Institutional SOP M0: Rodent Health Surveillance University Animal Care Committee (UACC) Standard Operating Procedure (SOP) SOP # M0 Title Rodent Health Surveillance Program. Purpose This Standard Operating Procedure (SOP) harmonizes the health surveillance practices of all rodent animal facilities at the University of Saskatchewan.. General Information A rodent health surveillance program is necessary to identify the presence of pathogens that may cause clinical or subclinical disease. Infection with such pathogens could jeopardize the validity and reproducibility of research data, or complicate its interpretation, or cause zoonotic concerns. Responsibility: The Facility Managers are responsible for the rodent health surveillance program with oversight from the Facility Veterinarian. The animal care staff must be properly trained and comfortable in performing the procedures.. Testing Schedule Surveillance is conducted quarterly and involves use of research animals, sentinel animals, and/or environmental monitoring of the IVC rack. To allow for flexibility in work schedules, samples for surveillance are collected within ± one week of the proposed testing day. Testing is tailored to the particular room and its occupancy or to a particular principal investigator and initiation of animal use. Month Pool fecal samples from up to 0 cages of research animals. The following tests are conducted: Charles River Enteric PRIA and Environmental Monitoring. Month Acquire sentinel mice ( mice per IVC rack) months in advance of this test date. The following tests are conducted: Charles River Assessment Plus serology panel and Charles River Prevalent PRIA. Month : Pool fecal samples from up to 0 cages of research animals. The following tests are conducted: Charles River Enteric PRIA and Environmental Monitoring. Month : Charles River Surveillance Plus PRIA Page of
Institutional SOP M0: Rodent Health Surveillance Note: Rodent housing for less than 0 days in a particular room does not require sentinel monitoring, if all in-all out approach is used and the animal room is disinfected prior to entry of new animals. If animals are housed between 0 days in a room then surveillance testing is required and involves pooling of fecal samples from up to 0 cages of research animals with test requests for Charles River Enteric PRIA and Environmental sampling.. Equipment/Materials. Virus and Antibody Free/Specific Pathogen Free (VAF/SPF) sentinels at - weeks of age a. Mouse female CD- (outbred) b. Rat male and/or female Sprague Dawley (outbred). Rodent Housing cages, water bottles, cage cards, feed, water, enrichment. PRIA sample collection supplies provided by Charles River (see Appendix ). Charles River Testing Kits ordered via the Animal Order Desk. BioMed bin. Procedure A. Sentinels. Obtain sentinels from an approved commercial supplier such as Charles River Canada. Adhere to Animal Receiving SOP (SOPA0) months in advance of the month testing date.. House mice per cage with one sentinel cage per single-sided rack or 0 mouse cages.. House rats per cage with one sentinel cage per 0-00 rat cages from one room.. Label each sentinel cage with a different colored cage card with SENTINEL clearly labeled on the card. The cage card should contain the protocol number and standard cage card information for the animal facility. B. Exposure and Testing. Testing schedule begins upon entrance of the first research animal in the animal room. Record in the health monitoring form the species/strain/location of animal and the test samples being submitted (see appendix ). Pooled fecal samples from up to 0 cages of research animals are used for months,, and of the testing schedule.. Sentinels must arrive months in advance of month of the testing schedule. For the sentinels, at each bedding change, transfer - ml of soiled bedding, ensuring to include fecal material, from each principal cage to the sentinel cage to expose sentinels to potential diseases carried by colony animals. Page of
Institutional SOP M0: Rodent Health Surveillance. Collect samples from one of the paired sentinels at month of the testing schedule. If test results are negative the sentinel animals are transferred to the UACC surplus animal/tissue share program. a. For serology testing: Collect a spot test blood sample (according to Charles River EZ-Spot whole blood spot sample collection method) using the facility-specific blood collection method (e.g. tail nick, facial vein venepuncture). b. For PRIA testing Collect fecal, fur/oral/skin swabs as per test requirements using the collection supplies provided by CRC (see Appendix ). If samples collected postmortem, also collect blood and freeze serum for future diagnostics, if needed. c. Dispose of carcass into BioMed bin.. The Facility Manager and the Facility Veterinarian reviews the sentinel reports.. Upon receipt of a positive test result use the second sentinel to confirm positive test result. If test results are negative transfer the animal to the UACC surplus animal/tissue share program.. Facility Managers maintain results of all sentinel evaluations. Copy of test results is made available to Facility Veterinarian. C. Level II Biocontainment Optional): Sentinels in Level II biocontainment are reverse tested NO dirty bedding is transferred from the principal cages to the sentinel cages. Animals