The accurate and rapid detection of Campylobacter spp. is critical for optimal surveillance throughout poultry processing in the United States. The further development of highly specific and sensitive assays to detect Campylobacter in poultry matrices has tremendous utility and potential for aiding the reduction of foodborne illness. The introduction and development of molecular methods such as polymerase chain reaction (PCR) have enhanced the diagnostic capabilities of the food industry to identify the presence of foodborne pathogens throughout poultry production. Further innovations in various methodologies, such as immune-based typing and detection as well as high throughput analyses, will provide important epidemiological data such as the identification of unique or region-specific Campylobacter. Comparable to traditional microbiology and enrichment techniques, molecular techniques/methods have the potential to have improved sensitivity and specificity, as well as speed of data acquisition. This review will focus on the development and application of rapid molecular methods for identifying and quantifying Campylobacter in U.S. poultry and the emergence of novel methods that are faster and more precise than traditional microbiological techniques.

Salmonella Enteritidis is responsible for a significant proportion of foodborne salmonellosis in the United States and continues to be attributable to table eggs despite increased federal oversight. Technologies, including feed additives, continue to be evaluated for preharvest application and their potential food safety benefits. Diamond V Original XPC, a Saccharomyces cerevisiae fermentation-based postbiotic (SCFP), was evaluated for its effectiveness in reducing Salmonella Enteritidis (SE) colonization in young layer pullets. A total of 40 day-old Hy-Line W-36 layer pullets were equally divided and randomly assigned to one of two dietary treatments, with SCFP or without SCFP (PCON), and orally gavaged on day 28 with SE at 106 CFU/mL. Another 20 day-old pullets were fed the same control feed without SCFP and blank inoculated on day 28 with 1 mL of sterile phosphate-buffered saline to serve as a negative control. Qualitative and quantitative analyses of cecal contents for Salmonella were performed for all birds on day 32. The prevalence of SE in the ceca of all directly challenged birds was 100%; however, the SE concentration in birds fed SCFP diet (3.35 log CFU/g) was significantly lower (P < 0.0001) than that of the PCON birds not fed SCFP (4.49 log CFU/g). The proportion of birds with enumerable SE concentrations was lower in SCFP-fed pullets (57.9%) than in the PCON pullets (95.0%). These data suggest that inclusion of SCFP in the diet may aid in the reduction of SE within the ceca of commercial laying hens and could serve as an additional preharvest food safety hurdle.

Poultry is a major reservoir for the pathogen Campylobacter jejuni. C. jejuni inhabits the poultry gastrointestinal tract as a part of the gut microbiota. The objective of this study was to evaluate both the survival of C. jejuni and the changes in the population dynamics of the cecal microbiome during an in vitro C. jejuni inoculation in the presence or absence of the functional metabolites of Diamond V Original XPC^TM (XPC). Two independent trials were conducted. Broiler chickens (n = 6 per Trial 1 and n = 3 per Trial 2) were raised according to standard industry guidelines and euthanized on Day 41. The ceca were collected aseptically, their contents removed independently and then used in an in vitro microaerobic model with 0.1% cecal contents + Campylobacter with or without 1% XPC (w/v). Before the inoculation with a chloramphenicol resistant marker strain of C. jejuni, the cecal contents were pre-incubated with XPC at 42°C for 24 h, in a shaking incubator (200 rpm) under microaerobic conditions, then experimentally inoculated with 10⁸/ml of C. jejuni into the appropriate treatment groups. At 0 and 24 h for Trial 1, and 48 h for Trial 2, sub-samples of the culture (n = 3 ceca, two technical replicates per ceca, XPC alone or ceca culture alone) were enumerated using a Petroff–Hausser counter, and the DNA was extracted for microbiome analysis. DNA was isolated using the Qiagen QIAamp Fast Stool DNA Mini Kit and sequenced using the Illumina MiSeq platform. The reads were filtered, normalized, and assigned taxonomical identities using the QIIME2 pipeline. The relative microbiota populations were identified via ANCOM. Altogether, evidence suggests that XPC alters the microbiome, and in turn reduces Campylobacter survival.

