Nonculturability Might Underestimate the Occurrence ofCampylobacterin Broiler Litter

Abstract

We investigated the contribution of litter to the occurrence of Campylobacter on three broiler farms, which were known to have low (LO) and high (HI-A and HI-B) Campylobacter prevalence. For this purpose, we collected litter samples (n = 288) during and after two rearing cycles from each farm. We evaluated the occurrence of Campylobacter (using selective enrichment and quantitative real-time polymerase chain reaction [q-PCR] analysis) in the litter samples as well as the litter's pH and moisture content. Ceca from each flock (n = 144) were harvested at slaughter age and used to quantify Campylobacter colony-forming units (CFUs). Campylobacter was only retrieved from 7 litter samples that were collected from HI-A and HI-B during the growing period, but no Campylobacter was isolated from LO farms. The q-PCR analysis detected Campylobacter in pooled litter samples from all three farms. However, in litter collected during the same rotation, Campylobacter levels were significantly higher (p < 0.05) in HI-A and HI-B litter samples in comparison to those in LO. Cecal samples from HI-A and HI-B yielded relatively high numbers of Campylobacter CFUs, which were undetectable in LO samples. Litter's pH and moisture did not affect the overall occurrence of Campylobacter in litter and ceca on any of the farms. Our data suggest that Campylobacter was generally more abundant in litter that was collected from farms with highly colonized flocks. Therefore, better approaches for assessing the occurrence of Campylobacter in litter might be warranted in order to reduce the dissemination of these pathogens on and off poultry farms.

Prevalence and Antimicrobial Resistance Profiles of Foodborne Pathogens Isolated from Dairy Cattle and Poultry Manure Amended Farms in Northeastern Ohio, the United States

“…One mL of the slurry was then ten-fold serially diluted using PBS, and 100 µL of each serial dilution was plated onto modified charcoal cefoperazone deoxycholate agar (mCCDA; Fisher Scientific, Waltham, MA, USA), RAPID’ L. mono (Bio Rad, Hercules, CA, USA), sorbitol MacConkey agar plates containing cefixine and tellurite (SMACct; Becton, Dickinson; BD, San Jose, CA, USA), and xylose lysine tergitol-4 agar (XLT-4; Bio Rad, Hercules, CA, USA) plates for the direct isolation of Campylobacter , L. monocytogenes , E. coli O157, and Salmonella , respectively. Samples that did not show any bacterial growth due to direct plating were subjected to enrichment using pathogen specific media [ 34 , 35 , 36 , 37 , 38 ].…”

“…To isolate Campylobacter spp., 1 mL of manure or soil slurry was suspended in 9 mL of Preston enrichment broth containing Campylobacter growth supplements (CM067, SR048, SR117, and SR232; Oxoid Ltd., Cambridge, UK) [ 38 ]. The suspensions were incubated under microaerobic conditions (5% O 2 , 10% CO 2 , and 85% N 2 ) at 42 °C for 48 h. After enrichment, 100 µL was plated on mCCDA containing a selective supplement (SR0155; Oxoid Ltd., Cambridge, UK) and incubated under microaerobic conditions at 42 °C for 48 h. The isolated colonies from mCCDA plate were then subcultured onto Mueller–Hinton (MH) agar containing a selective supplement (SR0117; Oxoid Ltd., Cambridge, UK) and incubated at 42 °C for 48 h under microaerobic conditions [ 39 , 40 , 41 ].…”

Association of Broiler Litter Microbiome Composition and Campylobacter Isolation

“…In addition, the outcome of this study relied on bacterial isolation, a method with a moderate sensitivity for Campylobacter detection that might lead to misclassification of controls. As an example, if Campylobacter cells entered a viable not culturable state ( 16 , 80 ), there is the possibility that flocks were assigned as controls due to lack of Campylobacter isolation in boot socks even though Campylobacter was present in the broiler house. However, the significant positive association between the relative abundance of Campylobacter ASVs in litter and the isolation of Campylobacter in boot socks supports the validity of the bacterial isolation approach.…”

“…Windrowing is performed to control poultry pathogens present in the broiler litter (13). However, there is evidence that Campylobacter can persist in reused broiler litter (14)(15)(16). Therefore, survival of Campylobacter in the broiler litter may play an important role in the ecology and colonization of this bacterium in broiler chickens.…”

Translating ‘big data’: better understanding of host-pathogen interactions to control bacterial foodborne pathogens in poultry

“…However, foodborne pathogens can persist for an extended period of time within hosts and agricultural environments despite decontamination efforts and therapeutic measures (Winfield and Groisman, 2003; Dinçer and Baysal, 2004; Foley et al ., 2011, 2013). Further, evidence of the association between antimicrobial use in animal production and the emergence of cross resistance, multi-drug resistant (MDR) isolates, and other evolutionary adaptations have been reported (Kassem et al ., 2017; Nisar et al ., 2017 a ; Deblais et al ., 2018 b ). Based on the National Outbreaks Reporting System (NORS) database, 54.7% of the total enteric zoonotic disease outbreaks recorded in the USA between 1996 and 2017 were associated with poultry; therefore, novel control and management practices must be established to assure food and public safety.…”