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Biological Laboratory Safety. REHS. Rutgers Environmental Health and Safety REHS supports Rutgers University by providing comprehensive and professional health, safety and environmental services to the University community. Protect Rutgers employees, students and visitors.
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REHS Rutgers Environmental Health and Safety • REHS supports Rutgers University by providing comprehensive and professional health, safety and environmental services to the University community. • Protect Rutgers employees, students and visitors. • REHS ensures compliance with government regulations
Regulatory Agencies • The safety standards and practices described here have been developed and are regulated by government agencies including • CDC (Center for Disease Control) • NIH (National Institutes of Health)
Agenda • Lab Safety • Biohazards • Biosafety Levels • Biosafety Cabinets
Lab Safety • Safe Laboratory Practices • Lab coats • Safety glasses • Proper footwear • Hair back • No food or drink in the laboratory
Biohazard • An agent of biological origin that can cause disease in humans • Microorganism • Toxin • Allergen
Biosafety The combined use of • laboratory practices, • laboratory facilities and • safety equipment to work with potentially infectious microorganisms.
Why use biosafety practices? To protect: • Workers/Students • Products/Experimental results • Environment/Laboratory classroom
Biosafety Levels • BL-1: agents are not known to cause disease • BL-2: agents are associated with human disease • BL-3: agents are associated with human disease and are potentially transmitted as aerosols • BL-4: agents of life threatening nature
Biosafety Level 1 (BL-1) Use BL-1 when working with: • Well characterized agents • Agents that are not known to cause disease in health humans • Agents that are of minimal hazard to lab personnel and the environment Examples of BL-1 Agents: • E. coli JM109, DH5a • Saccharomyces cerevisiae
Biosafety Level 1 (BL-1) Standard Work Practices • Use mechanical pipetting devices • Wash hands frequently • Minimize splashes and aerosols • Decontaminate work surfaces daily • Handle wastes properly • Maintain insect and rodent control program
Biosafety Level 1 (BL-1) Personal Protective Equipment (PPE) • Lab coat or apron • Safety glasses or goggles • Gloves as needed
Biosafety Level 2 (BL-2) Use BL-2 practices when working with: • Agents of moderate potential hazard to personnel and the environment Examples of BL-2 agents: • Human blood or body fluids • E. coli 0157:H7 • Clostridium botulinum • Retroviral vectors • Human cells in cell culture
Biosafety Level 2 (BL-2) • Adequate illumination • Eyewash facility • Negative air pressure • Autoclave available • Biological safety cabinet • Lab must be separated from public areas
Biosafety Level 2 (BL-2) Standard Work Practices • Use mechanical pipetting devices • Wash hands frequently • Minimize splashes and aerosols • Decontaminate work surfaces daily • Handle wastes properly • Maintain insect and rodent control program
Biosafety Level 2 (BL-2) Special Practices • Place used slides and coverslips in sharps containers, never in any other receptacle. • Sharps containers are: • Red in color • Marked with the biohazard symbol • Puncture resistant • Leak proof
Biosafety Level 2 (BL-2) Personal Protective Equipment (PPE) • Lab coat or apron • Safety glasses or goggles • Gloves • Biosafety cabinet • Aerosols or splashes • Large volumes • High concentrations
Biosafety Cabinets (BSCs) • Provide product, personal and environmental protection. • Various classes of BSCs are available • Amount of air exhausted • Amount of employee protection
Biosafety Cabinets (BSCs) • Sterile air is exhausted from BSCs through a HEPA filter. • HEPA filter can trap particles to 0.3 microns • Chemical vapors and fumes pass through HEPA filters
Biosafety Cabinets (BSCs) Helpful Hints • Enter straight into the cabinet, avoid sweeping motions • Don’t place materials on the grill • Keep discard pan or bag inside the cabinet • Decon materials before removal from the cabinet
Decontamination • Sterilization: destroy all microbial life, including spores • Disinfection: destroy a majority of microbial life, but not necessarily spores • Antiseptic: germistat used on skin to inhibit growth of microorganisms
Decontamination Agent Selection • Degree of killing required • Nature of item to be treated • Ease of use • Safety • Cost • Type and amount of organism
Decontamination Methods • Heat: steam heat, dry heat, incineration • Chemical: bleach, ethanol, hydrogen peroxide, ethylene oxide, paraformaldehyde • Radiation
Decontamination Disinfectants do not replace standard microbiological practices and good hygiene!
Biological Waste Types • Cultures, stocks • Sharps • Pipettes, tips and weigh boats • Other materials
Biological Waste BL-1 Waste • Solids • Collect and package in clear autoclave bags • Autoclave to sterilize • Dispose of in regular trash dumpster (as long as no chemical or radioactive contaminants are present) • Liquids • Collect in containers with lids • Autoclave or treat with 10% bleach to sterilize • Dispose of down the drain (as long as no chemical or radioactive contaminants are present)
Biological Waste Sharps (needles, syringes, scalpel blades, slides, blood vials, pasteur pipettes) • Collect in approved sharps container • Autoclave to sterilize • Dispose of in medical waste boxes
Biosafety and REHS • Protocol Approval • REHS provisionally approves experimental protocols. • All protocols must be approved if they involve: • recombinant DNA, • non-recombinant human, animal or plant pathogens, • human cell culture, • human tissue/blood experiments • Laboratory inspections • RU Biosafety Committee provides final approval to protocols