In silico prediction and expression analysis of vaccine candidate genes of Campylobacter jejuni.

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.

Campylobacter Biofilms: Potential of Natural Compounds to Disrupt Campylobacter jejuni Transmission.

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.

Campylobacter sp.: Pathogenicity factors and prevention methods-new molecular targets for innovative antivirulence drugs?

Infections caused by bacterial species from the genus Campylobacter are one of the four main causes of strong diarrheal enteritis worldwide. Campylobacteriosis, a typical food-borne disease, can range from mild symptoms to fatal illness. About 550 million people worldwide suffer from campylobacteriosis and lethality is about 33 million p.a. This review summarizes the state of the current knowledge on Campylobacter with focus on its specific virulence factors. Using this knowledge, multifactorial prevention strategies can be implemented to reduce the prevalence of Campylobacter in the food chain. In particular, antiadhesive strategies with specific adhesion inhibitors seem to be a promising concept for reducing Campylobacter bacterial load in poultry production. Antivirulence compounds against bacterial adhesion to and/or invasion into the host cells can open new fields for innovative antibacterial agents. Influencing chemotaxis, biofilm formation, quorum sensing, secretion systems, or toxins by specific inhibitors can help to reduce virulence of the bacterium. In addition, the unusual glycosylation of the bacterium, being a prerequisite for effective phase variation and adaption to different hosts, is yet an unexplored target for combating Campylobacter sp. Plant extracts are widely used remedies in developing countries to combat infections with Campylobacter. Therefore, the present review summarizes the use of natural products against the bacterium in an attempt to stimulate innovative research concepts on the manifold still open questions behind Campylobacter towards improved treatment and sanitation of animal vectors, treatment of infected patients, and new strategies for prevention. KEY POINTS: • Campylobacter sp. is a main cause of strong enteritis worldwide. • Main virulence factors: cytolethal distending toxin, adhesion proteins, invasion machinery. • Strong need for development of antivirulence compounds.

The effectiveness of a dietary direct-fed microbial and mannan oligosaccharide on ultrastructural changes of intestinal mucosa of turkey poults infected with Salmonella and Campylobacter.

Salmonella and Campylobacter are considered major public health burdens worldwide, and poultry are known to be one of the main reservoirs for these zoonotic pathogens. This study was conducted to evaluate the effect of a commercial probiotic or direct-fed microbial (DFM) Calsporin (CSP), and prebiotic or mannan oligosaccharide (MOS) (IMW50) on ultrastructural changes and the villous integrity of intestinal mucosa in turkey poults challenged with Salmonella and Campylobacter. A 21-day battery cage study was conducted using 4 dietary treatments including a basal diet (corn and soybean-based) nonsupplemented and uninfected as a negative control (NC); basal diet supplemented with 0.05% DFM (CSP); basal diet supplemented with 0.05% MOS (IMW50); and basal diet supplemented with 0.05% mixture of DFM and MOS at equal proportions. Female large white turkey poults aged 336 days were obtained from a local commercial hatchery and randomly distributed in electrically heated battery cages with 12 treatments of 4 replicates per treatment containing 7 poults per pen. The first 16 pens were not infected with bacteria, poults in pens 17-32 were orally challenged at day 7 with 105 cfu Salmonella Heidelberg, and the poults in pens 33-48 were orally challenged at day 7 with 105 cfu Campylobacter jejuni. Feed and water were provided ad libitum throughout the study. At day 21, ileal tissue samples from 1 bird per cage were collected for intestinal integrity and ultrastructural examination by scanning and electron microscopy. DFM and MOS supplementation was effective in both challenged and nonchallenged (not infected with Salmonella and Campylobacter) birds. Goblet cells and mucus were increased, with the presence of large numbers of segmented filamentous bacteria in DFM- and MOS-supplemented groups compared with birds in control treatments. The number and size of villi were reduced in poults exposed to Salmonella and Campylobacter. Results show that CSP and IMW50 provide protection of ileal mucosal integrity in poults exposed to Salmonella or Campylobacter.

