Research carried out in Europe and Asia has begun to address this question with various culture-based studies. Researchers from Taiwan, Finland, Sweden, Demark and the Netherlands have examined various dog populations and have been able to culture C. jejuni, C. coli, C. upsaliensis, C. helveticus, C. lari and other Campylobacter spp. from canine fecal samples using various growth conditions and media [13–17]. Reported carriage rates of Campylobacter spp. in domestic

dogs ranged from 2.7% to 100% of dogs tested [13, 16], with some studies reporting isolation of multiple species of Campylobacter from a single dog [15, 17]. A major influence on our understanding of Campylobacter ecology in dogs has been our reliance on culture-based methods. Tipifarnib nmr Various selective media have been used for Campylobacter isolation

[18], with most relying on a cocktail of antibiotics in a rich basal medium to selectively isolate Campylobacter. However, it has been recognized that Campylobacter 17-AAG chemical structure species other than C. coli, C. jejuni, and C. lari are often sensitive to the antibiotics in these media [19]. Filter-based methods, in combination with nonselective media, have been shown to result in the isolation of a greater diversity of Campylobacter species [20], but these approaches are more labour-intensive, less selective and prone to overgrowth of fecal contaminants [19]. As our understanding of campylobacters, both pathogenic and non-pathogenic, expands beyond C. jejuni and C. coli, so must our detection methods. The goal of this study was to take a culture-independent approach to the profiling of Campylobacter species in domestic pet dogs in an effort to evaluate this zoonotic reservoir and describe changes in fecal Campylobacter populations associated with diarrhea. Established species-specific

Megestrol Acetate quantitative PCR (qPCR) assays targeting the 60 kDa chaperonin (cpn60) gene of C. coli, C. concisus, C. curvus, C. fetus, C. gracilis, C. helveticus, C. hyointestinalis, C. jejuni, C. lari, C. mucosalis, C. rectus, C. showae, C. sputorum, and C. upsaliensis [21] were used to determine the Campylobacter profiles of 70 healthy dogs and 65 dogs with diarrhea. This study represents the largest culture-independent, quantitative investigation of Campylobacter in pet dogs conducted to date and is one of only a few studies to focus on North American animals. Results Campylobacter profiles from healthy and diarrheic dog fecal samples Total bacterial DNA was extracted from the feces of 70 healthy dogs (from 52 households) and 65 dogs with diarrhea (from 60 households) (Additional file 1: Table S1) and tested for the presence of 14 Campylobacter species. Each sample was tested for an individual species in four reactions (duplicate reactions within an assay and each assay run twice). If a sample did not yield three or four detectable test values (above the assay cut-off of 103 organisms/g of feces [21]), the sample was defined as undetectable for that test.