Thermotolerant Campylobacter Species Research Articles

Global systematic review and meta-analysis on foodborne thermotole­rant Campylobacter prevalence in different species of poultry birds and associated sources of contamination

Campylobacteriosis is one of the most prevalent foodborne bacterial diseases. Infected chickens and poultry products are a possible source of this illness in people all over the world. Therefore, the objective of this current meta-analysis was to summarise the available data on the prevalence of thermotolerant Campylobacter species in poultry and their products. A systematic literature search was conducted to gather pertinent articles from 2000 to 2021, using globally recognised four electronic databases, including Google Scholar, PubMed, Science Direct, and Scopus. The mean prevalence of campylobacteriosis in poultry species and their products (meat, eggs, and offal) was 44% (95% CI: 39–50%) with the highest prevalence in Australia (67%). Among the European countries, France had the maximum prevalence rate of 76%, while Japan had recorded 61% as the peak among Asian countries. Prevalence rates of 43% and 56% were calculated in United States and Brazil representing the American region. In the species-wise results, the mean prevalence rates of Campylobacter jejuni and Campylobacter coli were 29% and 16%, correspondingly. In case of live birds, chickens and turkeys possessed maximum prevalence rates of 47% and 40%, whereas 46% and 63% were recorded from poultry meat and liver. Besides, categorising the difference sources of contamination, the maximum prevalence rate of 62% was found in the poultry processing plants followed by 54% from supermarkets and 38% from farms. According to the current meta-analysis, Campylobacter jejuni was the most common bacterium worldwide, and poultry meat – the most frequent source of human infection. The predominance of Campylobacter species is a threat for public health, and national authorities must undertake strategies to control this disease in each country with the goal of establishing adequate risk management measures.

Prevalence and Distribution of Thermotolerant Campylobacter Species in Poultry: A Comprehensive Review with a Focus on the Factors Affecting the Detection and Enumeration of Campylobacter jejuni and Campylobacter coli in Chicken Meat

It is well known that the strong-evidence foodborne outbreaks of human campylobacteriosis are associated with the consumption of raw or incompletely thermally processed poultry meat, whereas broilers act as the main reservoir for Campylobacter species. Campylobacter jejuni and Campylobacter coli are the two main species of campylobacters detected in chicken meat, while they account for almost 90% of the reported cases of campylobacteriosis in humans. Over 80% of these cases are attributed to C. jejuni and about 10% of them are due to C. coli. Therefore, until recently the dominance of C. jejuni against all other Campylobacter spp. isolated from chicken meat samples was well-established and unquestionable. Lately, however, C. coli has been increasingly recovered from chicken meat to such an extent that it is now evident that it often comprises the dominant species among the identified campylobacters in the meat samples. This work attempts for the first time a detailed review of the literature to deepen into this noteworthy epidemiological swift in the prevalence of C. jejuni and C. coli, along with the distribution of Campylobacter spp. in chicken meat. Factors such as the sampling method followed for screening campylobacters in broiler carcasses (e.g., swabs or carcass rinsates, skinned or skinless meat excised samples) and part of the animal carcass from which the sample is obtained (e.g., neck, breast, leg), seasonality of sampling (summer vs. winter) and environmental conditions (e.g., rainfall, relative humidity) at the farm level, the isolation procedure (enumeration or detection) and pathogen identification (biochemical or molecular), the enrichment and plating isolation media (e.g., Bolton vs. Preston broth, charcoal-based vs. chromogenic agars), as well as the biofilm-forming ability of different campylobacters, highlight the multivariate dimension of the phenomenon and are thoroughly discussed in the present review.

Distribution of virulence and antibiotic resistance genes in Campylobacter jejuni and Campylobacter coli isolated from broiler chickens in Tunisia.

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. 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. 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 blaOXA-61 (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. 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.

Protective effect of Lactiplantibacillus plantarum LP5 in a murine model of colonisation by Campylobacter coli DSPV458.