Campylobacter: A Review of New Promising Remedies with Medicinal Plants and Natural Antioxidants.

Campylobacter (curved bacteria) is considered one of the most important and common zoonotic bacteria and the three leading bacterial causes of gastroenteritis and diarrhea. Antibacterial resistance is growing and expanding. The aim of this review article is to report anti-Campylobacter medicinal plants. For this purpose, the search terms consisting of Campylobacter, medicinal plants, essential oil, extract, and traditional medicine were used to retrieve the relevant articles published in the journals indexed in Information Sciences Institute, Web of Science, PubMed, Scopus, Google Scholar, and Scientific Information Databases. Then, the findings of eligible articles were analyzed. According to the analysis, 71 medicinal plants were found to exert anti-Campylobacter effect. The active compounds of these plants are possibly nature-based antibiotic agents that are effective on Campylobacter. If these compounds are isolated, purified, and studied in pharmaceutical investigations, they can be used to produce nature-based, anti-Campylobacter antibiotics.

Feed supplementation with biochar may reduce poultry pathogens, including Campylobacter hepaticus, the causative agent of Spotty Liver Disease.

Increased global regulation and restrictions on the non-therapeutic use of antibiotics in the poultry industry means that there is a need to identify alternatives that prevent infection while still conveying the growth and performance benefits afforded by their use. Biochars are produced by the incomplete pyrolysis of organic materials, with reports of use as a feed supplement and activity against pathogenic bacteria. In the current study the dose-dependent effects of biochar dietary inclusion in layer diets at 1%, 2% and 4% w/w were investigated to determine a) the efficacy of biochar as an anti-pathogenic additive on the intestinal microbiota and b) the optimal inclusion level. Biochar inclusion for anti-pathogenic effects was found to be most beneficial at 2% w/w. Poultry pathogens such as Gallibacterium anatis and campylobacters, including Campylobacter hepaticus, were found to be significantly lower in biochar fed birds. A shift in microbiota was also associated with the incorporation of 2% w/w biochar in the feed in two large scale trials on two commercial layer farms. Biochar inclusion for anti-pathogenic effects was found to be most beneficial at 2% w/w. Differential effects of the timing of biochar administration (supplementation beginning at hatch or at point of lay) were also evident, with greater impact on community microbial structure at 48 weeks of age when birds were fed from hatch rather than supplemented at point of lay.

In vitro efficacy of potentiated egg yolk powder against Campylobacter jejuni does not correlate with in vitro efficacy.

Campylobacter jejuni is a zoonotic agent responsible for the foodborne gastroenteritis campylobacteriosis. Control of C. jejuni load in the poultry primary production is recognized as an avenue to reduce human exposure to the pathogen. As for now, no commercially applicable control methods exist at the farm. Several studies tested egg yolk powders, potentiated or not against C. jejuni, as feed additives for chicken and suggested that the quantity and quality of the antibodies presence in the yolk are determinant factors for the full success of this approach. Unfortunately, data from these studies inconsistently showed a reduction of cecal C. jejuni carriage. Our first goal wwas to characterize (quantification by ELISA, agglutination test, bacterial antigen recognition profiles by Western blot, bactericidal effect by serum killing assays and C. jejuni mobility by soft agar migation) the antibodies extracted from egg yolk powders originating from different egg production protocols. Secondly, these powders were microencapsulated and recharacterized. Finally the protected powders were tested as a feed additive to destabilize C. jejuni colonization in an in vivo assay. Despite the in vitro results indicating the ability of the egg yolk powders to recognize Campylobacter and potentially alter its colonization of the chicken caecum, these results were not confirmed in the in vivo trial despite that specific caecal IgY directed toward Campylobacter were detected in the groups receiving the protected powders. More research is needed on Campylobacter in order to effectively control this pathogen at the farm.

Influence of a specific amino acid pattern in the diet on the course of an experimental Campylobacter jejuni infection in broilers.

