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Various species of campylobacters cause significant disease problems in both humans and animals. The continuing development of tools and methods for genetic and molecular manipulation of campylobacters enables the detailed study of bacterial virulence and disease pathogenesis. Campylobacter hepaticus is an emerging pathogen that causes spotty liver disease (SLD) in poultry. SLD has a significant economic and animal welfare impact as the disease results in elevated mortalities and significant decreases in egg production. Although potential virulence genes of C. hepaticus have been identified, they have not been further studied and characterized, as appropriate genetic tools and methods to transform and perform mutagenesis studies in C. hepaticus have not been available. In this study, the genetic manipulation of C. hepaticus is reported, with the development of novel plasmid vectors, methods for transformation, site-specific mutagenesis, and mutant complementation. These tools were used to delete the pglB gene , an oligosaccharyltransferase, a central enzyme of the N- glycosylation pathway, by allelic exchange. In the mutant strain, N -glycosylation was completely abolished. The tools and methods developed in this study represent innovative approaches that can be applied to further explore important virulence factors of C. hepaticus and other closely related Campylobacter species., Importance: Spotty liver disease (SLD) of layer chickens, caused by infection with Campylobacter hepaticus , is a significant economic and animal welfare burden on an important food production industry. Currently, SLD is controlled using antibiotics; however, alternative intervention methods are needed due to increased concerns associated with environmental contamination with antibiotics, and the development of antimicrobial resistance in many bacterial pathogens of humans and animals. This study has developed methods that have enabled the genetic manipulation of C. hepaticus . To validate the methods, the pglB gene was inactivated by allelic exchange to produce a C. hepaticus strain that could no longer N -glycosylate proteins. Subsequently, the mutation was complemented by reintroduction of the gene in trans , on a plasmid vector, to demonstrate that the phenotypic changes noted were caused by the mutation of the targeted gene. The tools developed enable ongoing studies to understand other virulence mechanisms of this important emerging pathogen., Competing Interests: D.M.A. is an employee of Bioproperties Pty Ltd, a veterinary vaccine business that manufactures and sells live vaccines for production animals. Therefore, he has a commercial interest in understanding the biology of veterinary pathogens such as Campylobacter hepaticus. All other authors declare no conflict of interest.

Background: Environmental Enteric Dysfunction (EED) is an acquired disorder of asymptomatic altered gut function, the etiology of which is unknown. EED is postulated to be a major contributor to growth faltering in early childhood in regions where early-life enteropathogenic carriage is prevalent. Few studies have examined the critical organ (the upper small bowel) with enteropathogens in the evolution of small bowel disease., Objectives: The objective of this study was to determine if fecal enteropathogenic detection predicts subsequent EED histology., Methods: Fecal samples were obtained from undernourished children aged <2 y without diarrhea enrolled in 3 cohort studies, who failed nutritional intervention and subsequently underwent endoscopy. Duodenal biopsies from 245 (Bangladesh n = 120, Pakistan n = 57, and Zambia n = 68) children were scored using a semiquantitative histologic grading protocol. Thirteen enteropathogens were sought in common across the 3 centers using TaqMan array cards (TAC) (Bangladesh and Pakistan) and the Luminex platform (Zambia). An additional 18 pathogens and 32 virulence loci were sought by TAC and included in sensitivity analyses restricted to TAC data., Results: Multivariable linear regressions adjusting for study center, age at stool collection, and stool-to-biopsy interval demonstrated the following: 1) an association of norovirus and Shigella detection with subsequent enterocyte injury [β 0.2 (95% CI: 0.1, 0.3); P = 0.002 and β 0.2 (95% CI: 0.0, 0.3); P = 0.008, respectively], 2) association of Campylobacter with intraepithelial lymphocytes [β 0.2 (95% CI: 0.0, 0.4); P = 0.046], and 3) association of Campylobacter and enterotoxigenic Escherichia coli with a summative EED histopathology index score [β 4.2 (95% CI: 0.8, 7.7); P = 0.017 and β 3.9 (95% CI: 0.5, 7.3); P = 0.027, respectively]. All but 2 of these associations (Shigella-enterocyte injury and Campylobacter-index score) were also demonstrated in TAC-only sensitivity analyses, which identified additional associations between other pathogens, pathogen burden, or virulence loci primarily with the same histologic parameters., Conclusions: The detection of some enteropathogens in asymptomatic infections is associated with subsequent EED histopathology. These novel findings offer a basis for future EED etiology and pathogenesis studies., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Introduction. Bacterial dysentery is one of the greatest causes of morbidity and mortality worldwide. Campylobacter spp. and diarrhoeagenic Escherichia coli (DEC) are recognised as the most common causes of bacterial enteritis in developing countries including India. Hypothesis/Gap statement. Rapid and accurate identification of dysentery causing organisms using molecular methods is essential for better disease management, epidemiology and outbreak investigations. Aim. In view of the limited information available on the dysentery causing agents like Campylobacter spp., enterohemorrhagic E. coli (EHEC)/enteropathogenic E. coli (EPEC) and enteroinvasive E. coli (EIEC)/ Shigella in India, this study was undertaken to investigate the presence of these pathogens in human and poultry stool samples by molecular methods. Methodology. In total, 400 human stool samples and 128 poultry samples were studied. Microaerophilic culture along with real-time multiplex PCR with the targets specific to the genus Campylobacter , Campylobacter jejuni , Campylobacter coli , EHEC, EPEC and EIEC/ Shigella was performed. Further species confirmation was done using MALDI-TOF MS. Results. On microaerophilic culture, C. coli was isolated in one human sample and two C. jejuni and one C. fetus in poultry samples. On PCR analysis, among human stool samples, typical EPEC (42%) was predominantly seen followed by Campylobacter spp. (19%) and EIEC/ Shigella (10%). In contrast, Campylobacter spp. (41%) was predominant in poultry samples, followed by typical EPEC (26%) and EIEC/ Shigella (9%). Poly-infections with Campylobacter spp. and DEC were also observed among both sources. Conclusion. The present study documented the increased prevalence of Campylobacter spp. in humans compared with the results of previous studies from India. Typical EPEC was found to be predominant in children less than 5 years of age in this study. The high prevalence of coinfections in the current study indicates that a multiple aetiology of diarrhoea is common in our settings.

Campylobacter concisus has been described as the etiological agent of periodontal disease, inflammatory bowel diseases, and enterocolitis. It is also detected in healthy individuals. There are differences between strains in healthy individuals and affected ones by production of two exototoxins. In this mini review authors discuss major facts about cultivation, isolation, virulence and immune response to C. concisus., Competing Interests: No Conflict of Interest is declared, (Copyright (c) 2021 Biljana Miljkovic-Selimovic, Tatjana Babic, Branislava Kocic, Ema Aleksic, Adam Malesevic, Zoran Tambur.)

Campylobacter is one of the most common causes of foodborne bacterial infections worldwide. Why poultry has been shown to be one of the most significant sources of these bacteria, ruminants, especially cattle, are also responsible for a high number of human Campylobacter jejuni , and to a lesser extent Campylobacter coli , infections. In this study, bovine and pig carcasses in Poland were investigated for the presence of Campylobacter and for their antimicrobial resistance. A total of 204 swabs from bovine carcasses and 355 swab samples from pig carcasses were tested during 2014-2018. Campylobacter was identified in 129 (36.3%) of the pig and in 11 (5.4%) of the bovine carcasses, respectively. The pig isolates were classified as C. coli (121; 34.1%) or C. jejuni (8; 2.3%), whereas the bovine Campylobacter were identified either as C. jejuni (8; 3.9% isolates) or C. coli (3; 1.5% strains). Resistance of the isolates ( n  = 140) to erythromycin, ciprofloxacin, nalidixic acid, streptomycin, and tetracycline revealed that the vast majority of C. coli was resistant to streptomycin (106 isolates; 85.5%), tetracycline (97; 78.2%), nalidixic acid (90; 72.6%), and ciprofloxacin (88; 71.0%). Among C. jejuni isolates ( n  = 16) the resistance rates to all antibiotics were lower than in C. coli , irrespective of the origin. A total of 74 of 121 (61.2%) C. coli isolates from the pig carcasses and one of three such isolates from the bovine samples were multiresistant. Most of the C. coli (64 isolates; 85.3%) had the ciprofloxacin+nalidixic acid+streptomycin+tetracycline resistance profile. The results suggest that pig and bovine carcasses may be an underestimated reservoir of Campylobacter , especially for C. coli in pigs. The high antimicrobial resistance rates of such strains to streptomycin, quinolones, and tetracyclines highlight the need for monitoring of these bacteria in such food and food products.

