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

• Principal Investigator (PI): Ensure personnel are made aware of the hazards associated with the infectious agent and receive training commensurate with their activities before commencing ABSL-2 experiments.
• Personnel: Comply with the safe work practices and procedures described within this ABP.

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/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).
• 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

• 4.2.2 Do not recap needles.
  • If recapping is required, receive approval from EHS and the IACUC. Once approved, use one-handed scoop or forceps/tongs to place the cap on the needle.
• 4.2.3 If a needle/scalpel must be set down during a procedure, place 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 potential allergens on your body.
  • 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 account for environment (organic load, surfaces), contact time, application, and safety: Center for Food Security and Public Health.
• 4.4.2 Cover spills with absorbent towels/pads and saturate with disinfectant. Allow 5–10 minutes of contact time. Segregate waste in red biohazard bags or sharps containers; re-apply disinfectant to the 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 the biosafety cabinet.
• 4.5.2 Dispose of sharps-related items (e.g., needles, syringes, Pasteur pipettes, blood tubes) directly into a sharps 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; allow at least 30 minutes’ contact time before disposal to sanitary drain; follow with copious 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 carcasses in bench diapers and place 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. Prefer 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 harvested tissue/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; use appropriate PPE and secondary containment when handling.
• 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 aerosol creation. Use a certified Class II biosafety cabinet for inoculation, necropsy/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 the 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)

• 6.2 Additional PPE when indicated (e.g., splash/agent risk, engineering controls not available): face shield, respiratory protection, cut/bite-resistant gloves.
• 6.3 Solid-toed shoes are required for entry into animal rooms.
• 6.4 Change gloves frequently (or decontaminate with disinfectant) to avoid contamination of equipment/surfaces. Replace any PPE that is contaminated or breached.
• 6.5 Remove PPE upon exiting the animal room and dispose of it in a red biohazard bag. Doffing order: remove outer gloves → remove gown (turn inside out) → remove shoe covers while stepping out (step-over technique) → remove inner gloves.
  • For more information, see ACUP 715: Personal Protective Equipment.

7. Response to Accidental Exposures

7.1 If a splash to mucous membranes, direct contact with open wounds, or sharps injury occurs:

1. Wash exposed skin with soap and water, or rinse eyes 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, proceed to Cayuga Medical Center.
   • Employees: evaluation/treatment at Cayuga Medical Center emergency room or local urgent care; personal physician as appropriate.
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: dial 911 from a campus phone, or call 607-255-1111 from a mobile 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)

9. Risk Assessment Overview

The following table summarizes key hazards, their likelihood and consequence, and the residual risk when controls described in this ABP are applied.