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Campylobacter and invasive non-typhoidal Salmonella (iNTS) are among the most common causative agents of gastroenteritis worldwide. As of now, no single combination licensed vaccine is available for public health use against both iNTS and Campylobacter species. Outer-membrane vesicles (OMVs) are nanoscale proteoliposomes released from the surface of gram-negative bacteria during log phase and harbor a variety of immunogenic proteins. Based on epidemiology of infections, we formulated a novel trivalent outer membrane vesicles (TOMVs)-based vaccine candidate against Campylobacter jejuni (CJ), Salmonella Typhimurium (ST) and Salmonella Enteritidis (SE). Isolated OMVs from CJ, ST and SE were combined in equal ratios for formulation of TOMVs and 5 µg of the developed vaccine candidate was used for intraperitoneal immunization of adult BALB/c mice. Immunization with TOMVs significantly activated both the humoral and cellular arm of adaptive immune response. Robust bactericidal effect was elicited by TOMVs immunized adult mice sera. TOMVs immunization induced long-term protective efficacy against CJ, ST and SE infections in mice. The study illustrates the ability of TOMVs-based combination immunogen in eliciting broad-spectrum protective immunity against prevalent Campylobacter and iNTS pathogens. According to the findings, TOMVs can work as a potent combination-based acellular vaccine candidate for amelioration of Campylobacter and iNTS-mediated gastroenteritis., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Campylobacter is a major foodborne and zoonotic pathogen, causing severe human infections and imposing a substantial economic burden on global public health. The ongoing spread and emergence of multidrug-resistant (MDR) strains across various fields exacerbate therapeutic challenges, raising the incidence of diseases and fatalities. Medicinal plants, renowned for their abundance in secondary metabolites, exhibit proven efficacy in inhibiting various foodborne and zoonotic pathogens, presenting sustainable alternatives to ensure food safety. This review aims to synthesize recent insights from peer-reviewed journals on the epidemiology and antimicrobial resistance of Campylobacter species, elucidate the in vitro antibacterial activity of medicinal plant compounds against Campylobacter by delineating underlying mechanisms, and explore the application of these compounds in controlling Campylobacter in food. Additionally, we discuss recent advancements and future prospects of employing medicinal plant compounds in food products to mitigate foodborne pathogens, particularly Campylobacter. In conclusion, we argue that medicinal plant compounds can be used as effective and sustainable sources for developing new antimicrobial alternatives to counteract the dissemination of MDR Campylobacter strains., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Background: Phytogenic additives would be helpful to alleviate the adverse effect of Campylobacter jejuni on the performance and physiological responses of broiler chickens., Objective: This experiment was carried out to investigate the effects of Echinacea purpurea (EP) and Thymbra spicata (TS) on the performance, nutrient digestibility, serum biochemistry, intestinal morphology, intestinal microbiota and immune responses of broilers challenged with C. jejuni from 0 to 42 days of age., Methods: A total of 240 male broiler chickens were divided into 6 groups and fed various diets: a control diet; the control diet supplemented with EP at 0.25% (EP25) or 0.50% (EP50); the control diet supplemented with TS at 0.25% (TS25) and 0.50% (TS50); or the control diet containing erythromycin at 55 ppm. Chicks were orally challenged with C. jejuni on Days 21 and 23 of age., Results: EP and TS25 diets enhanced European production efficiency factor, feed conversion ratio and digestibility of dry matter and organic matter. TS25 increased duodenal villous height (VH) and surface area on Day 42 of age. EP25 diet increased ileal VH compared to control and erythromycin diets. Diets containing certain EP25 and TS increased the Bifidobacterium population and decreased C. jejuni population on Day 39 of age. EP50 and TS50 diets increased antibody titration against Newcastle disease virus., Conclusions: In conclusion, EP and TS dietary supplementation improved performance, microflora, intestinal morphology and immune responses in C. jejuni-challenged broilers., (© 2024 The Author(s). Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

Background: Campylobacter is the most common food-borne pathogen in the European Union. In 2018, the crude incidence rate in Ireland was 63.6 per 100,000 population. Chicken is considered an important source of infection for humans. In 2015, the Campylobacter Stakeholders' Group (CSG) was established to reduce Campylobacter contamination levels in Irish broiler flocks., Aims: This work aimed to describe the Campylobacter monitoring programme that was established by the CSG, to analyse the results of this testing between 2019 and 2022, and to assess progress., Methods and Results: This paper describes the monitoring programme that was established by the CSG, which harmonized Campylobacter enumeration testing across all Irish broiler processors and allowed comparability of results for trend analysis. An analysis of the 2019-2022 data is presented here and compared to previous studies of Campylobacter levels in Irish broilers. An analysis of the 2019-2022 data showed a significant reduction in levels in both caeca and neck skin when the results from 2022 were compared to those from 2019 to 2020. Campylobacter spp. were detected in 37% of caecal samples from first depopulation (pre-thin) broilers and 30% of neck skin samples in 2022, with just 4% of carcases (in neck skin samples) with ≥1000 colony-forming units per gram detected in 2022. Campylobacter levels detected in Irish broilers, in the present monitoring programme were less than those reported in previous studies in both caecal and carcase samples, although not directly comparable for statistical significance because of differences in study methods., Conclusions: The cooperation between stakeholders and regulators of the Irish broiler chicken industry over the past decade has facilitated a coordinated approach to monitoring of Campylobacter levels in broilers, and implementation of control measures. This has enabled a steady reduction in the levels of Campylobacter in Irish chicken., (© 2024 Wiley‐VCH GmbH.)

Campylobacter jejuni ( C. jejuni ) is one of the most common causes of foodborne infections worldwide and a major contributor to diarrheal diseases. This study aimed to explore the ability of commensal gut bacteria to control C. jejuni infection. Bacterial strains from the intestinal mucosa of broilers were screened in vitro against C. jejuni ATCC BAA1153. The cell-free supernatant (CFS) of Ligilactobacillus salivarius UO.C249 showed potent dose-dependent antimicrobial activity against the pathogen, likely due to the presence of bacteriocin-like moieties, as confirmed by protease treatment. Genome and exoproteome analyses revealed the presence of known bacteriocins, including Abp118. The genome of Lg. salivarius UO.C249 harbors a 1.8-Mb chromosome and a 203-kb megaplasmid. The strain was susceptible to several antibiotics and had a high survival rate in the simulated chicken gastrointestinal tract (GIT). Post-protease treatment revealed residual inhibitory activity, suggesting alternative antimicrobial mechanisms. Short-chain fatty acid (SCFA) quantification confirmed non-inhibitory levels of acetic (24.4 ± 1.2 mM), isovaleric (34 ± 1.0 µM), and butyric (32 ± 2.5 µM) acids. Interestingly, extracellular vesicles (EVs) isolated from the CFS of Lg. salivarius UO.C249 were found to inhibit C. jejuni ATCC BAA-1153. Proteome profiling of these EVs revealed the presence of unique proteins distinct from bacteriocins identified in CFS. The majority of the identified proteins in EVs are located in the membrane and play roles in transmembrane transport and peptidoglycan degradation, peptidase, proteolysis, and hydrolysis. These findings suggest that although bacteriocins are a primary antimicrobial mechanism, EV production also contributes to the inhibitory activity of Lg. salivarius UO.C249 against C. jejuni ., Importance: Campylobacter jejuni ( C. jejuni ) is a major cause of gastroenteritis and a global public health concern. The increasing antibiotic resistance and lack of effective alternatives in livestock production pose serious challenges for controlling C. jejuni infections. Therefore, alternative strategies are needed to control this pathogen, especially in the poultry industry where it is prevalent and can be transmitted to humans through contaminated food products. In this study, Ligilactobacillus salivarius UO.C249 isolated from broiler intestinal mucosa inhibited C. jejuni and exhibited important probiotic features. Beyond bacteriocins, Lg. salivarius UO.C249 secretes antimicrobial extracellular vesicles (EVs) with a unique protein set distinct from bacteriocins that are involved in transmembrane transport and peptidoglycan degradation. Our findings suggest that beyond bacteriocins, EV production is also a distinct inhibitory signaling mechanism used by Lg. salivarius UO.C249 to control C. jejuni . These findings hold promise for the application of probiotic EVs for pathogen control., Competing Interests: R.H., S.M., and W.M. are co-inventors and co-applicants with the University of Ottawa on a patent application (US Patent Application Number 63/607,682, filed 08 December 2023) entitled "LIGILACTOBACILLUS PROBIOTICS, LIGILACTOBACILLUS EXTRACELLULAR VESICLES AND METHODS OF USING SAME" that covers the inhibitory activity of probiotic extracellular vesicles. The remaining authors declare no competing interests.

