTIBODY SECTION
  • 4.
    DEFINITION What isBiosafety? -the application of safety precautions that reduce a laboratorian’srisk of exposure to a potentially infectious microbe and limit contamination of the work environment and, ultimately, the community. What are BiosafetyLevels (BSLs)? -the level of safety from exposure to infectious agents; depends on work practices and safety equipment and facilities Prepared by: MONOCLONAL ANTIBODY SECTION
  • 5.
    BIOSAFETY LEVEL SPECIFICCONTAINMENT CONTROLS Each level are required for the following: 1. Laboratory practices (esp. GLP) 2. Safety equipment (e.gPPE) 3. Facility construction (BSC) Prepared by: MONOCLONAL ANTIBODY SECTION
  • 6.
    4LEVELS OF BIOSAFETY Low risk microbes High risk microbesEVERY LEVEL MUST FOLLOW THE STANDARD MICROBIOLOGICAL PRACTICES. Prepared by: MONOCLONAL ANTIBODY SECTION
  • 7.
    STANDARDMICROBIOLOGICALPRACTICES 1.Not eating,drinking, or applying cosmetics in the lab 2.Washing hands after working with infectious materials and before leaving the lab 3.Routinely decontaminating work surface Prepared by: MONOCLONAL ANTIBODY SECTION
  • 8.
    LEVELS OF CONTAINMENT? The primary risks: - 1.Infectivity 2.Severity of disease 3.Transmissibility 4.The nature of the work conducted 5.Origin of the microbe/agent in question 6.The root of exposure Prepared by: MONOCLONAL ANTIBODY SECTION
  • 9.
    BSL-1 (BIOSAFETYLEVEL1) •Well-characterizedagents with minimal potential hazard •Laboratories not necessarilyseparated from the general building •Work typically conducted on open bench tops •Specific training in the procedures conducted in the laboratory and must be supervised by a scientist with training in microbiology or a related science. •e.gnon-pathogenicE. Coli Prepared by: MONOCLONAL ANTIBODY SECTION
  • 10.
    BSL-1 (BIOSAFETYLEVEL1) 1.Laboratory practices Standard microbiological practices Work can be performed on an open lab bench or table. 2.Safety equipment Personnel protection equipment (e.glab coats, gloves, eye protection) are worn as needed. 3.Facility construction Not required special design on containment facility Laboratory doors to separate the working space with the rest of the facility. A sink for hand washing AB Prepared by: MONOCLONAL ANTIBODY SECTION
  • 11.
    BSL-2 (BIOSAFETYLEVEL2) buildsupon bsl-1. the microbes pose moderate hazardsto laboratoriansand the environment laboratory personnel have specific training in handling pathogenic agents access to the laboratory is restricted when work is being conducted personnel are supervised by scientists competent in handling infectious agents and associated procedures microbes are typically indigeneousand associated with diseases of varying severity (e.gStaphylococcus aureus) Prepared by: MONOCLONAL ANTIBODY SECTION
  • 12.
    BSL-2 (BIOSAFETYLEVEL2) BSL-2laboratories containment requirements: 1.Laboratory practices Access to the laboratory is restricted when work is being conducted. 2.Safety equipment Appropriate PPE is worn ( lab coats, gloves, eye protection, face shields, etc.) All procedures that can cause infection from aerosols performed in a biosafetycabinet (BSC) Autoclave/method of decontamination is available for proper disposals. 3.Facility construction The laboratory has self-closing doors. A sink and eyewash are readily available. BA Prepared by: MONOCLONAL ANTIBODY SECTION
  • 13.
    BIOHAZARD !!! •MandatoryWarning Sign •Designate BiosafetyLevel •Special Entry Procedures –Immunizations –PPE •Contact Information Prepared by: MONOCLONAL ANTIBODY SECTION
  • 14.
    BSL-3 (BIOSAFETYLEVEL3) buildsupon BSL-2. clinical, diagnostic, teaching, research, or production facilities. Work indigenous or exotic agents (serious or potentially lethal disease) ~ inhalation route exposure. Laboratory personnel MUST receive specific trainingin handling pathogenic & potentially lethal agents. Must be supervised by competent scientists. BSL-2 + procedures involving the manipulation of infectious materials must be conducted within BSCs. The microbes indigenous/exotic and can cause serious or potentially lethal disease through respiratory transmission. e.gMycobacterium tuberculosis, the bacteria that causes tuberculosis. Prepared by: MONOCLONAL ANTIBODY SECTION
  • 15.
