Experimental Issues

Scheduling a Procedure Room

1. All facilities have procedure room space for investigators and their staff to use. Some facilities have very little space, or the space is in high demand. Therefore, it is necessary to schedule the space. Do not block out large amounts of time, or list a block of time day after day if you really don't need it.

2. Investigators who do not show up within 15 minutes of their scheduled time forfeit the reservation.

3. Leave the area clean for the next user and put all sharp objects in the sharps container. If the area is soiled when you arrive, notify the Animal Facility Manager. When an investigator leaves the procedure room soiled and/or in disarray, there is a minimum $35.00 fee to clean the area.

4. The Mosier Hall Junior Surgery Laboratory and associated prep rooms are available for use by investigators, but are scheduled through the Clinical Sciences Department.

Using Biohazards

1. The main ARF facilities and the research Hill animal facilities have bio-containment housing available. Biohazard housing space is limited, and housing space must be assigned 3-4 weeks before the start of the experiment. Typically this space is assigned on a "first-come first served" basis. Space assignment in these facilities can be arranged by contacting the ARF Director, or facility manager.

2. Obtaining approval to use biohazardous agents

   1. Protocols involving the use of biohazardous agents in animals require approval by the Institutional Biosafety Committee in addition to ACUC approval. Information regarding the Biosafety Committee can be found on their website at: http://www.ksu.edu/research/comply/irb/index.html

   2. Agents that require biosafety approval include infectious agents (even replication-deficient vectors), toxins, carcinogens, recombinant DNA, and chemicals with known or suspected toxicity for humans or animals. The purpose of the review is to ensure the safety of both the animals and the animal care personnel.

   3. Questions regarding the need for biosafety review and approval should be directed to the Institutional Biosafety Committee (Dr. Jerry Jaax, 532-3233).

3. Personal protective equipment is mandatory whenever entering the bio-containment areas. Contact the Manager of the facility where your animals will be housed to answer any questions you might have and to receive proper training for accessing these areas. The PI is responsible for the additional costs associated with providing PPE for their experimental needs.

4. Investigators are required to provide a brief training session to ARF staff members so they understand the hazards of the agent being used, and the procedures required to follow when handling the animals and cages. The PI is to provide appropriate signage to be placed on the outside of any animal rooms detailing any project specific procedures. ARF will provide Biohazard signs for the immediate area.

Taking Animals to Your Lab

1. You may transport animals to your lab under the following conditions:

   1. Clean transport cages are obtained from the ARF corridor. Animals must be transferred into a transport cage located in the hall on a labeled cart. All transport cages must be covered with the provided bonnet or a micro-isolator lid. The appropriate lid choice will be need to be determined and discussed with the ARF veterinary staff or Facility Manager.

   2. Use approved routes through buildings to minimize contact with the public.

   3. Personal vehicles may not be used for the transport of any animals between facilities unless specifically approved by ARF.

2. Animals may not remain in laboratories for longer than 12 hours. The only exception is labs that have been approved by the ACUC as an animal holding facility

3. Animals may remain in satellite animal facilities for longer than 24 hours. To be a satellite you must have routine inspections of the facility and comply with requirements for the daily observation of the animals (including weekends and holidays) and the animal's environment (temperature, sanitation, humidity, ventilation, lighting, vermin control, etc.) within the laboratory. Contact the ARF Director for further information.

4. If animals are to undergo procedures in your lab, be sure that the lab room # is listed in your animal protocol. ARF is required to keep a list of these rooms and the IACUC is required to inspect these areas on a semi-annual basis.

5. Dogs, cats, rabbits, farm animals, nonhuman primates, and other large animals must be transported by ARF. Contact the ARF facility manager to make arrangements and please allow 24 hours advanced notice to make arrangements for transportation. Additional charges may apply for animal transportation.

Obtaining Training in Specific Procedures

1. It is absolutely critical that individuals performing procedures on animals be knowledgeable in the techniques. If you would like to perform procedures that are new to you, ARF has numerous resources to assist you.

2. The Clinical Veterinarian and ARF Director are responsible for the care of animals in ARF facilities and have their phone and pager numbers posted in the facility. Please contact them for questions or assistance with your animal needs. This includes but is not limited to: training, surgical procedures, diets, treatments, anesthesia, and contamination prevention.

3. A veterinary technician is available to teach you how to perform procedures such as proper restraint, blood collection, injections, and euthanasia techniques. The veterinary technicians can also be hired by investigators to perform routine procedures on their animals. Fees for this service are charged on an hourly basis. These services include drug administration (SQ, IP, or IV injections or gavage), blood collection (survival or nonsurvival), anesthesia monitoring, and sample preparation. These services can be arranged by contacting the facility manager or ARF veterinarian.

Euthanasia

1. Specific guidelines must be following in order to perform humane euthanasia of research animals. If these are not compatible with your experimental design, contact a veterinarian for advice.

2. It is critical that the animals are euthanized using the technique that was proposed by the investigator in the protocol and approved by the IACUC.

3. If using CO2 for animals, be aware that carbon dioxide is not a poison. It works by displacing the oxygen that the animals breathe. Although they lose consciousness rapidly, you must keep them in the CO2 chamber for a full 5 min to insure that they are completely euthanized.

4. For a complete step by step description of the CO2 euthanasia technique please contact a member of the ARF veterinary staff for a copy of ARF's IACUC approved SOP for CO2 euthanasia.

5. Carcasses should then be placed in a bag, labeled with investigator name, the date and placed in the appropriate freezer/refrigerator unit.

6. ARF will also euthanize animals for you on a fee for time basis.

7. Animals and animal waste products from biohazardous protocols (including those with genetically modified DNA) must be disposed of properly. Current FDA regulations covering biohazardous waste do not allow this material to enter the food chain therefore rendering or donation of these animals to the Raptor Club are not allowable.

Weaning and Overcrowding of Cages

1. Rats: No more than two adult rats may be housed per cage. Rats over 400g should be housed singly. Breeding pairs of rats are housed in large rat breeder boxes.

2. Mice: The maximum number that can be housed together depends on the mouse's weight. Mouse harem breeding schemes require the use of the large mouse boxes to prevent overcrowding when pups are born. For the small standard mouse cage, the guidelines are:

   <10 g	= 10 per cage (Unusually large litters are an obvious exception)
   10 - 15 g	= 5 per cage
   15 -25 g	= 7 per cage
   > 25 g	= 4 per cage

3. Mice and rats are generally weaned at 21 days of age.

4. If you wish to wean them at an age greater than 21 days, please add this request to your ARF 90 form and submit it to the ARF office. Weaning should never be delayed beyond 26 days unless you have specific justification for doing so.

If a Mouse or Rat Jumps Out of the Cage While You are Working in Any Animal Facility

1. Attempt to catch the animal if you can. If you are unable to do so, notify ARF immediately so they can help.

2. Do not place the animal back in the cage with the others. This animal may have acquired pathogens from being on the floor and could contaminate your whole colony as well as others.

3. Such animals should be placed in a new cage and then euthanized.

4. If you want to keep the animal, notify ARF so they can arrange to place the animal through the quarantine procedure appropriate for the facility in which the animal is housed.