Background: Preventing pathogens from entering the broiler premises is the main biosecurity measure at farm level. In conventional broiler production, chickens are kept indoors during the entire production period. Pathogens can enter the broiler-producing unit from sources such as water, equipment, personnel, insects, and rodents. The possible routes must be controlled, and corrective measures applied when necessary. The objective of this study was to (1) develop a hygiene protocol and test the scheme on 30 farms, and (2) compare the results to their Campylobacter-colonised status. A Hygiene Performance Rating protocol at farm level (HPR-F) was developed to systematically review the production to identify risk areas to biosecurity. The HPR-F consists of 13 categories with related questions. For each question, a score was given from 1 to 3, where 1 meant "acceptable", 2 was "potential for improvements", and 3 was "not acceptable". Scores for each question were multiplied with weight factors for hygienic impact and economic consequences describing whether the necessary improvement depends on a significant investment or is a cheap quick-fix and calculated into a percentage where 100% is perfect hygiene. The 30 farms in the study were selected from one county in Norway. The Campylobacter-results for each of the 30 farms in 2019-2021 were given according to rules in the Norwegian Action Plan against Campylobacter faecal sampling on-farm 3-6 days prior to slaughter., Results: The overall results from the HPR-F showed that the general hygiene level was high in all farms. The mean total hygiene score was 82% and varied from 70 to 92%. The category Handling dead chicken had the highest hygiene score (93%), and Ventilation had the lowest score (55%). The HPR-F results were compared to the Campylobacter-status for the 30 farms: Campylobacter-negative flocks had slightly higher total scores than Campylobacter-positive flocks (P = 0.19). Among others, the category Outdoor area (vegetation close to the premises' walls) was identified as the most stable factor in relation to be colonised with Campylobacter., Conclusions: The HPR-F tested in this research trial provides a tool for veterinarians, advisors, and poultry farmers to improve biosecurity at farm level and enhance the preventive animal health initiatives., (© 2024. The Author(s).)

Campylobacter jejuni continues to be a major public health issue worldwide. Poultry are recognized as the main reservoir for this foodborne pathogen. Implementing measures to decrease C. jejuni colonization on farms has been regarded as the most effective strategy to control the incidence of campylobacteriosis. The probiotics supplementation has been regarded as an attractive approach against C. jejuni in chickens. Here the inhibitory effects of one probiotic B. velezensis isolate CAU277 against C. jejuni was evaluated in vitro and in vivo. The in vitro antimicrobial activity showed that the supernatant of B. velezensis exhibited the most pronounced inhibitory effects on Campylobacter strains compared to other bacterial species. When co-cultured with B. velezensis, the growth of C. jejuni reduced significantly from 7.46 log 10 CFU/mL (24 h) to 1.02 log 10 CFU/mL (48 h). Further, the antimicrobial activity of B. velezensis against C. jejuni remained stable under a broad range of temperature, pH, and protease treatments. The in vivo experiments demonstrated that oral administration of B. velezensis significantly reduced the colonization of C. jejuni by 2.0 log 10 CFU/g of feces in chicken cecum at 15 d postinoculation. In addition, the supplementary of B. velezensis significantly increased microbial species richness and diversity in chicken ileum, especially enhanced the bacterial population of Alistipes and Christensenellaceae, and decreased the existence of Lachnoclostridium. Our study presents that B. velezensis possesses antimicrobial activities against C. jejuni and promotes microbiota diversity in chicken intestines. These findings indicate a potential to develop an effective probiotic additive to control C. jejuni infection in chicken., Competing Interests: DISCLOSURES The authors declare no conflict of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: Campylobacter jejuni (C. jejuni), a widely distributed global foodborne pathogen, primarily linked with contaminated chicken meat, poses a significant health risk. Reducing the abundance of this pathogen in poultry meat is challenging but essential. This study assessed the impact of Lactobacillus-fermented rapeseed meal (LFRM) on broilers exposed to C. jejuni-contaminated litter, evaluating growth performance, Campylobacter levels, and metagenomic profile., Results: By day 35, the litter contamination successfully colonized broilers with Campylobacter spp., particularly C. jejuni. In the grower phase, LFRM improved (P < 0.05) body weight and daily weight gain, resulting in a 9.2% better feed conversion ratio during the pre-challenge period (the period before artificial infection; days 13-20). The LFRM also reduced the C. jejuni concentration in the ceca (P < 0.05), without altering alpha and beta diversity. However, metagenomic data analysis revealed LFRM targeted a reduction in the abundance of C. jejuni biosynthetic pathways of l-tryptophan and l-histidine and gene families associated with transcription and virulence factors while also possibly leading to selected stress-induced resistance mechanisms., Conclusion: The study demonstrated that LFRM inclusion improved growth and decreased cecal Campylobacter spp. concentration and the relative abundance of pivotal C. jejuni genes. Performance benefits likely resulted from LFRM metabolites. At the molecular level, LFRM may have reduced C. jejuni colonization, likely by decreasing the abundance of energy transduction and l-histidine and l-tryptophan biosynthesis genes otherwise required for bacterial survival and increased virulence. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

Campylobacteriosis is a significant foodborne illness caused by Campylobacter bacteria. It is one of the most common bacterial causes of gastroenteritis worldwide, with poultry being a major reservoir and source of infection in humans. In poultry farms, Campylobacters colonize the intestinal tract of chickens and contaminate meat during processing. Vaccines under development against Campylobacters in poultry showed partial or no protection against their cecal colonization. Therefore, this review will elaborate on campylobacteriosis and emphasize the control strategies and recent vaccine trials against Campylobacters in poultry farms. The epidemiology, diagnosis, and treatment of Campylobacter infection, along with specific mention of poultry Campylobacter contamination events in Malaysia, will also be discussed.

For reducing Campylobacter (C.) in the food production chain and thus the risk to the consumer, the combined application of different measures as a multiple-hurdle approach is currently under discussion. This is the first study to investigate possible synergistic activities in vivo, aiming at reducing intestinal C. jejuni counts by administering (i) bacteriophages (phages) in combination with a competitive exclusion (CE) product and (ii) carvacrol combined with organic acids. The combined application of the two selected phages (Fletchervirus phage NCTC 12673 and Firehammervirus phage vB&#95;CcM-LmqsCPL1/1) and the CE product significantly reduced C. jejuni loads by 1.0 log 10 in cecal and colonic contents as well as in cloacal swabs at the end of the trial (33 and 34 days post hatch). The proportion of bacterial isolates showing reduced phage susceptibility ranged from 10.9% (isolates from cecal content) to 47.8% (isolates from cloacal swabs 32 days post hatch) for the Fletchervirus phage, while all tested isolates remained susceptible to the Firehammervirus phage. The use of carvacrol combined with an organic acid blend (sorbic acid, benzoic acid, propionic acid, and acetic acid) significantly reduced Campylobacter counts by 1.0 log 10 in cloacal swabs on day 30 only., (© 2024. The Author(s).)

Background: Bovine Genital Campylobacteriosis (BGC), a worldwide distributed venereal disease caused by Campylobacter fetus subsp. venerealis (Cfv), has a relevant negative economic impact in cattle herds. The control of BGC is hampered by the inexistence of globally available effective vaccines. The present in silico study aimed to develop a multi-epitope vaccine candidate against Cfv through reverse vaccinology., Results: The analysis of Cfv strain NCTC 10354 proteome allowed the identification of 9 proteins suitable for vaccine development. From these, an outer membrane protein, OmpA, and a flagellar protein, FliK, were selected for prediction of B-cell and T-cell epitopes. The top-ranked epitopes conservancy was assessed in 31 Cfv strains. The selected epitopes were integrated to form a multi-epitope fragment of 241 amino acids, which included 2 epitopes from OmpA and 13 epitopes from FliK linked by GPGPG linkers and connected to the cholera toxin subunit B by an EAAAK linker. The vaccine candidate was predicted to be antigenic, non-toxic, non-allergenic, and soluble upon overexpression. The protein structure was predicted and optimized, and the sequence was successfully cloned in silico into a plasmid vector. Additionally, immunological simulations demonstrated the vaccine candidate's ability to stimulate an immune response., Conclusions: This study developed a novel vaccine candidate suitable for further in vitro and in vivo experimental validation, which may become a useful tool for the control of BGC., (© 2024. The Author(s).)

