Thermophilic Campylobacter Species Research Articles

Role of Maternal Antibodies in the Protection of Broiler Chicks against Campylobacter Colonization in the First Weeks of Life.

Thermophilic Campylobacter species are the most common cause of bacterium-mediated diarrheal disease in humans globally. Poultry is considered the most important reservoir of human campylobacteriosis, but so far, no effective countermeasures are in place to prevent the bacterium from colonizing broiler flocks. This study investigated maternal antibodies' potential to offer protection against Campylobacter in broiler chicks via a field trial and an immunization trial. In the field trial, breeder flocks with high and low anti-Campylobacter antibody levels in the yolk were selected based on serological screening. Offspring were subsequently monitored for maternal antibodies and Campylobacter prevalence during early life. Although maternal antibodies declined rapidly in the serum of broilers, offspring from flocks with lower anti-Campylobacter antibody levels seemed to be more susceptible to colonization. In the immunization trial, breeders from a seropositive breeder flock were vaccinated with an experimental bacterin or subunit vaccine. Immunization increased antibody levels in the yolk and consequently in the offspring. Elevated maternal antibody levels were significantly associated with reduced Campylobacter susceptibility in broilers at 2 weeks old but not at 1 and 3 weeks old. Overall, the protective effect of maternal immunity should be cautiously considered in the context of Campylobacter control in broilers. Immunization of breeders may enhance resistance but is not a comprehensive solution.

Detection of multidrug-resistant Campylobacter species from food-producing animals and humans in Nigeria: Public health implications and one health control measures

Antimicrobial-resistant thermophilic Campylobacter species (TCS) pose tremendous public health problems because they are zoonotic, difficult to treat and usually harboured by food-producing animals (FPAs). This study ascertained the phenotypic antimicrobial resistance (AMR) in 56 phenotypically identified TCS from slaughtered cattle, poultry, and humans in Enugu State, Nigeria. The presence of selected AMR and virulence genes harboured by the animal and human isolates were also detected and compared in 36 PCR-confirmed Campylobacter species. All the 56 TCS were multidrug-resistant as none were susceptible to ampicillin, penicillin-G, amoxicillin-clavulanic acid, cephalothin and metronidazole. The isolates were 92.9 %, 62.5 %, 92.9 %, 42.9 %, 26.8 %, 25 %, 28.6 %, 53.7 %, 30.1 %, 32.1 % and 55.4 % resistant to ceftriaxone, nalidixic acid, cefotaxime, enrofloxacin, ciprofloxacin, streptomycin, gentamycin, erythromycin, azithromycin, chloramphenicol and tetracycline, respectively. The top four most effective classes of antimicrobials were aminoglycosides > macrolides > amphenicol > fluoroquinolones. The AMR genes detected and the percentage of the isolates that harboured them were: aadE-1 (33.3 %), aphA-3–1 (36.1 %), tetO (44.4%), Blaoxa-61 (61.1 %) and the multidrug efflux pump, cmeB (86.1%). Virulence genes detected and the corresponding percentage of TCS that harboured them were: cdtB (61.1 %), flaA (47.2 %), ciaB (38.9 %), and pldA (38.9 %). The cmeB was significantly detected in animal isolates (p = 0.018, OR = 5.1, CI = 0.7–6.6) while BlaOXA-61 predominated in human isolates (p = 0.019, OR = 6.2). Likewise, ciaB virulence gene was mostly detected (p = 0.019, OR = 6.4, CI = 1.3–25) in animal isolates. The findings underscore the roles of FPAs in the zoonotic dissemination of Campylobacter-associated AMR and virulence genes in the study area. This warrants the adoption of One Health control strategies to limit spread of the multidrug-resistant zoonotic Campylobacter species.

Whole genome characterization of thermophilic Campylobacter species isolated from dairy manure in small specialty crop farms of Northeast Ohio.

With more public interest in consuming locally grown produce, small specialty crop farms (SSCF) are a viable and growing segment of the food production chain in the United States. The goal of this study was to investigate the genomic diversity of Campylobacter isolated from dairy manure (n = 69) collected from 10 SSCF in Northeast Ohio between 2018 and 2020. A total of 56 C. jejuni and 13 C. coli isolates were sequenced. Multi-locus sequence typing (MLST) identified 22 sequence types (STs), with ST-922 (18%) and ST-61 (13%) predominant in C. jejuni and ST-829 (62%) and ST-1068 (38%) predominant in C. coli. Interestingly, isolates with similar genomic and gene contents were detected within and between SSCF over time, suggesting that Campylobacter could be transmitted between farms and may persist in a given SSCF over time. Virulence-associated genes (n = 35) involved in the uptake and utilization of potassium and organic compounds (succinate, gluconate, oxoglutarate, and malate) were detected only in the C. jejuni isolates, while 45 genes associated with increased resistance to environmental stresses (capsule production, cell envelope integrity, and iron uptake) were detected only in the C. coli isolates. Campylobacter coli isolates were also sub-divided into two distinct clusters based on the presence of unique prophages (n = 21) or IncQ conjugative plasmid/type-IV secretion system genes (n = 15). Campylobacter coli isolates harbored genes associated with resistance to streptomycin (aadE-Cc; 54%) and quinolone (gyrA-T86I; 77%), while C. jejuni had resistance genes for kanamycin (aph3'-IIIa; 20%). Both species harbored resistance genes associated with β-lactam (especially, blaOXA-193; up to 100%) and tetracycline (tetO; up to 59%). Our study demonstrated that Campylobacter genome plasticity associated with conjugative transfer might provide resistance to certain antimicrobials and viral infections via the acquisition of protein-encoding genes involved in mechanisms such as ribosomal protection and capsule modification.

