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ABSTRACT: The objective of this study was to determine prevalence and perform genomic analysis of Salmonella spp. and Campylobacter spp. isolated from different stages of an integrated NAE broiler complex. Environmental samples were screened with 3M-Molecular Detection System (MDS) and MDS positive samples were further processed for confirmation of results and identification. Core genome-based phylogenies were built for both bacteria isolated from this study along with selected NCBI genomes. The odds ratios and 95% confidence limits were compared among stages and sample types (α < 0.05) using multivariable model. Based on MDS results, 4% and 18% of total samples were positive for Salmonella spp. and Campylobacter spp. respectively. The odds of Salmonella detection in hatchery samples were 2.58 times as likely as compared to its detection in production farms’ samples (P = 0.151) while the odds of Campylobacter detection in production farms’ samples were 32.19 times as likely as its detection in hatchery (P = 0.0015). Similarly, the odds of Campylobacter detection in boot swabs, soil, water, and miscellaneous samples were statistically significant (P < 0.05) as compared with fly paper as reference group. The serovars identified for Salmonella were Typhimurium, Barranquilla, Liverpool, Kentucky, Enteritidis, Luciana, and Rough_O:r:1,5. For Campylobacter, the species identified were Campylobacter jejuni and Campylobacter coli. Phylogeny results show close genetic relatedness among bacterial strains isolated from different locations within the same stage and between different stages. The results show possibility of multiple entry points of such bacteria entering broiler complex and can potentially contaminate the final raw product in the processing plant. It suggests the need for a comprehensive control strategy with strict biosecurity measures and best management practices to minimize or eliminate such pathogens from the poultry food chain.

Poultry meat products are considered the major contributors to Campylobacteriosis in humans. The objective of this study was to determine the prevalence status, critical entry points, and movement patterns of Campylobacter spp. along different stages of an integrated broiler complex. To isolate bacteria and perform phylogenetic analysis, a total of 790 environmental samples were collected from 38 production houses, a hatchery, 6 transport trucks, and a processing plant of a commercial broiler complex. Odds ratio and 95% confidence intervals were compared among different stages and sample types (α = 0.05). Altogether 17% (137/790) of samples and 61% (23/38) of production houses were positive for Campylobacter spp. Similarly, 34% (46/135) of samples were identified as Campylobacter jejuni (C. jejuni), and 61% (83/135) were identified as Campylobacter coli (C. coli). The odds of Campylobacter spp. detection in broiler farms’ surroundings were 4 times (1.88–8.26; 95% CLs) more likely as compared to parent pullets and breeder farms’ surroundings (p = 0.0004). Similarly, among different sample types, the odds of Campylobacter spp. detection in boot swabs and sponge-stick swabs were more likely as compared to fly paper samples (p ≤ 0.0024). In addition, the odds of Campylobacter spp. detection in postpick whole carcass rinses were 4 times (1.99–7.59; 95% CLs) more likely as compared to postchill carcass rinses (p = 0.0004). The phylogeny results of both C. jejuni and C. coli indicate multiple critical entry points of bacterial strains along the chain and suggest the possibility of transmission of Campylobacter spp. from broiler grow-out flocks through transport and to final raw products (29%) in the processing plant. The results indicate potential risks of foodborne infections in consumers from ingestion of contaminated raw or undercooked poultry meat. Therefore, a comprehensive control strategy may be essential to reduce or eliminate Campylobacter spp. or other zoonotic pathogens from the poultry food chain.

