STANDARD OPERATING PROCEDURES

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1. Performance Standard: All rats are checked daily for health, feed and water, and clean cages. Rooms are stocked with in-date supplies. Health problems are reported to the Laboratory Animal Technologist, the ARC veterinary staff and the Principal Investigator (PI) or designated lab contact person.

2. Protective Equipment: The personal protective equipment (PPE) listed below is to be used as indicated.

Required for animal room entry:
- Disposable nitrile gloves
Suggested
- Safety glasses

3. Caging

Small mouse cage: 11" x 7" x 5" high, 77 sq. inches floor space, capacity 4 adult mice.
Large mouse cage: 19" x 10" x 6" high, 190 sq. inches floor space, capacity 8 adult mice. Use of large breeder cages needs to be approved by the Operations Manager or Associate Director.
Breeding animals:
- 1 male and 1 female should be housed in a small mouse cage. If a breeding trio (1 male, 2 females) is housed in a small mouse cage, the litter must be weaned at 21 days of age.
- 1 male and up to 3 females can be housed in a large mouse cage
- Litters are weaned at 21 days of age. Breeding pairs will be allowed a 3-day grace period after day 21 for weaning. If the litters are not weaned by 25 days of age, the ARC animal care tech responsible for the room that day should wean the mice on day 25. Complete a wean request form, temporary cage cards and a miscellaneous services slip. Litters from breeding trios housed in small mouse cages must be weaned when they reach 21 days of age. If the litters are not weaned by 22 days of age, the ARC animal care tech responsible for the room that day should wean the mice on day 22, and complete a wean request form and miscellaneous services slip. Send an e-mail to the lab contact person to let them know the litter has been weaned.

4. Environment

Light cycle: 12 hours light/12 hours dark (exceptions are noted on the room sign)
Humidity: 30%-70%
Temperature: 72°F +/- 2°
Report environmental problems in the room (i.e. room temperature variations >+/- 2°F from 72¡F, obvious ventilation changes) verbally to the Operations Manager. On weekends and holidays, also report these problems to DHMC Engineering for Borwell and FO&M for Vail and Moore. Phone numbers are posted in the Borwell facility breakroom and the Vail and Moore offices.

5. Water

Water is provided ad lib via bottle, stopper and sipper tube, unless otherwise indicated by an ARC special instruction card on the front of the cage.
Water bottles are changed once a week on the complete change ("cc") day, or more frequently if the water level becomes low.

6. Feed

Teklad Autoclavable Rodent Diet 7012 is fed ad lib, unless otherwise indicated by an ARC special instruction card on the front of the cage. Feed is added to the feed compartment of the wire bar lid at cage change time and whenever the feed level is less than one third of the feeder capacity. Feed should not be mounded, it should be level with the top of the feed compartment.
Feed bags are put directly into the feed bin. Keep the feed bin cover in place at all times when mice are not being fed.
Do not combine feed in one bag with feed in another bag within the same room, even if both bags of feed have the same milling date.
The feed arrives with the milling date printed on the bottom of the bag. The animal care tech assigned to the clean cage room writes the expiration date (6 months after the milling date) on the top of the bag when the feed is received, so that the expiration date is visible when the feed bin is opened.
Discard expired feed.

7. Bedding

Hardwood chips are used as contact bedding in Borwell and Vail, supplied approximately to a depth of 1/4" (approximately 1 3/4 cups of chips).
Aspen is used as contact bedding in Moore, supplied approximately to a depth of 1/2" (approximately 1 1/2 cups of aspen).
Exceptions need to be approved by the Operations Manager or ARC veterinary staff.
All cages are supplied with a Nestlet¨ or iso-BLOX¨.

8. Cage Changing

Cages are checked daily and changed if the bedding is wet or excessively soiled.
Gloved hands are wiped with Clidox 1:5:1 between cages at Borwell and Vail. Gloved hands do not need to be wiped with Clidox 1:5:1 between cages at Moore.
Cages are routinely changed once a week, on a "cc" day, unless they become soiled and require more frequent changing.
Small cages (11" x 7" x 5" high) containing litters are changed twice a week, on a "cc" day and breeder cage change ("BRC") day. The ARC tech identifies these cages by putting a large light blue colored dot on the cage card. NOTE: The ARC tech should remove the light blue colored dot when the litter has been weaned.
Changing cages of females with litters:
Transfer the litter to the clean cage by scooping it up with some of the nesting material (Nestlet¨ or iso-BLOX¨) and old bedding.
Avoid handling the pups.
Transfer the adult mice to the clean cage.
There is one cage of sentinel mice per rack. When doing a cage change, transfer 1 teaspoon-1 tablespoon of bedding from each soiled cage into an empty, clean cage. Avoid putting too much bedding into the cage. When all of the cages on the rack have been changed, place the sentinels into the cage containing the combined soiled bedding. Contact the Laboratory Animal Technologist if you do not find a sentinel cage on the rack.