Bovine genital campylobacteriosis, caused by the gram-negative bacteria Campylobacter fetus venerealis, and bovine trichomonosis, caused by the parasite protozoan Tritrichomonas foetus, are venereal diseases that occur with long intercalving periods and abortion. The control of both diseases relies on microbiological testing and culling infected bulls. Vaccination and antibiotic treatment may help in controlling campylobacteriosis but are not recommended for trichomonosis control. Several regions of the world have active control programs for trichomonosis, not campylobacteriosis. In Argentina, the state of La Pampa aims to eradicate trichomonosis and campylobacteriosis by imposing annual diagnostic testing of every bull and slaughtering positive animals. Prior studies indicated a declining trend in the prevalence of campylobacteriosis and trichomonosis in La Pampa. It was also proposed that the prevalence of one disease could be estimated from the prevalence of the other. The purpose of this retrospective analysis of data gathered from 2008 to 2021 was to determine the La Pampa program's efficacy. Descriptive statistics were employed to determine the reason behind the correlation between tricomonosis and campylobacteriosis diagnostic results. The outcomes refute the notion that this program of venereal eradication was a success. Furthermore, an excess of false positives in both diagnoses may have contributed to the correlation between the prevalences of campylobactriosis and trichomonosis. The practice of killing animals without verifying positive results hinders the determination of disease prevalence and results in the death of numerous healthy animals., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Campylobacter mitigation along the food production chain is considered effective for minimizing the public health burden of human campylobacteriosis. This study is the first combining different measures in a multiple-hurdle approach, using drinking water additives and feed additives in single and combined application schemes in commercial broiler plants. Broiler chickens in the study groups were naturally contaminated with Campylobacter. Application of an organic acid blend via drinking water, consisting of sodium propionate, potassium sorbate, and sodium diacetate, resulted in significant reductions of up to 4.9 log 10 CFU/mL in fecal samples and in cecal samples at slaughter. The application of a phage mixture, consisting of Fletchervirus phage NCTC 12673 and Firehammervirus phage vB&#95;CcM-LmqsCPL1/1, resulted in reductions of up to 1.1 log 10 CFU/mL in fecal samples 1 day after dosing. The sole administration of curcumin via feed resulted in small and inconsistent reductions. In the group receiving a combination of all tested measures, reductions of up to 1.1 log 10 CFU/mL were observed. Based on the results of our field trials, it was shown that both the sole application and the combined application of mitigation measures in primary production can reduce the Campylobacter load in broiler chickens, while no synergism could be observed., (© 2024. The Author(s).)

The incidence of human Campylobacter jejuni infections is increasing worldwide. It is highly desirable to prevent campylobacteriosis in individuals at risk for severe disease with antibiotics-independent non-toxic compounds. Activated charcoal (AC) has long been used as an anti-diarrheal remedy. Here, we tested the disease-mitigating effects of oral AC versus placebo in human gut microbiota-associated (hma) IL-10 -/- mice starting a week prior to C. jejuni infection. On day 6 post-infection, the gastrointestinal C. jejuni loads were comparable in both infected cohorts, whereas campylobacteriosis symptoms such as wasting and bloody diarrhea were mitigated upon AC prophylaxis. Furthermore, AC application resulted in less pronounced C. jejuni -induced colonic epithelial cell apoptosis and in dampened innate and adaptive immune cell responses in the colon that were accompanied by basal concentrations of pro-inflammatory mediators including IL-6, TNF-α, IFN-γ, and nitric oxide measured in colonic explants from AC treated mice on day 6 post-infection. Furthermore, C. jejuni infection resulted in distinct fecal microbiota shift towards higher enterobacterial numbers and lower loads of obligate anaerobic species in hma mice that were AC-independent. In conclusion, our pre-clinical placebo-controlled intervention study provides evidence that prophylactic oral AC application mitigates acute murine campylobacteriosis.

Campylobacter is the main cause of human gastroenteritis worldwide, with 50 to 80% of the cases related to consumption of poultry products. Maternal antibodies (MAB) from commercial breeder flocks may protect their progeny against infection during the first few weeks of life. We here studied the prevalence of Campylobacter antibody titers in broiler breeder flocks and to which extent immunization of broiler breeders increases maternal anti-Campylobacter titers in their progeny and protects the offspring against Campylobacter colonization. Two vaccines were used: a bacterin mix of 13 Campylobacter strains and a subunit vaccine comprising 6 immunodominant Campylobacter antigens. All sampled on-farm breeder flocks were positive for anti-Campylobacter antibodies, yet in some breeder flocks only very low titers were detected. Vaccination of SPF broiler breeder flocks with both subunit and bacterin vaccines resulted in a prolonged presence of anti-Campylobacter antibodies in the serum and intestinal mucus of chicks. These bacterin- or subunit vaccine-induced MAB conferred protection against Campylobacter colonization in chicks until 7 and 21 d of age, respectively, but only at a low challenge dose (10 2.5 cfu). The concentration of MAB in the mucus is probably too low to sufficiently capture Campylobacter when higher challenge doses are used. In conclusion, vaccinating broiler breeders protects their offspring against Campylobacter colonization under low pathogen exposure conditions., (Copyright © 2023. Published by Elsevier Inc.)

Controlling Campylobacter jejuni during broiler production is a topic of interest from a public health standpoint, as colonized birds can contaminate poultry products during processing and sicken humans if not properly cooked or handled before consumption. The aim of this study was to evaluate dietary yeast cell wall (YCW) as a potential alternative to antibiotic growth promoters with or without a C. jejuni challenge. A total of 2240 day-old Ross 708 males were randomly assigned within 8 experimental groups with a 4 × 2 factorial design, with 4 diets (negative control [CTL-], positive control [CTL+, bacitracin, 50 g/ ton], YCW constant dose [400 g/ton], and YCW step-down dose [SD, 800, 400, and 200 g/ton in the starter, grower, and finisher periods, respectively]) and with or without a Day-16 C. jejuni oral gavage challenge at a 10 3 -colony-forming-units (CFU)/ml dose. Body weights and feed consumption were measured on Days 0, 14, 28, and 41 to determine broiler performance. Ileum tissue samples were collected from 24 birds per treatment on Days 17 and 24 (1 and 8 days postinoculation [PI]) for relative gene expression (RGE) analysis. Cecal content samples were collected from 24 birds per treatment on Days 24, 34, and 42 for C. jejuni enumeration and prevalence calculation. A total of 80 birds per treatment were processed to determine carcass yield on Day 44, and on Day 45, 16 carcass rinsates per treatment were collected for C. jejuni enumeration and prevalence calculation. The interaction between diet and inoculation did not influence growth performance ( P > 0.05). However, a diet effect was observed in the starter period where birds fed SD diet had a lower feed conversion ratio than birds fed CTL- diet ( P = 0.0165). Additionally, the treatment of birds inoculated with C. jejuni fed with SD had a trend to a lower feed conversion ratio during the grower period ( P = 0.0550). The RGE of interleukin 1β and interleukin 10 was similar in all treatments 1 and 8 days PI. The RGE of avian beta defensin 10 was similar in all treatments on Day 1 PI, but different on Day 8 PI ( P = 0.0476). All birds inoculated with C. jejuni had similar CFU per milliliter counts in the cecal contents at Days 24, 34, and 42 ( P > 0.05), and all birds inoculated with phosphate-buffered saline were negative for C. jejuni after prevalence testing. After processing 1) carcass yield was similar in all treatments ( P > 0.05); 2) C. jejuni -inoculated birds fed CTL- had lower CFU per milliliter counts than birds provided CTL+ and constant-dose diets ( P = 0.0383); and 3) all birds inoculated with PBS were negative for Campylobacter . Overall, under the conditions of this study, the addition of YCW during a C. jejuni challenge did not have an impact on growth performance, innate immune response, cecal colonization, carcass yield, or carcass colonization after processing.

Background: Effective strategies are urgently needed to control Campylobacteriosis, one of the most important foodborne gastrointestinal diseases worldwide. Administering bacteriophages (phages) is under evaluation as a possible intervention strategy in primary poultry production to reduce the public health risk of human infection. A major challenge is the translation of results from small-scale animal studies to large broiler flocks. In this study, the in vitro lytic activity of 18 Campylobacter-specific group II phages and 19 group III phages were examined singly, and in different combinations from the same group and from both groups using a planktonic killing assay. Based on these results, a combination of phage NCTC 12,673 (group III) and vB&#95;CcM-LmqsCPL1/1 (group II) was selected for in vivo application in a seeder bird model to study its effectiveness under conditions as close as possible to field conditions. One hundred eighty Ross 308 broiler chickens were divided into a control and a treatment group. Ten days post hatch, seeder birds were orally inoculated with the C. jejuni target strain. Phages were administered via drinking water at a total concentration of 10 7 PFU/mL four, three, and two days before necropsy., Results: Combining group II and group III phages resulted in significantly higher in vitro growth inhibition against the C. jejuni target strain BfR-CA-14,430 than single application or combinations of phages from the same group. The results of the animal trial showed that the application of the two phages significantly reduced Campylobacter counts in cloacal swabs. At necropsy, Campylobacter counts in colonic content of the treatment group were significantly reduced by 2 log 10 units compared to the control group., Conclusions: We demonstrated that combining phages of groups II and III results in significantly increased lytic activities. The in vitro results were successfully translated into practical application in a study design close to field conditions, providing new data to apply phages in conventional broiler flocks in the future. Phage application reduced the fecal Campylobacter excretion and Campylobacter concentrations in the colon of broilers., (© 2023. BioMed Central Ltd., part of Springer Nature.)

