ABP Infectious Agents in Rodents

1. Purpose and Scope

This Animal Biosafety Procedure (ABP) describes prudent practices, procedures, and equipment to reduce risk when inoculating infectious agents in rodents at Animal Biosafety Level 2 (ABSL-2). The practices and procedures follow those described 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 before commencing ABSL-2 experiments. In addition, 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 Receive laboratory-specific training for safe manipulation of the infectious agent, symptoms associated with accidental exposure, and spill and exposure response procedures.
  • 3.1.2 Complete the ABSL-2 online training module and CARE training for handling rodents.
  • 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 the infectious agent in rodents.
  • 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 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.

• 3.2.4 Post a hazard ID card on cages that contain infected animals. The card will include the biohazard symbol, the name of the infectious agent, and the date of inoculation.

• 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 Sections 5.1, engineering controls, and 6.1, personal protective equipment (PPE).
  • 4.1.2 Use an appropriate manual or physical restraint device. If the procedure or inoculation conditions pose too high a risk with an awake animal (e.g., retro-orbital injections, inexperienced individual performing the procedure), sedate the animal before inoculations.
  • 4.1.3 Use a disinfectant-soaked cloth to wipe away excess inoculum leaking from the inoculation site.

• 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 (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. 
    • If recapping is required, receive approval from EHS and the IACUC. Once approved, only use one of the following two methods: one-handed scoop technique, forceps, or tongs to place the cap on the needle.
  • 4.2.3 If researchers must temporarily set down a needle/scalpel during a procedure, set the sharp end inside a clean 50-mL conical tube or equivalent to act as a protective sheath (rather than recapping) and prevent injury.

• 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 soap and water are not readily available.
  • 4.3.3 If working long hours in a rodent room, consider taking a full-body shower to reduce the amount of potential allergens present on your body.

    • For more information, see ACUP 713: Hygiene – Hand Washing.

• 4.4 Decontamination and Spill Response

  • 4.4.1 Decontaminate work surfaces and equipment (e.g., inside of biosafety cabinet, animal cages) with a suitable disinfectant- allow at least 5-10 minutes of contact time. Appropriate 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 and Public Health
  • 4.4.2 Cover spills with absorbent towels/pads and saturate with disinfectant. Allow at least 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 Inside a biosafety cabinet, place a water-soaked paper towel in the dirty cage to generate steam when autoclaved or leave a water bottle in the cage. Wipe exterior of the cage with disinfectant before removing from biosafety cabinet.
  • 4.5.2 Dispose of sharps-related items (e.g., needles, syringes, Pasteur pipettes, blood tubes) directly in a sharps disposal container.
  • 4.5.3 Dispose of non-sharps items (e.g., gloves, intact plasticware) in a red biohazard bag.
  • 4.5.4 Treat infectious liquid waste 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.
  • 4.5.5 Place carcasses (no gloves, plastic, etc.) in compostable bags suitable for disposal in the Waste Management digester. Wipe bags with an appropriate disinfectant and store all bags in a larger biohazard bag or biohazard-labeled drawer in the refrigerator. Alternatively, roll up carcasses in bench diapers and place them directly in biohazard bags.
  • 4.5.6 Animal care staff will dispose of waste and carcasses unless other arrangements are made.

• 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 Perform tissue harvest in a certified class II biosafety cabinet- use a tray or bench diaper to collect fluids. Preferably, use tape instead of pins to secure carcasses.
  • 4.7.2 When possible, use only one sharps item (e.g., scalpel, 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, and transport as per section 4.6. Fixed tissues (e.g., 10% buffered formalin) are no longer considered biohazardous. When handling these samples, use appropriate personal protective equipment and transport them in a secondary container.
  • 4.7.4 Follow the sharps handling practices outlined in section 4.2.

5. Engineering Controls

• 5.1 Use of Biosafety Cabinets

  • 5.1.1 Perform all procedures carefully to minimize the creation of aerosols. Use a certified class II biosafety cabinet for inoculation, necropsy and tissue harvest, cage changing, and manipulating high concentrations or large volumes of infectious agents.
  • 5.1.2 Wipe cages with appropriate disinfectant when moving out of biosafety cabinet.

• 5.2 Housing and Handling of Infected Animals

  • 5.2.1 House animals in a primary containment device appropriate for the rodent species, such as a ventilated micro-isolator cage or static micro-isolator cage with a filter top.
  • 5.2.2 Conduct inoculations, cage changing, and other procedures in a biosafety cabinet.
  • 5.2.3 Whenever possible, use forceps to transfer infected animals between cages.

6. Personal Protective Equipment (PPE)

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

  • Disposable solid front gown

  • Disposable gloves (nitrile- avoid latex when possible) - Use double gloves when handling infectious agents or inoculating animals. Outer glove should overlay the cuff of gown.
  • Shoe covers
  • Safety glasses

• 6.2 Wear additional PPE (e.g., face shield, respiratory protection, cut/bite-resistant gloves) when appropriate engineering controls are not available or indicated by the agent, hazards, or experimental conditions.

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

• 6.4 Change gloves frequently (or decontaminate with disinfectant) during activities to avoid contamination of equipment and surfaces. Remove and replace other PPE if contaminated or breached.

• 6.5 Remove PPE upon exiting the animal room and dispose of it in a red biohazard bag. First, remove outer gloves, gown (turning inside out), 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:

1. Wash the exposed area with soap and water or rinse in eyewash for at least 5 minutes.
2. Perform first aid, if applicable
3. Notify supervisor
   • Students seek medical evaluation at Cornell Health. After hours seek evaluation at Cayuga Medical Center.
   • Evaluation or treatment for employees is provided in the Cayuga Medical Center emergency room or several urgent care clinics in Ithaca. Employees may also see their personal care physician.
4. Document exposures, injuries, and illnesses in the Cornell University Injury/Illness/Exposure Report

5. Incidents involving risk group 2 or 3 biological agents or recombinant materials will be reported by the Biosafety Officer to the Institutional Biosafety Committee.

• 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

• Environmental Health & Safety (EHS): 607-255-8200 (off-hours 607-255-1111)
• Cornell Health, 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)