Review

Authors: Samuel K Sheppard, Martin C J Maiden

The global significance of Campylobacter jejuni and Campylobacter coli as gastrointestinal human pathogens has motivated numerous studies to characterize their population biology and evolution. These bacteria are a common component of the intestinal microbiota of numerous bird and mammal species and cause disease in humans, typically via consumption of contaminated meat products, especially poultry meat. Sequence-based molecular typing methods, such as multilocus sequence typing (MLST) and whole genome sequencing (WGS), have been instructive for understanding the epidemiology and evolution of these bacteria and how phenotypic variation relates to the high degree of genetic structuring in C. coli and C. jejuni populations. Here, we describe aspects of the relatively short history of coevolution between humans and pathogenic Campylobacter, by reviewing research investigating how mutation and lateral or horizontal gene transfer (LGT or HGT, respectively) interact to create the observed population structure. These genetic changes occur in a complex fitness landscape with divergent ecologies, including multiple host species, which can lead to rapid adaptation, for example, through frame-shift mutations that alter gene expression or the acquisition of novel genetic elements by HGT. Recombination is a particularly strong evolutionary force in Campylobacter, leading to the emergence of new lineages and even large-scale genome-wide interspecies introgression between C. jejuni and C. coli. The increasing availability of large genome datasets is enhancing understanding of Campylobacter evolution through the application of methods, such as genome-wide association studies, but MLST-derived clonal complex designations remain a useful method for describing population structure.

Topics: Adaptation, Physiological; Campylobacter coli; Campylobacter jejuni; Evolution, Molecular; Recombination, Genetic

PubMed: 26101080
DOI: 10.1101/cshperspect.a018119

Authors: Shuxin Zhang, Jiahua Shi, Xuan Li...

Campylobacter spp. is one of the most frequent pathogens of bacterial gastroenteritis recorded worldwide. Campylobacter jejuni (C. jejuni) and Campylobacter coli (C. coli) are the two major disease-associated species, accounting for >95 % of infections, and thus have been selected for disease surveillance. Monitoring temporal variations in pathogen concentration and diversity excreted from community wastewater allows the early detection of outbreaks. Multiplex real-time/quantitative PCR (qPCR) enables multi-target quantification of pathogens in various types of samples including wastewater. Also, an internal amplification control (IAC) is required for each sample when adopting PCR-based methods for pathogen detection and quantification in wastewater to exclude the inhibition of the wastewater matrix. To achieve reliable quantification of C. jejuni and C. coli towards wastewater samples, this study developed and optimized a triplex qPCR assay by combining three qPCR primer-probe sets targeting Campylobacter jejuni subsp. jejuni, Campylobacter coli, and Campylobacter sputorum biovar sputorum (C. sputorum), respectively. This triplex qPCR assay not only can directly and simultaneously detect the concentration of C. jejuni and C. coli in wastewater but also can achieve the PCR inhibition control using C. sputorum primer-probe set. This is the first developed triplex qPCR assay with IAC for C. jejuni and C. coli, to be used in the wastewater-based epidemiology (WBE) applications. The optimized triplex qPCR assay enables the detection limit of the assay (ALOD and wastewater (PLOD) as 10 gene copy/μL and 2 log10 cells/mL (2 gene copies/μL of extracted DNA), respectively. The application of this triplex qPCR to 52 real raw wastewater samples from 13 wastewater treatment plants demonstrated its potential as a high-throughput and economically viable tool for the long-term monitoring of C. jejuni and C. coli prevalence in communities and the surrounding environments. This study provided an accessible methodology and a solid foundation for WBE-based monitoring of Campylobacter spp. relevant diseases and paved the road for future WBE back-estimation of C. jejuni and C. coli prevalence.

Topics: Campylobacter jejuni; Campylobacter coli; Wastewater; Sensitivity and Specificity; Campylobacter; DNA, Bacterial

PubMed: 37268129
DOI: 10.1016/j.scitotenv.2023.164574

Authors: Jai W Mehat, Roberto M La Ragione, Arnoud H M van Vliet...