Campylobacter jejuni (C. jejuni) is one of the most important zoonotic pathogens worldwide. In Europe, the majority of the cases are caused by consuming contaminated poultry meat. The objective of the present study was to investigate potential effects of different crude protein levels in complete diets for broilers on infection dynamics of C. jejuni after experimental infection. In total, 300 commercial broilers line Ross 308 were divided into 4 different groups, including 5 replications of 15 chickens each. The chickens were fed a conventional diet (212 g CP/kg DM) and a protein-reduced test diet (190 g CP/kg DM) supplemented with essential amino acids. This resulted simultaneously in lower amino-acid concentrations preferentially utilized by C. jejuni, such as aspartate, glutamate, proline, and serine. One group of each feeding concept was infected artificially with C. jejuni at day 21 by applying an oral C. jejuni inoculum containing 4.17 ± 0.09 log10 cfu of C. jejuni to 3 of 15 chickens, called "seeders." Feeding the test diet resulted in a significant reduction (P < 0.001) in CP intake (31.5 ± 1.20 g CP/broiler/day and 27.7 ± 0.71 g CP/broiler/day, respectively), a significant decrease (P < 0.05) in crude mucin in excreta (55.7 ± 8.23 g/kg DM and 51.9 ± 7.62 g/kg DM, respectively), and in goblet cell number in cecal crypts (P < 0.05; 15.1 ± 5.71 vs. 13.6 ± 5.91 goblet cells/crypt). In groups receiving the test diet, the excretion of C. jejuni was significantly reduced in seeders by 1.9 log10 cfu/g excreta at day 23 (3.38a ± 2.55 vs. 1.47b ± 2.20; P = 0.033). At day 25, prevalence of C. jejuni in cloacal swabs amounted to 53.3% in the group fed the test diet and 75.7% in the control group, respectively (P < 0.05). In summary, a definite amino acid pattern in the broiler diets could contribute to a development of an effective feeding strategy to reduce the prevalence of C. jejuni infection in chickens (Patent No 17187659.2-1106).

Rifaximin Fails to Prevent Campylobacteriosis in the Human Challenge Model: A Randomized, Double-Blind, Placebo-Controlled Trial.

Background: Campylobacter species are a leading cause of diarrheal disease globally with significant morbidity. Primary prevention efforts have yielded limited results. Rifaximin chemoprophylaxis decreases rates of travelers' diarrhea and may be suitable for high-risk persons. We assessed the efficacy of rifaximin in the controlled human infection model for Campylobacter jejuni. Methods: Twenty-eight subjects were admitted to an inpatient facility and randomized to a twice-daily dose of 550 mg rifaximin or placebo. The following day, subjects ingested 1.7 × 105 colony-forming units of C. jejuni strain CG8421. Subjects continued prophylaxis for 3 additional days, were followed for campylobacteriosis for 144 hours, and were subsequently treated with azithromycin and ciprofloxacin. Samples were collected to assess immunologic responses to CG8421. Results: There was no difference (P = 1.0) in the frequency of campylobacteriosis in those receiving rifaximin (86.7%) or placebo (84.6%). Additionally, there were no differences in the clinical signs and symptoms of C. jejuni infection to include abdominal pain/cramps (P = 1.0), nausea (P = 1.0), vomiting (P = .2), or fever (P = 1.0) across study groups. Immune responses to the CG8421 strain were comparable across treatment groups. Conclusions: Rifaximin did not prevent campylobacteriosis in this controlled human infection model. Given the morbidity associated with Campylobacter infection, primary prevention efforts remain a significant need. Clinical Trials Registration: NCT02280044.

Effects of dry whey powder and calcium butyrate supplementation of corn/soybean-based diets on productive performance, duodenal histological integrity, and Campylobacter colonization in broilers.

