ABP Infectious Agents in Non-Rodents

1. Purpose and Scope

This Animal Biosafety Procedure (ABP) describes prudent practices, procedures, and equipment to reduce risk when inoculating infectious agents in non-rodent animal species that cannot be manipulated inside a biosafety cabinet (e.g., goats, cows, dogs, cats). The practices and procedures outlined in this document are by those described for Animal Biosafety Level 2 (ABSL-2) in the CDC/NIH Biosafety in Microbiological and Biomedical Laboratories (BMBL), 6th edition

2. Responsibilities

The Principal Investigator will ensure that personnel are made aware of the hazards associated with the infectious agent and that they receive training commensurate with their activities, including handling techniques for the particular animal species, before commencing ABSL-2 experiments. Personnel will comply with the safe work practices and procedures described within this Animal Biosafety Procedure.

3. Administrative Controls

• 3.1 Training

  • 3.1.1 The PI is responsible for provision of laboratory-specific training for safe manipulation of the infectious agent, symptoms associated with accidental exposure, and spill and exposure-response procedures. Document all training.
  • 3.1.2 Receive training in safe handling techniques for specific animal species.
  • 3.1.3 CARE and EHS will provide additional on-site training, as necessary.

• 3.2 Access and Signage

  • 3.2.1 Inform the facility manager before introducing infectious agents in animals.
  • 3.2.2 Review the hazards and potential risks of the experiment, and complete IACUC module 2 before accessing the animal facility.
  • 3.2.3 The facility supervisor will post a hazard sign at the entrance to the animal room. Research, EHS, and CARE staff will develop information contained in the sign, which will include:
    • The biohazard symbol and ABSL-2 designation
    • Name of the infectious agent, as well as human and animal clinical signs of infection
    • Potential shedding of the agent by the animal
    • Personal protective equipment
    • Disinfectant(s)
    • Contact information for CARE, EHS, and Cornell Health Services

• 3.3 Medical Surveillance

  • 3.3.1 Participate in the Animal Users Health and Safety Program (AUHSP).

4. Work Practice and Procedure Controls

• 4.1 Inoculation of animals

  • 4.1.1 See Section 6.1 for specific personal protective equipment.
  • 4.1.2 Use an appropriate manual restraint or physical restraint device. If the procedure or conditions of inoculation pose too high a risk with an awake animal (e.g., animal species difficult to handle, inexperienced individual performing the procedure), sedate the animal before inoculation.

• 4.2 Sharps Handling

  • 4.2.1 Substitute plasticware for glassware whenever possible, and implement the following practices described in the table:

    • Safe Practices for Handling Sharps
      Limit the use of sharps to when no other alternatives are available
      Keep all sharps in full view at all times
      Use only Luer-lock syringes and needles or units where the needle is integral to the syringe
      Implement safety-engineered sharps where practical (retractable needles, needle tip shields, self-sheathing scalpels, etc)
      Dispose of sharps directly, without manipulation (i.e., do not bend, shear, break, recap, or use hands to remove needles from syringes or blades from scalpels), in an approved sharps disposal container. Maintain disposal container in the animal room within arm’s reach
      Handle broken glass or other sharps with a secondary device such as forceps or broom and dustpan- not your hands

  • 4.2.2 Do not recap needles. However, if recapping must be done first receive approval by EHS and the IACUC, and use one of the following two methods: one-handed scoop technique; forceps, or tongs to place the cap on the needle.

• 4.3 Hygiene

  • 4.3.1 Eating, drinking, smoking, handling contact lenses, applying cosmetics, storing food for human consumption, and mouth pipetting are strictly prohibited in animal facilities.
  • 4.3.2 Wash hands thoroughly with soap and water after removing gloves. Use an alcohol-based hand sanitizer if a sink is not readily available.
    • For more information, see ACUP 713: Hygiene – Hand Washing

• 4.4 Decontamination and Spill Response

  • 4.4.1 Decontaminate work surfaces and equipment with a suitable disinfectant- allow at least 5-10 minutes of contact time. Suitable disinfectants must be active against the targeted agent, and address factors such as environment (e.g., organic load, surfaces) contact time, application, and safety:  The Center for Food Security & Public Health
  • 4.4.2 Cover spills of inoculum with absorbent towels/pads and saturate with disinfectant. Allow 5-10 minutes of contact time to achieve adequate disinfection. Appropriately segregate waste in red biohazard bags or sharps disposal containers and re-apply disinfectant to spill area.

