Maintaining Pathogen-Free Facilities

Personnel Access

1. Contact the Animal Facility Manager or ARF Veterinary staff to ask questions about proper facility/room entry order if you have animals located in more than one room within the ARF facilities.

2. All facilities and animal rooms require personal protective equipment (PPE) to enter. Experience shows that groups who strictly adhere to procedures avoid contamination of their animals. Contamination most often occurs when there is a failure to use proper technique. In the majority of ARF facilities, the PPE consists of your lab coat, which you should wear whenever you are in the vivarium. It is best if you have a spare lab coat that you reserve to wear only in the vivarium; this minimizes spread of animal dander and fur from your clothes to the rest of the facility. Closed-toes shoes will avoid injury from rolling equipment and sharps. Wear gloves whenever you are handling rodents and small animals.

3. Do not bring equipment into a facility if it has been in contact with animals from another facility. Contact the ARF staff for advice for decontamination of equipment if needed.

4. Rooms requiring additional PPE will have instructions listed on the door. In these instances, PPE will be provided at the room entrance and you will need to dispose of all PPE when leaving the room into the provided trash receptacle.

5. Animals are not to be housed in the laboratories over night. Investigators that require their animals to be removed from the animal facility for time periods greater than 12 hours or during the overnight period, should request prior approval from the IACUC.

6. Rodents are extremely sensitive to alterations in the daily light/ dark cycle therefore Do Not override the automated timers and interrupt the dark cycle in the animal rooms, except in an emergency situation.

Rodent Animal Room Procedures and Animal Transfers

1. When animals or biological materials are moved from one room or facility to another, contamination may result. ARF maintains a rodent sentinel program in which rodents, provided by ARF, are housed in the same room as research animals. These sentinels are exposed to bedding from the other animals in the room. At pre-determined intervals they are euthanized and tested for evidence of infection. The results of these tests determine the disease status of the room.

2. Under no circumstances are animals to be transferred between rooms of an animal facility or into another facility without proper approval. ARF tracks all animal movements to be sure that animals do not carry pathogens from one room into another. In case of a breach in containment, ARF uses these records to track animal movements and quickly determine the rooms at risk that need special containment, monitoring, or quarantine.

3. When an IACUC approved protocol requires animal transportation to an investigators lab, the animal must be transported in the designated transport cages that are provided by ARF. Animals should never be transported in the elevator on the north end of Coles Hall as this is for human traffic only. All used transportation cages should be returned to the designated drop-off sites for processing.

4. Transportation cages are located in the corridors where rodents are housed, and labeled as "Transport Cages." Animals should be transferred into the transport cages using the cage change station, where available, or a counter top, never on the floor. Cages are fitted with filter tops to protect the animals, environment and the public from aerosolized allergens. Cages are turned in for cleaning by taking them to the cage wash area (basement of Coles, South end, or Mosier cage wash room, P213). There are also dirty cage bins on the 2nd floor of Coles Hall, North alcove near the freezer room, where cages can be placed.

5. Any cages labeled as "Biohazardous material" are to be appropriately bagged, labeled and taken directly to the Coles Hall cage wash area and left by the autoclave on the South side. These bags must be tied shut.

6. The floor areas are considered to be dirty, potentially contaminated surfaces; therefore, only empty used cages should be placed on the floor. Animals that make contact with the floor are considered to be contaminated and normal policy is to euthanize those animals. Exceptions may be granted by the ARF director; however contaminated animals should not be reintroduced into the colony and need to be maintained in isolation until a determination is made.

7. Once animals leave the animal facility, they should not be returned to the facility unless they are part of an approved protocol that includes survival procedures.

Animal Quarantine Program

Background
When new animals are introduced into the animal facilities, there is an inherent risk of introducing pathogens into the resident animal population. Every time animals are introduced it puts the resident animal population at risk. The inadvertent introduction of even a single infected animal could cause a disease outbreak, which could result in lost time, lost research data, and the need for expensive eradication efforts. Eradicating a pathogen from an animal facility may take years.

The highest risk for introducing infectious agents into an animal colony is through the introduction of live animals from non-approved vendors. The increase in transgenic and knockout mouse strains available for research over the past few years and the rise in national and international collaborations have resulted in a growing number of animals being introduced from non-commercial sources. This would include other universities, research institutions or private biotechnology companies. While most of them maintain a health-monitoring program similar to that of ARF many do not. To assure that no pathogens are introduced along with these animals, ARF keeps them in quarantine until the health status of every single shipment is confirmed. The majority of animals at the KSU CVM are purchased from commercial vendors, which routinely screen their colonies for pathogens. ARF maintains a list of these "approved vendors"; animals from these sources can be introduced into the animal rooms without going through the quarantine process.