ABSTRACT: Transportation is a potential point of cross-contamination before broiler chickens arrive at the processing plant for slaughter. Previous studies have associated the use of uncleaned transport containers with the introduction of pathogenic bacteria onto uncontaminated broilers. The objective of this study was to quantify the transfer of Salmonella from transport drawer perforated flooring to broiler chickens during different holding times. For traceability, the flooring of each drawer was inoculated with fecal content slurry containing a marker strain of Salmonella Infantis. Three drawers per treatment were used, and each drawer was subjected to one of the following treatments: pressure wash, disinfectant, and pressure wash (A), pressurized steam followed by forced hot air (B), or no cleaning (C). Drawers were classified as top, middle, or bottom based on their relative position with each other. After treatment, broilers were introduced to each drawer and held for 2, 4, or 6 h. At each timepoint, broilers were removed from drawers, euthanized, and carcasses rinsed to obtain Salmonella counts. Samples under the limit of direct plating detection were enriched, plated, and later confirmed positive or negative. Differences were observed per treatment, holding time, and drawer relative position (P < 0.0001). Broilers placed in transport containers that underwent a cleaning procedure (A or B) had lower levels of Salmonella when compared to broilers placed in noncleaned containers (C). However, most of the samples below the limit of detection were positive after enrichment, indicating that both procedures evaluated need improvement for efficient pathogen inactivation. A decrease in Salmonella transfer was observed after 6 h in rinsates obtained from broilers placed in noncleaned containers (C). Rinsates obtained from top drawers had less Salmonella than the middle or bottom drawers when broilers were placed in transport containers that underwent a cleaning procedure (A and B). The application of pressurized steam and forced hot air was comparable to the use of water washes and disinfectant indicating a potential role in cleaning poultry transport containers.

ABSTRACT: In the United States, cleaning poultry transport containers prior to arrival at the broiler grow-out farm is not currently a widely adopted practice in the industry. However, previous studies have shown that transport containers have an important role in cross-contamination before the broilers arrive at the processing plant. The objective of this study was to evaluate the efficacy of pressurized steam followed by forced hot air to clean transport container flooring and compare it to conventional cleaning procedures. Fiberglass and plastic flooring were cut into even pieces and inoculated with chicken intestinal contents containing Salmonella Infantis or Campylobacter jejuni. The cleaning treatments were pressurized steam, forced hot air, pressurized steam followed by forced hot air, water pressure washing, water pressure washing before and after disinfectant, and no cleaning. Counts for Salmonella, Campylobacter, Escherichia coli, coliforms, and aerobic bacteria were assessed. All reductions were made in comparison to noncleaned samples. Forced hot air applied by itself was not efficient in reducing Campylobacter, coliforms, and E. coli; and limited reductions (less than 1 log10 CFU/cm2) were observed for Salmonella and aerobic bacteria. Then, for all bacteria types evaluated, pressurized steam by itself showed reductions of 2.4 to 3.5 log10 CFU/cm2. Samples that were cleaned with a single-pressure water wash showed reductions of 4.0 to 4.6 log10 CFU/cm2 for all bacteria types. For Salmonella, Campylobacter, and E. coli, the greatest reductions were observed when samples were cleaned with pressurized steam followed by forced hot air (4.3–6.1 log10 CFU/cm2) or water washed before and after disinfectant (4.5–6.2 log10 CFU/cm2), and these treatments did not differ from each other. Pressurized steam followed by forced hot air was shown to be an efficient cleaning procedure to reduce poultry-associated pathogens on transport cage flooring, with the benefit of using less water than conventional water cleaning. Processors may be able to adapt this process to reduce potential cross-contamination and lessen the level of pathogens entering the processing plant.

Various culture-based methods to detect Salmonella in animal feed have been developed due to the impact of this bacterium on public and animal health. For this project, tris phosphate carbonate (TPC) and buffered peptone water (BPW) buffering capacities were compared as pre-enrichment mediums for the detection of Salmonella in feed ingredients. A total of 269 samples were collected from 6 feed mills and mixed with the pre-enrichments; pH was measured before and after a 24 h incubation. Differences were observed when comparing pH values by sample type; DDGS and poultry by-product meal presented lower initial pH values for TPC and BPW compared to the other samples. For both TPC and BPW, meat and bone meal presented higher final pH values, while soybean meal and peanut meal had lower final pH values. Furthermore, for BPW, post cooling, pellet loadout, and wheat middlings reported lower final pH values. Additionally, most feed ingredients presented significant differences in pH change after 24 h of incubation, except DDGS. From meat and bone meal samples, four Salmonella isolates were recovered and identified: three using BPW and one using TPC. TPC provided greater buffer capacity towards neutral pH compared to BPW, but BPW was more effective at recovering Salmonella.