A spatial-genomic analysis reveals that bird species living closer to humans have higher diversity of the pathogen Campylobacter and its antimicrobial resistance genes. This suggests that urbanization could promote pathogen transmission among wild animals and, potentially, humans.

Enumeration of Campylobacter from environmental waters can be difficult due to its low concentrations, which can still pose a significant health risk. Spectrophotometry is an approach commonly used for fast detection of water-borne pollutants in water samples, but it has not been used for pathogen detection, which is commonly done through a laborious and time-consuming culture or qPCR Most Probable Number enumeration methods (i.e., MPN-PCR approaches). In this study, we proposed a new method, MPN-Spectro-ML, that can provide rapid evidence of Campylobacter detection and, hence, water concentrations. After an initial incubation, the samples were analysed using a spectrophotometer, and the spectrum data were used to train three machine learning (ML) models (i.e., supported vector machine - SVM, logistic regression-LR, and random forest-RF). The trained models were used to predict the presence of Campylobacter in the enriched water samples and estimate the most probable number (MPN). Over 100 stormwater, river, and creek samples (including both fresh and brackish water) from rural and urban catchments were collected to test the accuracy of the MPN-Spectro-ML method under various scenarios and compared to a previously standardised MPN-PCR method. Differences in the spectrum were found between positive and negative control samples, with two distinctive absorbance peaks between 540-542nm and 575-576nm for positive samples. Further, the three ML models had similar performance irrespective of the scenario tested with average prediction accuracy (ACC) and false negative rates at 0.763 and 13.8%, respectively. However, the predicted MPN of Campylobacter from the new method varied from the traditional MPN-PCR method, with a maximum Nash-Sutcliffe coefficient of 0.44 for the urban catchment dataset. Nevertheless, the MPN values based on these two methods were still comparable, considering the confidence intervals and large uncertainties associated with MPN estimation. The study reveals the potential of this novel approach for providing interim evidence of the presence and levels of Campylobacter within environmental water bodies. This, in turn, decreases the time from risk detection to management for the benefit of public health.

Although MALDI-TOF mass spectrometry (MS) is considered as the gold standard for rapid and cost-effective identification of microorganisms in routine laboratory practices, its capability for antimicrobial resistance (AMR) detection has received limited focus. Nevertheless, recent studies explored the predictive performance of MALDI-TOF MS for detecting AMR in clinical pathogens when machine learning techniques are applied. This chapter describes a routine MALDI-TOF MS workflow for the rapid screening of AMR in foodborne pathogens, with Campylobacter spp. as a study model.

Campylobacter is highly associated with poultry and frequently causes foodborne illness worldwide. Thus, effective control measures are necessary to reduce or prevent human infections. In this review, Campylobacter control methods applicable at postharvest level for poultry meat during production, storage, and preparation are discussed. Drying and temperature are discussed as general strategies. Traditional strategies such as steaming, freezing, sanitizing, organic acid treatment, and ultraviolet light treatment are also discussed. Recent advances in nanotechnology using antibacterial nanoparticles and natural antimicrobial agents from plants and food byproducts are also discussed. Although advances have been made and there are various methods for preventing Campylobacter contamination, it is still challenging to prevent Campylobacter contamination in raw poultry meats with current methods. In addition, some studies have shown that large strain-to-strain variation in susceptibility to these methods exists. Therefore, more effective methods or approaches need to be developed to substantially reduce human infections caused by Campylobacter.

Campylobacter is the leading bacterial cause of infectious intestinal disease, but the pathogen typically accounts for a very small proportion of the overall stool microbiome in each patient. Diagnosis is even more difficult due to the fastidious nature of Campylobacter in the laboratory setting. This has, in part, driven a change in recent years, from culture-based to rapid PCR-based diagnostic assays which have improved diagnostic detection, whilst creating a knowledge gap in our clinical and epidemiological understanding of Campylobacter genotypes - no isolates to sequence. In this study, direct metagenomic sequencing approaches were used to assess the possibility of replacing genome sequences with metagenome sequences; metagenomic sequencing outputs were used to describe clinically relevant attributes of Campylobacter genotypes. A total of 37 diarrhoeal stool samples with Campylobacter and five samples with an unknown pathogen result were collected and processed with and without filtration, DNA was extracted, and metagenomes were sequenced by short-read sequencing. Culture-based methods were used to validate Campylobacter metagenome-derived genome (MDG) results. Sequence output metrics were assessed for Campylobacter genome quality and accuracy of characterization. Of the 42 samples passing quality checks for analysis, identification of Campylobacter to the genus and species level was dependent on Campylobacter genome read count, coverage and genome completeness. A total of 65% (24/37) of samples were reliably identified to the genus level through Campylobacter MDG, 73% (27/37) by culture and 97% (36/37) by qPCR. The Campylobacter genomes with a genome completeness of over 60% (n=21) were all accurately identified at the species level (100%). Of those, 72% (15/21) were identified to sequence types (STs), and 95% (20/21) accurately identified antimicrobial resistance (AMR) gene determinants. Filtration of stool samples enhanced Campylobacter MDG recovery and genome quality metrics compared to the corresponding unfiltered samples, which improved the identification of STs and AMR profiles. The phylogenetic analysis in this study demonstrated the clustering of the metagenome-derived with culture-derived genomes and revealed the reliability of genomes from direct stool sequencing. Furthermore, Campylobacter genome spiking percentages ranging from 0 to 2% total metagenome abundance in the ONT MinION sequencer, configured to adaptive sequencing, exhibited better assembly quality and accurate identification of STs, particularly in the analysis of metagenomes containing 2 and 1% of Campylobacter jejuni genomes. Direct sequencing of Campylobacter from stool samples provides clinically relevant and epidemiologically important genomic information without the reliance on cultured genomes.