BACKGROUND

Campylobacter jejuni and Campylobacter coli are major global causes of bacterial gastroenteritis. Whilst several individual colonisation and virulence factors have been identified, our understanding of their role in the transmission, pathogenesis and ecology of Campylobacter has been hampered by the genotypic and phenotypic diversity within C. jejuni and C. coli. Autotransporter proteins are a family of outer membrane or secreted proteins in Gram-negative bacteria such as Campylobacter, which are associated with virulence functions. In this study we have examined the distribution and predicted functionality of the previously described capC and the newly identified, related capD autotransporter gene families in Campylobacter.

RESULTS

Two capC-like autotransporter families, designated capC and capD, were identified by homology searches of genomes of the genus Campylobacter. Each family contained four distinct orthologs of CapC and CapD. The distribution of these autotransporter genes was determined in 5829 C. jejuni and 1347 C. coli genomes. Autotransporter genes were found as intact, complete copies and inactive formats due to premature stop codons and frameshift mutations. Presence of inactive and intact autotransporter genes was associated with C. jejuni and C. coli multi-locus sequence types, but for capC, inactivation was independent from the length of homopolymeric tracts in the region upstream of the capC gene. Inactivation of capC or capD genes appears to represent lineage-specific gene decay of autotransporter genes. Intact capC genes were predominantly associated with the C. jejuni ST-45 and C. coli ST-828 generalist lineages. The capD3 gene was only found in the environmental C. coli Clade 3 lineage. These combined data support a scenario of inter-lineage and interspecies exchange of capC and subsets of capD autotransporters.

CONCLUSIONS

In this study we have identified two novel, related autotransporter gene families in the genus Campylobacter, which are not uniformly present and exhibit lineage-specific associations and gene decay. The distribution and decay of the capC and capD genes exemplifies the erosion of species barriers between certain lineages of C. jejuni and C. coli, probably arising through co-habitation. This may have implications for the phenotypic variability of these two pathogens and provide opportunity for new, hybrid genotypes to emerge.

Topics: Campylobacter coli; Campylobacter jejuni; Gene Deletion; Genome, Bacterial; Phylogeny; Type V Secretion Systems; Virulence Factors

PubMed: 32306949
DOI: 10.1186/s12864-020-6704-z

Authors: Lucia Rivas, Pierre-Yves Dupont, Brent Gilpin...

ABSTRACT

A pilot survey was performed to determine the prevalence of Campylobacter jejuni and Campylobacter coli on three age classes (lamb, hogget, and mutton) of ovine carcass trim postdressing and prechill. Sampling of hogget carcasses was undertaken 6 months before sampling of lamb and mutton carcasses. A total of 120 trim samples were collected from 11 processing plants across New Zealand. All samples were enriched and screened using PCR for the presence of C. jejuni and C. coli, and isolation was attempted for all screen-positive samples. Enumeration of Campylobacter from lamb trim samples showed that Campylobacter bacteria were present in very low numbers (<10 CFU/g). The overall prevalence of Campylobacter for ovine trim based on PCR detection was 33% (39 of 120 samples), with prevalences for hogget, lamb, and mutton carcass trim of 56% (28 of 50), 11% (4 of 35), and 20% (7 of 35), respectively. Whole genome sequencing was performed on a selection of C. jejuni and C. coli isolates, and the data were used to subtype using multilocus sequence typing (MLST) and whole genome MLST. Twenty-five MLST sequence types (STs) were identified among 44 isolates, including ST42, ST50, ST3222, and ST3072, which have been previously reported to be associated with ruminant sources. Four novel STs were also identified. Whole genome MLST analysis further discriminated isolates within a single ST type and demonstrated a genetic diversity among the ovine isolates collected. Genes associated with the oxacillinase class of β-lactamase enzymes were identified in 41 of 44 Campylobacter isolates. This study provides preliminary data that can be incorporated into existing source attribution models to assist in determining the potential contribution of ovine sources to the burden of campylobacteriosis in New Zealand.

