NOTE: Level 2 housing additionally excludes Murine Norovirus (MNV) and Level 3 housing excludes Helicobacter. The Specific Pathogen Free Rodent Barrier Facility only receives mice from approved vendors or via rederivation from the Vanderbilt Transgenic Mouse Service Core and excludes both Murine Norovirus (MNV) and Helicobacter.

Housing Service level:
Basic Plus Service: PPE required: gown and gloves, rodents are housed in IVC cages that are washed and autoclaved, food and bedding is autoclaved; auto water is hyper chlorinated, cage change performed in biosafety cabinet or animal transfer station, gloved hands are dipped in disinfectant between cages.

Quality assurance monitoring for endo and ectoparasites and viral agents.

Yale's Sentinel Program and Testing Procedures:
Sentinels: Obtained from commercial source, immune competent 4–6-week-old CD-1 females from Charles River or, if unavailable, SW from Taconic. Sentinels are housed two per cage on the bottom shelf of each rack or side of a ventilated rack.
Sentinels are changed every two weeks into a cage containing dirty bedding from two rows of soiled cages. Cages are sampled systematically so that every cage on the rack is sampled over a 4–6-week continuous cycle; the sentinel cage is changed after investigators' animals.

Lab Animal Health Environmental Monitoring:
Exhaust Air Dust (EAD) or environmental PCR testing to screen for pathogens. To decrease the use of sentinel animals and increase the diagnostic sensitivity of monitoring rodent colonies for infectious agents, centralized whole rack level monitoring is being used as an adjunct to or a replacement for the use of soiled bedding sentinels.

Testing Schedule
Three months after placement, all sentinels are bled for viral serology for MPV, MHV, EDIM and TMEV. At the same time sentinels are tested for pinworms by adhesive tape to the perianal area and fecal flotation. Sentinel bedding is swabbed and tested by PCR for fur mites.

Six months after placement, sentinels are submitted for necropsy examination, parasitology, and virology. Sentinel bedding is swabbed and tested by PCR for fur mites.

List of Agents tested:

3-month Serology Quarterly (IFA)
Mouse parvovirus, Murine rotavirus, Mouse hepatitis virus, Theiler's encephalomyelitis virus, tape test for Syphacia and fecal float for Aspicularis.
Sentinel bedding is swabbed and tested by PCR for fur mites.

6-month Serology Biannual (IFA)
Ectromelia virus, Lymphocytic chorimeningitis virus, Minute virus of mice, Mouse hepatitis virus, Mouse parvovirus, Murine rotavirus, Mycoplasma pulmonis, Pneumonia virus of mice, Sendai Virus,
Theiler's encephalomyelitis virus.

Necropsy: gross examination of tissue; if suspicious lesions detected, tissues are processed for histology
Examination of pelt for ectoparasites, Sentinel bedding is swabbed and tested by PCR for fur mites.

Examination of cecal contents for internal parasites:
Parasitologic agents All rooms (agent only reported if present)
Syphacia obvelata
Aspicularis tetraptera
Spironucleus
Giardia muris
Fur Mites


*We currently do not screen for Helicobacter or Norovirus (MNV) nor do we exclude these two agents from basic plus service with the exception of specifically designated rooms.

Housing Service level:

• Full Service: Autoclaved cages, food and bedding is autoclaved, water is hyper chlorinated for bottles or ultra-filtered for hydro pacs. Cage change performed in biosafety cabinet, cages are wiped down with a disinfectant-soaked towel before opening, gloved hands are dipped in disinfectant between cages.
• Cages are systematically changed with decontamination of the biosafety cabinet and PPE change between groups of investigators animals. Quality assurance is monitored environmentally with Interceptor filters for endo and ecto parasites, viral agents and opportunistic bacteria. PPE required: gown, gloves, shoe covers, hair bonnet, mask and sleeve covers.

Agents (screened in Full-Service rooms only) Quarterly:
Encephalitozoon cuniculi, Aspiculuris tetraptera, Bordetella bronchiseptica, Bordetella hinzii, Campylobacter coli, Campylobacter jejuni, Campylobacter spp., CAR bacillus, Citrobacter rodentium, Clostridium piliforme, Corynebacterium bovis, Corynebacterium kutscheri, Corynebacterium sp. (HAC2), Cryptosporidium spp., Demodex musculi, Ectromelia, EDIM, Encephalitozoon cuniculi, Entamoeba muris, Giardia muris, Hantaan, Helicobacter bilis, Helicobacter ganmani, Helicobacter hepaticus, Helicobacter mastomyrinus, Helicobacter rodentium, Helicobacter spp., Helicobacter typhlonius, Klebsiella oxytoca, Klebsiella pneumoniae, LCMV, MAV1, MAV2, MHV, MNV, Mouse kidney parvovirus (MKPV), MPV, MVM, Mycoplasma pulmonis, Myocoptes, Ornithonyssus spp., Pasteurella pneumotropica biotype Heyl, Pasteurella pneumotropica biotype Jawetz, Pneumocystis spp., Proteus mirabilis, Pseudomonas aeruginosa, PVM, Radfordia/Myobia, REO3, Salmonella spp., Sendai, Spironucleus muris, Staphylococcus aureus, Staphylococcus xylosus, Streptobacillus moniliformis, Streptococcus pneumoniae, Streptococcus sp. beta hemolytic Group A, Streptococcus sp. beta hemolytic Group B, Streptococcus sp. beta hemolytic Group C, Streptococcus sp. beta hemolytic Group G, Syphacia muris, Syphacia obvelata, TMEV, Tritrichomonas muris, Chlamydia muridarum, Orinithonyssus bacoti.

TREATMENT REGIMENS /OUTBREAK MANAGEMENT

Pinworm
Outbreak pinworm treatment consists of feeding Fenbendazole medicated chow on a
"Week-on/week-off" schedule for five weeks (three treatment weeks). Entire room is placed under quarantine with PPE requirements enhanced and placed lowest on the facility room order. Follow up testing begins at least 24 days post treatment: 2 rounds of perineal tape testing or fecal exams- 24 and 48 days apart (~ 30% or more of cages) depending on species of pinworm detected. Additionally, all exhaust air dust of the vent racks is swabbed and tested simultaneously. The room is not cleared until the subsequent 6-month sentinel screening is performed and negative.

Fur Mites
Outbreak Mite treatment regimen consists of administration of rodent chow medicated with 12 ppm Ivermectin. This is given for 4 weeks consecutively. Entire room is placed under quarantine with PPE efforts enhanced and placed lowest on the facility room order. Follow up testing begins at least 24 days post treatment. Direct testing of animals is performed (3 rounds of fur plucks- 24 and 48 days apart (~ 30% or more of cages). Additionally, all exhaust air dust of the vent racks is swabbed and tested simultaneously. If all tests are negative the room is cleared of its fur mite positive status.

Mouse Parvovirus
The room is placed under quarantine, placed lowest on the facility and additional PPE implemented. The plan to eradicate the virus involves placement of additional sentinels in the room and systematic testing and culling of positive animals. Targeted sentinels are housed one per cage and placed every two rows on the rack found MPV positive where they receive soiled bedding transfers at cage change. The first round of testing is performed by serology three weeks after sentinels receive soiled bedding transfers. The second and third rounds of testing are performed at two-week intervals. Additionally, all exhaust air dust of the vent racks is swabbed and tested simultaneously. The room is not considered cleared of the virus until three consecutive rounds of testing with negative results have been achieved.