9. Lid Changing

Wire bar lids are changed every 2 weeks.
Fill the feed compartment when the lid is changed, at cage change time and when the food level is less than one third of the feeder's capacity.
Food may be transferred from the old lid to the new lid.
Food fed to one cage of mice is never fed to a different cage.

10. Rack Changing

Shelf racks are changed and sanitized once a month.

11. Room Sanitation

Room sanitation, including cleaning implements (broom, Doodlebug pad & holder) is to be done every 6 months and when room is emptied.
Animals, cages and racks may be moved from the room into the corridor or another empty, clean room during room sanitation.
See SOP K.2 for details.

12. Cage Cards

Each cage is required to have an ARC cage card on it, attached to the cage by a card holder.

13. Animal Health

Observe mice for health status, adequate feed and water.
Check the Clinical Incidence Log for active Clinical Incidence Forms (CIFs). Perform treatments as indicated in the CIF and record observations.
Report ill animals using the CIF, place a 3/4" green dot on the cage card and e-mail the PI or lab contact person, Laboratory Animal Technologist and ARC veterinary staff. If immediate attention is needed, verbally report this to the Laboratory Animal Technologist or ARC veterinary staff. If immediate attention is needed on weekends, call the on-call vet.
Place a "Please Wean" sticker on the cage card of cages containing litters that are 21 days old. Send an e-mail to the PI and lab contact person to let them know litters need to be weaned (include the room number in the e-mail).
Report overcrowded cages to the PI or lab contact person and put an "Overcrowded" sticker on the cage card. Refer to section 3 of this SOP for cage capacities.
When you find a dead animal and the animal is not part of a biohazardous study, place the carcass in a black plastic bag or glove and seal the bag or glove. Place an "Animal Found Dead" sticker on the cage card. Write on the cage card that an animal was found dead and the date the animal was found dead. Contact the PI or lab contact person to let them know an animal was found dead in the cage. Document dead animals using the "Mortality Tracking Form" card. Attach the white copy to the bag or glove containing the carcass and contact the lab. If the mouse is the last one in the cage, date out the cage card and put it and the beige copy of the "Mortality Tracking Form" in the cage card drop box. If the carcass isn't from the Borwell quarantine area, place the carcass in the appropriate drawer in the 48-hour bin in the carcass cooler. Carcasses from animals found dead in the Borwell quarantine area are put into the refrigerator in the quarantine suite hallway. Carcasses from Moore are placed in the freezer located in room B52. See SOP H.3 for biohazardous studies procedure.

14. Daily Activity Log

Using a pen with permanent black or blue ink, record all animal care and room care activities on the Daily Activity Log.
Comments can be made on the back of the log. Using a pen with permanent black or blue ink, put your initials next to the appropriate date on the back of the Daily Activity Log to indicate you've checked for comments.
Using a pen with permanent black or blue ink, record the current ("now"), maximum ("high") and minimum ("low") temperatures and reset the min-max thermometer.

15. Housekeeping and Room Maintenance

Animal room floors are sanitized with freshly prepared bleach solution twice each week, on "cc" and mop ("m") or "BRC" days. Clean the animal room floor sooner if needed. To prepare bleach solution: mix 4oz. (1/2 cup) bleach with 1 gallon of warm (not hot) water. Turn off the hose water supply when you're finished sanitizing the floor.
Drain baskets are emptied when the floors are sanitized.
Feed bins are taken to the cagewash room and sanitized at least monthly.
Burned out fluorescent light tubes are replaced as needed by the animal care technician in charge of the room at Borwell. For burned out fluorescent light tubes in the Vail facility, technicians should contact DMS Facilities. For burned out fluorescent light tubes in the Moore facility, technicians should contact FO&M Work Control.
Door frames and door windows are cleaned and disinfected at least monthly.