Campylobacter jejuni is a leading global cause of bacterial gastroenteritis in humans, and poultry are a major reservoir. Glycoconjugate vaccines containing the conserved C. jejuni N-glycan have previously been reported to be effective at reducing caecal colonisation of chickens by C. jejuni. These include recombinant subunit vaccines, live E. coli strains expressing the N-glycan on the surface as well as outer membrane vesicles (OMVs) derived from these E. coli strains. In this study, we evaluated the efficacy of live E. coli expressing the C. jejuni N-glycan from a plasmid and glycosylated OMVs (G-OMVs) derived from them against colonisation by different C. jejuni strains. Despite the C. jejuni N-glycan being expressed on the surface of the live strain and the OMVs, no reduction in caecal colonisation by C. jejuni was observed and N-glycan-specific responses were not detected., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Campylobacter-associated enteric disease is estimated to be responsible for more than 160 million cases of gastroenteritis each year and is linked to growth stunting of infants living under conditions of poor sanitation and hygiene. Here, we examine naturally occurring Campylobacter-associated diarrhea among rhesus macaques as a model to determine if vaccination could reduce severe diarrheal disease and infant growth stunting. Compared to unvaccinated controls, there are no Campylobacter diarrhea-associated deaths observed among vaccinated infant macaques and all-cause diarrhea-associated infant mortality is decreased by 76% (P = 0.03). By 9 months of age, there is a 1.3 cm increase in dorsal length that equaled a significant 1.28 LAZ (Length-for-Age Z score) improvement in linear growth among vaccinated infants compared to their unvaccinated counterparts (P = 0.001). In this work, we show that Campylobacter vaccination not only reduces diarrheal disease but also potentially serves as an effective intervention that improves infant growth trajectories., (© 2023. The Author(s).)

Campylobacter jejuni (C. jejuni) is the most common food-borne pathogen that causes human gastroenteritis in the United States. Consumption of contaminated poultry products is considered as the major source of human Campylobacter infection. An effective vaccine would be a promising alternative to antibiotic supplements to curb C. jejuni colonization in poultry gastrointestinal (GI) tract. However, the genetic diversity among the C. jejuni isolates makes vaccine production more challenging. Despite many attempts, an effective Campylobacter vaccine is not yet available. This study aimed to identify suitable candidates to develop a subunit vaccine against C. jejuni, which could reduce colonization in the GI tract of the poultry. In the current study, 4 C. jejuni strains were isolated from retail chicken meat and poultry litter samples and their genomes were sequenced utilizing next-generation sequencing technology. The genomic sequences of C. jejuni strains were screened to identify potential antigens utilizing the reverse vaccinology approach. In silico genome analysis predicted 3 conserved potential vaccine candidates (phospholipase A [PldA], TonB dependent vitamin B12 transporter [BtuB], and cytolethal distending toxin subunit B [CdtB]) suitable for the development of a vaccine. Furthermore, the expression of predicted genes during host-pathogen interaction was analyzed by an infection study using an avian macrophage-like immortalized cell line (HD11). The HD11 was infected with C. jejuni strains, and the RT-qPCR assay was performed to determine the expression of the predicted genes. The expression difference was analyzed using ΔΔCt methods. The results indicate that all 3 predicted genes, PldA, BtuB, and CdtB, were upregulated in 4 tested C. jejuni strains irrespective of their sources of isolation. In conclusion, in silico prediction and gene expression analysis during host-pathogen interactions identified 3 potential vaccine candidates for C. jejuni., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Enteropathogens, namely Salmonella and Campylobacter, are a concern in global public health and have been attributed in numerous risk assessments to a poultry source. During the last decade, a large body of research addressing this problem has been published. The literature reviewed contains review articles on certain aspects of poultry production chain; however, in the past decade there has not been a review on the entire chain-farm to fork-of poultry production. For this review, a pool of 514 articles were selected for relevance via a systematic screening process (from >7500 original search articles). These studies identified a diversity of management and intervention strategies for the elimination or reduction of enteropathogens in poultry production. Many studies were laboratory or limited field trials with implementation in true commercial operations being problematic. Entities considering using commercial antienteropathogen products and interventions are advised to perform an internal validation and fit-for-purpose trial as Salmonella and Campylobacter serovars and biovars may have regional diversity. Future research should focus on nonchemical application within the processing plant and how a combination of synergisticinterventions through the production chain may contribute to reducing the overall carcass burden of enteropathogens, coupled with increased consumer education on safe handling and cooking of poultry., (© 2023 Institute of Food Technologists®.)

Campylobacter infections traced mainly to poultry products are major bacterial foodborne zoonoses. Among the many control strategies evaluated at primary poultry level to reduce these infections, vaccination could be a solution, but no effective vaccines are available to date. A better understanding of the immune mechanisms involved in protection against Campylobacter would be helpful for designing novel vaccine strategies. The present study was designed to analyze in more depth the immune responses developed in broilers in order to potentially identify which immune parameters may be important for establishing protection against Campylobacter by comparing the immune responses obtained here with those obtained in a previous study performed on vaccinated specific-pathogen-free Leghorn chickens that presented a partial reduction of Campylobacter after experimental challenge. The protection against Campylobacter colonization was evaluated at different time points over 40 d of rearing, by measuring specific IgY levels in serum and IgA antibodies in bile reflecting the systemic and mucosal humoral responses respectively and the relative expressions of 9 cecal immune marker genes (cytokines and antimicrobial peptides), which reflect the innate and cellular immune responses. Despite no reduction of Campylobacter in the cecum, a systemic immune response over time characterized by the production of specific anti-flagellin IgY was observed, in addition to upregulation of the antimicrobial peptide avian β-defensin (AvBD) 12 gene expression in the cecum of vaccinated broilers compared with the placebo group. However, the levels of specific anti-flagellin mucosal IgA antibodies in the bile as well as the relative expression of other cecal cytokines studied was underexpressed in the vaccinated group or similar in both groups., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Campylobacter jejuni is a leading cause of foodborne bacterial gastroenteritis worldwide, and raw or undercooked chicken meat is considered the major source of human campylobacteriosis. In this study, we identified 36 compounds that showed inhibitory effects on C. jejuni growth at low concentrations by screening a chemical compound library. Three of the 36 compounds were herbal compounds, including tryptanthrin (TRP), an indoloquinazoline alkaloid. TRP has been reported to have a variety of biological properties, such as antimicrobial, anti-inflammatory, and antitumor activities, but there was previously no information about its anti-C. jejuni activity. We further conducted in vitro and in vivo experiments to evaluate the potential of TRP for the control of C. jejuni in chicken farms. The MIC of TRP for C. jejuni was much lower than that of 13 other herbal compounds that were previously reported to have anti-C. jejuni activities. Time-kill assays under growing and nongrowing conditions demonstrated that TRP has bactericidal activity against C. jejuni. In addition, TRP showed a narrow-spectrum antimicrobial effect against C. jejuni, and there was little potential for the development of TRP-resistant C. jejuni during serially passaged culture. In chick infection experiments, the administration of TRP in drinking water significantly reduced the cecal colonization of C. jejuni when TRP was used either before or after C. jejuni infection. These data suggest that TRP is effective for the control of C. jejuni in chicken farms. IMPORTANCE Campylobacter is a widespread pathogen in the food chain of chickens. Once chickens become infected, large numbers of Campylobacter cells are excreted in their feces. The development of an effective material for reducing the amount of Campylobacter in the chicken intestinal tract will make it possible to reduce the contamination of the food chain with Campylobacter and to produce safe and secure chicken meat. In the present study, in vivo experiments revealed that the use of an herbal compound, tryptanthrin, significantly reduced the number of Campylobacter cells in the chicken gut by a bactericidal mechanism. Furthermore, our in vitro experiments demonstrated that, compared with the other herbal compounds, tryptanthrin achieved antimicrobial activity against C. jejuni at the lowest concentration. The use of tryptanthrin may lead to the development of a novel control measure for reducing the colonization of C. jejuni in the food chain.

Application of organic acids via feed or drinking water is under discussion as a possible intervention strategy to reduce Campylobacter (C.) load in primary poultry production. A previous in vitro study showed that reduced concentrations of sorbic acid, benzoic acid, propionic acid, and acetic acid were required for antibacterial activity against Campylobacter when using a mixture of these 4 acids compared to when using the single acids. The present study aimed at determining the antibacterial efficiency of this combination in vivo as a drinking water additive for reducing shedding and intestinal C. jejuni colonization in broilers. Furthermore, we assessed whether the inoculated C. jejuni strain BfR-CA-14430 adapted in vivo to the applied organic acids. Results of this study showed that adding the organic acids consistently reduced Campylobacter loads in cloacal swabs. While significant reductions were observed within the entire study period, a maximum 2 log reduction occurred at an age of 18 d. However, after dissection at the end of the trial, no significant differences were detected in Campylobacter loads of cecal and colon contents compared to the control group. Susceptibility testing of re-isolates from cloacal swabs and cecal content revealed equal minimum inhibitory concentration (MIC) values compared to the inoculated test strain, suggesting that C. jejuni remained susceptible throughout the trial., (Copyright © 2022. Published by Elsevier Inc.)