Development of a simple, rapid and specific assay for the simultaneous detection of Campylobacter spp. and Salmonella spp. based on duplex loop-mediated isothermal amplification (d-LAMP), combined with lateral-flow biosensor (LFB) is reported herein. LAMP amplicons of both pathogens were simultaneously amplified and specifically differentiated by LFB. The specificity of the d-LAMP-LFB was evaluated using a set of 68 target and 12 non-target strains, showing 100% inclusivity and exclusivity. The assay can simultaneously detect Campylobacter and Salmonella strains as low as 1 ng and 100 pg genomic DNA per reaction, respectively. The lowest inoculated detection limits for Campylobacter and Salmonella species in artificially contaminated chicken meat samples were 103 CFU and 1 CFU per 25 grams, respectively, after enrichment for 24 h. Furthermore, compared to culture-based methods using field chicken meat samples, the sensitivity, specificity and accuracy of d-LAMP- LFB were 95.6% (95% CI, 78.0%-99.8%), 71.4% (95% CI, 29.0%-96.3%) and 90.0% (95% CI, 73.4%-97.8%), respectively. The developed d-LAMP-LFB assay herein shows great potentials for the simultaneous detection of the Campylobacter and Salmonella spp. and poses a promising alternative approach for detection of both pathogens with applications in food products., Competing Interests: The authors have declared that no competing interests exist.

Campylobacter spp. are the most common bacterial pathogens associated with human gastroenteritis in industrialized countries. Contaminated chicken is the food vehicle associated with the majority of reported cases of campylobacteriosis, either by the consumption of undercooked meat or via cross- contamination of ready-to-eat (RTE) foods during the handling of contaminated raw chicken parts and carcasses. Our results indicate that cooking salt (used for seasoning) is a potential vehicle for Campylobacter spp. cross-contamination from raw chicken to lettuce, through unwashed hands after handling contaminated chicken. Cross-contamination events were observed even when the chicken skin was contaminated with low levels of Campylobacter spp. (ca. 1.48 Log CFU/g). The pathogen was recovered from seasoned lettuce samples when raw chicken was contaminated with levels ≥ 2.34 Log CFU/g. We also demonstrated that, once introduced into cooking salt, Campylobacter spp. are able to survive in a culturable state up to 4 hours. After six hours, although not detected following an enrichment period in culture medium, intact cells were observed by transmission electron microscopy. These findings reveal a "novel" indirect cross-contamination route of Campylobacter in domestic settings, and a putative contamination source to RTE foods that are seasoned with salt, that might occur if basic food hygiene practices are not adopted by consumers when preparing and cooking poultry dishes., Competing Interests: No authors have competing interests.

Spotty liver disease (SLD) is a bacterial disease of chicken, causing mortalities and reduction in egg production, hence, contributing to economic loss in the poultry industry. The causative agent of SLD has only recently been identified as a novel Campylobacter species, Campylobacter hepaticus. Specific primers were designed from the hsp60 gene of Campylobacter hepaticus and PCR followed by high-resolution melt curve analysis was optimised to detect and differentiate three species of Campylobacter (Campylobacter coli, Campylobacter jejuni and Campylobacter hepaticus). The three Campylobacter species produced a distinct curve profile and was differentiated using HRM curve analysis. The potential of the PCR-HRM curve analysis was shown in the genotyping of 37 Campylobacter isolates from clinical specimens from poultry farms. PCR-HRM curve analysis of DNA extracts from bile samples or cultures from bile samples, were identified as Campylobacter hepaticus and confirmed by DNA sequencing. The DNA sequence analysis of selected samples from each of the three HRM distinctive curves patterns showed that each DNA sequence was associated with a unique melt profile. The potential of the PCR-HRM curve analysis in genotyping of Campylobacter species was also evaluated using faecal specimens from 100 wild birds. The results presented in this study indicate that PCR followed by HRM curve analysis provides a rapid and robust technique for genotyping of Campylobacter species using either bacterial cultures or clinical specimens., Competing Interests: The authors have declared that no competing interests exist.

Diarrhoea remains a major cause of childhood morbidity and mortality worldwide. This study aimed to monitor the aetiology of acute diarrhoea in children in Shanghai. Paediatric outpatients with acute diarrhoea were enrolled in the study from Jan 2015 to Dec 2018. Faecal samples were collected for testing. Enteric bacteria were identified and typed by culture and serotyping, respectively. Enteric viruses were identified by real-time PCR. Enteric pathogens were identified in 1572 (58.4%) of the 2692 enrolled children with acute diarrhoea. Viruses were detected more frequently than bacteria (41.3% versus 25.0%). Nontyphoidal Salmonella spp. (NTS) was the most common (10.3%) bacteria isolated, followed by enteropathogenic Escherichia coli (EPEC) (6.5%), enteroaggregative Escherichia coli (EAEC) (6.2%), Campylobacter spp. (3.6%), enterotoxigenic Escherichia coli (ETEC) (1.1%), Shigella spp. (0.2%), and enterohemorrhagic Escherichia coli (EHEC) (0.1%). Rotavirus was the most common (16.0%) virus detected, followed by norovirus (15.5%), adenovirus (7.2%), sapovirus (3.0%) and astrovirus (2.7%). Rotavirus, norovirus and NTS were the major pathogens responsible for diarrhoea in Shanghainese children. Improving uptake of the rotavirus vaccine and strengthening foodborne-pathogen prevention will aid in reducing the burden of diarrhoeal disease in children in Shanghai., Competing Interests: The authors have declared that no ompeting interests exist.

Campylobacteriosis is an important contributor to the global burden of acute gastroenteritis (AGE). In Nicaragua, the burden, risk factors, and species diversity for infant campylobacteriosis are unknown. Between June 2017 and December 2018, we enrolled 444 infants from León, Nicaragua, in a population-based birth cohort, conducting weekly household AGE surveillance. First, we described clinical characteristics of symptomatic Campylobacter infections, and then compared clinical characteristics between Campylobacter jejuni/coli and non-jejuni/coli infections. Next, we conducted a nested case-control analysis to examine campylobacteriosis risk factors. Finally, we estimated the population attributable fraction of campylobacteriosis among infants experiencing AGE. Of 296 AGE episodes in the first year of life, Campylobacter was detected in 59 (20%), 39 were C. jejuni/coli, and 20 were non-jejuni/coli species, including the first report of Campylobacter vulpis infection in humans. Acute gastroenteritis symptoms associated with C. jejuni/coli lasted longer than those attributed to other Campylobacter species. In a conditional logistic regression model, chickens in the home (odds ratio [OR]: 3.8, 95% CI: 1.4-9.8), a prior AGE episode (OR: 3.3; 95% CI: 1.4-7.8), and poverty (OR: 0.4; 95% CI: 0.2-0.9) were independently associated with campylobacteriosis. Comparing 90 infants experiencing AGE with 90 healthy controls, 22.4% (95% CI: 11.2-32.1) of AGE episodes in the first year of life could be attributed to Campylobacter infection. Campylobacter infections contribute substantially to infant AGE in León, Nicaragua, with non-jejuni/coli species frequently detected. Reducing contact with poultry in the home and interventions to prevent all-cause AGE may reduce campylobacteriosis in this setting.

Campylobacter spp. represents the most common cause of gastroenteritis worldwide with the potential to cause serious sequelae. The ability of Campylobacter to survive stressful environmental conditions has been directly linked with food-borne illness. Toxin-antitoxin (TA) modules play an important role as defense systems against antimicrobial agents and are considered an invaluable strategy harnessed by bacterial pathogens to survive in stressful environments. Although TA modules have been extensively studied in model organisms such as Escherichia coli K12 , the TA landscape in Campylobacter remains largely unexplored. Therefore, in this study, a comprehensive in silico screen of 111 Campylobacter (90 C. jejuni and 21 C. coli ) isolates recovered from different food and clinical sources was performed. We identified 10 type II TA systems belonging to four TA families predicted in Campylobacter genomes. Furthermore, there was a significant association between the clonal population structure and distribution of TA modules; more specifically, most (12/13) of the Campylobacter isolates belonging to ST-21 isolates possess HicB-HicA TA modules. Finally, we observed a high degree of shared synteny among isolates bearing certain TA systems or even coexisting pairs of TA systems. Collectively, these findings provide useful insights about the distribution of TA modules in a heterogeneous pool of Campylobacter isolates from different sources, thus developing a better understanding regarding the mechanisms by which these pathogens survive stressful environmental conditions, which will further aid in the future designing of more targeted antimicrobials.