BACKGROUND: Campylobacter is the main cause of gastroenteritis in humans in industrialized countries, and poultry is its principal reservoir and source of human infections. Dietary supplementation of broiler feed with additives could improve productive performance and elicit health benefits that might reduce Campylobacter contamination during primary production. The aim of this study was to assess the effect of dietary supplementation with whey (a prebiotic) and calcium butyrate (a salt of a short-chain fatty acid) on productive traits, duodenal histological integrity, and Campylobacter colonization and dissemination in broiler chickens during the 42-day rearing period. RESULTS: Six hundred one-day-old Ross-308 chickens were placed into 20 ground pens and assigned to one of 4 corn/soybean-based dietary treatments (5 replicates of 30 chicks per treatment) following a randomized complete block design: 1) basal diet with no supplementation as the control, 2) diet supplemented with 6% dry whey powder, 3) diet containing 0.1% coated calcium butyrate, and 4) diet containing 6% whey and 0.1% calcium butyrate. At age 15 days, 6 chickens per pen were experimentally inoculated with Campylobacter jejuni. The results showed that supplementation of the corn/soybean-based diet with 6% whey alone or, preferably, in combination with 0.1% coated calcium butyrate improved growth and feed efficiency, had a beneficial effect on duodenal villus integrity, and decreased mortality. These favourable effects were particularly significant during the starter period. Six days after oral challenge, Campylobacter was widespread in the flock, and the birds remained positive until the end of the rearing period. Although Campylobacter was not isolated from environmental samples, it was detected by real-time polymerase chain reaction (PCR) in dust, air filters, and drinkers while birds shed culturable C. jejuni cells. No differences (p > 0.050) in colonization or shedding levels that could be attributed to the diet were observed during the assay. CONCLUSIONS: Beneficial effects on performance and intestinal health were observed, particularly during the starter period, when chickens were fed a diet supplemented with both whey and coated calcium butyrate. However, none of the tested diets provided the chicks any differential degree of protection against Campylobacter infection.

The In Vitro and In Vivo Effect of Carvacrol in Preventing Campylobacter Infection, Colonization and in Improving Productivity of Chicken Broilers.

The current trend in reducing the antibiotic usage in animal production imposes urgency in the identification of novel biocides. The essential oil carvacrol, for example, changes the morphology of the cell and acts against a variety of targets within the bacterial membranes and cytoplasm, and our in vitro results show that it reduces adhesion and invasion of chicken intestinal primary cells and also biofilm formation. A trial was conducted to evaluate the effects of dietary supplementation of carvacrol at four concentrations (0, 120, 200, and 300 mg/kg of diet) on the performance of Lactobacillus spp., Escherichia coli, Campylobacter spp., and broilers. Each of the four diets was fed to three replicates/trial of 50 chicks each from day 0 to 35. Our results show that carvacrol linearly decreased feed intake, feed conversion rates and increased body weight at all levels of supplementation. Plate count analysis showed that Campylobacter spp. was only detected at 35 days in the treatment groups compared with the control group where the colonization occurred at 21 days. The absence of Campylobacter spp. at 21 days in the treatment groups was associated with a significant increase in the relative abundance of Lactobacillus spp. Also, carvacrol was demonstrated to have a significant effect on E. coli numbers in the cecum of the treatment groups, at all supplementation levels. In conclusion, this study shows for the first time that at different concentrations, carvacrol can delay Campylobacter spp., colonization of chicken broilers, by inducing changes in gut microflora, and it demonstrates promise as an alternative to the use of antibiotics.

Biochar, Bentonite and Zeolite Supplemented Feeding of Layer Chickens Alters Intestinal Microbiota and Reduces Campylobacter Load.

A range of feed supplements, including antibiotics, have been commonly used in poultry production to improve health and productivity. Alternative methods are needed to suppress pathogen loads and maintain productivity. As an alternative to antibiotics use, we investigated the ability of biochar, bentonite and zeolite as separate 4% feed additives, to selectively remove pathogens without reducing microbial richness and diversity in the gut. Neither biochar, bentonite nor zeolite made any significant alterations to the overall richness and diversity of intestinal bacterial community. However, reduction of some bacterial species, including some potential pathogens was detected. The microbiota of bentonite fed animals were lacking all members of the order Campylobacterales. Specifically, the following operational taxonomic units (OTUs) were absent: an OTU 100% identical to Campylobacter jejuni; an OTU 99% identical to Helicobacter pullorum; multiple Gallibacterium anatis (>97%) related OTUs; Bacteroides dorei (99%) and Clostridium aldenense (95%) related OTUs. Biochar and zeolite treatments had similar but milder effects compared to bentonite. Zeolite amended feed was also associated with significant reduction in the phylum Proteobacteria. All three additives showed potential for the control of major poultry zoonotic pathogens.

