Biological Laboratory Safety

1. Biological Laboratory Safety

2. 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

3. 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)

4. Agenda • Lab Safety • Biohazards • Biosafety Levels • Biosafety Cabinets

5. Lab Safety • Safe Laboratory Practices • Lab coats • Safety glasses • Proper footwear • Hair back • No food or drink in the laboratory

6. Biohazard • An agent of biological origin that can cause disease in humans • Microorganism • Toxin • Allergen

7. Biosafety The combined use of • laboratory practices, • laboratory facilities and • safety equipment to work with potentially infectious microorganisms.

8. Why use biosafety practices? To protect: • Workers/Students • Products/Experimental results • Environment/Laboratory classroom

9. 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

10. 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

11. 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

12. Biosafety Level 1 (BL-1) Personal Protective Equipment (PPE) • Lab coat or apron • Safety glasses or goggles • Gloves as needed

13. 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

14. Biosafety Level 2 (BL-2) • Adequate illumination • Eyewash facility • Negative air pressure • Autoclave available • Biological safety cabinet • Lab must be separated from public areas

15. 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

16. 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

17. 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

18. Biosafety Cabinets (BSCs) • Provide product, personal and environmental protection. • Various classes of BSCs are available • Amount of air exhausted • Amount of employee protection

19. Biosafety Cabinets (BSCs)

20. 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

21. 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

22. 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

23. Decontamination Agent Selection • Degree of killing required • Nature of item to be treated • Ease of use • Safety • Cost • Type and amount of organism

24. Decontamination Methods • Heat: steam heat, dry heat, incineration • Chemical: bleach, ethanol, hydrogen peroxide, ethylene oxide, paraformaldehyde • Radiation

25. Decontamination Disinfectants do not replace standard microbiological practices and good hygiene!

26. Biological Waste Types • Cultures, stocks • Sharps • Pipettes, tips and weigh boats • Other materials

27. 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)

28. 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

29. 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