Background: Despite the significant reductions in under-five mortality, campylobacteriosis has emerged as one of the most common causative agents of bacterial foodborne gastroenteritis in humans. We performed this systematic review and meta-analysis to estimate the pooled prevalence of Campylobacter species and associated risk factors among children less than 5 years of age in Ethiopia., Methods: A systematic search was conducted on PubMed, Web of Science, EMBASE, Google Scholar and the Cochrane Library. All identified observational studies reporting the prevalence and determinants of diarrhea among children under 5 years of age in Ethiopia were included. Two authors independently extracted data and analyzed them using STATA Version 13 statistical software. A random-effects model was computed to estimate the pooled prevalence and the associations between determinant factors and campylobacteriosis., Results: Out of 166 papers reviewed, 8 studies fulfilled the inclusion criteria and were included in the meta-analysis. The pooled prevalence of Campylobacter species among children under 5 years of age in Ethiopia was 10% (95% CI: 7, 13). Contact with domestic animals (OR: 3.2, 95% CI: 2.0, 5.1), illiterate mothers (OR: 2.1, 95% CI: 1.1, 3.8), consumption of animal products (OR: 1.7, 95% CI: 0.7, 4.5), and status of mothers' personal hygiene (OR: 1.1, 95% CI: 0.7, 1.8) were significantly associated with the prevalence of Campylobacter species., Conclusion: In our study, Campylobacter species among children under 5 years of age in Ethiopia were significantly high. Contact with domestic animals, illiterate mothers and consumption of animal products were significantly associated with prevalence of Campylobacter species.

The aim of this study was to determine the pathogenic markers associated with Campylobacter infection in humans. A total of 104 Campylobacter isolates obtained from poultry and humans were examined for the presence of nine virulence genes and their ability to adhere to, invade and produce cytotoxin were defined using HeLa cells. The diversity of the Campylobacter spp. isolates was studied based on sequencing of the SVR-region of flaA gene. Altogether 45 flaA-SVR alleles were identified among 104 Campylobacter isolates of poultry and human origin. All Campylobacter isolates possessed flaA, cadF and racR genes involved in adherence. Accordingly, all poultry and human isolates exhibited adherence towards HeLa cells at mean levels of 0.95% and 0.82% of starting viable inoculum, respectively. The genes involved in invasion (iam and pldA) and cytotoxin production (cdtA, cdtB and cdtC) were also widely distributed among the human and poultry Campylobacter isolates. Significantly higher invasiveness was observed for poultry isolates (mean level of 0.002% of starting bacterial inoculum) compared to human isolates (0.0005%). Interestingly the iam gene, associated with invasion, was more common in human (100%) than poultry (84%) isolates, and the poultry isolates lacking the iam gene showed a marked reduction in their ability to invade HeLa cells. Moreover, virB11 was present in 22% of the poultry and 70.4% of the human isolates. Strains lacking virB11 showed a slight reduction in invasion, however in the absence of iam even the poultry isolates containing virB11 were unable to invade HeLa cells. The mean cytotoxicity of Campylobacter isolates from poultry and human was 26.7% and 38.7%, respectively. Strains missing both the cdtB and cdtC genes were non-cytotoxic compared to strains containing all three cdtABC genes, which were the most cytotoxic among the C. jejuni and C. coli isolates from both sources. No cytotoxic effect was observed in only 4% of poultry and 5.6% of human isolates., 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. Declaration of competing interest The authors have no conflict of interest to declare., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)

Campylobacter jejuni is a prevalent foodborne pathogen mainly transmitting through poultry. It remains unknown how chicken-transmitted C. jejuni and microbiota impact on human campylobacteriosis. Campylobacter jejuni AR101 (Cj-P0) was introduced to chickens and isolated as passage 1 (Cj-P1). Campylobacter jejuni Cj-P1-DCA-Anaero was isolated from Cj-P0-infected birds transplanted with DCA-modulated anaerobic microbiota. Specific pathogen free Il10 -/- mice were gavaged with antibiotic clindamycin and then infected with Cj-P0, Cj-P1, or Cj-P1-DCA-Anaero, respectively. After 8 days post infection, Il10 -/- mice infected with Cj-P1 demonstrated severe morbidity and bloody diarrhea and the experiment had to be terminated. Cj-P1 induced more severe histopathology compared to Cj-P0, suggesting that chicken transmission increased C. jejuni virulence. Importantly, mice infected with Cj-P1-DCA-Anaero showed attenuation of intestinal inflammation compared to Cj-P1. At the cellular level, Cj-P1 induced more C. jejuni invasion and neutrophil infiltration into the Il10 -/- mouse colon tissue compared to Cj-P0, which was attenuated with Cj-P1-DCA-Anaero. At the molecular level, Cj-P1 induced elevated inflammatory mediator mRNA accumulation of Il17a, Il1β, and Cxcl1 in the colon compared to Cj-P0, while Cj-P1-DCA-Anaero showed reduction of the inflammatory gene expression. In conclusion, our data suggest that DCA-modulated anaerobes attenuate chicken-transmitted campylobacteriosis in mice and it is important to control the elevation of C. jejuni virulence during chicken transmission process.

Background: Campylobacter infections in HIV positive patients often present with substantial mortality and morbidity when compared to HIV negative patients., Aim: This study assessed the prevalence of Campylobacter, antibiotic resistance phenotypes and genetic factors, and risk of Campylobacter infection associated with living in close proximity to domestic animals in HIV patients with gastric enteritis at Korle-Bu Teaching Hospital, Accra, Ghana., Methods: Resistance to different antibiotics was assessed with Kirby-Bauer disk diffusion method. In addition, all the Campylobacter isolates were tested for ampicillin (blaOXA-61), erythromycin (aph-3-1), tetracycline tet(O), streptomycin (aadE), and the energy-dependent multi-drug efflux pump (cmeB) resistance genes using multiplex polymerase chain reaction., Results: Out of a total of 140 (97 females and 43 males) tested patients, 71 (50.7%) patients were positive for Campylobacter coli. Female patients aged within 31-40 years (31.6%) and 41-50 years (31.6%) had high frequency of Campylobacter infection. Most of the infected patients lived in close proximity to chickens (53.5%), however, some patients (14.1%) lived in close proximity to goats. Phenotypic resistance evaluation revealed widespread resistance to ampicillin (100%), tetracycline (100%), ciprofloxacin (71.8%), erythromycin (69%), and gentamicin (49.3%). However, limited no of isolates contained blaOXA-61 (1.41%), cmeB (7.0%) and tet(O (7.0%) resistance genes., Conclusion: HIV patients with gastric enteritis were infected with resistant Campylobacter coli. Further studies are required to examine correlation of infected patients with C. coli and risk of living in close proximity to poultry birds. There is the need for routine investigation of Campylobacter in patients with gastroenteritis in order to assist in the development of strategies for combating diseases involving resistant zoonotic bacteria strains., Competing Interests: The authors have declared that no competing interests exist.

Campylobacteriosis is the leading bacterial cause of human diarrheal illness worldwide. Campylobacteriosis incidence exhibits seasonality and has been attributed to ambient temperature. However, the role of ambient temperature on campylobacteriosis remains poorly understood. To examine the impact of ambient temperature on local campylobacteriosis in Germany, weekly incidences on NUTS-3 level were analysed using a novel small-scaled approach, regression and time lags. Campylobacteriosis incidence correlated positively with temperatures between - 5 and 28 °C. The sigmoid regression model estimated an incidence increase of 0.52 per 5 °C temperature rise in the observation period. The weekly average of daily minimum temperature was most significant at a time lag of two weeks and showed the steepest incidence increase of 0.13 per 1 °C temperature increase in a temperature corridor of 5.1 to 12.2 °C. The impact of average minimum temperatures on campylobacteriosis incidence is crucial, likely to be indirect and especially relevant in the recent part of the infection chain. Vectors or human behaviour are presumably more directly linked with temperature than the pathogen's microbiology and should be examined. These variables outweigh the direct temperature-pathogen relationship when the whole chain of infection is considered. In the context of climate change, campylobacteriosis is likely to increase in Germany due to an increased temperature effect.

Background: Over one-third of the population of Havelock North, New Zealand, approximately 5500 people, were estimated to have been affected by campylobacteriosis in a large waterborne outbreak. Cases reported through the notifiable disease surveillance system (notified case reports) are inevitably delayed by several days, resulting in slowed outbreak recognition and delayed control measures. Early outbreak detection and magnitude prediction are critical to outbreak control. It is therefore important to consider alternative surveillance data sources and evaluate their potential for recognizing outbreaks at the earliest possible time., Objective: The first objective of this study is to compare and validate the selection of alternative data sources (general practice consultations, consumer helpline, Google Trends, Twitter microblogs, and school absenteeism) for their temporal predictive strength for Campylobacter cases during the Havelock North outbreak. The second objective is to examine spatiotemporal clustering of data from alternative sources to assess the size and geographic extent of the outbreak and to support efforts to attribute its source., Methods: We combined measures derived from alternative data sources during the 2016 Havelock North campylobacteriosis outbreak with notified case report counts to predict suspected daily Campylobacter case counts up to 5 days before cases reported in the disease surveillance system. Spatiotemporal clustering of the data was analyzed using Local Moran's I statistics to investigate the extent of the outbreak in both space and time within the affected area., Results: Models that combined consumer helpline data with autoregressive notified case counts had the best out-of-sample predictive accuracy for 1 and 2 days ahead of notified case reports. Models using Google Trends and Twitter typically performed the best 3 and 4 days before case notifications. Spatiotemporal clusters showed spikes in school absenteeism and consumer helpline inquiries that preceded the notified cases in the city primarily affected by the outbreak., Conclusions: Alternative data sources can provide earlier indications of a large gastroenteritis outbreak compared with conventional case notifications. Spatiotemporal analysis can assist in refining the geographical focus of an outbreak and can potentially support public health source attribution efforts. Further work is required to assess the location of such surveillance data sources and methods in routine public health practice., (©Mehnaz Adnan, Xiaoying Gao, Xiaohan Bai, Elizabeth Newbern, Jill Sherwood, Nicholas Jones, Michael Baker, Tim Wood, Wei Gao. Originally published in JMIR Public Health and Surveillance (http://publichealth.jmir.org), 17.09.2020.)