The Campylobacter genus is the leading cause of human gastroenteritis, with the consumption of contaminated poultry meat as the main route of infection. Probiotic bacteria, such as Lactobacillus , Bacillus , Escherichia coli Nissle, and Bifidobacterium species, have a great immunomodulatory capacity and exhibit antipathogenic effects through various molecular mechanisms. Reducing Campylobacter levels in livestock animals, such as poultry, will have a substantial benefit to humans as it will reduce disease transmissibility through the food chain. Moreover, probiotic-based strategies might attenuate intestinal inflammatory processes, which consequently reduce the severity of Campylobacter disease progression. At a molecular level, probiotics can also negatively impact on the functionality of various Campylobacter virulence and survival factors (e.g., adhesion, invasion), and on the associated colonization proteins involved in epithelial translocation. The current review describes recent in vitro , in vivo , and preclinical findings on probiotic therapies, aiming to reduce Campylobacter counts in poultry and reduce the pathogen's virulence in the avian and human host. Moreover, we focused in particular on probiotics with known anti- Campylobacter activity seeking to understand the biological mechanisms involved in their mode of action.

Campylobacter spp. are some of the most important food-borne zoonoses in Europe and broiler meat is considered the main source of Campylobacter infections. Organic and free-range broilers have access to outdoor reservoirs of Campylobacter and are more frequently infected at slaughter than the conventional broiler flocks. Limitations to biosecurity and treatment options in these production types calls for additional solutions. This review examines intervention methods with sufficient strength and quality, which are able to reduce the load of Campylobacter safely and efficiently and discuss their applicability in organic and free-range broiler production. Four different products passed the inclusion criteria and their quality examined: ferric tyrosine chelate, a prebiotic fermentation product of Saccharomyces cerevisiae, short-chain fatty acid butyrate coated on microbeads added to feed, and a mix of organic acids added to the drinking water. Though potential candidates for reducing Campylobacter in broilers were identified, there is a lack of large scale intervention studies that demonstrate an effect under field conditions of a free-range broiler production., (© 2022. The Author(s).)

Poultry is generally recognized as the main source of human campylobacteriosis and Campylobacter is highly prevalent at the farm level. To reduce the relative risk of human campylobacteriosis attributable to broiler meat, it is necessary to reduce Campylobacter loads in broiler ceca but to date, no effective, reliable and practical strategy is available. The marine environment is a rich source of original natural compounds exhibiting different biological activities. The objective of this study was to test a phlorotannin extract of the brown seaweed Ascophyllum nodosum as a potential control strategy against Campylobacter in broilers. Bactericidal activity has been demonstrated in vitro, on several Campylobacter spp. strains at a range of 0.06 to 0.47 mg/mL. Therefore, an in vivo trial in experimental facilities was performed to evaluate addition of 0.2% (w/w) of an A. nodosum extract to feed distributed at the end of rearing from day 31 to day 35, and to assess the effect on artificial Campylobacter jejuni colonization. No statistical differences in Campylobacter enumeration were observed between the treated and control groups. Another trial was performed in a commercial broiler flock. Feed containing the extract at 0.2% (w/w) (2 kg/t) was distributed during the last 5 days of rearing (day 33-day 38). No significant effects on Campylobacter colonization and on growth parameters were observed compared to the control group. Additional studies are needed to assess whether active polyphenols are found in the cecum., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

The aim of this study was to evaluate the effect of control strategies (probiotic supplementation and vaccination) at farm on thermotolerant Campylobacter infection due to the consumption of salad cross-contaminated from broiler meat. The broiler food chain was modelled considering the most common practices applied in Argentina (baseline model) and their effect on the prevalence and counts of Campylobacter. Probiotic supplementation and vaccination against Campylobacter spp. were included in different models to evaluate their effectiveness. The parameter distributions of each intervention were obtained based on a systematic review and meta-analysis previously described. The control measures applied at farms were evaluated considering their effectiveness in reducing both the prevalence and the count of thermotolerant Campylobacter in comparison with the baseline model estimation and expressed as relative change in campylobacteriosis risk. Additionally, the identification of the most important input parameters for the model was performed by sensitivity analysis. The model estimated a risk of campylobacteriosis per consumed serving of salad contaminated with poultry meat of 4.99 x 10 -3 (95% CI: 6.12 x 10 -6 -1.13 x 10 -2 ), corresponding to an annual incidence risk estimated of 1,876,009 persons. Scenario analysis indicated that the application of vaccines against Campylobacter (probability of campylobacteriosis = 9.55 x 10 -4 ; 95% CI: 5.31 x 10 -4 -1.29 x 10 -3 ) and the supplementation of broilers with probiotics (probability of campylobacteriosis = 1.32 x 10 -3 ; 8.55 x 10 -4 -1.69 x 10 -3 ) can offer a modest reduction in risk estimates. The intervention efficacy was 80.86% and 73.54% for vaccination and probiotic supplementation, respectively. On-farm interventions were effective to mitigate the risk of campylobacteriosis., (© 2022 Wiley-VCH GmbH.)

Campylobacteriosis is one of the most common types of bacterial gastroenteritis affecting humans, and poultry is considered a major source of the causative organism, Campylobacter spp. Broilers may arrive contaminated at slaughterhouses, and transport crates could be considered a potential source of contamination. Thus, cleaning and disinfection procedures are crucial to avoid cross-contamination among flocks. Despite its public health importance in Latin American countries, virulence factors of Campylobacter jejuni remain poorly studied in this region. Thus, this study aimed to: 1) determine the occurrence of contaminated crates at a poultry slaughterhouse, 2) compare the contamination before and after the cleaning and disinfection procedures, and 3) detect virulence-associated genes in C. jejuni strains by PCR. Campylobacter spp. were recovered from 8 of the 10 flocks evaluated, and C. jejuni was detected as the main species. There was no significant difference in the Campylobacter detection or quantification between crates at the reception platform and crates after the cleaning/disinfection processes. However, crates after 24 h of natural drying, presented a significant (P < 0.05) lower amount of Campylobacter cells than before the cleaning and disinfection processes. A negative relationship (R 2  = 0.210, P = 0.045) between environmental conditions and Campylobacter quantification was found for transport crates after 24 h of natural drying. There was no significant difference (P > 0.05) in the detection of two C. jejuni virulence genes, flaA (encode a major flagellin protein) and cadF (encode an adhesion and fibronectin-binding protein), among various stages of the cleaning and disinfection processes. Our results demonstrate the high contamination levels of Campylobacter strains in broiler flocks and the potential involvement of poultry transport crates in transmitting these bacteria. This study also suggests that ineffective cleaning and disinfection procedures can increase Campylobacter contamination and facilitate the spread of bacteria in poultry establishments., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

To maintain food safety and flock health in broiler chicken production, biosecurity approaches to keep chicken barns free of pathogens are important. Canadian broiler chicken producers must deep clean their barns with chemical disinfectants at least once annually (full disinfection [FD]) and may wash with water (water wash [WW]) throughout the year. However, many producers use FD after each flock, assuming a greater efficacy of more stringent cleaning protocols, although little information is known regarding how these two cleaning practices affect pathogen population and gut microbiota. In the present study, a crossover experiment over four production cycles was conducted in seven commercial chicken barns to compare WW and FD. We evaluated the effects of barn cleaning methods on commercial broiler performance, cecal microbiota composition, Campylobacter and Salmonella occurrence, and Campylobacter jejuni and Clostridium perfringens abundance, as well as on short-chain fatty acid (SCFA) concentrations in the month-old broiler gut. The 30-day body weight and mortality rate were not affected by the barn cleaning methods. The WW resulted in a modest but significant effect on the structure of broiler cecal microbiota (weighted-UniFrac; adonis P =  0.05, and unweighted-UniFrac; adonis P =  0.01), with notable reductions in C. jejuni occurrence and abundance. In addition, the WW group had increased cecal acetate, butyrate, and total SCFA concentrations, which were negatively correlated with C. jejuni abundance. Our results suggest that WW may result in enhanced activity of the gut microbiota and reduced zoonotic transmission of C. jejuni in broiler production relative to FD in the absence of a disease challenge. IMPORTANCE We compared the effects of barn FD and WW methods on gut microbial community structures and pathogen prevalence of broiler chickens in a nonchallenging commercial production setting. The results revealed that barn cleaning methods had little impact on the 30-day body weight and mortality rate of broiler chickens. In addition, the FD treatment had a subtle but significant effect on the broiler cecal microbiota with increased abundances of Campylobacter and decreased SCFA concentrations, which would support the adoption of WW as a standard practice. Thus, compared to FD, WW can be beneficial to broiler chicken production by inhibiting zoonotic pathogen colonization in the chicken gut with reduced cost and labor of cleaning.