The influence of feeding crimped kernel maize silage on growth performance and intestinal colonization with Campylobacter jejuni of broilers.

An infection trial and a production trial over 35 days were conducted in parallel to study the influence of feeding crimped kernel maize silage (CKMS) on the intestinal Campylobacter jejuni colonization and broiler performance, respectively. The CKMS was used at dietary inclusion levels of 15% and 30% in maize-based diets. Broilers were orally inoculated with 2 × 10(5) log cfu/ml C. jejuni on day 14. Four birds from each pen were randomly selected and killed by cervical dislocation on days 3, 6, 9, 14 and 21 post infection and intestinal contents from ileum, caeca and rectum as well as liver samples were taken. Body weight and feed consumption of broilers were registered on days 13, 22 and 35. On day 35, litter dry matter (DM) was measured and the condition of the foot pads was evaluated. There was no significant effect of CKMS on the colonization of C. jejuni. Body weight of the broilers supplemented with 15% CKMS was comparable with the control maize-based feed, whereas addition of 30% CKMS reduced broiler body weight (P < 0.001). However, DM intake and feed conversion ratio were the same in all three dietary treatments. Furthermore, the foot pad condition of broilers significantly improved with the inclusion of CKMS on broiler diets as a result of a higher DM content in the litter material. It is concluded that CKMS did not influence intestinal Campylobacter colonization, but improved the foot pad health of broilers.

Efficacy of feed additives against Campylobacter in live broilers during the entire rearing period.

Poultry meat is the major source of human campylobacteriosis, the most frequently reported zoonosis in the EU. The prevalence of Campylobacter colonization in European broiler flocks is 71%. Despite considerable efforts, there is still no effective strategy available to prevent or reduce Campylobacter colonization in broilers. This study tested a wide variety of feed additives to reduce Campylobacter shedding in primary poultry production. Twelve additives containing organic or fatty acids, monoglycerides, plant extracts, prebiotics, or probiotics were tested. For each additive, broilers contaminated with Campylobacter jejuni were fed with an additive free diet (control group) or with a supplemented diet (treated group) and Campylobacter loads compared at three sampling times. No treatment was able to prevent broiler colonization by Campylobacter, and there was a high degree of variation in contamination among the birds. At 14 d of age, eight treatments significantly decreased the colonization level compared to the control group by a maximum of 2 log10 CFU/g. At 35 d of age, three of these treatments still had a significant effect with a maximum reduction of 1.88 log10 CFU/g for a probiotic. At 42 d of age, only one short-chain fatty acid was still significantly efficient with a mean reduction over 2 log10 CFU/g. In addition, a probiotic and a prebiotic-like compound significantly decreased the contamination by a maximum of 3 log10 CFU/g, only at the 42-d sampling period. This study gives promising results regarding the use of feed additives to reduce Campylobacter infection in flocks. Nevertheless, a global approach, combining intervention measures at the different steps of the broiler meat production chain could have a greater impact on the reduction of public health risk.

Effects of feeding plant-derived agents on the colonization of Campylobacter jejuni in broiler chickens.