Contaminated poultry meat is considered to be the main source of human infection with Campylobacter spp., a pathogen that asymptomatically colonizes broiler chickens during fattening and contaminates carcasses during slaughter. To prevent or reduce the colonization of broiler flocks with Campylobacter spp., applying different organic acids, especially in combinations, via feed or drinking water seems to be a promising approach. However, only very few combinations of organic acids have been tested for their antibacterial efficacy against Campylobacter spp. Therefore, the in vitro susceptibility of 30 Campylobacter spp. isolates (20 C. jejuni and ten C. coli) to ten organic acids and ten combinations was determined. The testing of minimum inhibitory concentration (MIC) values was performed at pH 6.0 and 7.3 by using the broth microdilution method and included the following organic acids: Caprylic acid, sorbic acid, caproic acid, benzoic acid, ascorbic acid, propionic acid, acetic acid, formic acid, fumaric acid and tartaric acid and combinations thereof. The lowest MIC values were seen for caprylic acid (MIC range at pH 7.3: 0.5-2 mmol/L) and sorbic acid (MIC range at pH 7.3: 1-4 mmol/L). One to two dilution steps lower MIC values were determined at the lower pH value of 6.0. Furthermore, ten combinations consisting of three to five organic acids were developed. In addition to the tested antibacterial activity, other criteria were included such as approval as feed additives, reported synergistic effects and chemical properties. For nine of ten combinations, the MIC90 values of the organic acids decreased 1.25- to 241.5-fold compared to the MIC90 values for the individual substances. Furthermore, nine of ten combinations exhibited synergistic activities against two or more of the tested C. jejuni and C. coli isolates. A combination of caprylic acid, sorbic acid and caproic acid exhibited synergistic activities against the largest number of Campylobacter spp. isolates (six C. jejuni and four C. coli) with fractional inhibitory concentration (FIC) indices (∑FIC) ranging from 0.33 to 1.42. This study shows in vitro synergistic activities of different organic acids in combinations against the major Campylobacter species and could therefore be a promising basis for reducing Campylobacter spp. in vivo., Competing Interests: The authors have declared that no competing interests exist.

Infectious diarrhea affects over four billion individuals annually and causes over a million deaths each year. Though not typically prescribed for treatment of uncomplicated diarrheal disease, antimicrobials serve as a critical part of the armamentarium used to treat severe or persistent cases. Due to widespread over- and misuse of antimicrobials, there has been an alarming increase in global resistance, for which a standardized methodology for geographic surveillance would be highly beneficial. To demonstrate that a standardized methodology could be used to provide molecular surveillance of antimicrobial resistance (AMR) genes, we initiated a pilot study to test 130 diarrheal pathogens ( Campylobacter spp., Escherichia coli , Salmonella , and Shigella spp.) from the USA, Peru, Egypt, Cambodia, and Kenya for the presence/absence of over 200 AMR determinants. We detected a total of 55 different determinants conferring resistance to ten different categories of antimicrobials: genes detected in ≥ 25 samples included bla TEM , tet (A), tet (B), mac (A), mac (B), aadA1/A2 , strA , strB , sul1 , sul2 , qacE Δ1, cmr , and dfrA1 . The number of determinants per strain ranged from none (several Campylobacter spp. strains) to sixteen, with isolates from Egypt harboring a wider variety and greater number of genes per isolate than other sites. Two samples harbored carbapenemase genes, bla OXA-48 or bla NDM . Genes conferring resistance to azithromycin ( ere (A), mph (A)/ mph (K), erm (B)), a first-line therapeutic for severe diarrhea, were detected in over 10% of all Enterobacteriaceae tested: these included >25% of the Enterobacteriaceae from Egypt and Kenya. Forty-six percent of the Egyptian Enterobacteriaceae harbored genes encoding CTX-M-1 or CTX-M-9 families of extended-spectrum β-lactamases. Overall, the data provide cross-comparable resistome information to establish regional trends in support of international surveillance activities and potentially guide geospatially informed medical care.

Campylobacter is the causal agent of campylobacteriosis in humans, a self-limiting gastroenteritis. Campylobacteriosis is a zoonosis, commonly transmitted from contaminated chicken meat by either direct consumption or cross contamination during food manipulation. Presence of plasmids encoding for resistance to antibiotics such as tetracycline is common among Campylobacter isolates. In this report, we studied the effect of the temperature in the conjugation frequency of several tet(O) carrying plasmids, providing tetracycline resistance to the recipient cells. The conjugation frequency from donor cells carrying three previously characterized plasmids (pCjA13, pCjA9 and pTet) and from two clinical isolates was determined. Two temperatures, 37 and 42 °C, mimicking the conditions encountered by C. jejuni in the human and broiler chicken gastrointestinal tracts, respectively, were assessed. Our results clearly indicate that the conjugation process is promoted at high temperature. Accordingly, the transcriptional expression of some putative conjugative apparatus genes is thermoregulated, being induced at 42 °C. The two plasmids present in the clinical isolates were sequenced and assembled. Both plasmids are highly related among them and to the pTet plasmid. The high identity of the genes putatively involved in the conjugation process among the plasmids is in agreement with the similar behavior regarding the temperature dependency of the conjugative process. This report suggest that conjugation of plasmids carrying antibiotic resistance genes occurs preferentially at temperatures that resemble the gastrointestinal tract of birds, the main reservoir of C. jejuni., Competing Interests: Declaration of Competing Interest None., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The annual number of outbreaks of Vibrio parahaemolyticus and Salmonella food poisoning and that of patients in Japan, from 2000 to 2018, decreased exponentially even though the size of the individual outbreaks (the number of patients per outbreak) tended to become larger. For food poisonings caused by Campylobacter, the annual number of outbreaks increased exponentially while outbreak size became smaller and the annual number of patients remained almost unchanged. For food poisoning caused by norovirus, both the number of outbreaks and that of patients remained high throughout. Over time, the geographical and seasonal distribution of food poisonings became narrower for Vibrio parahaemolyticus and Salmonella, while they became wider for Campylobacter and norovirus. Further analyses using the attack rate-patient number plots suggested that the number of the outbreaks was determined mainly by the levels of microbial contamination of foods before they were brought into the facilities for consumption.