Background: Campylobacteriosis is currently the most frequently reported zoonosis. Dogs, especially puppies or those with diarrhea, are considered a possible source of human infection. Probiotic bacteria, such as Lactobacillus species, seem to be a valuable tool in controlling of intestinal pathogenic microorganisms in dogs. The main purpose of this study was to assess the anti-Campylobacter activity and some probiotic properties, like ability to produce H 2 O 2, bile salt and low pH tolerance of Lactobacillus strains isolated from gastrointestinal tract of healthy dogs., Results: A total of 39 rectal swabs derived from healthy dogs and 19 from dogs with diarrhea were examined to detect Lactobacillus and Campylobacter bacteria respectively. In total, 30 strains of Lactobacillus genus and four strains of Campylobacter genus were isolated and identified. Of the 30 strains of Lactobacillus, 22 showed an inhibitory effect towards Campylobacter. Four strains with the strongest antagonism towards Campylobacter bacteria (L. salivarius 25 K/L/1, L. rhamnosus 42 K/L/2, L. sakei 50 K/L/1 and L. agilis 55 K/L/1) were selected to assess their potential probiotic traits. Three out of four analyzed strains produced extracellular H 2 O 2. All displayed very good or moderate survival at pH 3.0 and 2.0 and showed high tolerance to 0.5% and 1% bile salts., Conclusions: Among selected Lactobacillus strains, all may have a potential probiotic application in reducing Campylobacter spp. in dogs and thus prevent transmission of infection to humans, although the best candidate for probiotic seems to be L. sakei 50 K/L/1. Further in vitro and in vivo studies are needed., (© 2022. The Author(s).)

The MORDOR trial in Niger, Malawi, and Tanzania found that biannual mass distribution of azithromycin to children younger than 5 years led to a 13.5% reduction in all-cause mortality (NCT02048007). To help elucidate the mechanism for mortality reduction, we report IgG responses to 11 malaria, bacterial, and protozoan pathogens using a multiplex bead assay in pre-specified substudy of 30 communities in the rural Niger placebo-controlled trial over a three-year period (n = 5642 blood specimens, n = 3814 children ages 1-59 months). Mass azithromycin reduces Campylobacter spp. force of infection by 29% (hazard ratio = 0.71, 95% CI: 0.56, 0.89; P = 0.004) but serological measures show no significant differences between groups for other pathogens against a backdrop of high transmission. Results align with a recent microbiome study in the communities. Given significant sequelae of Campylobacter infection among preschool aged children, our results support an important mechanism through which biannual mass distribution of azithromycin likely reduces mortality in Niger., (© 2022. The Author(s).)

Campylobacter jejuni is the leading bacterial cause of human gastroenteritis worldwide and handling or consumption of contaminated poultry meat is the key source of infection. Glycoconjugate vaccines containing the C. jejuni N-glycan have been reported to be partially protective in chickens. However, our previous studies with subunit vaccines comprising the C. jejuni FlpA or SodB proteins with up to two or three C. jejuni N-glycans, respectively, failed to elicit significant protection. In this study, protein glycan coupling technology was used to add up to ten C. jejuni N-glycans onto a detoxified form of Pseudomonas aeruginosa exotoxin A (ExoA). The glycoprotein, G-ExoA, was evaluated for efficacy against intestinal colonisation of White Leghorn chickens by C. jejuni strains M1 and 11168H relative to unglycosylated ExoA. Chickens were challenged with the minimum dose required for reliable colonisation, which was 10 2 colony-forming units (CFU) for strain M1 and and 10 4 CFU for strain 11168H. Vaccine-specific serum IgY was detected in chickens vaccinated with both ExoA and G-ExoA. However, no reduction in caecal colonisation by C. jejuni was observed. While the glycan dose achieved with G-ExoA was higher than FlpA- or SodB-based glycoconjugates that were previously evaluated, it was lower than that of glycoconjugates where protection against C. jejuni has been reported, indicating that protection may be highly sensitive to the amount of glycan presented and/or study-specific variables., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: [Brendan Wren, Jon Cuccui reports a relationship with ArkVax that includes: equity or stocks.]., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Microbial biofilms occur naturally in many environmental niches and can be a significant reservoir of infectious microbes in zoonotically transmitted diseases such as that caused by Campylobacter jejuni , the leading cause of acute human bacterial gastroenteritis world-wide. The greatest challenge in reducing the disease caused by this organism is reducing transmission of C. jejuni to humans from poultry via the food chain. Biofilms enhance the stress tolerance and antimicrobial resistance of the microorganisms they harbor and are considered to play a crucial role for Campylobacter spp. survival and transmission to humans. Unconventional approaches to control biofilms and to improve the efficacy of currently used antibiotics are urgently needed. This review summarizes the use plant- and microorganism-derived antimicrobial and antibiofilm compounds such as essential oils, antimicrobial peptides (AMPs), polyphenolic extracts, algae extracts, probiotic-derived factors, d-amino acids (DAs) and glycolipid biosurfactants with potential to control biofilms formed by Campylobacter , and the suggested mechanisms of their action. Further investigation and use of such natural compounds could improve preventative and remedial strategies aimed to limit the transmission of campylobacters and other human pathogens via the food chain.

Using data on rearing and welfare metrics of multiple commercial broiler flocks, we investigate how welfare measures such as hock burn, mortality, and pododermatitis, among others, impact the likelihood of a flock becoming colonized by Campylobacter. Using both logistic regression and Bayesian networks, we show that, while some welfare metrics were weakly related to Campylobacter colonization, evidence could not be found to suggest that these metrics directly exacerbated Campylobacter colonization, rather that they were both symptoms of the same parent variable - the managing company. Observed dependency on the management of the flock suggested that yet-undiscovered differences in rearing practice were the principal factor explaining both poor bird welfare and increased risk of Campylobacter, suggesting that action can be taken to improve both these factors simultaneously., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: Poultry is the world's most popular animal-based food and global production has tripled in the past 20 years alone. Low-cost vaccines that can be combined to protect poultry against multiple infections are a current global imperative. Glycoconjugate vaccines, which consist of an immunogenic protein covalently coupled to glycan antigens of the targeted pathogen, have a proven track record in human vaccinology, but have yet to be used for livestock due to prohibitively high manufacturing costs. To overcome this, we use Protein Glycan Coupling Technology (PGCT), which enables the production of glycoconjugates in bacterial cells at considerably reduced costs, to generate a candidate glycan-based live vaccine intended to simultaneously protect against Campylobacter jejuni, avian pathogenic Escherichia coli (APEC) and Clostridium perfringens. Campylobacter is the most common cause of food poisoning, whereas colibacillosis and necrotic enteritis are widespread and devastating infectious diseases in poultry., Results: We demonstrate the functional transfer of C. jejuni protein glycosylation (pgl) locus into the genome of APEC χ7122 serotype O78:H9. The integration caused mild attenuation of the χ7122 strain following oral inoculation of chickens without impairing its ability to colonise the respiratory tract. We exploit the χ7122 pgl integrant as bacterial vectors delivering a glycoprotein decorated with the C. jejuni heptasaccharide glycan antigen. To this end we engineered χ7122 pgl to express glycosylated NetB toxoid from C. perfringens and tested its ability to reduce caecal colonisation of chickens by C. jejuni and protect against intra-air sac challenge with the homologous APEC strain., Conclusions: We generated a candidate glycan-based multivalent live vaccine with the potential to induce protection against key avian and zoonotic pathogens (C. jejuni, APEC, C. perfringens). The live vaccine failed to significantly reduce Campylobacter colonisation under the conditions tested but was protective against homologous APEC challenge. Nevertheless, we present a strategy towards the production of low-cost "live-attenuated multivalent vaccine factories" with the ability to express glycoconjugates in poultry., (© 2021. The Author(s).)