Salmonella enterica and Campylobacter spp. cause a considerable number of human illnesses each year, and the vast majority of cases are foodborne. The purpose of this study was to establish the baseline of Salmonella and Campylobacter in beef products purchased from U.S. retail markets. Sampling was carried out in 38 American cities. Retail raw ground and whole-muscle beef (n = 2,885) samples were purchased and examined for the presence of Salmonella. Samples testing positive for Salmonella were identified with the commercial BAX System, which is a real-time PCR-based system. Of the original samples purchased, 1,185 were selected and tested for the presence of Campylobacter. Positive samples were isolated via direct plating and confirmed via agglutination and biochemical testing. Salmonella was detected in 0.66% of the total samples purchased. The prevalence of Salmonella in ground beef packages was 0.42% for modified atmosphere packaging, 0.63% for chub packaging, and 0.59% for overwrapped packages. Salmonella was detected in 1.02% of whole-muscle cuts. There was no relationship (P = 0.18) between product type (ground or whole muscle) and the percentage of positive samples. Campylobacter was recovered from 9.3% of samples. A greater percentage (17.24%, P < 0.01) of whole-muscle cuts tested positive for Campylobacter compared with ground beef samples (7.35%). Estimating pathogen baselines in U.S. retail beef is essential for allotting resources and directing interventions for pathogen control. These data can be utilized for a more complete understanding of these pathogens and their impact on public health from the consumption of beef products.

Postbiotic feed additives may aid foodborne pathogen reduction during poultry rearing. The study objective was to evaluate a postbiotic additive in parallel to an industry control diet and the subsequent associated burden of Salmonella enterica on a single, commercial broiler farm in Honduras. Twelve houses were matched and assigned the standard diet (CON) or standard diet plus postbiotic (SCFP). New litter was placed in each house and retained across flock cycles with sampling prior to each chick placement and three consecutive rearing cycles. At ~33–34 days, 25 ceca were collected on-farm from each house, treatment, and cycle. Salmonella prevalence in litter for CON (30.6%) and SCFP (27.8%) were equivalent; however, Salmonella load within positive samples was lower (p = 0.04) for SCFP (3.81 log₁₀ MPN/swab) compared to CON (5.53 log10 MPN/swab). Cecal prevalence of Salmonella was lower (p = 0.0006) in broilers fed SCFP (3.4%) compared to CON (12.2%). Salmonella load within positive ceca were numerically reduced (p = 0.121) by 1.45 log₁₀ MPN/g for SCFP (2.41 log₁₀ MPN/g) over CON (3.86 log₁₀ MPN/g). Estimated burden was lower (p = 0.003) for SCFP flocks (3.80 log₁₀ MPN) compared to CON (7.31 log₁₀ MPN). These data demonstrate the preharvest intervention potential of postbiotics to reduce Salmonella enterica in broiler chickens.

Determining the efficacy of feed-additive technologies utilized as pre-harvest food-safety interventions against Salmonella enterica may be influenced by factors including, but not limited to, mechanism of action, experimental design variables, Salmonella serovar(s), exposure dose, route, or duration in both controlled research and real-world field observations. The purpose of this study was to evaluate the dietary inclusion of a Saccharomyces cerevisiae fermentation-derived postbiotic (SCFP) additive (Diamond V, Original XPC^®) on the colonization of cecal and ovarian tissues of commercial pullets directly and indirectly exposed to Salmonella Enteritidis (SE). Four hundred and eighty commercial, day-of-age W-36 chicks were randomly allotted to 60 cages per treatment in two identical BSL-2 isolation rooms (Iowa State University) with four birds per cage and fed control (CON) or treatment (TRT) diets for the duration of study. At 16 weeks, two birds per cage were directly challenged via oral gavage with 1.1 × 10⁹ CFU of a nalidixic-acid-resistant SE strain. The remaining two birds in each cage were thus horizontally exposed to the SE challenge. At 3, 7, and 14 days post-challenge (DPC), 20 cages per group were harvested and sampled for SE prevalence and load. No significant differences were observed between groups for SE prevalence in the ceca or ovary tissues of directly challenged birds. For the indirectly exposed cohort, SE cecal prevalence at 7 DPC was significantly lower for TRT (50.0%) vs. CON (72.5%) (p = 0.037) and, likewise, demonstrated significantly lower mean SE cecal load (1.69 Log₁₀) vs. CON (2.83 Log₁₀) (p = 0.005). At 14 DPC, no significant differences were detected but ~10% fewer birds remained positive in the TRT group vs. CON (p > 0.05). These findings suggest that diets supplemented with SCFP postbiotic may be a useful tool for mitigating SE colonization in horizontally exposed pullets and may support pre-harvest food-safety strategies.