Draft genome sequences of Campylobacter species isolated from the cecal contents of broiler chickens.

Here, we report the draft genomes of 10 Campylobacter strains isolated from the cecal contents of market-age broiler chickens naturally colonized with Campylobacter. Through a comprehensive analysis of these draft genomes, we have unveiled their core genetic elements and several antimicrobial resistance genes.

Inclusion of trans-cinnamaldehyde and caprylic acid in feed results in detectable concentrations in the chicken gut and reduces foodborne pathogen carriage.

Poultry act as a major reservoir host for Salmonella and Campylobacter spp., the 2 leading causes of foodborne illnesses globally and in the United States. Preharvest stage interventions to reduce foodborne pathogen carriage in poultry are increasingly informed by consumer preference for antibiotic-free poultry production. The in-feed inclusion of plant-derived antimicrobial compounds is a promising antibiotic alternative strategy to reduce foodborne pathogen load in the broiler chicken gut. Yet, the fate of these phytochemicals through the broiler chicken gastrointestinal tract is unknown. Likewise, while in-feed phytochemicals have been widely demonstrated in challenge models to reduce foodborne pathogen carriage, little is known regarding efficacy to curb natural routes of infection. As such, the aim of the present study was 2-fold. We sought to determine the concentrations of 2 phytochemicals, trans-cinnamaldehyde and caprylic acid, in each region of the chicken gastrointestinal tract following their in-feed inclusion over a 6-wk production period. In addition, we investigated how the in-feed provision of these phytochemicals may protect against environmental acquisition of Campylobacter jejuni and Salmonella spp. Trans-cinnamaldehyde and caprylic acid were detected in crop, gizzard, duodenal, jejunal, and ileal contents. Crop and gizzard concentrations were not significantly (P > 0.05) different. A significant (P < 0.05) decrease in phytochemical concentration was observed in intestinal regions compared to crop and gizzard. Trans-cinnamaldehyde was consistently identified in cecal and colon contents, while caprylic acid was not detectable in these regions. Trans-cinnamaldehyde and caprylic acid were found to reduce (P < 0.05) Salmonella load. Together, our data establish that the in-feed addition of trans-cinnamaldehyde and caprylic acid, 2 phytochemicals that have previously been shown to exert antimicrobial activity against poultry-associated foodborne pathogens, results in detectable concentrations in the broiler chicken gastrointestinal tract. By providing researchers with a gastrointestinal region-by-region map of phytochemical concentrations, the present study is expected to inform the choice of in-feed phytochemicals targeting foodborne pathogen carriage in the broiler chicken gastrointestinal tract.

A Multiomic Analysis of Chicken Serum Revealed the Modulation of Host Factors Due to Campylobacter jejuni Colonization and In-Water Supplementation of Eugenol Nanoemulsion.

Campylobacter jejuni is a foodborne pathogen that causes campylobacteriosis globally, affecting ~95 million people worldwide. Most C. jejuni infections involve consuming and/or handling improperly cooked poultry meat. To better understand chicken host factors modulated by Campylobacter colonization, we explored a novel LCMS-based multiomic technology using three experimental groups: (1) negative control, (2) positive control, and (3) eugenol nanoemulsion (EGNE) treatment (supplemented with 0.125% EGNE in the water) of broiler chickens (n = 10 birds/group). Birds in groups two and three were challenged with C. jejuni on day 7, and serum samples were collected from all groups on day 14. Using this multiomic analysis, we identified 1216 analytes (275 compounds, seven inorganics, 407 lipids, and 527 proteins). The colonization of C. jejuni significantly upregulated CREG1, creatinine, and 3-[2-(3-Hydroxyphenyl) ethyl]-5-methoxyphenol and downregulated sphingosine, SP d18:1, high mobility group protein B3, phosphatidylcholines (PC) P-20:0_16:0, PC 11:0_26:1, and PC 13:0_26:2. We found that 5-hydroxyindole-3-acetic acid significantly increased with the EGNE treatment when compared to the positive and negative controls. Additionally, the treatment increased several metabolites when compared to the negative controls. In conclusion, this study revealed several potential targets to control Campylobacter in broiler chickens.

Ability of Garlic and Ginger Oil to Reduce Salmonella in Post-Harvest Poultry.

Approximately 1.35 million human salmonellosis cases are reported in the United States every year, resulting in over 26,000 hospitalizations and 400 deaths. Consumption of contaminated poultry products is one of the leading causes of human salmonellosis. Poultry meat becomes contaminated when feces from an infected bird comes into contact with the carcass during processing. Additional carcasses can then become cross-contaminated along the processing line. While chemicals such as peracetic acid are currently used to kill microbes such as Salmonella, consumers are increasingly calling for more natural alternatives. Our objective for this study was to determine the ability of the phytochemicals garlic and ginger oil to reduce Salmonella prevalence in the processing environment. In a simulated scalding tank environment, dipping contaminated chicken skin samples in a solution containing both garlic and ginger oil reduced Salmonella by up to 2 log CFU. Furthermore, the oils prevented Salmonella growth in the tank solution. The mechanism of action of garlic and ginger was evaluated using the sub-inhibitory concentration of each oil individually. While both were found to decrease autoinducer-2 (AI-2) levels, no effect was seen on expression of 10 genes involved in Salmonella virulence and survival. In total, this work demonstrates the potential of garlic and ginger to reduce Salmonella prevalence in the post-harvest environment. However, more work remains to be done to understand the mechanism of action.

Effect of Salmonella Typhimurium Colonization on Microbiota Maturation and Blood Leukocyte Populations in Broiler Chickens.

Reducing Salmonella in commercial chickens is vital to decreasing human salmonellosis infections resulting from contact with contaminated poultry and poultry products. As the intestinal microbiota plays an important role in preventing pathogen colonization, we sought to understand the relationship between Salmonella infection and the cecal microbiota and the host immune system. Day-of-hatch broiler chicks were assigned to three treatments: control, artificial (SA), and natural (SN) Salmonella infection. At seven days of age, control and SA birds were inoculated with PBS or Salmonella Typhimurium, respectively. Five SA birds were transferred to SN cages to facilitate natural infection. Cecal content and blood samples were collected at 0, 8, 14, and 21 days of age for microbiota and leukocyte analysis, respectively. A significant change in microbiota composition was observed in both groups as noted by a decrease in Lactobacillus and Escherichia and an increase in Bacteroides. Leukocyte analysis revealed a decrease in the percentage of circulating monocytes at 7 days post-infection while a decrease in thrombocyte and an increase in heterophil percentages were seen at 14 days post-infection. Taken together, these results demonstrate the ability of Salmonella to modulate the intestinal microbiota to facilitate colonization. Additionally, results indicated an early role of monocytes and thrombocytes during colonization, followed by heterophils.

A cathepsin L-like cysteine proteinase gene from the protozoan parasite, Cryptobia salmositica.

The present study describes the identification of a cathepsin L-like cysteine proteinase gene (CYS) from the hemoflagellate Cryptobia salmositica. Genomic DNA sequence of cysteine proteinase was obtained by genome walking using degenerate primers. Specific primers were designed to amplify the cDNA of cysteine proteinase from mRNA by rapid amplification of cDNA ends-PCR. The open reading frame of CYS is 1,329 bp, with 443 deduced amino acids. Based on the sequence analysis, cysteine proteinase of C. salmositica is similar to the cathepsin L-like cysteine proteinase of kinetoplastid parasites such as Leishmania spp. and Trypanosoma spp. The identification of CYS proteinase gene could help to design cysteine proteinase specific inhibitors. Further studies are required to characterize the complete genomic organization of the cysteine proteinase.