Kathryn Mariucci, CPIA, RBP
Senior IACUC Manager
UM Biosafety Officer
SPF Mouse Serology
Last Updated: May 4, 2017
Specific pathogen-free (SPF) mouse sentinel serology is done as part of a comprehensive preventive health care and disease surveillance program.
It is the responsibility of the AV to provide oversight and for the LAR staff to understand, to be proficient in, and to participate in the serology testing program.
Examination and Sample Collection
- On a quarterly basis, collect SPF sentinel mice from each mouse room into clean shoebox cages.
- All mouse rooms in Skaggs Building (SB), SB 066, 061B, and HSB004 mouse quarantine.
- Take one sentinel mouse from every rack and place into a clean cage.
- Label each box with a new cage card that includes room number, rack number, and investigator.
- Move mice on a clean cart to HSB016-LAR Clinical Lab
- Fill out University of Missouri Research Animal Diagnostic Laboratory (MU RADIL) serology paperwork (print from computer or get hard copy from file cabinet).
- The order in which the mice are listed on the submission form will be the order in which animals are necropsied. List smallest room number first and proceed to larger room numbers.
- Pre-label all Opti-spot dried blood spot cards that will hold samples.
- Items needed
- scissors o clear cellophane tape o 25 gauge needles
- forceps o Opti-spot dried blood sport card o 1-mL syringes
- Rescue |RTU o fecal flotation tubes o sharps container
- slides o sterile scalpel blades o paper towels
- sharpie® markers o Microscopes (optical and dissecting)
- Place mouse (proper order of serology paperwork) in CO2 chamber and euthanize using 20% volume displacement per minute inside the chamber.
- Retrieve blood via cardiac puncture.
- Use a 1 cc syringe with 25 ga needle.
- Place one large drop (approximately 25 microliters) of blood in the designated circle of the dried blood spot card.
- Allow sample to dry for 1 hour before folding protective upper tab over the blood spot and tuck under the lower tab.
- Label tail with the number from serology paperwork using a Sharpie® marker.
- Use dissecting microscope to check body for ectoparasites and abnormalities.
- Collect a fur tape sample and a perianal sample using cellophane tape. Press the tape firmly against the skin. For perianal samples, place tape over anus and draw a large circle around the sample area using a Sharpie® marker.
- Place fur and perianal tapes on separate slides and label.
- 3 samples of each specific type can fit on a slide.
- Do not mix fur tapes and perianal tapes on the same slide.
- Use optical microscope to check the sample for the presence of pinworm eggs and other parasites.
- Incise the mouse along the ventral midline of the body.
- Check for abnormalities of internal organs (location, size, texture, color).
- Collect one fecal pellet from rectum.
- Place in pre-labeled fecal flotation tube.
- 3 samples from different mice may go into one tube, in the order written on the submission form.
- Push contents of cecum onto a slide.
- Label slide with mouse number(s); 3 samples can fit on one slide.
- Use dissecting microscope to check the sample for the presence of adult pinworms
- Double check that all procedures were finished and all necessary samples were collected before discarding each carcass.
- Once all necropsies are completed, prepare fecal floats for parasite eggs.
- There should be a maximum of three fecal samples in one tube.
- Use a transfer pipet to add a small amount of "Vetus Feca Test solution (a 1:18 dilution of Vetus Feca Test Dry Concentrate" diluted with water) to tube and mix well with feces. (This 1:18 solution and concentrate are located in HSB016 metal cabinet).
- Use a transfer pipet to fill the tube to top with Vetus Feca solution, creating a meniscus.
- Place square coverslip on top of tube.
- Let stand for 10 minutes.
- Place the cover slip on a clean slide; create minimum air bubbles.
- Use compound microscope to examine the samples for eggs of internal parasites.
Packaging Samples for IDEXX RADIL
- Place Opti-spot samples into a ziplock bag.
- Place the IDEX RADIL paperwork together with the ziplock bag of samples into an appropriately sized shipping envelope for priority mail and seal.
- Mail from campus post office.
- 2-day service (either UPS or USPS, whichever is cheaper).
- Use IDXX RADIL address listed on paperwork.
- Charge to UM credit card.