Abstract: New Zealand (NZ) has a high rate of reported campylobacteriosis cases. Cross-contamination in home kitchens during poultry handling is considered the main factor in campylobacteriosis transmission. The main aim of this study was to measure NZ consumers' food safety awareness and self-reported food safety practices associated with handling raw poultry. This study will contribute to the existing knowledge to explain the reasons behind the increase of campylobacteriosis incidents. Findings can help inform future consumer education campaigns to help reduce the incidence of campylobacteriosis in NZ. A cross-sectional survey composed of 31 multiple-choice questions was designed, piloted, and used to collect information about the last time consumers purchased and prepared raw poultry at home. A street-intercept survey in public places, such as supermarkets in the Canterbury region, was used to recruit respondents for this study. A descriptive and inferential data analysis was performed, including a one-way analysis of variance test used to compare the mean scored responses of the respondents among different sociodemographics. Overall, 301 valid responses were obtained. Scores representing reported safe food practices ranged between 2 and 19 (maximum 21), with a mean score of 9.83 (standard deviation of 3.50 with a standard error of 0.20). There was some variation in correctly answered questions by respondents for food hygiene (25%), cross-contamination prevention (55%), temperature control and storage practices (49%), and food safety (52%). Approximately 30% of the respondents reported symptoms of a foodborne disease experienced once to four times during the past 12 months. The study identified low adherence to current recommended food safety practices, including safe food storage and temperature control. The findings can be used to inform a communication campaign regarding food safety needs to be designed urgently in NZ to reduce the rate of campylobacteriosis., (Copyright ©, International Association for Food Protection.)

Campylobacter, a leading cause of foodborne diseases, is well recognized worldwide. Poultry and poultry products are considered as major sites for Campylobacter infection in humans. The extensive uses of antibiotics mostly as growth promoters and for therapeutic purposes have led to the emergence of antibiotic-resistant strains of foodborne pathogens including Campylobacter. A key tenet of this paper is the need for reviewing the previous studies conducted around the globe on the prevalence and antimicrobial resistance of Campylobacter spp. isolates in duck to better understand the sources and trends of infection. Based on published data, the prevalence of Campylobacter spp. in duck and duck-related samples ranged from 0% to 100% and was largely influenced by the isolation method. Among Campylobacter spp., C. jejuni was the predominant cause of campylobacteriosis, followed by C. coli. Campylobacter spp. from ducks were mostly resistant to fluoroquinolones and tetracycline and a lesser extent to gentamicin, chloramphenicol, and erythromycin. Some studies showed that ducks may pose a risk for acquiring campylobacteriosis because they had genotypes quite similar to human isolates detected previously. A continued monitoring approach is needed, at national and international levels, with enhanced surveillance and reporting of trends, as well as harmonization of surveillance systems toward a one-health approach to monitoring antimicrobial resistance in animal production particularly if increased resistance rates are being demonstrated., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: Water, sanitation and hygiene (WASH) interventions should support infant growth but trial results are inconsistent. Frequently, interventions do not consider behaviours or transmission pathways specific to age. A household playspace (HPS) is one intervention component which may block faecal-oral transmission. This study was a two-armed, parallel-group, randomised, controlled feasibility trial of a HPS in rural Ethiopia. It aimed to recommend proceeding to a definitive trial. Secondary outcomes included effects on infant health, injury prevention and women's time., Methods: November 2019-January 2020 106 households were identified and assessed for eligibility. Recruited households (N = 100) were randomised (blinded prior to the trial start) to intervention or control (both n = 50). Outcomes included recruitment, attrition, adherence, and acceptability. Data were collected at baseline, two and four weeks., Findings: Recruitment met a priori criteria (≥80%). There was no loss to follow-up, and no non-use, meeting adherence criteria (both ≤10%). Further, 48.0% (95% CI 33.7-62.6; n = 24) of households appropriately used and 56.0% (41.3-70.0; n = 28) cleaned the HPS over four weeks, partly meeting adherence criteria (≥50%). For acceptability, 41.0% (31.3-51.3; n = 41) of infants were in the HPS during random visits, failing criteria (≥50%). Further, the proportion of HPS use decreased during some activities, failing criteria (no decrease in use). A modified Barrier Analysis described good acceptability and multiple secondary benefits, including on women's time burden and infant injury prevention., Interpretation: Despite failing some a priori criteria, the trial demonstrated mixed adherence and good acceptability among intervention households. A definitive trial to determine efficacy is warranted if recommended adjustments are made., Funding: People In Need; Czech Development Agency., Trial Registration: RIDIE-ID-5de0b6938afb8., Competing Interests: The authors have declared that no competing interests exist.

Acute diarrheal illness and gastroenteritis caused by Campylobacter jejuni infection remain significant public health risks in developing countries with substantial mortality and morbidity in humans, particularly in children under the age of five. Genetic diversities among Campylobacter jejuni and limited understanding of immunological correlations of host protection remain primary impediments for developing an effective measure to control Campylobacter infection. Moreover, the lack of a reliable in vivo model to mimic natural infection against Campylobacter jejuni has substantially delayed the vaccine-development process. Given the role of bacterial outer membrane associated proteins in intestinal adherence and invasion as well as modulating dynamic interplay between host and pathogens, bacterial outer-membrane vesicles have emerged as a potential vaccine target against a number of gut pathogens, including Campylobacter jejuni . Here, we describe a mucosal vaccine strategy using chitosan-coated outer-membrane vesicles to induce specific immune responses against Campylobacter jejuni in mice. To overcome the challenges of mucosal delivery of outer membrane vesicles in terms of exposure to variable pH and risk of enzymatic degradation, we preferentially used chitosan as a nontoxic, mucoadhesive polymer. We show that intragastric delivery of chitosan-coated outer-membrane vesicles imparts significant immune protection against Campylobacter jejuni with high level local and systemic antibody production. Further, immunization with the outer membrane vesicles resulted in potent cellular responses with an increased CD4 + and CD8 + T cell population. Moreover, significant upregulation of IFN-γ and IL-6 gene expression suggests that mucosal delivery of outer membrane vesicles promotes a Th1/Th2 mixed-type immune response. Together, as an acellular and nonreplicating canonical end product of bacterial secretion, mucosal delivery of outer membrane vesicles may represent a promising platform for developing an effective vaccine against Campylobacter jejuni .

As one of the main causes of bacterial diarrhea and a major risk factor for triggering Guillain-Barré autoimmune syndrome, campylobacteriosis, that is, Campylobacter spp. infections, represents a major health issue worldwide. There is thus a pressing need for developing an effective and broad-coverage campylobacteriosis vaccine. Campylobacter jejuni , an encapsulated, multidrug resistant Gram-negative bacterium, expresses virulence-associated capsular polysaccharides (CPSs), which constitute exquisite targets for the design of glycoconjugate vaccines. In that context, synthetic carbohydrate chemistry acts as a crucial enabling technology for the preparation of homogeneous constructs while allowing antigenic epitopes to be deciphered and probed at the molecular level. This review aims at covering recent developments in CPS-based campylobacteriosis vaccines as well as in the total syntheses of C. jejuni -related mono- and oligosaccharide mimics.

In many countries campylobacteriosis ranks as one of the most frequently reported foodborne illnesses and poultry is the commodity that is most often associated with these illnesses. Nevertheless, efforts to reduce the occurrence of pathogen contamination on poultry are often more focused on Salmonella. While some control measures are pathogen specific, such as pre-harvest vaccination for Salmonella, improvements in sanitary dressing and interventions applied during the slaughter process can be effective against all forms of microbial contamination. To investigate the potential effectiveness of these non-specific pathogen reduction strategies in the United States, it is helpful to assess if, and by how much, Campylobacter contamination of chicken meat has changed across time. This study assesses change considering data collected in both slaughter and retail establishments and comparing observed trends in contamination with trends in human surveillance data. The results support the assertion that substantial reductions in Campylobacter contamination of chicken meat in the late 1990s and early 2000s contributed to a reduction in the human case rate of campylobacteriosis. Further reductions in chicken meat contamination between 2013 and 2018 are more difficult to associate with trends in human illnesses, with one contributing factor being the inclusion of culture independent diagnostic test results in the official case counts during that time. Other contributing factors are discussed., (Published by Elsevier B.V.)

Poultry is seen as the main reservoir for Campylobacter. Control of this zoonotic pathogen in primary production could potentially reduce the colonization in broiler flocks and consequently reduce the number of human infections. In the present study, 20 broiler flocks from 10 farms, were sampled immediately before and 5 to 7 d after partial depopulation (thinning) for the presence of Campylobacter using cecal droppings and overshoes. At the time of thinning, the catching crew, transportation vehicles, forklift, and transport containers were sampled for the presence of Campylobacter. Samples were cultivated; presumed positive isolates were confirmed by PCR. The isolates were molecularly typed by flaA restriction analysis and pulsed field gel electrophoresis. Results show that all flocks were thinned using Campylobacter-contaminated equipment and materials. One-third of the broiler flocks became colonized after thinning. In 67% of the colonization cases, identical strains were found matching those of container systems, transport trucks, and/or forklifts. This identifies thinning as an important risk factor for Campylobacter introduction into broiler houses. Setup and compliance with biosecurity practices during thinning is essential to prevent Campylobacter colonization of broiler flocks., (Copyright © 2020. Published by Elsevier Inc.)

