Prevalence of Enteropathogens in Dogs Attending 3 Regional Dog Parks in Northern California

Hascall, K.L.; Kass, P.H.; Saksen, J.; Ahlmann, A.; Scorza, A.V.; Lappin, M.R.; Marks, S.L.

Fecal centrifugation flotations were performed on all samples at the veterinary commercial reference laboratory. c Fresh feces were examined for parasite ova, cysts, and oocysts by use of a zinc sulfate single centrifugation flotation technique as previously described; 11 however, the technician scanned the slide at 109 magnification in a grid pattern for approximately 60-120 seconds. A PCR diarrhea panel was performed on each sample for the following 11 enteropathogens and toxin genes: Cryptosporidium spp., Giardia spp., Salmonella spp., Campylobacter jejuni, Campylobacter coli, canine enteric coronavirus, canine distemper virus, canine parvovirus 2, canine circovirus, Clostridium difficile toxin A (TcdA) and toxin B (TcdB) genes, and Clostridium perfringens alpha toxin gene and enterotoxin gene (cpe). Fecal samples were processed by a previously validated protocol. 12, 13 Analysis was performed on a Roche LightCycler 480 e and raw data analyzed by the 2nd derivative maximum method to generate crossing points (CP). Real-time PCR was run with 7 quality controls including (1) PCR-positive controls, (2) PCR negative controls, (3) negative extraction controls, (4) DNA pre-analytical quality control targeting the host ssr rRNA (18S rRNA) gene complex, (5) RNA pre-analytical quality control targeting the host ssr rRNA gene complex, (6) an internal positive control spiked into the lysis solution, and (7) an environmental contamination monitoring control.