Topics: Animals; Campylobacter; Campylobacter Infections; Campylobacter coli; Campylobacter jejuni; Chickens; Genotype; Multilocus Sequence Typing; New Zealand; Prevalence; Sheep

PubMed: 32766835
DOI: 10.4315/JFP-20-220

Review

Authors: K N Eberle, A S Kiess

Human campylobacteriosis, an infection caused by the bacterium Campylobacter, is a major issue in the United States food system, especially for poultry products. According to the Center for Disease Control, campylobacterosis is estimated to affect over 2.4 million people annually. Campylobacter jejuni and Campylobacter coli are 2 species responsible for the majority of campylobacterosis infections. Phenotypic and genotypic typing methods are often used to discriminate between bacteria at the species and subspecies level and are often used to identify pathogenic organisms, such as C. jejuni and C. coli. This review describes the design as well as advantages and disadvantages for 3 current phenotypic techniques (biotyping, serotyping, and multilocus enzyme electrophoresis) and 6 genotypic techniques (multilocus sequence typing, PCR, pulse-field gel electrophoresis, ribotyping, flagellin typing, and amplified fragment length polymorphisms) for typing pathogenic Campylobacter spp.

Topics: Animals; Bacterial Typing Techniques; Campylobacter Infections; Campylobacter coli; Campylobacter jejuni; Chickens; Phenotype; Poultry Diseases; Reproducibility of Results

PubMed: 22184452
DOI: 10.3382/ps.2011-01414

Authors: Anna Nilsson, Astrid Skarp, Cecilia Johansson...

Campylobacter jejuni and Campylobacter coli are important bacterial enteropathogens. Poultry is the best-known reservoir for Campylobacter infection but natural bodies of water have also been shown to be important pathways for transmission. Campylobacter can survive in cold water but most of the studies have focused on C. jejuni only. In this paper, we take a closer look at the biology and water survival strategies of C. coli. Eight C. coli isolates cultivated from raw (incoming) surface water at water plants in Sweden were characterized using whole-genome sequencing and phenotypical assays. Phylogenetic analysis assigned the Swedish water isolates to clades 2 and 3, known to include C. coli of environmental origin. In addition, 53 earlier published sequences of C. coli clade 2 and 3 from environmental waters were included for in silico analyses. Generally, clade 2 isolates had larger genomes, which included a functional tricarballylate utilization locus, while clade 3 isolates contained different genes involved in oxidative stress as well as putative virulence factors. The Swedish water isolates of clade 2 formed large, blurry bacterial colonies on agar, whereas clade 3 colonies were smaller. All Swedish isolates were motile, but clade 3 isolates formed larger motility zones on soft agar, and none of these isolates produced biofilm. Although water survival varied between the analyzed isolates, there were hardly any clade-specific significant differences. Our results highlight the diversity of C. coli in general, and show differences in metabolic capabilities and ways to handle oxidative stress between clade 2 and 3 water isolates.

Topics: Bacterial Proteins; Campylobacter coli; Fresh Water; Genome, Bacterial; Phenotype; Phylogeny; Sweden; Water Microbiology

PubMed: 29424055
DOI: 10.1002/mbo3.583

Authors: Anastasia-Lisa Dieckmann, Thomas Riedel, Boyke Bunk...