The aim of this study was to investigate the occurrence and diversity of Campylobacter species in chelonians. From July 2016 to September 2017, a total of 452 individuals from a large variety of tortoises (n = 366) and turtles/terrapins (n = 86) kept in private collections and breeding centres, wildlife rescue centres, zoos, pet shops, and veterinary clinics from Northern Italy was sampled and subjected to microbiological examination. Campylobacter genus and species confirmation was performed by single and multiplex PCRs. Out of 452 samples, five (1.1%) tested positive: three for C. iguaniorum (two Testudo graeca and one Testudo hermanni), one for C. fetus subsp. testudinum (Stigmochelys pardalis) and one for C. geochelonis (Testudo hermanni). This study suggests that Campylobacter spp. are not common in chelonians, but a variety of species can be detected in these hosts, including those potentially pathogenic for humans. Further studies are needed to understand the epidemiology and the pathogenic potential for both animals and humans of reptile-associated Campylobacter spp., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The aim of this study was to investigate the frequency of Campylobacter species, to detect the antibiotic resistance profiles and the virulence genes and to determine the clonal proximity of the isolates in the samples of cutting board, slaughterhouse waste water, wall, knife and carcass from three different slaughterhouses in Kayseri region. For this purpose, a total of 150 samples, 10 of each from knife, wall, cutting board, carcass smear sample and slaughterhouse wastewater were collected from each of the three types of slaughterhouses in 2018 in Kayseri. For the isolation of the Campylobacter species, following preenrichment, the suspensions were inoculated onto modified charcoal cefoperazone desoxycholate (CCD) agar and were incubated at 37°C under microaerophilic condition for 48-72 hours. Suspicious colonies with gray-white color were recovered and subjected to phenotypical (Gram staining, oxidase, catalase test, and motion test) tests. Multiplex polymerase chain reaction (mPCR) was used for the molecular identification of the Campylobacter species. Antimicrobial susceptibilities of the isolates identified at the species level were detected by using the disk diffusion test and antibiotic gradient test. Virulence genes (iam, cadF, cdtA, flaA, ceuE, cdtC, cdtB and virB11) among the isolates were evaluated by PCR. The molecular typing of the isolates determined at species level was performed by Enterobacterial Repetitive Intergenic Consensus PCR (ERIC-PCR). In the study, 17 (11.3%) of the 150 samples taken from the slaughterhouse were found to be suspicious in terms of Campylobacter spp. and as a result of phenotypic identification tests, all of the isolates were verified as Campylobacter spp.. As a result of mPCR; eight of the isolates were identified as Campylobacter jejuni, eight as Campylobacter fetus and one as Campylobacter coli. The isolation of the Campylobacter species from different sources was found to be higher in slaughterhouse wastewater than those of others (p<0.001) and the difference in the proportional distribution of the Campylobacter species obtained from various sources was statistically significant (p<0.05). As a result of the disk diffusion test, while, all C.jejuni isolates were resistant to ciprofloxacin, 87.5%, 25%, 25% and 12.5% of C.jejuni isolates were resistant to enrofloxacin, neomycin, amoxicillin/clavulanic acid, and erythromycin, respectively. In addition, 25%, 25% and 12.5% of C.fetus isolates were resistant to amoxicillin/clavulanic acid, neomycin and gentamicin, respectively. C.coli isolate was not resistant to any of the antibiotics tested. Antibiotic gradient test results were found to be compatible with the disc diffusion test results. One of the virulence genes examined, virB11, was not detected in any of the isolates. Moreover, iam gene was not present in C.fetus and C.coli isolates, but only in one C.jejuni isolate. The flaA gene was detected in six C.jejuni isolates. C.coli isolate and seven C.jejuni and seven C.fetus isolates were positive in terms of the cdtC gene. The cdtA, cdtB, ceuE and cadF genes were found to be positive in all C.jejuni isolates. All isolates analyzed in the study demonstrated different ERIC-PCR profiles. In conclusion, it was shown that Campylobacter strains isolated from slaughterhouses were resistant to the most of the current antibiotics. Moreover, the presence of highly virulent Campylobacters in the slaughterhouse environment threatens public health due to the risk of contamination of the humans via carcasses and foods. Therefore, it is recommended that strict hygiene rules should be followed to reduce Campylobacter species contamination in slaughterhouses.

In the presented review of the book N.R. Efimochkina "Bacterial food pathogens of the Campylobacter genus" (Moscow, 2019) attention is drawn to the safety of food products and food raw materials, especially poultry products for consumers. In general, the problematic issues of campylobacteriosis and its causative agent are highlighted in a new way: the main reservoirs, sources and factors of pathogen transmission are indicated, theoretical issues are analyzed and generalized data of screening studies of food products and environmental objects in the poultry industry for bacteria of the genus Campylobacter are presented. Particular attention is paid to laboratory diagnostics of the pathogen for the presence of uncultivated forms of these pathogens, the existence of Campylobacter in biofilms, and the resistance of isolated cultures to antibiotics. Laboratory methods that improve the detection of these pathogens are proposed. The developed methodological documents that accelerate the processes of Campylobacter isolation and antibiotic resistance assessment are relevant, which is important for the efficiency of production control and sanitary inspections of poultry and meat processing enterprises at control critical points. The introduction of the methods described in the monograph for analyzing the content of Campylobacter in food products, on the surface of equipment and inventory will help to effectively manage the risks associated with contamination of food products by these pathogens. The monograph will be of great importance for improving the knowledge of a number of medical staff, including sanitary and hygienic profile, that is important for environmental protection, the prevention of food poisoning and intestinal infections with alimentary transmission factor., Competing Interests: The authors declare no overt and potential conflict of interest related to the publication of this article., (Copyright© GEOTAR-Media Publishing Group.)

Aquatic recreation in urban estuaries worldwide is often restricted by fecal pollution. Variability in the occurrence of fecal pathogens and their differential virulence potentials within these estuaries may result in variable public health risks. To address this hypothesis, Campylobacter were isolated from the Yarra River estuary, Australia and then characterized via HeLa cell cytotoxicity and attachment to and the invasion of Caco-2 monolayers. Overall, 54% (n=216) of estuarine samples (water and sediment combined) yielded biochemically confirmed culturable Campylobacter; higher detection was recorded in water (92%, n=90) than in the bank and bed sediments combined (27%, n=126). The seasonality of occurrence was not significant. HeLa cell cytotoxicity revealed that estuarine Campylobacter had low cytotoxin titers; the 95% confidence interval (CI) ranged between 61 and 85, which was markedly lower than the mean value (~386) for the C. jejuni 11168 reference pathogenic strain. The Caco-2 attachment of estuarine Campylobacter isolates (n=189) revealed that the 95%CI for the attachment efficiency of the test strains ranged between 0.09 and 0.1%, with only 3.7% having a higher efficiency than the 5 th percentile value for C. jejuni 11168. None of the estuarine strains exhibited Caco-2 invasion capabilities. In contrast to the common assumption during quantitative microbial/risk assessments (QMRAs) that all environmental strains are pathogenic, the present results revealed that Campylobacter within the Yarra River estuary had very low virulence potential. Since this is the first study to use human epithelial cell lines to characterize estuary-borne pathogens, these results generate valuable insights for a better understanding of the public health risks in urban estuaries that will underpin more robust QMRAs.

World Health Organization (WHO) defined zoonotic diseases as diseases transmitted from animals to humans and vice versa with or without vector, where more than 75% of the human's diseases have been transmitted from animals or their products. Nevertheless, campylobacteriosis is still one of the most important food borne zoonotic diseases that is likely to challenge global public health all over the world. In Egypt, campylobacteriosis causes severe losses in comparison with other food borne pathogens like Salmonella or Escherichia coli. The potential sources of Campylobacter transmission are poultry backyards and meat, cattle meat, raw milk and water. The main clinical manifestations of human's campylobacteriosis are gastroenteritis and later on nervous signs. Prevention and control strategic plans of campylobacteriosis are very crucial., Competing Interests: No Conflict of Interest is declared, (Copyright (c) 2019 Wafaa A Abd El-Ghany.)

Background: The early detection of enteric infections in older adults is challenging because typical signs and symptoms of disease may be less common, absent, or overlooked. Understanding illness characteristics of enteric infections among older adults could improve the timeliness and accuracy of clinical diagnoses, thereby improving patient outcomes and increasing cases reported to surveillance., Methods: Here, we describe illness characteristics (percentage reporting bloody diarrhea, fever, vomiting, abdominal pain; percentage hospitalized; duration of hospitalization; and duration of illness) among older adults (≥65 years) with acute gastroenteritis and culture-confirmed Campylobacter and nontyphoidal Salmonella infections in Australia, Canada, and the United States and compare these characteristics with those among younger people (<5 years, 5-24 years, and 25-64 years)., Results: A significant negative correlation was found between all symptoms and increasing age group, except for bloody diarrhea in cases of acute gastroenteritis. Adults aged ≥85 years reported bloody diarrhea in only 9% of nontyphoidal Salmonella and 4% of Campylobacter infections compared with 59% and 55% among children aged <5 years. Conversely, a greater percentage of older adults (≥65) than younger persons (<5, 5-24, 25-64) reported being hospitalized, with an increasing linear relationship in age groups 65 years and older., Conclusions: Although older adults are more likely to have severe illness and be hospitalized, we found that the proportion of persons reporting symptoms typically associated with enteric infections decreases with age. These findings have implications for clinical recognition and treatment of gastrointestinal illness, as well as for public health research., (© The Author(s) 2019. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.)

The Campylobacter and Arcobacter genera encompass closely related species that are ubiquitous in nature and are harboured in the gastrointestinal tract of many animals, including food-producing animals (cattle, sheep, pigs and poultry). In humans Campylobacter spp. is the cause of most of the gastroenteritis cases worldwide and in more severe cases the infection can result in Guillian Barré syndrome. Similarly, Arcobacter species can cause gastroenteritis as well as bacteraemia. Infections in humans can be induced by the consumption of contaminated vegetables, meat, milk and water. However, food originating from animals, especially meat, has been recognised as a source of infection, in fact, poultry meat and meat products have been globally reported as the main source of infection. It is clear that food-producing animals are important reservoirs for Campylobacter and Arcobacter species, which implies successful colonisation of the gastrointestinal tract at primary production and contamination during the slaughter process. During slaughter the evisceration step has been recognised as the most likely point of contamination, as accidental spillage of intestinal fluid and rapture of gastrointestinal tract can occur. Therefore, improper hygienic practices can ultimately allow for the contamination of finished/retail products intended for human consumption. This literature review will seek to explore the infection of food-producing animals with Campylobacter and Arcobacter species at primary production and contamination during the slaughter of food-producing animals.