• 4.5 Handling of Waste

  • 4.5.1 Dispose of sharps-related items (e.g., needles, syringes, blood tubes) directly in a sharps disposal container.
  • 4.5.2 Dispose of non-sharps items (e.g., gloves, intact plasticware) in a red biohazard bag.
  • 4.5.3 Treat any remaining inoculum with concentrated household bleach to a final volume of 10% bleach and allow at least 30 minutes contact time before disposal in the sanitary waste drain follow with copious amounts of water.
  • 4.5.4 Coordinate handling of infectious carcasses with animal care staff. Appropriate disposal may include packaging in red biohazard bags or covered carts.

• 4.6 Transport of Biohazardous Materials

  • 4.6.1 Transport infectious agents and contaminated samples between the laboratory and animal facility in a sealed, secondary container with absorbent toweling, and labeled with the biohazard symbol.

• 4.7 Tissue Harvest

  • 4.7.1 Arrange with animal care staff for transport of animals to an appropriate room or facility to euthanize animals and harvest tissues (e.g., necropsy floor at the Animal Health Diagnostic Center). Alternatively, perform euthanasia and tissue harvest in the animal room.
  • 4.7.2 When possible, use only one sharp item (e.g., knife, scissors) at a time and keep it in full view.
  • 4.7.3 Place any harvested tissue or fluids in appropriate primary containers (e.g., screw-top vial, sealable plastic bag), decontaminate exterior of the container, and transport as per section 4.6. Fixed tissues (e.g., 10% buffered formalin) are no longer considered biohazardous. Use appropriate personal protective equipment when handling these samples and transport them in a secondary container.
  • 4.7.4 Follow the sharps handling practices outlined in section 4.2.

5. Housing and Handling of Infected Animals

• 5.1 House animals in a holding area with an adjacent anteroom. Prepare inoculum and supplies, and don personal protective equipment (as indicated in section 6) within the anteroom.

• 5.2 When possible, have someone assist you when inoculating, euthanizing animals, or collecting samples.

6. Personal Protective Equipment (PPE)

• 6.1 Don the following minimum PPE before entering ABSL-2 animal rooms:

  • Disposable fluid-resistant coveralls
  • Disposable gloves (nitrile- avoid latex when possible) - Use double gloves when handling the infectious agent or when inoculating animals. The outer glove should overlay the cuff of the coverall
  • Boots or shoe covers

• 6.2 Wear additional PPE when indicated by the agent, animal species, hazards, or experimental conditions:

  • Face mask and eye protection (e.g., safety glasses or face shield)
  • Hairnet
  • Cut/bite-resistant gloves
  • Respiratory protection

• 6.3 Solid toed shoes are required for entry into animal rooms.

• 6.4 Remove PPE upon exiting the animal room and dispose of it in a red biohazard bag. First remove outer gloves, gown (turning inside out), hair net, facial protection, boots or shoe covers while stepping out of the room (step-over technique), and finally inner gloves.

  • For more information, see ACUP 715: Personal Protective Equipment

7. Response to Accidental Exposures

• 7.1 Personnel who sustain an overt exposure such as a splash to mucous membranes, direct contact with open wounds, or a sharps injury should: 

  • Wash exposed area with soap and water or rinse in eyewash for at least 5 minutes
  • Perform first aid, if applicable
  • Notify supervisor
  • Seek medical evaluation at Cornell Health Services, Occupational Medicine (607-255-6960) as soon as possible after exposure or injury. After hours seek evaluation at Cayuga Medical Center.
  • Contact Cornell Health Services, Occupational Medicine if you develop symptoms suggestive of exposure to the hazardous agent.
  • Document exposures, injuries, and illnesses in the Cornell University Injury/Illness/Exposure Report, Incident Reporting.
    • For more information, see ACUP 707: Animal Related Injury

8. Emergency Phone Numbers

• Police, Fire, and Medical Emergencies: calling 911 from an on-campus phone or by calling 607-255-1111 from a cellular phone
• Environment, Health and Safety (EHS): 607-255-8200 (off-hours 607-255-1111)
• Cornell Health Services, Occupational Medicine: 607-255-6960 (off-hours 607-255-5155)
• Cornell Animal Resources and Education (CARE): 607-253-4378 (off-hours 1-800-349-2456 for veterinary medical emergencies)