Detection of
            <i>Campylobacter jejuni</i>
            and
            <i>Campylobacter coli</i>
            in Environmental Waters by PCR Enzyme-Linked Immunosorbent Assay

Contamination of retail poultry by Campylobacter spp. and Salmonella enterica is a significant source of human diarrheal disease. Isolation and identification of these microorganisms require a series of biochemical and serological tests. In this study, Campylobacter ceuE and Salmonella invA genes were used to design probes in PCR-enzyme-linked immunosorbent assay (ELISA), as an alternative to conventional bacteriological methodology, for the rapid detection of Campylobacter jejuni, Campylobacter coli, and S. enterica from poultry samples. With PCR-ELISA (40 cycles), the detection limits for Salmonella and Campylobacter were 2 ؋ 10 2 and 4 ؋ 10 1 CFU/ml, respectively. ELISA increased the sensitivity of the conventional PCR method by 100-to 1,000-fold. DNA was extracted from carcass rinses and tetrathionate enrichments and used in PCR-ELISA for the detection of Campylobacter and S. enterica, respectively. With PCR-ELISA, Salmonella was detected in 20 of 120 (17%) chicken carcass rinses examined, without the inclusion of an enrichment step. Significant correlation was observed between PCR-ELISA and cultural methods (kappa ‫؍‬ 0.83; chi-square test, P < 0.001) with only one false negative (1.67%) and four false positives (6.67%) when PCR-ELISA was used to screen 60 tetrathionate enrichment cultures for Salmonella. With PCR-ELISA, we observed a positive correlation between the ELISA absorbance (optical density at 405 nm) and the campylobacter cell number in carcass rinse, as determined by standard culture methods. Overall, PCR-ELISA is a rapid and cost-effective approach for the detection and enumeration of Salmonella and Campylobacter bacteria on poultry.

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Rapid Detection of Campylobacter coli , C. jejuni , and Salmonella enterica on Poultry Carcasses by Using PCR-Enzyme-Linked Immunosorbent Assay

Yang

Berrang

Liu³

et al. 2003

“…The presence of C. coli in samples of environmental water has been reported previously (33,39), and there have been a small number of reports linking cases of human disease to water supplies (11,21). It has also been reported that a laboratory strain of C. coli is less well adapted to survival in water than C. jejuni or C. lari is (37).…”

Section: Vol 70 2004 Characteristics Of C Coli From Cheshire Studymentioning

confidence: 95%

Genotypic and Antibiotic Susceptibility Characteristics of a Campylobacter coli Population Isolated from Dairy Farmland in the United Kingdom

Leatherbarrow

Kemp

Williams

et al. 2004

Campylobacter infections are the most common cause of bacterial enteritis in humans, and nearly 8% of such infections are caused by Campylobacter coli. Most studies have concentrated on Campylobacter jejuni, frequently isolated from intensively farmed poultry and livestock production units, and few studies have examined the spread and relatedness of Campylobacter across a range of geographical and host boundaries. Systematic sampling of a 100-km 2 area of mixed farmland in northwest England yielded 88 isolates of C. coli from a range of sample types and locations, and water was heavily represented. Screening for antibiotic resistance revealed a very low prevalence of resistance, while genotyping performed by using three methods (flaA PCR restriction fragment length polymorphism [RFLP], pulsed-field gel electrophoresis [PFGE], and fluorescent amplified fragment length polymorphism [fAFLP]) provided insights into the genomic relatedness of isolates from different locations and hosts. Isolates were classified into 23 flaA groups, 34 PFGE groups, and five major fAFLP clusters. PFGE banding analysis revealed a high level of variability and no clustering by sample type. fAFLP and flaA analyses successfully grouped the isolates by sample type. We report preliminary findings suggesting that there is a strain of C. coli which may have become adapted to survival or persistence in water and that there is a group of mainly water-derived isolates from which unusual flaA PCR fragments were recovered.

“…Other reported sources of infection include contaminated water and raw milk (22,32); however, in most cases the exact source of disease is not determined (43). Environmental loading and transmission may be factors associated with nonoutbreak cases, particularly those related to contamination of ambient waters (4,28,41,42).…”

mentioning

confidence: 99%

Distribution and Ecology of Campylobacters in Coastal Plain Streams (Georgia, United States of America)

Vereen

Lowrance

Cole

et al. 2007

Campylobacter is the leading cause of bacterium-associated diarrhea in the United States and most developed countries. While this disease is considered a food-borne disease, many clinical cases cannot be linked to a food source. In rural and agrarian areas environmental transmission may be an important factor contributing to case loads. Here we investigated the waterborne prevalence of campylobacters in a mixed-use rural watershed in the coastal plain of southern Georgia (United States). Six sites representing various degrees of agricultural and human influence were surveyed biweekly to monthly for 1 year for the presence of culturable thermophilic campylobacters and other measures of water quality. Campylobacters were frequently present in agricultureand sewage-impacted stretches of streams. The mean campylobacter counts and overall prevalence were highest downstream from a wastewater treatment plant that handled both human and poultry slaughterhouse waste (<595 CFU ml ؊1 ; 100% positive); the concentrations were significantly higher than those for the four upstream sites (P < 0.05). The counts were significantly correlated with the number of fecal coliform bacteria, conductivity, pH, and concentrations of nutrients (NO 3 ؊ , PO 4 3؊ , and NH 3 ). Campylobacters were isolated more frequently and larger numbers were present during the summer months, similar to the occurrence of clinical cases of campylobacteriosis in this region. A multivariate model showed that the levels were significantly influenced by increasing precipitation, which also peaked in the summer. The results indicate that loading from both human and domestic animal waste may be high in the watershed studied during the summer months. Mixed-use watersheds supporting agriculture production, human populations, and wildlife may be at risk for contamination by campylobacters and may be an important route for human exposure.