Laboratory Biosafety Lecture Notes PDF

Laboratory Biosafety 1 laboratory Biosafety Definition: Application of safety principles to a laboratory in which biological organisms are manipulated Protection: workers. products. environment 2 :General Lab Requirements Knowledgeable supervisor Knowle...

Laboratory Biosafety 1 laboratory Biosafety Definition: Application of safety principles to a laboratory in which biological organisms are manipulated Protection: workers. products. environment 2 :General Lab Requirements Knowledgeable supervisor Knowledgeable personnel – Aware of potential hazards – Proficient in practices & – techniques Lab specific biosafety manual Basic Laboratory Biosafety 1.Biohazard warning sign for laboratory doors 2. only authorized personnels should be allowed to enter laboratory working areas. 3. laboratory doors should be kept closed. 4.children shouldn't be authorized to enter lab areas. 5. Animals should not be admitted other than those involved in the work of the laboratory. 4 Personnel protection 1. Laboratory coat, gowns, overshoes, should be worn at all the times for work in lab. 2. Gloves should be worn while handling blood or any body fluids. 3. Safety glasses(Goggles) should be used to protect eyes from splashes and artificial sources of UV 4. Opened toe (sandals) must not be worn in laboratories 5 Personnel protection 5. Eating, drinking, smoking are prohibited in the laboratory areas 6.Storing human foods or drinks anywhere in the laboratory working areas is prohibited 7. Lab coat should not be stored in the same locker with street coat. 8. Immunization program. Standard Practices in All LAB.Levels 1. Pipetting by mouth must be strictly forbidden. 2. All technical procedures should be performed in a way to prevent aerosols or droplets, handle live agents inside biosafety cabinet. 3. Limiting use of needles not to substitute pipettes, substitute plastic to glass. 4. All spills, accidents, exposure to infectious materials should be reported and signed by the supervisor. 7 Standard Practices in All LAB.Levels 5. Read the procedure of cleaning up of all spills and follow to use proper disinfectant. 6 Contaminated liquids should be decontaminated before discharge to sanitary sewer. 7. Wear applicable Personal Protection Equipment. 8. Reported documents that are removed from the laboratory should be protected from contamination.  Biosafety levels Four Biosafety levels provide increasing degrees of protection (BSL-1, 2, 3, 4) 9  Biosafety level 1 Suitable for work involving agents not known to cause disease in healthy humans and of minimal potential hazard to laboratory personnel and environment. Ex. Bacillus Subtilis E.coli 10 Biosafety level 1 Safety Equipment (Primary Barriers) biological safety cabinet are generally not required for Biosafety Level 1. laboratory coats, gowns, or uniforms should be worn to prevent contamination. Gloves should be worn if the skin on the hands is broken or if a rash is present. Protective eyewear should be worn for splashes of microorganisms or other hazardous materials. Biosafety level 1 Laboratory Facilities (Secondary Barriers) Laboratories should have doors for access control. Each laboratory contains a sink for handwashing. The laboratory is designed so that it can be easily cleaned. Bench tops are resistant to water and moderate heat and the organic solvents, acids, alkalis, and chemicals. Spaces between benches, cabinets, and equipment are accessible for cleaning.  Biosafety level 2 Suitable for work involving agents of moderate potential hazard to personnel and the environment. Ex. Measles virus - HBV Salmonella Biosafety level 2 Safety Equipment (Primary Barriers) : 1. Properly maintained biological safety cabinets, preferably Class II. 2. Sealed rotor heads or centrifuge safety cups are used, 3. Face protection (goggles, mask, face shield or other splatter guard) is used. 5. Protective laboratory coats, gowns, or uniforms designated for lab use are worn while in the laboratory. 6. Gloves are worn when hands may contact potentially infectious materials, contaminated surfaces or equipment. Biosafety level 2 Laboratory Facilities (Secondary Barriers): 1. Install biological safety cabinets in such a manner that fluctuations of the room supply and exhaust air do not cause the biological safety cabinets to operate outside their parameters for containment. 2.Locate biological safety cabinets away from doors, from windows that can be opened, and from other potentially disruptive equipment. 3. An eyewash station is readily available. 4. Illumination is adequate for all activities.  Biosafety levels 3 Suitable for work with infectious agents which cause serious or potentially lethal disease as a result of exposure by the inhalation route. Ex. TB - HIV 16 Biosafety levels 3 Safety Equipment (Primary Barriers) : 1. Protective laboratory clothing such as solid-front or wrap- around gowns, scrub suits, or coveralls should be worn. 2. Gloves must be worn when handling infectious materials, infected animals, and when handling contaminated equipment. 3. Disposable gloves are not reused. 4. All manipulations of infectious materials, harvesting of tissues or fluids are conducted in a Class II or Class III biological safety cabinet. 5. Respiratory and face protection are used when in rooms containing infected animals. Biosafety levels 3 Laboratory Facilities (Secondary Barriers) 1.The laboratory is separated and access to the laboratory is restricted. Passage through a series of two self-closing doors is the basic requirement. 2. The interior surfaces of walls, floors, and ceilings of areas where BSL-3 agents are handled are constructed for easy cleaning and decontamination. 3.All windows in the laboratory are closed and sealed. 4. A method for decontaminating all laboratory wastes is available (i.e., autoclave, chemical disinfection, incineration). Biosafety levels 3 5. Biological safety cabinets are required and are located away from doors. 6. A ducted exhaust air ventilation system is provided. 7. Continuous any equipment that may produce aerosols are contained in devices that exhaust air through HEPA filters before discharge into the laboratory. 8. Facilities should be re-verified, at least annually, against these procedures as modified by operational experience. BSL-3 Gowning Area BSL-3 Practices  Biosafety levels 4 Suitable for work with dangerous agents that pose a high individual risk of aerosol transmitted laboratory infectious and life threatening disease. Ex. Ebola Zaire virus - Rift valley fever virus 22 Biosafety levels 4 Safety Equipment (Primary Barriers): All procedures within the facility are conducted in the Class III biological safety cabinet. Biosafety levels 4 Laboratory Facility (Secondary Barriers): 1.Either a separate building or a clearly isolated zone within a building. 2. Walls, floors, and ceilings of the cabinet room and inner change room are constructed to form a sealed internal shell which facilitates fumigation and is resistant to entry and exit of animals and insects. 3. Automatically operated hand washing sink is provided. 4. Access doors to the laboratory are self-closing and lockable. Biosafety levels 4 5.Any windows are breakage-resistant and sealed. 6. A dedicated non-recirculating ventilation system is provided. 7. The supply air to and exhaust air from the cabinet room, inner change room, and anteroom pass through HEPA filters. 8. The facility must be tested for verification that the design and operational parameters have been met prior to operation. 9. Appropriate communication systems are provided between the laboratory and the outside (e.g., voice, fax, computer).  References Biosafety in Microbiological and Biomedical Laboratories”,5th Ed nationmaster.com/encyclopedia/Biosafety bsafe.us researchcompliance.uc.edu National Institutes of Health Bethesda, MD 20892 http://www.cdc.gov/OD/ohs/pdffiles/bsl123. http://bmbl.od.nih.gov/sect3bsl4.htm Thank you