    BSL-3 (BIOSAFETYLEVEL3) 1.Laboratory practices 1.Under medical surveillance and might receive immunizations for microbes they work with. Access to the laboratory is restricted and controlled at all times. 2.Safety equipment Appropriate PPE must be worn, and respirators might be required. All work with microbes must be within an appropriate BSC. 3.Facility construction A hands-free sink and eyewash near the exit. The laboratory must have special engineering and design features directional airflow (drawing air from clean areas towards potentially contaminated areas). Entrance = two sets of self-closing and locking doors. A B C C Prepared by: MONOCLONAL ANTIBODY SECTION
  • 16.
    BSL-4 (BIOSAFETYLEVEL4) buildsupon BSL-3 (the highest level of biological safety) = very few. The microbes are dangerous and exotic, high risk of aerosol- transmitted infections (frequently fatal and without treatment or vaccines). Identical agents must be handled at BSL-4 until sufficient data obtained. All laboratory staff and supervisor must: 1. Have specific and thorough training (handling extremely hazardous infectious agents). 2. Understand the primary and secondary containment functions of standard and special practices, containment equipment, and laboratory design characteristics. 3. Competent in handling agents and procedures requiring BSL-4 containment. e.gMarburgand Ebola viruses (high individual risk of life- threatening disease, aerosol transmission, or related agent with unknown risk of transmission). c Prepared by: MONOCLONAL ANTIBODY SECTION
  • 17.
    BSL-4 (BIOSAFETYLEVEL4) 1.Laboratorypractices Change clothing before entering,shower upon exiting, decontaminate all materials before exiting. 2. Safety equipment •Must be performed within an appropriate Class III BSC, or by wearing a full body, air-supplied, positive pressuresuit. 3.Facility construction The laboratory is isolated and restricted zone. The laboratory has dedicated supply and exhaust air, vacuum lines and decontamination systems. Access to the laboratory is controlled by the laboratory supervisor (Institutional Policies). Two types of laboratory providing absolute separation of the worker from the infectious agents = Suit Laboratory & Cabinet Laboratory A Prepared by: MONOCLONAL ANTIBODY SECTION
  • 18.
    SUMMARY BIOSAFETY LEVEL LABORATORY TYPE LABORATORY PRACTICES SAFETY EQUIPMENT RISK GROUP EXAMPLE Basic: BSL-1 Basic teaching, research GMT None; open bench work 1 canine hepatitis, non- pathogenic E. coli, some cell cultures & non- infectious bacteria Basic: BSL-2 Primary health services; diagnostic services, research GMT plus protective clothing, biohazards sign Open bench plus BSC for aerosols 2 Staphylococcus aureus, Lyme disease, Salmonella, mumps, measles Containment: BSL-3 Special diagnostic services, research Level 2 + special clothing, access control, directed airflow BSC and/or other primary devices for all activities 3 Mycobacterium tuberculosis, Brucellasp., rabies virus, Chikungunya Max. containment: BSL-4 Dangerous pathogen units Level 3 + airlock entry, shower exit, special waste disposal Class III BSC, or positive pressure suites with class II BSCs, double ended autoclave 4 Marburg virus, Ebola virus, Nipahvirus Relation of biosafetylevels, practices and equipments to the risk group: - GMT= Good microbiological techniques ; BSC= Biosafetycabinet Prepared by: MONOCLONAL ANTIBODY SECTION
  • 19.
    REFERENCES: CDC website:http://www.cdc.gov/ (e-learning, quick lessons, many interactive resources) Wikipedia: http://en.wikipedia.org/wiki/Biosafety_level Biosafetyin Microbiological and Biomedical Laboratories (BMBL) 5th Edition: e-book http://www.cdc.gov/biosafety/publications/bmbl5/index.htm Prepared by : DebbraMarcel, Monoclonal Antibody Section, Veterinary Research Institute (VRI)