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. 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 Dont 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