Thermotolerant Campylobacter species are the leading cause of foodborne bacterial diarrheal disease worldwide. Campylobacter coli, abundant in pigs and pork products, have been identified as a source of human infection. In this study, we propose the use of Lactiplantibacillus plantarum LP5 as a probiotic to reduce colonisation of this intestinal pathogen in a murine colonisation model of C. coli DSPV458. Six-week-old adult female Balb/cCmedc mice were housed in groups: Control, Campy and Pro-Campy. Control and Pro-Campy groups received antibiotics for 5 days and the Campy group for 12 days. Pro-Campy group was inoculated for 7 days with 8.78 log10 cfu total of L. plantarum LP5 suspended in De Man, Rogosa and Sharpe broth. All groups were inoculated with 6.72 log10 cfu of C. coli DSPV458 suspended in brain heart infusion broth. L. plantarum LP5 was recovered only in the Pro- Campy group. C. coli DSPV458 was recovered at higher levels in the Control and Campy groups. The differences with the Pro-Campy group were significant. As regards faeces, Control and Campy groups reached 7.41 and 7.84 log10 cfu/g, respectively, and the Pro-Campy group only 4.62 log10 cfu/g. In the caecum, Control and Campy groups reached 8.01 and 9.26 log10cfu/g, respectively, and the Pro-Campy group only 4.51 log10 cfu/g. In the ileum, Control and Campy groups reached 3.43 and 3.26 log10 cfu/g, respectively, and the Pro-Campy group did not show detectable levels. The reduction of C. coli DSPV458 in the Pro-Campy group compared to the Control group in faeces, caecum and ileum was 99.55, 99.98 and 100%, respectively. Animals were maintained under normal health conditions, and haematological parameters were within the standard values for Balb/cCmedc. The incorporation of a probiotic generated a protective effect in the mice colonisation model. The protective effect would also apply to intestinal colonisation by indigenous enterobacteria. Therefore, the strategy used in this study is of great importance to understand the protection mechanisms in a murine model, as well as its application in food-producing animals.

Detection of Campylobacter species in different types of samples from dairy farms

BackgroundLivestock, domestic pets and wildlife can be intestinal carriers of thermotolerant Campylobacter species. These reservoirs can in turn contaminate the environment and food products, thus creating pathways to campylobacteriosis in...

Ant(6)-I Genes Encoding Aminoglycoside O-Nucleotidyltransferases Are Widely Spread Among Streptomycin Resistant Strains of Campylobacter jejuni and Campylobacter coli.

Thermotolerant Campylobacter species C. jejuni and C. coli are actually recognized as the major bacterial agent responsible for food-transmitted gastroenteritis. The most effective antimicrobials against Campylobacter are macrolides and some, but not all aminoglycosides. Among these, susceptibility to streptomycin is reduced by mutations in the ribosomal RPSL protein or by expression of ANT(6)-I aminoglycoside O-nucleotidyltransferases. The presence of streptomycin resistance genes was evaluated among streptomycin-resistant Campylobacter isolated from humans and animals by using PCR with degenerated primers devised to distinguish ant(6)-Ia, ant(6)-Ib and other ant-like genes. Genes encoding ANT(6)-I enzymes were found in all possible combinations with a major fraction of the isolates carrying a previously described ant-like gene, distantly related and belonging to the new ant(6)-I sub-family ant(6)-Ie. Among Campylobacter isolates, ant(6)-Ie was uniquely found functional in C. coli, as shown by gene transfer and phenotype expression in Escherichia coli, unlike detected coding sequences in C. jejuni that were truncated by an internal frame shift associated to RPSL mutations in streptomycin resistant strains. The genetic relationships of C. coli isolates with ANT(6)-Ie revealed one cluster of strains presented in bovine and humans, suggesting a circulation pathway of Campylobacter strains by consuming contaminated calf meat by bacteria expressing this streptomycin resistance element.

Antimicrobial Resistance of Thermotolerant Campylobacter Species Isolated from Humans, Food-Producing Animals, and Products of Animal Origin: A Worldwide Meta-Analysis.

The objective of this meta-analysis was to summarize available information on the prevalence of antimicrobial-resistant Campylobacter species in humans, food-producing animals, and products of animal origin. A number of multilevel random-effect meta-analysis models were fitted to estimate mean occurrence rate of antimicrobial-resistant thermotolerant Campylobacter and to compare them throughout the years and among the species, food-producing animals (i.e., bovine, pigs, broilers, hen, goat, and sheep), country of origin, sample type, methodology to determine the antimicrobial susceptibility, and the species of Campylobacter. Among the considered antibiotics, thermotolerant Campylobacter showed the highest resistance to tetracycline (pool estimate [PE] = 0.493; 95% CI 0.466-0.519), nalidixic acid (PE = 0.385; 95% CI 0.348-0.423), and ciprofloxacin (PE = 0.376; 95% CI 0.339-0.415). In general, the prevalence of antimicrobial-resistant Campylobacter spp. was higher in hen, broilers, and swine. Campylobacter coli showed a higher prevalence of antimicrobial resistance than Campylobacter jejuni. Independent of the antimicrobial evaluated, the disk diffusion method showed higher prevalence of antimicrobial-resistant Campylobacter than the methods based on the minimum inhibitory concentration estimation. The meta-analysis showed that the prevalence of antimicrobial-resistant Campylobacter is relevant essentially in foods derived from hens and broilers, and it was observed worldwide. The prevalence of this pathogen is of public health importance and the increase in the prevalence of Campylobacter strains resistant to the antimicrobial of choice worsens the situation, hence, national authorities must monitor the situation in each country with the aim to establish the appropriate risk management measures.