The intriguing recent discovery of strains, especially of clade 1, that (i) possess mosaic / alleles, (ii) demonstrate mixed multilocus sequence types (MLSTs) and (iii) have undergone genome-wide introgression has led to the speculation that these two species may be involved in an accelerated rate of horizontal gene transfer that is progressively leading to the merging of both species in a process coined 'despeciation'. In an MLST-based neighbour-joining tree of a number of and isolates of different clades, three prominent isolates formed a seemingly separate cluster besides the previously described and clades. In the light of the suspected, ongoing genetic introgression between the and species, this cluster of isolates is proposed to present one of the hybrid clonal complexes in the despeciation process of the genus. Specific DNA methylation as well as restriction modification systems are known to be involved in selective uptake of external DNA and their role in such genetic introgression remains to be further investigated. In this study, the phylogeny and DNA methylation of these putative / hybrid strains were explored, their genomic mosaic structure caused by introgression was demonstrated and basic phenotypic assays were used to characterize these isolates. The genomes of the three hybrid strains were sequenced using PacBio SMRT sequencing, followed by methylome analysis by Restriction-Modification Finder and genome analysis by Parsnp, Smash++ and blast. Additionally, the strains were phenotypically characterized with respect to growth behaviour, motility, eukaryotic cell invasion and adhesion, autoagglutination, biofilm formation, and water survival ability. Our analyses show that the three hybrid strains are clade 1 . strains, which have acquired between 8.1 and 9.1 % of their genome from . The genomic segments acquired are distributed over the entire genome and do not form a coherent cluster. Most of the genes originating from are involved in chemotaxis and motility, membrane transport, cell signalling, or the resistance to toxic compounds such as bile acids. Interspecies gene transfer from has contributed 8.1-9.1% to the genome of three isolates and initiated the despeciation between and . Based on their functional annotation, the genes originating from enable the adaptation of the three strains to an intra-intestinal habitat. The transfer of a fused type II restriction-modification system that recognizes the CAYNNNNNCTC/GAGNNNNNRTG motif seems to be the key for the recombination of the genetic material with genomes.

Topics: Bacterial Typing Techniques; Campylobacter Infections; Campylobacter coli; Campylobacter jejuni; Chromosomes, Bacterial; DNA, Bacterial; Epigenome; Gene Transfer, Horizontal; Genome, Bacterial; Genomics; Multilocus Sequence Typing; Phylogeny; Sequence Analysis, DNA

PubMed: 34661518
DOI: 10.1099/mgen.0.000679

Review

Authors: Manel Gharbi, Awatef Béjaoui, Cherif Ben Hamda...

BACKGROUND

Thermo-tolerant Campylobacter species are the major cause of foodborne diseases worldwide. This study aimed to evaluate the prevalence of virulence genes and antibiotic resistance determinants in Campylobacter jejuni and Campylobacter coli isolates, and to investigate the relationship between these two traits.

METHODS

A total of 132 Campylobacter isolates from poultry were tested for the presence of 13 virulence genes; flaA, cadF, racR, virB11, pldA, dnaJ, cdtA, cdtB, cdtC, ciaB, wlaN, cgtB and ceuE. The mechanisms underlying antibiotic resistance phenotypes were also studied by PCR and MAMA-PCR.

RESULTS

PCR results revealed the presence of antimicrobial resistance genes in C. jejuni and C. coli as follows: cmeB (80% and 100%), tet(O) (100% and 80%), and the bla (81% and 93%), respectively. None of these strains harbored the aphA-3 gene. The Thr-86-Ile mutation associated with resistance to quinolones was found in 90% of C. jejuni and 80% of C. coli isolates. While the A2075G and A2074C mutations linked to the erythromycin resistance were detected in 100% of both species. Virulence genes were prevalent and ranged from 40 to 100%. A positive relationship was revealed between cadF, racR, and ciaB genes and resistance to ampicillin, amoxicillin/clavulanic acid, chloramphenicol, and nalidixic acid, in C. jejuni. However, no association was observed for C. coli isolated strains.

CONCLUSION

This study provides for the first time an overview of antibiotic resistance mechanisms and pathogenic profiles of Campylobacter isolates, which emphasizes the potential risk for consumer health.