1. This review explores current and proposed on-farm interventions and assess the potential of these interventions against Campylobacter spp. 2. Interventions such as vaccination, feed/water-additives and, most importantly, consistent biosecurity, exhibit potential for the effective control of this pathogen and its dissemination within the food chain. 3. Due to the extensive diversity in the Campylobacter spp. genome and surface-expressed proteins, vaccination of poultry is not yet regarded as a completely effective strategy. 4. The acidification of drinking water through the addition of organic acids has been reported to decrease the risk of Campylobacter spp. colonisation in broiler flocks. Whilst this treatment alone will not completely protect birds, use of water acidification in combination with in-feed measures to further reduce the level of Campylobacter spp. colonisation in poultry may be an option meriting further exploration. 5. The use of varied types of feed supplements to reduce the intestinal population and shedding rate of Campylobacter spp. in poultry is an area of growing interest in the poultry industry. Such supplements include pro - and pre-biotics, organic acids, bacteriocins and bacteriophage, which may be added to feed and water. 6. From the literature, it is clear that a distinct, albeit not unexpected, difference between the performance of in-feed interventions exists when examined in vitro compared to those determined in in vivo studies. It is much more likely that pooling some of the discussed approaches in the in-feed tool kit will provide an answer. 7. Whilst on-farm biosecurity is essential to maintain a healthy flock and reduce disease transmission, even the most stringent biosecurity measures may not have sufficient, consistent and predictable effects in controlling Campylobacter spp. Furthermore, the combination of varied dietary approaches and improved biosecurity measures may synergistically improve control.

Campylobacter, Gram-negative bacteria, is the most common cause of acute bacterial enteritis in human beings, both in developing and developed countries. It is believed that poultry, in particular broiler chickens, is the main host of human infection with Campylobacter. Handling and consumption of contaminated chicken meat are the usual modes of transmission. Prevention and reduction of Campylobacter colonization in poultry farms will cut off the road of infection transmission to humans throughout the food chain. With the incidence of antibiotic resistance and with growing concern about superbugs, the search for natural and safe alternatives will considerably increase in the coming years. In this review, we will discuss the prevalence and risk factors of Campylobacter colonization in broiler chickens and sources of infection. This review also provides extensive and recent approaches to prevent and control Campylobacter colonization in broiler chickens, including biosecurity measures, natural feed/drinking water additives with antimicrobial properties, bacteriocins, bacteriophages, antimicrobial peptides, and vaccination strategies to prevent and control the incidence of human campylobacteriosis.

Human infections by Campylobacter species are among the most reported bacterial gastrointestinal diseases in the European Union and worldwide with severe outcomes in rare cases. Considering the transmission routes and farm animal reservoirs of these zoonotic pathogens, a comprehensive One Health approach will be necessary to reduce human infection rates. Bacteriophages are viruses that specifically infect certain bacterial genera, species, strains or isolates. Multiple studies have demonstrated the general capacity of phage treatments to reduce Campylobacter loads in the chicken intestine. However, phage treatments are not yet approved for extensive use in the agro-food industry in Europe. Technical inconvenience is mainly related to the efficacy of phages, depending on the optimal choice of phages and their combination, as well as application route, concentration and timing. Additionally, regulatory uncertainties have been a major concern for investment in commercial phage-based products. This review addresses the question as to how phages can be put into practice and can help to solve the issue of human campylobacteriosis in a sustainable One Health approach. By compiling the reported findings from the literature in a standardized manner, we enabled inter-experimental comparisons to increase our understanding of phage infection in Campylobacter spp. and practical on-farm studies. Further, we address some of the hurdles that still must be overcome before this new methodology can be adapted on an industrial scale. We envisage that phage treatment can become an integrated and standardized part of a multi-hurdle anti-bacterial strategy in food production. The last part of this chapter deals with some of the issues raised by legal authorities, bringing together current knowledge on Campylobacter-specific phages and the biosafety requirements for approval of phage treatment in the food industry.

Numerous studies point out that at present, a complete elimination of Campylobacter species in the poultry food chain is not feasible. Thus, the current aim should be to establish control measures and intervention strategies to minimize the occurrence of Campylobacter spp. in livestock (esp. poultry flocks) and to reduce the quantitative Campylobacter burden along the food chain in animals and subsequently in foods. The most effective measures to mitigate Campylobacter focus on the primary production stage. Nevertheless, measures applied during slaughter and processing complement the general meat hygiene approaches by reducing fecal contamination during slaughtering and processing and as a consequence help to reduce Campylobacter in poultry meat. Such intervention measures at slaughter and processing level would include general hygienic improvements, technological innovations and/or decontamination measures that are applied at single slaughter or processing steps. In particular, approaches that do not focus on a single intervention measure would need to be based on a thorough process of evaluation, and potential combinatory effects have to be modeled and tested. Finally, the education of all stakeholders (including retailers, food handlers and consumers) is required and will help to increase awareness for the presence of foodborne pathogens in raw meat and meat products and can thus aid in the development of the required good kitchen hygiene.

Although extensive research has been carried out to describe the transmission pathways of Campylobacter entering livestock farms, the role of livestock farms as source of Campylobacter contamination of the environment is still poorly investigated. It is assumed that Campylobacter-positive livestock farms contribute to an environmental contamination, depending on the animal species on the farm, their Campylobacter status, the housing system, manure management as well as their general farm hygienic and biosecurity management. Different emission sources, like manure, air, water, insects and rodents as well as personnel, including equipment and vehicles, contribute to Campylobacter emission into the environment. Even though Campylobacter are rather fastidious bacteria, they are able to survive in the environment for even a longer period of time, when environmental conditions enable survival in specific niches. We conclude that a significant reduction of Campylobacter emission in the environment can be successfully achieved if various intervention strategies, depending on the farm type, are applied simultaneously, including proper general and personal hygiene, establishing of hygienic barriers, insect controls, manure management and hygienization of stables, barns and exhaust air.

Zoonotic Campylobacter species-mainly C. jejuni and C. coli-are major causes of food-borne bacterial infectious gastroenteritis worldwide. Symptoms of intestinal campylobacteriosis include abdominal pain, diarrhea and fever. The clinical course of enteritis is generally self-limiting, but some infected individuals develop severe post-infectious sequelae including autoimmune disorders affecting the nervous system, the joints and the intestinal tract. Moreover, in immunocompromised individuals, systemic spread of the pathogens may trigger diseases of the circulatory system and septicemia. The socioeconomic costs associated with Campylobacter infections have been calculated to several billion dollars annually. Poultry meat products represent major sources of human infections. Thus, a "One World-One Health" approach with collective efforts of public health authorities, veterinarians, clinicians, researchers and politicians is required to reduce the burden of campylobacteriosis. Innovative intervention regimes for the prevention of Campylobacter contaminations along the food chain include improvements of information distribution to strengthen hygiene measures for agricultural remediation. Given that elimination of Campylobacter from the food production chains is not feasible, novel intervention strategies fortify both the reduction of pathogen contamination in food production and the treatment of the associated diseases in humans. This review summarizes some current trends in the combat of Campylobacter infections including the combination of public health and veterinary preventive approaches with consumer education. The "One World-One Health" perspective is completed by clinical aspects and molecular concepts of human campylobacteriosis offering innovative treatment options supported by novel murine infection models that are based on the essential role of innate immune activation by bacterial endotoxins.

The development of interventions to reduce human foodborne pathogens in the gastrointestinal (GI) tract of chickens will be important for improving the microbial food safety of poultry. Saccharomyces-derived prebiotic refined functional carbohydrates (RFC), composed primarily of β-glucans, mannanoligosaccharides (MOS), and D-mannose have been demonstrated to reduce GI colonization of Salmonella and Campylobacter when administered to poultry. Although they are presumed to inhibit adhesion of pathogens to the GI epithelium, this functionality of RFC has not been well characterized. In this study, we investigated the effects of RFC and other prebiotics on the adhesion of Salmonella Typhimurium and Campylobacter jejuni to the LMH chicken epithelial cell line in vitro. The reduction of adherent pathogens was observed to be dose-dependent with C. jejuni being more sensitive than Salmonella to inhibition by RFC. Comparison of the primary constituent carbohydrates of RFC found D-mannose to inhibit both pathogens less effectively than β-glucan and MOS, suggesting that it contributes less to inhibition of pathogen adhesion than the other carbohydrates. Finally, the reduction of adherent pathogens by RFC was compared with that of fructooligosaccharides (FOS), galactooligosaccharides (GOS), and raffinose. All 4 prebiotics inhibited adhesion of both pathogens to chicken epithelial cells. Reduction of adherent Salmonella was greatest with FOS and lowest with GOS, whereas reduction of adherent C. jejuni was greater with RFC and raffinose than with FOS and GOS. These results will inform future research elucidating mechanisms important to adhesion inhibition of pathogens by RFC and other prebiotics., (Copyright © 2020. Published by Elsevier Inc.)