The MORDOR I trial 1 , conducted in Niger, Malawi and Tanzania, demonstrated that mass azithromycin distribution to preschool children reduced childhood mortality 1 . However, the large but simple trial design precluded determination of the mechanisms involved. Here we examined the gut microbiome of preschool children from 30 Nigerien communities randomized to either biannual azithromycin or placebo. Gut microbiome γ-diversity was not significantly altered (P = 0.08), but the relative abundances of two Campylobacter species, along with another 33 gut bacteria, were significantly reduced in children treated with azithromycin at the 24-month follow-up. Metagenomic analysis revealed functional differences in gut bacteria between treatment groups. Resistome analysis showed an increase in macrolide resistance gene expression in gut microbiota in communities treated with azithromycin (P = 0.004). These results suggest that prolonged mass azithromycin distribution to reduce childhood mortality reduces certain gut bacteria, including known pathogens, while selecting for antibiotic resistance.

Background: In hospitalised patients with diarrhoea a positive campylobacter stool Polymerase Chain Reaction (PCR) test with negative culture results as well as Enteropathogenic Escherichia coli (EPEC) positive stool PCRs, challenges the clinician and may lead the unexperienced clinician astray. The aim of the study was to elucidate the clinical significance of positive Campylobacter and/or EPEC test results in hospitalised patients with diarrhoea., Methods: We conducted a retrospective case-case study. Case groups with 1) EPEC only and 2) EPEC in combination with any other pathogen in the PCR multiplex array, 3) PCR positive/culture negative Campylobacter, and 4) PCR positive/culture positive Campylobacter were compared. Medical records were reviewed and cases classified according to pre-specified clinical criteria as infectious gastroenteritis or non-infectious causes for diarrhoea. We analyzed the association between laboratory findings (the 4 subgroups) and the pre-specified clinical classification. We further sequenced culture negative campylobacter samples and tested EPEC for bundle forming pilus A (bfpA) gene, distinguishing typical from atypical EPEC., Results: A total of 291 patients were included, 169 were PCR positive for Campylobacter and 122 for EPEC. For both pathogens, co-infections were more common in culture negative/PCR positive samples than in culture positive samples. Clinical characteristics differed significantly in and between groups. Campylobacter culture positive patients had very high prevalence of characteristics of acute infectious gastroenteritis, whereas patients with PCR positive test results only often had an alternative explanation for their diarrhoea. Culture positives were almost exclusively C. jejuni/coli, whereas in culture negatives, constituting a third of the total PCR positives, C. concisus was the most frequent species. The vast majority of EPEC only positives had documented non-infectious factors that could explain diarrhoea. The EPEC co-infected group mimicked the culture positive campylobacter group, with most patients fulfilling the infectious gastroenteritis criteria., Conclusions: In hospitalised patients, positive PCR results for campylobacter and EPEC should be interpreted in a clinical context after evaluation of non-infectious diarrhoea associated conditions, and cannot be used as a stand-alone diagnostic tool.

Objectives: We characterized clinical isolates of Campylobacter using whole-genome sequencing (WGS) for detection of virulence genes, antimicrobial resistance markers and phylogenetic analysis in order to increase the knowledge on the molecular epidemiology of Campylobacter in Ireland, where there are significant gaps due to the widespread in the use of culture independent methods for the diagnosis of campylobacteriosis., Methods: WGS was applied to 122 Campylobacter human isolates collected over a 10-years period, from diarrhoeal stool samples submitted for routine enteric screening., Results: Genes associated with cytotoxin production such as cdtA, cdtB and cdtC were found in 88%, 89% and 89% isolates, respectively; adherence, colonization and invasion genes such as cadF, dnaJ, racR, iam, virB11 and ciaB were found in 99%, 99%, 98%, 99%, 1% and 80% isolates, respectively. Genetic markers associated with resistance to quinolones (C257T in gyrA), beta-lactams (blaoxa-61) and tetracycline (tet(O)) were present in 43%, 71% and 25% isolates, respectively. The cmeABC operon was present in 94% of isolates. No macrolide or aminoglycoside resistance markers were detected. Phylogenetic analysis showed that 112 isolates were assigned to 29 sequence types grouped into 17 clonal complexes. Four clusters previously unidentified were detected. These results shown the similarity of Irish data compared to what has been described globally., Conclusions: WGS has shown a high discriminatory power for cluster detection, demonstrating that its integration in routine laboratory surveillance could improve the detection and management of outbreaks. In addition we were able to demonstrate that virulence genes in clinical Campylobacter infections in Ireland were similar to those known previously. High prevalence of quinolone resistance markers has been found, which has implications for antimicrobial stewardship., Competing Interests: The authors have declared that no competing interests exist.

Campylobacter is among the four main causes of gastroenteritis worldwide and has increased in both developed and developing countries over the last 10 years. The vast majority of reported Campylobacter infections are caused by Campylobacter jejuni and, to a lesser extent, C. coli ; however, the increasing recognition of other emerging Campylobacter pathogens is urgently demanding a better understanding of how these underestimated species cause disease, transmit, and evolve. In parallel to the enhanced clinical awareness of campylobacteriosis due to improved diagnostic protocols, the application of high-throughput sequencing has increased the number of whole-genome sequences available to dozens of strains of many emerging campylobacters. This has allowed for comprehensive comparative pathogenomic analyses for several species, such as C. fetus and C. concisus These studies have started to reveal the evolutionary forces shaping their genomes and have brought to light many genomic features related to pathogenicity in these neglected species, promoting the development of new tools and approaches relevant for clinical microbiology. Despite the need for additional characterization of genomic diversity in emerging campylobacters, the increasing body of literature describing pathogenomic studies on these species deserves to be discussed from an integrative perspective. This review compiles the current knowledge and highlights future work toward deepening our understanding about genome dynamics and the mechanisms governing the evolution of pathogenicity in emerging Campylobacter species, which is urgently needed to develop strategies to prevent or control the spread of these pathogens., (Copyright © 2019 American Society for Microbiology.)

Bacteria of the genus Campylobacter spp. are one of the most common causes of gastroenteritis and can lead to serious sequelae. Several studies have estimated the disease burden of Campylobacter spp. with the quantitative metric of disability-adjusted life years (DALY). The aim of this systematic review is to give an overview of the information available about different countries and periods for which DALYs were calculated and how the different results are comparable. One of the most important transmission pathways for Campylobacter spp. is food. Therefore, special attention was given to studies that only estimated the foodborne disease burden of Campylobacter bacteria. With a systematic search for the period 1/1996-6/2016, one worldwide and 21 country-specific publications of the WHO were identified. Because of the different methods and the quality of the different data sets, the estimated results of all Campylobacter health outcomes of the country-specific studies vary from 0.4 DALYs per 100000 people in France to 109 DALY per population in Poland. The calculation of the attributable foodborne disease burden was based on the estimations of the incidences of all Campylobacter health outcomes with the associated uncertainty for each result. So the estimations of the foodborne disease burden show a large range from 0.5 DALYs per 100000 people in Greek to 21.2 DALYs per 100000 people in New Zealand. This span can only be partially explained by the country-specific variability in the food production, the consumption behavior and the incidence of Campylobacter bacteria., Competing Interests: Die Autoren geben an, dass kein Interessenkonflikt besteht., (© Georg Thieme Verlag KG Stuttgart · New York.)

Antibiotic-resistant C ampylobacter could adversely affect treatment outcomes, especially in children. We investigated the antibiotic susceptibility profiles, virulence potentials and genetic relatedness of Campylobacter spp. from paediatric and water samples in the North West Province, South Africa. Overall, 237 human and 20 water isolates were identified using culture and real-time polymerase chain reaction (PCR). The antibiotic susceptibility profiles were determined using the disk diffusion method. Gradient strips were used to determine the minimum inhibitory concentration of each antibiotic. Antibiotic resistance ( gryA , tetO and 23S rRNA 2075G and 2074C) and virulence ( cadF and ciaB ) genes were also investigated using PCR. A phylogenetic tree to ascertain the clonality between water and clinical isolates was constructed using MEGA 7. Overall, 95% (water) and 64.7% (human) of the isolates were resistant to at least one antibiotic tested. The highest resistance was against clarithromycin (95%) for water and ampicillin (60.7%) for human isolates. The 23S rRNA 2075G/2074C mutation was the most expressed resistance gene. Phylogenetic reconstruction revealed eight intermixed clades within water and human Campylobacter isolates. This study suggests the possible circulation of potentially pathogenic antibiotic-resistant Campylobacter in the Northwest Province, South Africa with drinking water being a possible vector for disease transmission in this area., Competing Interests: The authors declare no conflict of interest.

