Biosafety Practices and Procedures

Safe Practices and Procedures: BSL 1 | BSL 2 | ABSL 1 | ABSL 2 | Field Safety

The practices and procedures used are selected based on the risk assessment. Signs outside the laboratory doors indicate the biosafety risk level and include the laboratory supervisor’s name and phone number.

Biosafety Level I

Laboratory coats or gowns are recommended. Non-latex gloves should be worn if the skin on the hands is broken or a rash is present. Goggles or faceshields should be used if splashes of microorganisms are likely.

General Practices:

  1. During periods of active manipulation of biologically active materials, access to the laboratory is limited or restricted. Doors to the laboratory are kept closed.
  2. Hands are washed after viable materials are handled and before leaving the laboratory.
  3. No eating, drinking, applying cosmetics, or handling of contact lenses is permitted in the work area. Contact lens wearers should wear goggles or faceshields.
  4. Food for human consumption cannot be stored in any laboratory refrigerator or freezer.
  5. No mouth pipetting.
  6. Sharps are disposed appropriately. Biohazardous sharps containers are available for use in each laboratory.
  7. Work practices are designed to minimize aerosols and splashes.
  8. Work surfaces are decontaminated at least once a day during active use of a laboratory and after the spill of any viable materials. See Appendix III for a discussion of appropriate disinfectants.
  9. All cultures, stocks and regulated waste are decontaminated by an autoclave or appropriate chemical decontaminates.
    • Tissue culture waste (culture media) can be inactivated by placing it in a solution of 10,000 ppm hypochlorite for 8 hours. Sewer with copious water.
    • Tissue-culture contaminated pipettes can be placed in a solution of 2500 ppm hypochlorite for 8 hours before disposal by autoclaving or incineration.

Biosafety Level II

The “General Practices” outlined for Biosafety Level I also apply to the increased risk setting of Biosafety Level II. Additionally, special practices are applied to ensure the increased risk does not result in injury or illness to students, faculty and staff. Bloodborne pathogen protocols supercede Biosafety Level II activities when human blood or body fluids are present.

Special Practices

  1. Individuals entering the laboratory are advised of the risks. Biosafety procedures are incorporated into the standard operating procedures for the laboratory, and personnel are required to read and follow these procedures.
  2. Traffic flow or movement is restricted when a biological safety cabinet is in use.
  3. Procedures that are likely to produce an aerosol or splash should be conducted in a biological safety cabinet or if that is not feasible a faceshield should be worn. Examples of activities that might produce aerosols or splashes include. centrifugation, grinding, blending, vigorous shaking or mixing, sonic disruption, opening containers of infectious materials whose internal pressures might be different than ambient pressures, and harvesting infected tissues from animals or embryonate eggs.
  4. Biological safety cabinets should be used if either manipulating high concentrations of infectious material or large quantities.
  5. Protective clothing is either disposed in the laboratory or laundered by the institution. It is never taken out of the laboratory by individuals.
  6. Gloves are worn when hands might encounter potentially infectious materials, contaminated surfaces or equipment. Hands are washed following removal of gloves.
  7. Contaminated sharps are handled with extreme caution:
    • Use of needles and other sharp objects is discouraged unless absolutely necessary for the procedure in question.
    • Do not recap needles—discard the needle and syringe in a biohazard sharps container.
    • Broken glassware is never cleaned up by hand. Instead dustpans and brushes are used to collect the broken glass and it is disposed in a biohazard sharps container.
  8. Materials contaminated with human body fluids are decontaminated by autoclaving and placed in a container with a cover that prevents leakage during storage, transport or shipping. Other potentially infectious waste is placed in a container with a cover that prevents leakage before decontamination by autoclave.
  9. Laboratory equipment and surfaces are decontaminated routinely after work with infectious materials is completed and especially after any spill.
  10. Animals not involved in the work are not allowed into the laboratory.
  11. Safety training including identification of risks, safe handling and emergency procedures is required for all employees.
  12. Medical surveillance:
    • Spills that might results in overt exposure are reported in the appropriate Accident/Injury form and medical evaluation, surveillance and treatment are provided. Records of exposures are maintained.
    • Individuals at higher risk for infection (e.g. immunosuppression) are not allowed in the laboratory. The responsibility for assessing each circumstance and establishing policies lies with the laboratory supervisor.
    • The laboratory supervisor is also responsible for determining whether appropriate immunizations or tests for the agents handled are available.