in Level II suite are experimentally contaminated with a pathogen, therefore testing of sentinel animals will validate adequate cage changing procedures by showing whether or not pathogen transfer occurs between cages. Sentinels should be present in the Level II suite at all times to allow for the reverse testing. To allow for flexibility in work schedules, samples for surveillance are collected within ± one week of the proposed testing day as follows: Month Pool fecal samples from up to 0 cages of research animals. The following tests are conducted: Charles River Enteric PRIA and Environmental sampling. Month Acquire sentinel mice ( mice per IVC rack) months in advance of this test date. The following tests are conducted: Charles River Assessment Plus serology panel, Charles River Enteric PRIA, and Charles River Prevalent PRIA. Month : Pool fecal samples from up to 0 cages of research animals. The following tests are conducted: Charles River Enteric PRIA and Environmental sampling. Month : Charles River Surveillance Plus PRIA Page of
Institutional SOP M0: Rodent Health Surveillance. Safety Take care to avoid injury such as needle sticks and animal bites/scratches. Report any injury to Safety Resources.. Potential Complications and Troubleshooting Sentinel animals should remain healthy. An ill sentinel animal could indicate a disease outbreak in the facility; therefore, the animal must be humanely euthanized and samples collected and submitted for diagnostic testing.. References. Companion guide to Rodent Health Surveillance for Research Facilities, Charles River. PCR Rodent Infectious Agent (PRIA) Panels (http://www.criver.com/files/pdfs/researchmodels/rm_ld_r_pria_guidebook.aspx). Rodent EZ-Spot Serology (http://www.criver.com/products-services/basicresearch/health-monitoring-diagnostic-services/ezspot) (http://www.criver.com/files/pdfs/research-models/rm_ld_d_ez_spot.aspx). Environmental Monitoring (http://www.criver.com/products-services/basicresearch/health-monitoring-diagnostic-services/environmental-monitoring). Revision History. Updated testing schedule and tests performed to account for new diagnostic testing procedures offered by Charles River.. Updated reference list to reflect changes in diagnostic tests from Charles River. Date Created: September 0 Written by: Michele Moroz SOP Review and Revision History Revision Number Revision Date Reviewer May, 0 Michele Moroz, Melanie van der Loop, Jane Alcorn, Monique Burmester Page of
Institutional SOP Appendix M0.0: Rodent Health Surveillance Appendix : PRIA Sample Collection Procedures and Supplies NOTE: Do NOT combine different types of sample sets (i.e. tissues, fecal, and swabs) in the same vial.. Here are the three types of swabs used for PRIA testing. For individual fur mite PRIA testing, use the pink sticky swab. The pink end is sticky and should be used to swab the entire animal. a. To obtain a sample for fur mite testing, swab the entire surface of the animal with the fur swab, going against the direction of hair growth. Cover as much surface area as possible, as fur mites may be few in number and like to hide at the base of the hairs. Ideally the entire body should be swabbed firmly; however, the most important areas to swab include the following: Around ears Base of neck Under chin Inguinal areas Base of tail b. Once swab has been used to swab the animal(s), cut off the tip quite close to the pink end. Place the pink end into the.0 ml snap top vial. Page of
Institutional SOP Appendix M0.0: Rodent Health Surveillance. For pooled fur mite PRIA testing, please use the Hydraflock (non-sticky) swab. a. To obtain a sample for fur mite testing, swab the entire surface of the animal with the fur swab, going against the direction of hair growth. Cover as much surface area as possible, as fur mites may be few in number and like to hide at the base of the hairs. Ideally the entire body should be swabbed firmly; however, the most important areas to swab include the following: Around ears Base of neck Under chin Inguinal areas Base of tail b. Once swab has been used to swab the animal(s), cut off the tip quite close to the swab tip. Place the tip into the.0 ml snap top vial. Samples can be pooled up to 0 swab tips.. For pooled or individual oral swabs, use the Oral swabs (sterile pack of ). Page of
Institutional SOP Appendix M0.0: Rodent Health Surveillance a. To obtain an oral swab sample, swab oral cavities of animals (-0 animals). Either use swab for up to animals, or you can swab each animal with a new swab and pool the swab tips. Once swab has been used to swab all animal(s), cut off the tip quite close to the swab tip and place into the.0 ml snap top vial.. For fecal samples: a. Mouse fecal samples individual or pooled up to 0 pellets use the.0 ml snap top vial. b. Rat fecal samples individual or pooled up to pellets use the.0 ml snap top vial. For pooled - pellets, use the.0 ml snap top vial.. For lung tissues, nasal aspirate, and lung wash, use the.0 ml snap top vial. ml snap top vial ml snap top vial. For pooled rat fecal samples (- 0 pellets), use the ml screw top vial. Page of
Institutional SOP Appendix M0.0: Rodent Health Surveillance Date samples taken: PRIA Sample Submission Record Technicians: 0 0 0 Page of
Institutional SOP Appendix M0.0: Rodent Health Surveillance Date samples shipped: 0 0 0 Page of