The aim of this work was to test the potential use of plant-derived extracts and compounds to control Campylobacter jejuni in broiler chickens. Over a 7-wk feeding period, birds were fed a commercial diet with or without plant extracts (Acacia decurrens, Eremophila glabra), essential oil [lemon myrtle oil (LMO)], plant secondary compounds [terpinene-4-ol and α-tops (including α-terpineol, cineole, and terpinene-4-ol)], and the antibiotic virginiamycin. Traditional culture and real-time quantitative PCR techniques were used to enumerate the numbers of C. jejuni in chicken fecal and cecal samples. In addition, BW and feed intake were recorded weekly for the calculation of BW gain and feed conversion ratio. The mean log10 counts of C. jejuni were similar (P > 0.05) across treatments. However, significantly lower levels of fecal Campylobacter counts (P < 0.05) were recorded at d 41 for the α-tops treatment by culture methods. No differences (P > 0.05) in BW gain were obtained for dietary supplementation, except for the E. glabra extract, which had a negative impact (P < 0.001) on BW, resulting in sporadic death. Results from this study suggest that supplemental natural compounds used in the current study did not reduce the shedding of C. jejuni to desired levels.

Use of phages to control Campylobacter spp.

The use of phages to control pathogenic bacteria has been investigated since they were first discovered in the beginning of the 1900s. Over the last century we have slowly gained an in-depth understanding of phage biology including which phage properties are desirable when considering phage as biocontrol agents and which phage characteristics to potentially avoid. Campylobacter infections are amongst the most frequently encountered foodborne bacterial infections around the world. Handling and consumption of raw or undercooked poultry products have been determined to be the main route of transmission. The ability to use phages to target these bacteria has been studied for more than a decade and although we have made progress towards deciphering how best to use phages to control Campylobacter associated with poultry production, there is still much work to be done. This review outlines methods to improve the isolation of these elusive phages, as well as methods to identify desirable characteristics needed for a successful outcome. It also highlights the body of research undertaken so far and what criteria to consider when doing in-vivo studies, especially because some in-vitro studies have not been found to translate into to phage efficacy in-vivo.

Is allicin able to reduce Campylobacter jejuni colonization in broilers when added to drinking water?

Reducing Campylobacter shedding on the farm could result in a reduction of the number of human campylobacteriosis cases. In this study, we first investigated if allicin, allyl disulfide, and garlic oil extract were able to either prevent C. jejuni growth or kill C. jejuni in vitro. Allyl disulfide and garlic oil extract reduced C. jejuni numbers in vitro below a detectable level at a concentration of 50 mg/kg (no lower concentrations were tested), whereas allicin reduced C. jejuni numbers below a detectable level at a concentration as low as 7.5 mg/kg. In further experiments we screened for the anti-C. jejuni activity of allicin in a fermentation system closely mimicking the broiler cecal environment using cecal microbiota and mucus isolated from C. jejuni-free broilers. During these fermentation experiments, allicin reduced C. jejuni numbers below a detectable level after 24 h at a concentration of 50 mg/kg. In contrast, 25 mg/kg of allicin killed C. jejuni in the first 28 h of incubation, but anti-C. jejuni activity was lost after 48 h of incubation, probably due to the presence of mucin in the growth medium. This had been confirmed in fermentation experiments in the presence of broiler cecal mucus. Based on these results, we performed an in vivo experiment to assess the prevention or reduction of cecal C. jejuni colonization in broiler chickens when allicin was added to drinking water. We demonstrated that allicin in drinking water did not have a statistically significant effect on cecal C. jejuni colonization in broilers. It was assumed, based on in vitro experiments, that the activity of allicin was thwarted by the presence of mucin-containing mucus. Despite promising in vitro results, allicin was not capable of statistically influencing C. jejuni colonization in a broiler flock, although a trend toward lower cecal C. jejuni numbers in allicin-treated broilers was observed.

Campylobacter bacteriophages and bacteriophage therapy.

Members of the genus Campylobacter are frequently responsible for human enteric disease with occasionally very serious outcomes. Much of this disease burden is thought to arise from consumption of contaminated poultry products. More than 80% of poultry in the UK harbour Campylobacter as a part of their intestinal flora. To address this unacceptably high prevalence, various interventions have been suggested and evaluated. Among these is the novel approach of using Campylobacter-specific bacteriophages, which are natural predators of the pathogen. To optimize their use as therapeutic agents, it is important to have a comprehensive understanding of the bacteriophages that infect Campylobacter, and how they can affect their host bacteria. This review will focus on many aspects of Campylobacter-specific bacteriophages including: their first isolation in the 1960s, their use in bacteriophage typing schemes, their isolation from the different biological sources and genomic characterization. As well as their use as therapeutic agents to reduce Campylobacter in poultry their future potential, including their use in bio-sanitization of food, will be explored. The evolutionary consequences of naturally occurring bacteriophage infection that have come to light through investigations of bacteriophages in the poultry ecosystem will also be discussed.