Presencia de serotipos de Campylobacter jejuni O:19 en la cadena avícola Argentina

Thermotolerant species of Campylobacter have been focus of attention in the last years because they are the major agent causing zoonotic foodborne diseases. In addition, Campylobacter jejuni O:19 serotype was associated with Guillain Barré syndrome. The aim of this study was to determine the proportion of C. jejuni O:19 serotype isolated at different stages of three poultry meat supply chain in Santa Fe, Argentina. The analysis showed that 18% of isolated C. jejuni belong to serotype O:19. It was also determined that the presence of these strains is given in almost all production stages. These results reflect a significant risk to public health of consumers. Epidemiological studies of Campylobacter should be considered to establish a risk manager policy.

Recovery of Campylobacter spp. from Food and Environmental Sources.

The recovery of Campylobacter species from food and environmental sources is challenging due to the slow growth of these bacteria and the need to suppress competing organisms during the isolation procedures. The addition of multiple selective antimicrobials to growth media can negatively impact recovery of some Campylobacter spp. Here, we describe our current method for the isolation of thermotolerant Campylobacter species, mainly C. jejuni and C. coli, from food and environmental samples. We emphasize the use of membrane filtration during plating for the specific isolation of Campylobacter spp. and a reduced use of antimicrobial supplements throughout the whole isolation process.

Development of a monoclonal antibody-based colony blot immunoassay for detection of thermotolerant Campylobacter species

Campylobacter species, particularly thermotolerant Campylobacter spp., such as C. jejuni, are major human foodborne pathogens. Culture methods have been routinely used for the detection of this organism in various types of samples. An alternative, simple and rapid confirmation test(s) without further tedious biochemical tests would be useful. Meanwhile, Campylobacter-like colonies can be difficult to identify on agar plates overgrown with competitive bacteria, which can lead to false-negative results. This study was to develop a simple colony blot immunoassay using a new monoclonal antibody (Mab) produced in the present study for rapid screening, confirmation and quantification of campylobacters on culture agar plates. The procedure developed in this study was able to specifically detect thermotolerant Campylobacter spp., but not other non-thermotolerant Campylobacter and non-Campylobacter reference strains tested. This assay could detect 105 cells in a single dot. This assay showed 100% correlation with the culture method for the blotted membranes from 21 either chicken meat or vegetable samples experimentally inoculated with thermotolerant campylobacters. Among 101 natural samples of chicken meat (n=44), chicken feces (n=20) and vegetables (n=37), this assay also showed positive for 23 chicken meat and 14 fecal samples that were positive for thermotolerant campylobacters by culture method, and identified four additional suspects that were culture negative. Membranes stored at 4°C for at least 4years could also be used for this assay. The assay developed in this study can be used in quantitative study for immediate or archival usage, and for diagnostic test to preliminarily confirm the presence of thermotolerant Campylobacter on agar plates.

Concentrations of faecal coliforms, Escherichia coli, enterococci and Campylobacter spp. in equine faeces

AIMS: To determine the concentration of Campylobacter spp. as well as faecal indicator bacteria; faecal coliforms, Escherichia coli and enterococci in the faeces of healthy adult horses in a sample of properties in the Canterbury region of New Zealand.METHODS: The faeces of healthy adult horses (n=59), including ponies, pleasure horses and Thoroughbreds, were collected from eight properties around Christchurch, New Zealand. The faeces were analysed for concentrations of Campylobacter spp and faecal indicator bacteria; faecal coliforms, Escherichia coli and enterococci. The presence of other animals on the properties sampled as well as the age, feed and health of the horses at the time of sampling was recorded.RESULTS: Enterococci and faecal coliforms were isolated from all samples, and E. coli was isolated from 58/59 samples. Mean concentrations of faecal coliforms and E. coli did not differ between properties, but there was a significant difference in mean concentration of enterococci between properties. Campylobacter spp. were detected in two faecal samples with one isolate being determined by PCR analysis to be a thermotolerant Campylobacter species, the other C. jejuni.CONCLUSIONS: This is the first known report quantifying the concentration of Campylobacter spp. present in healthy adult horses in New Zealand. The presence of equine faecal material in water could elevate concentrations of faecal bacteria and therefore needs to be considered as a source of water contamination. The access of horses to waterways and coastal environments may also need to be restricted to prevent transmission of faecal indicator bacteria and potentially zoonotic agents.