Animal Biosafety Level I

This policy only applies to animals being deliberately infected with biologically active agents. Facilities for laboratory animals are physically separate from other activities whenever possible. Lab coats and gowns are recommended. Lab coats must remain in the animal rooms. Disposable gowns are not worn outside of the animal suite. The animal suite which houses permanent colonies of animals is restricted. Its external doors are self-closing and locking and doors to animal rooms are kept closed when experimental animals are present. Animal rooms have negative pressure compared to hallways. Hand washing sinks are available in the animal suite and cages are washed manually.

General Practices

  1. The laboratory director establishes polices and procedures for emergency situations.
  2. Only authorized personnel advised of the risks are allowed to enter the animal suite.
  3. No eating, drinking, smoking, applying of cosmetics, handling contact lenses, or storage of human food is allowed in the animal suite.
  4. Procedures are designed to minimize aerosols and spills.
  5. Work surfaces are decontaminated after use or after a spill of viable material.
  6. All wastes (including unpreserved animal tissue, carcasses and contaminated bedding) are transported from the animal suite in leak-proof containers and disposed appropriately.
  7. Sharps are properly disposed.
  8. Employees wash their hands after handling animals, after removing gloves and before exiting the animal suite.

Animal Biosafety Level II

This policy only applies to animals being deliberately infected with biologically active agents and animals captured in the wild. The General Practices of Animal Biosafety Level I are utilized as well as the special practices listed below. Appropriate eye and face protection are used if risk assessment indicates that would be prudent. Biological safety cabinets, other physical containment devices, or personal protective equipment are used when the procedure has a high risk for creating aerosols.

Special practices

  1. Access to the animal suite is restricted to the fewest number of people possible. Individuals entering the room are advised of the risks.
  2. An appropriate medical surveillance program is in place. Vaccinations are up-to-date.
  3. Needles, syringes or other sharp instruments are used only when no other viable alternatives exist. Needles are not recapped. Used needles and syringes are placed in biohazard sharps containers.
  4. Employees receive appropriate training on the hazards, safe handling and emergency procedures. Training records are maintained. Individuals at increased risk for infection are not allowed in the animal suite unless special precautions are taken to protect them.
  5. All equipment must be decontaminated before removal from the room.
  6. Spills and accidents that expose individuals to infectious materials are reported in the appropriate Accident/Injury form. Medical evaluations, surveillance and treatment are provided as needed.

Biosafety and Field Work

Wild animals pose special hazards. Latent infections are more common in field-collected animals, necessitating Animal Biosafety Level II practices. Some animals normally possess bacteria that are pathogenic in humans. The laboratory supervisor is responsible for assessing the risk and taking appropriate protective measures. For example, many amphibians and reptiles carry Salmonella spp as part of their normal flora. Precautions to be taken include providing for hand-washing or hand-decontamination procedures in the field. Hand-decontamination procedures should consider the susceptibility of the organisms potentially present. For example, Purell© is not effective against Salmonella. However, Sani-dex© Antimicrobial Hand Wipes containing 40% alcohol are effective against gram negative organisms and are routinely supplied in field safety kits. In Table A, some organisms that might be encountered only during field work are included to assist laboratory supervisors in identifying risks and protective measures.

During field work, mosquito and tick bites can expose individuals to serious disease. Employees are strongly advised to wear appropriate protective clothing and use insect repellent. Upon return, employees should examine themselves for deer ticks. If a deer tick is attached, it should be carefully removed and frozen, and dated with the employees’ names. They should complete the Accident/Injury form and check the “incident” box and submit the form. Thus if symptoms later occur, this form will be on-file.