Effect of coated and non-coated fatty acid supplementation on broiler chickens experimentally infected with Campylobacter jejuni.

The aim of this study was to examine whether and to what extent the supplementation of feed with a coated or non-coated mixture of fatty acids (caprylic and capric acid) affects broiler chickens experimentally infected with Campylobacter jejuni. The study was carried out using 48 chickens divided into four experimental groups. Throughout the whole rearing period (1-42 days), the chickens were fed a diet supplemented with 0.25% caprylic and capric acid (1:1), coated or non-coated. At the age of 14 and 28 days, chickens were orally challenged with C. jejuni. At regular time intervals post-inoculation, the shedding of C. jejuni was assayed using quantitative real-time PCR. Both supplements significantly decreased faecal C. jejuni counts by 1.2-4.1 log(10) CFU/g 4 days post-inoculation; after this time period, the effect of medium-chain fatty acids (MCFA) was less pronounced or absent. Campylobacter jejuni counts in excreta samples were significantly lower in chickens fed coated MCFA than in those fed non-coated MCFA. No effect of MCFA on feed intake or growth of chickens was observed. In conclusion, (i) MCFA are active against C. jejuni and (ii) the encapsulation enhanced the efficacy of the acids. These results allow the recommendation of using MCFA as feed additives in chickens, preferably 2-3 days before slaughter.

Extracts of edible and medicinal plants in inhibition of growth, adherence, and cytotoxin production of Campylobacter jejuni and Campylobacter coli.

UNLABELLED: Campylobacter spp. is recognized as one of the most common cause of food-borne bacterial gastroenteritis in humans. Campylobacter infection causes campylobacteriosis, which can range from asymptomatic to dysentery-type illnesses with severe complications, such as Guillian-Barre syndrome. Epidemiological studies have revealed that consumption of poultry products is an important risk factor of this disease. Adherence and cytotoxic activity of the bacteria to host mucosal surfaces have been proposed to be critical steps in pathogenesis. Innovative tools for controlling Campylobacter, such as natural products from plants, represent good alternatives for use in foods or as therapeutic agents. In this study, 28 edible or medicinal plants species were analyzed for their bactericidal effects on the growth of Campylobacter jejuni and C. coli. The extracts of Acacia farnesiana, Artemisia ludoviciana, Opuntia ficus-indica, and Cynara scolymus were the most effective against these microorganisms at minimal bactericidal concentrations (MBCs) of 0.3, 0.5, 0.4, and 2.0 mg/mL, respectively. No effect on growth was detected with lower concentrations of extract (25%, 50%, or 75% of the MBC) added to the media. The effect of each extract (75% of the MBC) on adherence and cytotoxicity of C. jejuni and C. coli was evaluated in Vero cells. Adherence of Campylobacter to Vero cells was significantly affected by all the extracts. Cytotoxic activity of bacterial cultures was inhibited by A. farnesiana and A. ludoviciana. These plant extracts are potential candidates to be studied for controlling Campylobacter contamination in foods and the diseases associated with this microorganism. PRACTICAL APPLICATION: Innovative tools for controlling Campylobacter, such as natural products from plants, represent good alternatives for use in foods or as therapeutic agents. The extracts of Acacia farnesiana, Artemisia ludoviciana, Opuntia ficus-indica, and Cynara scolymus were the most effective against these microorganisms. Adherence and cytotoxic activity of the bacteria to host mucosal surfaces which are critical steps in pathogenesis were decreased by these extracts. Our results point to these plants as potential candidates for the control of Campylobacter contamination in foods, the treatment of the diseases associated with this microorganism, and as feed supplements to reduce on-farm prevalence of Campylobacter.