Divergent Distribution of the Sensor Kinase CosS in Non-Thermotolerant Campylobacter Species and Its Functional Incompatibility with the Response Regulator CosR of Campylobacter jejuni

Two-component signal transduction systems are commonly composed of a sensor histidine kinase and a cognate response regulator, modulating gene expression in response to environmental changes through a phosphorylation-dependent process. CosR is an OmpR-type response regulator essential for the viability of Campylobacter jejuni, a major foodborne pathogenic species causing human gastroenteritis. Although CosR is a response regulator, its cognate sensor kinase has not been identified in C. jejuni. In this study, DNA sequence analysis of the cosR flanking regions revealed that a gene encoding a putative sensor kinase, which we named cosS, is prevalent in non-thermotolerant Campylobacter spp., but not in thermotolerant campylobacters. Phosphorylation assays indicated that C. fetus CosS rapidly autophosphorylates and then phosphorylates C. fetus CosR, suggesting that the CosRS system constitutes a paired two-component signal transduction system in C. fetus. However, C. fetus CosS does not phosphorylate C. jejuni CosR, suggesting that CosR may have different regulatory cascades between thermotolerant and non-thermotolerant Campylobacter species. Comparison of CosR homolog amino acid sequences showed that the conserved phosphorylation residue (D51), which is present in all non-thermotolerant Campylobacter spp., is absent from the CosR homologs of thermotolerant Campylobacter species. However, C. jejuni CosR was not phosphorylated by C. fetus CosS even after site-directed mutagenesis of N51D, implying that C. jejuni CosR may possibly function phosphorylation-independently. In addition, the results of cosS mutational analysis indicated that CosS is not associated with the temperature dependence of the Campylobacter spp. despite its unique divergent distribution only in non-thermotolerant campylobacters. The findings in this study strongly suggest that thermotolerant and non-thermotolerant Campylobacter spp. have different signal sensing mechanisms associated with the CosR regulation.

Human and animal Campylobacteriosis in Tanzania: A review.

The thermotolerant species of Campylobacter have become very important in public health, particularly as agents of infectious diarrhoea in human beings. Though the mechanism by which they cause disease is yet to be fully explained, they have been recognized as the leading cause of bacterial enteritis in both developed and developing countries. The organisms colonize different animal species without causing any symptoms of disease; and humans acquire infections through contact with or consumption of contaminated meat especially raw/undercooked poultry meat. The growing trend of antibiotic resistant Campylobacter isolates continues to pose significant public health challenges. In this review we present the available information generated in Tanzania about Campylobacter infections in humans and animals. We conducted a structured literature search of PUBMED and ScienceDirect electronic databases and identified 15 articles. Studies on humans reported Campylobacter infections in both symptomatic and asymptomatic subjects; with higher prevalence in children under the age of five years. Studies on animals found colonization of both domestic and wild species. Among isolates, some demonstrated antimicrobial resistance. The available information for both human and animal Campylobacteriosis in the country is sparse. It however provides an insight of the bacteriological and epidemiological aspects of Campylobacter infections in the country and eventually creates more awareness on the need to develop control strategies. Since the organism is zoonotic its control strategies should adopt the "One Health" approach involving collaborative efforts from veterinary and human medicine.

In vivo broiler experiments to assess anti-Campylobacter jejuni activity of a live Enterococcus faecalis strain

Bacterial gastroenteritis caused by thermotolerant Campylobacter species, mainly Campylobacter jejuni, has been the most reported zoonotic disease in many developed countries in recent years. Reducing Campylobacter shedding on the farm could result in a reduction of the number of campylobacteriosis cases. In 2 independent broiler seeder experiments, in which broiler chickens were orally inoculated with 2 amounts of Enterococcus faecalis MB 5259, we established whether a live E. faecalis strain was capable of reducing cecal Campylobacter colonization in broiler chickens. In previous in vitro experiments it has been demonstrated that this E. faecalis MB 5259 displays anti-Campylobacter activity. The effect of pH and bile salts on E. faecalis MB 5259 showed that growth and survival of E. faecalis MB 5259 can be impaired during passage through the gastrointestinal tract of broiler chickens. Despite these results E. faecalis MB 5259 was capable of colonizing the broiler ceca. Contrary to the in vitro experiments, in which E. faecalis MB 5259 inhibited C. jejuni MB 4185 growth, no inhibition was observed in the in vivo experiments independent of the inoculum size.