Objective: Acute gastroenteritis (AGE) is a risk factor for post-infectious irritable bowel syndrome (PI-IBS). This systematic review evaluates the prevalence and risk-factors of PI-IBS after AGE by specific pathogens. Materials and methods: Medline (1966-2019) and Embase (1974-2019) were searched for studies evaluating PI-IBS minimum 3 months after AGE with Campylobacter spp., Salmonella spp., Shigella spp., Escherischia coli , Clostridium difficile , norovirus, rotavirus, Cryptosporidium spp. or Giardia intestinalis using validated criteria for IBS. Pooled prevalence (PP), odds ratios (OR) and risk factors were determined for single pathogens, groups of bacteria, viruses and parasites, and overall for AGE caused by any pathogen. Random-effect models were used for meta-analyses. Results: A total of 34 articles were included. PP of PI-IBS after Campylobacter spp. was 12% (confidence interval 95% [CI]: 10-15%), Salmonellosis 12% (CI: 9-15%), Shigellosis 11% (CI: 8-15%), C. difficile 14% (CI: 4-29%) and E. coli spp. 12% (CI: 5-20%). OR of PI-IBS after salmonellosis was 5.5 (CI: 2.3-12.8) and after shigellosis 13.8 (CI: 4.2-45.4). Bacterial AGE overall showed OR 5.8 (CI: 4.0-8.3) and AGE caused by any pathogen OR 4.9 (CI: 3.9-6.1). Few studies exist on viral and parasitic gastroenteritis. Conclusions: Current literature show similar risks for bacterial pathogens. Studies are limited for viral and parasitic pathogens. The evaluated risk-factors for PI-IBS varied among the included studies and the existing evidence is insufficient to identify pathogen-specific risk factors.

Background: The clinical course of Campylobacter infection varies in symptoms and severity depending on host factors, virulence of the pathogen and initiated therapy. The type VI secretion system (T6SS) has been identified as a novel virulence factor, which mediates contact-dependent injection of enzymes and toxins into competing bacteria or host cells and facilitates the colonisation of a host organism. We aimed to compare the clinical course of Campylobacter infection caused by strains with and without the T6SS and identify possible associations between this putative virulence factor and the clinical manifestations of disease., Methods: From April 2015 to January 2017, patients with detection of Campylobacter spp. were identified at the University Hospital of Basel and the University Children's Hospital of Basel and included in this case-control study. Presence of the T6SS gene cluster was assayed by PCR targeting the hcp gene, confirmed with whole genome sequencing. Pertinent clinical data was collected by medical record review. Differences in disease- and host-characteristics between T6SS-positive (case) and -negative (control) were compared in a uni- and multi-variable analysis. Hospital admission, antibiotic therapy, admission to intensive care unit, development of bacteraemia and in-hospital mortality were considered as clinical endpoints., Results: We identified 138 cases of Campylobacter jejuni infections and 18 cases of Campylobacter coli infections from a paediatric and adult population. Analyses were focused on adult patients with C. jejuni (n = 119) of which 16.8% were T6SS-positive. Comparisons between T6SS-positive and -negative C. jejuni isolates did not reveal significant differences regarding clinical manifestations or course of disease. All clinical endpoints showed a similar distribution in both groups. A higher score in the Charlson Comorbidity Index was associated with T6SS-positive C. jejuni isolates (p < 0.001) and patients were more likely to have a solid organ transplant and to be under immunosuppressive therapy., Conclusions: Our study does not provide evidence that T6SS is associated with a more severe clinical course. Interestingly, T6SS-positive isolates are more commonly found in immunocompromised patients: an observation which merits further investigation.

The introduction of biogas plants is a promising way to recycle organic wastes with renewable energy production and reducing greenhouse gas. Application of anaerobic digestate as a fertilizer reduces the consumption of chemical fertilizers. In this study, the survival of pathogenic bacteria and plant growth promoting bacteria (PGPB) in two full-scale biogas plants operated at mesophilic condition were investigated. Feedstock and anaerobic digestate samples were collected from biogas plants and bacteria load in samples were detected using standard dilution plate method. Pathogenic bacteria were reduced to not detected level through mesophilic digestion tank except for Campylobacter. However, it could be reduced by 98.7% through a sterilization tank. Bacillus was detected at 8.00 and 7.81 log 10  CFU/g dry matter in anaerobic digestates, and it was also resistant to sterilization tank. Bacillus spp. is considered to be the safe bacteria that hold remarkable abilities for promoting plant growth. The results showed that treatment at biogas plants is effective to reduce pathogenic bacteria in dairy manure, and sterilization could further reduce the sanitary risks of pathogenic bacteria relating to anaerobic digestate application. Anaerobic digestates could also be utilized as bio-fertilizer as the high load of plant growth promoting bacteria., (© 2018 Japanese Society of Animal Science.)

Preventing and controlling zoonoses through the design and implementation of public health policies requires a thorough understanding of transmission pathways. Modelling jointly the epidemiological data and genetic information of microbial isolates derived from cases provides a methodology for tracing back the source of infection. In this paper, the attribution probability for human cases of campylobacteriosis for each source, conditional on the extent to which each case resides in a rural compared to urban environment, is estimated. A model that incorporates genetic data and evolutionary processes is applied alongside a newly developed genetic-free model. We show that inference from each model is comparable except for rare microbial genotypes. Further, the effect of 'rurality' may be modelled linearly on the logit scale, with increasing rurality leading to the increasing likelihood of ruminant-sourced campylobacteriosis.

Campylobacter showae a bacterium historically linked to gingivitis and periodontitis, has recently been associated with inflammatory bowel disease and colorectal cancer. Our aim was to generate genome sequences for new clinical C. showae strains and identify functional properties explaining their pathogenic potential. Eight C. showae genomes were assessed, four strains isolated from inflamed gut tissues from paediatric Crohn's disease patients, three strains from colonic adenomas, and one from a gastroenteritis patient stool. Genome assemblies were analyzed alongside the only 3 deposited C. showae genomes. The pangenome from these 11 strains consisted of 4686 unique protein families, and the core genome size was estimated at 1050 ± 15 genes with each new genome contributing an additional 206 ± 16 genes. Functional assays indicated that colonic strains segregated into 2 groups: adherent/invasive vs. non-adherent/non-invasive strains. The former possessed Type IV secretion machinery and S-layer proteins, while the latter contained Cas genes and other CRISPR associated proteins. Comparison of gene profiles with strains in Human Microbiome Project metagenomes showed that gut-derived isolates share genes specific to tongue dorsum and supragingival plaque counterparts. Our findings indicate that C. showae strains are phenotypically and genetically diverse and suggest that secretion systems may play an important role in virulence potential.

We investigated the efficacy of syringe filters and membrane filters with different pore sizes for recovering Campylobacter. A syringe filter with a 0.45 μm pore size achieved the highest recovery rate (0.29%), while polycarbonate membrane filters with pore sizes of 0.6 μm and 0.4 μm recovered less Campylobacter, at 0.01% and 1.3 × 10 -3 %, respectively. We also tested 601 diarrheic stool samples using membrane and syringe filtration methods. A total of 23 Campylobacter jejuni/coli were isolated from both syringe and membrane filtration; nine and one were isolated from only syringe or only membrane filtration, respectively (p < .05). The syringe filtration technique was better than membrane filtration for the isolation of Campylobacter., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Background: Barrett’s oesophagus (BO) is a pre-malignant condition in which normal squamous epithelium of the lower oesophagus and gastresophageal junction is replaced by columnar cells and progress to oesophageal adenocarcinoma. The increase burden of oesophagus cancer morbidity and mortality worldwide make study of factors involved in the pathogenesis of BO essential. However, most of studies that examine the environmental risk factors associated with increased incidence and prevalence of BO have largely ignored the potential role of bacteria in disease aetiology. Aims: This study examined the role of Campylobacter concisus isolated from Barrett’s and adenocarcinoma patient samples as one of possible environmental factors in the progression of Barrett’s oesophagus to oesophagus adenocarcinoma. Methods: We focused on the effect of C. concisus on the expression caudal type homeobox 1 gene (CDX1) and cyclooxygenase-2 (COX-2) in three BO cell lines using quantitative real-time PCR. In addition, the attachment and invasion characteristics of C. concisus were also tested. Results: Results showed that C. concisus had a strong attachment to the cell lines and induce the expression of CDX1 in Barrett’s cell lines in a time-dependent manner. Conclusion: Findings indicate that C. concisus could be as a new challenge in the progression of BO to adenocarcinoma., (Creative Commons Attribution License)

