Biological Laboratory Safety

REHSRutgers 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)

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 WasteBL-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