GyrB versus 16s rRna sequencing for the identification of Campylobacter jejuni, Campylobacter coli, and Campylobacter lari

Species of the genus Campylobacter are responsible for the largest number of bacterial food-borne illness cases occurring yearly in the developed world. The majority of disease is caused by three of the thermotolerant Campylobacter species: Campylobacter jejuni, Campylobacter coli, and Campylobacter lari. An inability to differentiate these three species using the commonly employed 16S rRNA sequencing procedure has led to the development of alternative methods to identify these bacteria. Some of these methods include the utilization of the gyrB gene. A reliable method was developed for the differentiation of C. jejuni, C. coli, and C. lari employing the gyrB gene. It involves amplification and sequencing of a species-variable region of the gene with a single pair of DNA primers. The method works well for the separation and organization of the three Campylobacter strains as well as satisfying the suggested guidelines for sequence based identification for most strains investigated.

GyrB versus 16s rRna sequencing for the identification of Campylobacter jejuni, Campylobacter coli, and Campylobacter lari

Species of the genus Campylobacter are responsible for the largest number of bacterial food-borne illness cases occurring yearly in the developed world. The majority of disease is caused by three of the thermotolerant Campylobacter species: Campylobacter jejuni, Campylobacter coli, and Campylobacter lari. An inability to differentiate these three species using the commonly employed 16S rRNA sequencing procedure has led to the development of alternative methods to identify these bacteria. Some of these methods include the utilization of the gyrB gene. A reliable method was developed for the differentiation of C. jejuni, C. coli, and C. lari employing the gyrB gene. It involves amplification and sequencing of a species-variable region of the gene with a single pair of DNA primers. The method works well for the separation and organization of the three Campylobacter strains as well as satisfying the suggested guidelines for sequence based identification for most strains investigated.

Campylobacter y campylobacteriosis: una mirada desde América del Sur

The thermotolerant species of Campylobacter have become very important in public health, especially as agents of infectious diarrhea in human beings. In this brief revision we present part of the available information generated in South America about epidemiological, clinical and bacteriological aspects of campylobacteriosis and we identify some differences between the observed and documented campylobacteriosis in South America compared to those described in industrialized countries.

Campylobacter y campylobacteriosis: una mirada desde América del Sur

The thermotolerant species of Campylobacter have become very important in public health, especially as agents of infectious diarrhea in human beings. In this brief revision we present part of the available information generated in South America about epidemiological, clinical and bacteriological aspects of campylobacteriosis and we identify some differences between the observed and documented campylobacteriosis in South America compared to those described in industrialized countries.

Occurrence of thermotolerant Campylobacter species in surface waters of a Mediterranean area and in its prevailing pollution sources

To determine the prevalence of Campylobacter in surface waters of a highly populated Mediterranean area. Surface water and wastewater samples were collected from an area in the north-east of Spain during a 2-year study. All the samples were analysed using the MPN method and Multiplex PCR to quantify and identify Campylobacter. It was detected in 82% of the samples from the Llobregat River with a mean of 1·3 MPN 100 ml(-1). The lowest counts were obtained in summer. Campylobacter coli was the predominant species in this river. The bacteria were isolated from marsh water but not from seawater samples. The highest counts of campylobacters were found in poultry wastewater where Camp. jejuni was the predominant species, as in urban sewage. In pig slurry, Camp. coli was the only species detected. Campylobacter jejuni and Camp. coli are present and widely distributed in the surface water of the studied area. The two species co-exist, with Camp. coli being predominant. In river water, campylobacter counts presented a seasonal distribution. No relationship with faecal indicators was found. This study provides the first data on the occurrence and concentrations of thermotolerant campylobacter species in surface water in a Mediterranean area.

Enhanced enrichment and detection of thermotolerant Campylobacter species from water using the Portable Microbe Enrichment Unit and real-time PCR

An enhanced enrichment using the Portable Microbe Enrichment Unit (PMEU) with the microaerobic bubbling of broths was applied for the detection of thermotolerant Campylobacter species from water. This PMEU enrichment was compared with the conventional static enrichment of the international standard ISO 17995:2005. In addition, Campylobacter detection after enrichment using a real-time PCR detection was compared with colony counts. The tests with stressed Campylobacter jejuni cells in drinking water indicated that the PMEU enrichment yielded a significantly higher number of Campylobacter cells in the Bolton broth compared with the conventional static incubation. Application of the real-time PCR technique shortened the Campylobacter detection time. This combination of method modifications can be used for Campylobacter detection from water and adds methodological repertoire for the rapid survey and management of waterborne outbreaks.