Background: Infectious aneurysm is a rare complication in intracranial aneurysm. Moreover, giant aneurysm is a rare entity in intracranial aneurysm. In the great majority of infectious intracranial aneurysms, vasculitis and/or endocarditis is associated., Case Description: Here, we report the case of an 83-year-old man who developed a giant infectious intracranial aneurysm on the anterior communicating artery. This patient had never shown any intracranial bleeding. A surgery was performed, and purulent liquid and aneurysm wall were removed during procedure. The bacteriologic analyses reported Campylobacter fetus., Conclusions: This bacterium seems to be more and more frequent in North America and Europe, according to the current literature. Moreover, cases of mycotic extracranial aneurysm were reported. Here we report the first case of intracranial giant infectious aneurysm in an adult patient, without any endocarditis or vasculitis. This new entity could be the cause of a bacterial graft on an asymptomatic giant preexisting intracranial aneurysm or a consequence of campylobacteriosis., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The pathogenicity of Campylobacter concisus , increasingly found in the human gastrointestinal (GI) tract, is unclear. Some studies indicate that its role in GI conditions has been underestimated, whereas others suggest that the organism has a commensal-like phenotype. For the enteropathogen C. jejuni , the lipooligosaccharide (LOS) is a main driver of virulence. We investigated the LOS structure of four C. concisus clinical isolates and correlated the inflammatory potential of each isolate with bacterial virulence. Mass spectrometric analyses of lipid A revealed a novel hexa-acylated diglucosamine moiety with two or three phosphoryl substituents. Molecular and fragment ion analysis indicated that the oligosaccharide portion of the LOS had only a single phosphate and lacked phosphoethanolamine and sialic acid substitution, which are hallmarks of the C. jejuni LOS. Consistent with our structural findings, C. concisus LOS and live bacteria induced less TNF-α secretion in human monocytes than did C. jejuni Furthermore, the C. concisus bacteria were less virulent than C. jejuni in a Galleria mellonella infection model. The correlation of the novel lipid A structure, decreased phosphorylation, and lack of sialylation along with reduced inflammatory potential and virulence support the significance of the LOS as a determinant in the relative pathogenicity of C. concisus .

Campylobacter is the most common cause of bacterial food-borne diarrhoeal disease worldwide. Chicken meat is considered the main source of human infection; however, C. jejuni and C. coli have also been reported in a range of livestock and wildlife species, including pheasants. Wild pheasant meat reaches the consumer's table because of hunting but there is a lack of information concerning the risk of Campylobacter infection in humans. This study aimed to determine the prevalence of Campylobacter in wild game pheasants in Scotland, to identify the main sequence types (STs) present and to evaluate their impact on public health. A total of 287 caecal samples from five Scottish regions were collected during the hunting season 2013/2014. Campylobacter was detected and enumerated using standard culture methods. PCR and High Throughput Multi Locus Sequence Typing (HiMLST) were used for species identification and sequence typing. In total, 36.6% of 287 caecal samples (n = 105; 95% CI: 14-59.2) were Campylobacter positive. Using PCR, 62.6% of samples (n = 99) were identified as C. coli and 37.4% as C. jejuni. HiMLST (n = 80) identified 19 different STs. ST-828 (n = 19) was the most common, followed by ST-827 (n = 12) and ST19 (n = 7). Sixteen of the 19 STs isolated are present in humans and eight are C. coli STs that account for 6.96% of human infections, although the overall risk to public health from pheasant meat is still considered to be low., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Historically, Campylobacteriosis has been considered to be zoonotic; the Campylobacter species that cause human acute intestinal disease such as Campylobacter jejuni and Campylobacter coli originate from animals. Over the past decade, studies on human hosted Campylobacter species strongly suggest that Campylobacter concisus plays a role in the development of inflammatory bowel disease (IBD). C. concisus primarily colonizes the human oral cavity and some strains can be translocated to the intestinal tract. Genome analysis of C. concisus strains isolated from saliva samples has identified a bacterial marker that is associated with active Crohn's disease (one major form of IBD). In addition to C. concisus , humans are also colonized by a number of other Campylobacter species, most of which are in the oral cavity. Here we review the most recent advancements on C. concisus and other human hosted Campylobacter species including their clinical relevance, transmission, virulence factors, disease associated genes, interactions with the human immune system and pathogenic mechanisms.

In recent years, an increasing number of Campylobacter species have been associated with human gastrointestinal (GI) diseases including gastroenteritis, inflammatory bowel disease, and colorectal cancer. Campylobacter concisus, an oral commensal historically linked to gingivitis and periodontitis, has been increasingly detected in the lower GI tract. In the present study, we generated robust genome sequence data from C. concisus strains and undertook a comprehensive pangenome assessment to identify C. concisus virulence properties and to explain potential adaptations acquired while residing in specific ecological niche(s) of the GI tract. Genomes of 53 new C. concisus strains were sequenced, assembled, and annotated including 36 strains from gastroenteritis patients, 13 strains from Crohn's disease patients and four strains from colitis patients (three collagenous colitis and one lymphocytic colitis). When compared with previous published sequences, strains clustered into two main groups/genomospecies (GS) with phylogenetic clustering explained neither by disease phenotype nor sample location. Paired oral/faecal isolates, from the same patient, indicated that there are few genetic differences between oral and gut isolates which suggests that gut isolates most likely reflect oral strain relocation. Type IV and VI secretion systems genes, genes known to be important for pathogenicity in the Campylobacter genus, were present in the genomes assemblies, with 82% containing Type VI secretion system genes. Our findings indicate that C. concisus strains are genetically diverse, and the variability in bacterial secretion system content may play an important role in their virulence potential.

It has been reported that patients with chronic heart failure exhibit an intestinal overgrowth of primary gut bacterial pathogens, such as Shigella spp., Salmonella spp., Campylobacter spp., and Yersinia enterocolitica. We failed to reproduce these findings in a cohort of 39 patients admitted to the hospital with decompensated heart failure by means of conventional stool bacterial cultures and a multiplexed polymerase chain reaction assay., (Copyright © 2018. Published by Elsevier Inc.)

Modified Cary-Blair medium derived devices have been implemented in many laboratories to optimize culture recovery of common bacterial enteric pathogens. Our analysis constitutes the first report of routine laboratory experience supporting the idea that the use of such devices enhances Campylobacter recovery from stools., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Campylobacteriosis is an important, worldwide public health problem with numerous socio-economic impacts. Since 2015, approximately 230,000 cases have been reported annually in Europe. In the United States, Australia and New Zealand, campylobacteriosis is the most commonly reported disease. Poultry and poultry products are considered important sources of human infections. Poultry meat can become contaminated with Campylobacter during slaughter if live chickens are intestinal carriers. Campylobacter spp. can be transferred from animals to humans through consumption and handling of contaminated food products, with fresh chicken meat being the most commonly implicated food type. Regarding food-borne disease, the most important Campylobacter species are Campylobacter jejuni and Campylobacter coli. In humans, clinical signs of campylobacteriosis include diarrhoea, abdominal pain, fever, headache, nausea and vomiting. Most cases of campylobacteriosis are sporadic and self-limiting, but there are post-infection complications, for example, Guillain-Barrés syndrome. This review summarizes an analysis undertaken by the DISCONTOOLS group of experts on campylobacteriosis. Gaps were identified in: (i) knowledge of true number of infected humans; (ii) mechanisms of pathogenicity to induce infection in humans; (iii) training to prevent transfer of Campylobacter from raw to ready-to-eat food; (iv) development of effective vaccines; (v) understanding transmission routes to broiler flocks; (vi) knowledge of bacteriocins, bacteriophages and antimicrobial peptides as preventive therapies; (vii) ration formulation as an effective preventive measure at a farm level; (viii) development of kits for rapid detection and quantification of Campylobacter in animals and food products; and (ix) development of more effective antimicrobials for treatment of humans infected with Campylobacter. Some of these gaps are relevant worldwide, whereas others are more related to problems encountered with Campylobacter in industrialized countries., (© 2018 Blackwell Verlag GmbH.)

Campylobacter concisus is an oral bacterium that is associated with inflammatory bowel disease (IBD) including Crohn's disease (CD) and ulcerative colitis (UC). C. concisus consists of two genomospecies (GS) and diverse strains. This study aimed to identify molecular markers to differentiate commensal and IBD-associated C. concisus strains. The genomes of 63 oral C. concisus strains isolated from patients with IBD and healthy controls were examined, of which 38 genomes were sequenced in this study. We identified a novel secreted enterotoxin B homologue, Csep1. The csep1 gene was found in 56% of GS2 C. concisus strains, presented in the plasmid pICON or the chromosome. A six-nucleotide insertion at the position 654-659 bp in csep1 (csep1-6bpi) was found. The presence of csep1-6bpi in oral C. concisus strains isolated from patients with active CD (47%, 7/15) was significantly higher than that in strains from healthy controls (0/29, P = 0.0002), and the prevalence of csep1-6bpi positive C. concisus strains was significantly higher in patients with active CD (67%, 4/6) as compared to healthy controls (0/23, P = 0.0006). Proteomics analysis detected the Csep1 protein. A csep1 gene hot spot in the chromosome of different C. concisus strains was found. The pICON plasmid was only found in GS2 strains isolated from the two relapsed CD patients with small bowel complications. This study reports a C. concisus molecular marker (csep1-6bpi) that is associated with active CD.