Topics: Animals; Anti-Bacterial Agents; Campylobacter; Campylobacter coli; Campylobacter Infections; Campylobacter jejuni; Chickens; Drug Resistance, Microbial; Tunisia; Virulence

PubMed: 34340908
DOI: 10.1016/j.jmii.2021.07.001

Authors: Mark E Berrang, Richard J Meinersmann, Nelson A Cox...

Campylobacter can be difficult to recover from complex samples due to overgrowth by background bacteria. A 0.45- or 0.65-μm-pore-size filter overlaid on agar plates can be used as a means to separate Campylobacter from confounding non-Campylobacter cells, facilitating detection on solid plating media. It is unclear what percentage of cells in a Campylobacter suspension passes through a filter and results in visible colonies. The objective of this study was to compare the number of Campylobacter cells detected by the filter method with those detected by direct plating and determine if the filter method can be used to estimate cellular density of an unknown Campylobacter in suspension. Overnight liquid cultures of six subtypes of Campylobacter jejuni and six of Campylobacter coli, all originally detected in chicken samples, were used for this study. Motility of isolates was tested by using a stab into soft agar, incubating plates, and measuring colony size. Each subtype was applied to Campy-Cefex agar directly and through a 0.45- or 0.65-μm-pore-size filter. Filters were removed, plates were incubated, and colonies were counted; three replications were conducted. Mean recovery by direct plating was 8.3 log CFU/mL. Regardless of pore size, the overall mean number of Campylobacter detected by using the filter method was significantly less than that using direct plating (P < 0.05). The mean difference between direct plating and plating though a 0.65-μm-pore-size filter for motile Campylobacter was log 2.4 CFU/mL, with a 95% confidence interval of ±0.2 log CFU/mL.

Topics: Agar; Animals; Bacteria; Campylobacter; Campylobacter coli; Campylobacter jejuni; Chickens; Collodion; Culture Media; Filtration

PubMed: 29140745
DOI: 10.4315/0362-028X.JFP-17-211

Authors: Shuxin Zhang, Jiahua Shi, Elipsha Sharma...

Campylobacter jejuni and coli are two main pathogenic species inducing diarrhoeal diseases in humans, which are responsible for the loss of 33 million lives each year. Current Campylobacter infections are mainly monitored by clinical surveillance which is often limited to individuals seeking treatment, resulting in under-reporting of disease prevalence and untimely indicators of community outbreaks. Wastewater-based epidemiology (WBE) has been developed and employed for the wastewater surveillance of pathogenic viruses and bacteria. Monitoring the temporal changes of pathogen concentration in wastewater allows the early detection of disease outbreaks in a community. However, studies investigating the WBE back-estimation of Campylobacter spp. are rare. Essential factors including the analytical recovery efficiency, the decay rate, the effect of in-sewer transport, and the correlation between the wastewater concentration and the infections in communities are lacking to support wastewater surveillance. This study carried out experiments to investigate the recovery of Campylobacter jejuni and coli from wastewater and the decay under different simulated sewer reactor conditions. It was found that the recovery of Campylobacter spp. from wastewater varied with their concentrations in wastewater and depended on the detection limit of quantification methods. The concentration reduction of Campylobacter. jejuni and coli in sewers followed a two-phase reduction model, and the faster concentration reduction during the first phase is mainly due to their partitioning onto sewer biofilms. The total decay of Campylobacter. jejuni and coli varied in different types of sewer reactors, i.e. rising main vs. gravity sewer. In addition, the sensitivity analysis for WBE back-estimation of Campylobacter suggested that the first-phase decay rate constant (k) and the turning time point (t) are determining factors and their impacts increased with the hydraulic retention time of wastewater.

Topics: Humans; Wastewater; Wastewater-Based Epidemiological Monitoring; Campylobacter coli; Campylobacter jejuni; Biofilms; Campylobacter Infections

PubMed: 36801582
DOI: 10.1016/j.watres.2023.119737