Campylobacter is a globally important pathogen with well-studied risk factors, but the burden of risk factors has not been quantified. We quantified the cost of illness attributable to specific domestic risk factors for C. jejuni and C. coli in Australia. We used data from a 2018-2019 case-control study to estimate odds ratios and attributable fractions for risk factors. We used data on national incidence, hospitalization, and premature mortality to quantify burden. We then applied costs related to healthcare utilization, pain and suffering, premature mortality, and lost productivity to each risk factor. In Australia, C. jejuni caused 83.0% of campylobacteriosis infections and chicken consumption resulted in the highest attributable fraction (30.0%), costing approximately US$110 million annually. The excess burden of campylobacteriosis associated with the use of proton-pump inhibitors (PPIs) was US$45 million, with almost half these costs due to disease in adults over 65 years of age. Contact with young dogs (US$30 million) and chicken feces (US$10 million) also contributed to costs and burden. Campylobacteriosis is a significant cost to Australia, particularly because of lost productivity. Effective cross-sectoral interventions to improve chicken meat safety and reduce inappropriate use of PPIs might have substantial economic and human benefits.

The persistence of non-typhoidal Salmonella and Campylobacter in chicken meat is a considerable public health risk and a future challenge. This study aimed to determine the prevalence of Salmonella and Campylobacter in poultry processing lines where different chlorine concentrations were used in the chill tank. The samples were collected from four types of processing plants in Sri Lanka, considering the chlorine concentration used in the chill tank, which ranged from 2 ppm to 50 ppm. Salmonella and Campylobacter were isolated from whole carcass washings, neck skin, and cecal samples. Subsequently, an antimicrobial susceptibility test was performed for the isolates. The results revealed the overall prevalence of Salmonella and Campylobacter was 78.25% and 63.5%, respectively. Positive percentages of Salmonella and Campylobacter were high in the carcasses compared to the neck skin and ceca. The Campylobacter counts on the whole carcasses were significantly low (p < 0.001), at higher chlorine concentrations ranging from 20 to 30 ppm and 40 to 50 ppm. The pathogen prevalence in the whole carcasses was 84.7% Campylobacter coli, 39.1% Campylobacter jejuni, 71.1% Salmonella Typhimurium, and 28.8% Salmonella Infantis. The highest resistance was observed for tetracycline (63.8%) in Salmonella, while it was for gentamicin (87.8%) in Campylobacter. The prevalence percentage of multidrug-resistant Campylobacter was 51.2%, while it was 2.12% for Salmonella. The persistence of multidrug-resistant Salmonella and Campylobacter on the post-chill carcasses was highlighted in the present study as a significant public health threat that has to be addressed urgently.

This study was undertaken to evaluate and compare the efficacy of different multi-antigen vaccines, including heat-inactivated, whole lysate, and subunit (outer membrane proteins [OMPs]) C. jejuni vaccines along with the immunostimulant CpG ODN in controlling Campylobacter colonization in chickens. In the first trial, 125 μg of C. jejuni OMPs and 50 μg of CpG ODN were administered individually or in combination, either in ovo to chick embryos or subcutaneously (SC) to one-day-old chicks. In the second trial, different concentrations of C. jejuni antigens (heat-killed, whole lysate, and OMPs) were administered SC to one-day-old chicks. The results of the first trial revealed that SC immunization with the combination of CpG ODN and C. jejuni OMPs elevated interferon (IFN)-γ, interleukin (IL)-1β, and IL-13 gene expression in the spleen, significantly increased serum IgM and IgY antibody levels, and reduced cecal C. jejuni counts by approximately 1.2 log10. In contrast, in ovo immunization did not elicit immune responses or confer protection against Campylobacter. The results of the second trial showed that SC immunization with C. jejuni whole lysate or 200 μg OMPs reduced C. jejuni counts by approximately 1.4 and 1.1 log10, respectively. In conclusion, C. jejuni lysate and OMPs are promising vaccine antigens for reducing Campylobacter colonization in chickens.

Antimicrobial resistance (AMR) is a growing global health challenge, compromising bacterial infection treatments and necessitating robust surveillance and mitigation strategies. The overuse of antimicrobials in humans and farm animals has made them hotspots for AMR. However, the spread of AMR genes in wildlife and the environment represents an additional challenge, turning these areas into new AMR hotspots. Among the AMR bacteria considered to be of high concern for public health, Campylobacter has been the leading cause of foodborne infections in the European Union since 2005. This study examines the prevalence of AMR genes and virulence factors in Campylobacter isolates from wild birds and surface waters in Luxembourg. The findings reveal a significant prevalence of resistant Campylobacter strains, with 12% of C. jejuni from wild birds and 37% of C. coli from surface waters carrying resistance genes, mainly against key antibiotics like quinolones and tetracycline. This study underscores the crucial role of the environment in the spread of AMR bacteria and genes, highlighting the urgent need for enhanced surveillance and control measures to curb AMR in wildlife and environmental reservoirs and reduce transmission risks to humans. This research supports One Health approaches to tackling antimicrobial resistance and protecting human, animal, and environmental health.

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