Sensitivity of Direct Culture, Enrichment and PCR for Detection of Campylobacter jejuni and C. coli in Broiler Flocks at Slaughter

Abstract

Broiler chicken flocks are a significant source of Campylobacter jejuni and Campylobacter coli that result in the major public health problem of campylobacteriosis. Accurate estimates of the prevalence of both C. coli and C. jejuni in flocks would enhance epidemiological understanding, risk assessment and control options. This study combined results from a panel of 10 detection tests (direct culture, enrichment and PCR) on caecal samples from flocks at slaughter. A parallel interpretation approach was used to determine the presence of Campylobacter spp. and for C. jejuni and C. coli individually. The sample was considered positive if at least one method detected the target and this interpretation was taken to represent a 'proxy gold standard' for detection in the absence of a gold standard reference test. The sensitivity of each individual method to detect Campylobacter spp., C. jejuni and C. coli was then estimated relative to the proxy gold standard. Enrichment in adapted Exeter broth (deficient in polymyxin B) with a resuscitation step was 100% sensitive, whilst direct culture on modified charcoal cefoperazone deoxycholate agar (mCCDA) was highly sensitive (97.9%). Enrichment methods using Preston broth and Bolton broth were significantly less sensitive. Enrichment in Exeter broth promoted the recovery of C. jejuni, whilst enrichment in Bolton broth favoured C. coli. A RT-PCR detection test could identify 80% of flocks that were co-colonised with both species. This study found that 76.3% (n = 127) of flocks were colonised with Campylobacter spp. The majority (95.9%) of Campylobacter-positive flocks were colonised with C. jejuni; however, approximately one-third of positive flocks were simultaneously colonised with both C. jejuni and C. coli. The findings highlight the impact of different detection methodologies on the accuracy of the estimated incidence of both C. jejuni and C. coli entering the abattoir within broiler flocks and the associated public health risks.

Molecular Detection, Typing, and Quantification of Campylobacter spp. in Foods of Animal Origin

“…This methodology can readily be applied to Campylobacter spp. and C jejuni (Frasao and others ) as well as C. coli , as has been described by various authors (Bratz and others ; Zendehbad and others ; Kashoma and others ; Rodgers and others ; Shams and others ); C. jejuni, C. coli, C. lari , and C. upsaliensis can also be identified using this method (Girgis and others ) (Table ). In addition, Raja and Rao () used the technique to detect 2 different pathogens, C. jejuni and Listeria monocytogenes , in chicken meat during processing.…”

“…The choice of method will depend on the relevant conditions. Some authors use the boiling method (Shams and others ; Zang and others ), others prefer the kit (Frasao and others , b; Ayaz and others ; Gosselin‐Theberge and others ; Rodgers and others ), whereas others employ phenol‐chloroform‐isoamyl alcohol (25: 24: 1) extraction (Raja and others ).…”

Co-occurrence of Campylobacter Species in Children From Eastern Ethiopia, and Their Association With Environmental Enteric Dysfunction, Diarrhea, and Host Microbiome

“…was attempted; however, due to technical and logistical challenges faced in Ethiopia, the results generated were deemed unreliable (data not shown). The fastidious nature of Campylobacter and difficulty of isolation, rendered culture and isolation unreliable for estimation of prevalence (32). By consequence, other studies utilized different approaches to estimate the prevalence of Campylobacter in stool samples, including enzyme immunoassay (EIA), PCR, and shotgun metagenomics (7,33,34).…”

Isolation and Whole‐Genome Sequencing of Environmental Campylobacter

“…A common method for determining the species Campylobacter is by performing PCR with genomic DNA and primers specific for either C. jejuni or C. coli. To identify C. jejuni, primers for the mapA gene are commonly used, while primers for the ceuE gene are used to identify C. coli (Rodgers, Simpkin, Lee, Clifton-Hadley, & Vidal, 2017;Vidal et al, 2016). These primers are then used in a standard PCR along with a species standard.…”