Procedure for Ordering Animals from Non-Approved Vendors

1. Request Form
   For each animal shipment an Animal Requisition Form (ARF-90) must be submitted. The request form is found on-line on the ARF website at: http://www.vet.ksu.edu/depts/arf/index.htm. If multiple shipments from the same source are expected, please submit a request for each shipment. This is necessary because each group is tested and evaluated separately and is assigned a unique number to avoid confusion. Please fill out the form completely; all information is necessary for completion of the introduction. A current approved KSU ACUC protocol number must be present on every Animal Requisition Form (ARF-90) before animals can be ordered. In most situations, the time from when an Animal Requisition Form (ARF-90) is submitted until the animal shipment is approved for shipment to KSU is two weeks.

2. Health Report
   To determine the necessary quarantine procedure, a health monitoring report from the institution of origin must be obtained. ARF can obtain the health report directly from the veterinarian at the institution of origin. Please provide the name and telephone number or e-mail address of the facility veterinarian on the request form.

3. Approval
   Once both the request form and the health report have been submitted the request is processed, usually within 24 hours. The investigator is notified of approval by e-mail. Copies of this e-mail will also go to the shipping contact at the sending institution (if an e-mail address was provided).
   The approval will list the quarantine process required for this group.

4. Shipping Arrangements
   Once an Animal Requisition Form (ARF-90) has been approved, the ARF staff will contact the shipping contact at the other institution to make arrangements for the shipment. The investigator will be informed as soon as the animals arrive at ARF. The investigator should inform the sending investigator not to have the animals shipped before the introduction request has been approved. For safety reasons all non-approved shipments will be returned to the sender.

Quarantine
Once the animals arrive at ARF they will be placed in quarantine. Access to quarantine is limited and special arrangements should be made if you need to observe or work with the animals prior to their release.

Standard Quarantine Procedure
Most groups will undergo the standard quarantine procedure. On arrival, the animals are transported into the quarantine room and housed in cages under BSL 2 conditions to exclude transmission of infectious agents. A physical examination as well as a "tape test" to detect evidence of pinworms is performed on each shipment as they are transferred to their quarantine cages. Two (2) sentinel animals are placed with each shipment. After 5 weeks, a sentinel is removed, and submitted to for health monitoring, were it is necropsied and samples are submitted for histopathology, microbiology, parasitology and serology for all relevant animal pathogens. This testing usually takes about 2 weeks. Total duration of quarantine for a pathogen-free group is ~7 weeks. Animals may be fed fenbendazole-medicated feed during the quarantine period.

If technical services are required during the quarantine period (DNA collection, dosing, bleeding, etc), they are billed at the usual ARF technical services rate of $18/hour. If an investigator requires access to their animals during the quarantine period, arrangements should be made in advance with the Animal Facility Manager (Susan Rose. 532-5647). Investigator access to quarantine rooms is limited to between the hours of 8am to 11:30am and 1:00pm to 4:30pm Mon.- Fri. Once an investigator or their staff has been in a quarantine room, they will not be permitted into another animal room that same day. They are therefore encouraged to enter quarantine rooms late in the day so there is less risk of them inadvertently entering another animal room.

Quarantine Procedure for Animals Known to be Infected with a Pathogen:
Animals that are known to harbor pathogens will either be treated or rederived to eliminate the pathogen(s) prior to entering the quarantine period. Investigators wishing to import animals known to be infected should contact the ARF veterinary staff for a more detailed description of the quarantine program and a list of additional costs that may be incurred during the quarantine process.

Acute Use
If animals will be used acutely within a few weeks after arrival, no quarantine process will be started. Acute use animals are intended to be used within 6 weeks of their arrival at ARF. These animals will not be released from quarantine into the general animal population. Prior to ordering the animals, a meeting with the ARF staff is required in which the intended procedure is discussed in the context of protecting the ARF colonies from pathogens. If the investigator decides later to keep these animals, the quarantine process will start at that time. The duration of quarantine will be 6-8 weeks, provided the animals prove to be pathogen-free. Arrangements may be requested to transport animals from quarantine for acute use in the lab.

Definitions

Approved commercial vendors
Currently the list of approved animal vendors for the ARF includes the following: Charles River Breeding Laboratories, Harlan Sprague Dawley, Hilltop Lab Animals, Jackson Laboratories, National Cancer Institute, Frederick Cancer Research & Development Center, Sasco, and Taconic.

Health Report
Report on the health monitoring of an animal colony is usually obtained by periodic testing of sentinel animals placed into an animal room. Ideally it includes viral serology, microbiology and parasitology. The health report reflects the health status of an entire colony not necessarily of individual animals. Not to be confused with a health certificate which is based on the clinical examination of an animal or a small group of animals.

Non-Approved Source
Usually a non-commercial research institution, university, or biotech company, which is not on the list of approved vendors. Some approved vendors maintain contract breeding colonies for research institutions. Sometimes these colonies are housed separate from the commercial colonies. In this case they are considered non-approved sources (e.g. research colonies at Jackson Laboratories).

Rederivation
Animals infected with undesirable animal pathogens may need to be rederived. Rederivation procedures depend on the pathogen involved and may include treatment with antimicrobial agents, "burning out" of acute infections, cesarean rederivation, or embryo transfer. Once the animals are determined to be free of relevant pathogens, the animals can be moved to the investigator's animal room.