Campylobacter spp. isolated from poultry in Iran: Antibiotic resistance profiles, virulence genes, and molecular mechanisms.

Campylobacter spp. genera is one of the most common causes of microbial enteritis worldwide. The objective of this work was to investigate the antimicrobial resistance (AMR) patterns, virulence genes, and genetic variation of thermophilic Campylobacter species collected from chicken meat samples in Iran. A total of 255 meat specimens were taken and transferred to the laboratory. Culture methods were utilized to identify the Campylobacter genus, and PCR and sequencing were performed to confirm the organisms. Antimicrobial susceptibility evaluation was performed using broth microdilution for six antimicrobials [ciprofloxacin (CIP), nalidixic acid (NAL), sitafloxacin (SIT), erythromycin (ERY), tetracycline (TET), and gentamicin (GEN)]. By using PCR, AMR and virulence genes were detected. The detection rate of Campylobacter spp. was 64 (25.09%) out of 255 meat samples, with C.jejuni and C.coli accounting for 41 (64.06%) and 14 (21.87%), respectively. Other Campylobacter isolates accounted for 14.06% of the total (nine samples). The antibiotic susceptibility of all Campylobacter isolates was tested using six antibiotics, and all (100%) were resistant to CIP and NAL. However, TET resistance was observed in 93.9% and 83.3% of C.jejuni and C.coli isolates, respectively. Four (8.2%) C.jejuni isolates were multidrug-resistant (MDR), while none of the C.coli isolates were MDR. Two of the four MDR isolates were resistant to CIP, NAL, TET, and ERY, whereas the other two isolates were resistant to CIP, NAL, TET, and GEN. The values of the Minimum Inhibitory Concentration (MIC) were as follows: CIP, 64-256 μg/ml; NAL, 128-512 μg/ml; TET, 2-1024 μg/ml; SIT, 0.25-1μg/ml; ERY, 1-32 μg/ml; and GEN, 1-256 μg/ml. recR, dnaJ, cdtC, cdtB, cdtA, flaA, ciaB, cadF, and pidA were discovered in more than 50% of C.jejuni isolates, although wlaN, virbll, cgtB, and ceuE were found in <50%. flaA, cadF, pidA, and ciaB were discovered in more than 50% of the C.coli samples, whereas recR, cdtC, cdtB, cdtA, and cgtB were found in less than half. For C.coli, the percentages for wlaN, dnaJ, virbll, and ceuE were all zero. The results of this study show Campylobacter isolates obtained from poultry have higher resistance to quinolones and TET, pathogenicity potential, and varied genotypes.

Isolation, identification and antibiotic resistance profile of thermophilic Campylobacter species from Bovine, Knives and personnel at Jimma Town Abattoir, Ethiopia.

Thermophilic Campylobacter species are common cause of animal and human bacterial diseases with growing resistance to antimicrobials. The aim of this study was to determine the prevalence and antimicrobial susceptibility pattern of Campylobacter species from bovine, knives and personnel in Jimma Town, Ethiopia. Faecal samples and carcasses swabs were collected from cattle systematically selected from the annual plan of Jimma Municipal Abattoir. Personnel hand and knife swabs were collected after slaughtering each selected cattle. A cross-sectional study with systematic sampling method was conducted from October 2019 to September 2020 for the isolation, identification and antimicrobial susceptibility pattern of thermophilic Campylobacter species. Isolation and identification of Campylobacter species were performed according to the techniques recommended by the International Organization for Standardization, and in vitro antibiotic susceptibility testing was screened using the standard agar disc diffusion method as recommended by Clinical and Laboratory Standards Institutions. A total of 684 samples (171 samples from faeces, carcasses, knives and personnel hands, were collected independently). The overall prevalence of thermophilic Campylobacterspecies was 5.6% (38/684). Majority of the isolates were from faecal samples (12.9%, n = 22) followed by carcass swabs(4.1% n = 7), knife swabs(3.5% n = 6) and personnel hand swabs(1.8% n = 3). Isolated and identified species of C.jejuni, C. coli and C. lari accounted for 63.2%, 23.7% and 13.2%, respectively. The isolated Campylobacter species were found to be resistant to Cephalothin (100%), Ampicillin (60.5%), Cefotaxime (60.5%), Chloramphenicol (47.4%) and Tetracycline (42.1%). On the other hand, the isolates were susceptible to Nalidixic acid (86.8%), Ciprofloxacin (86.8%), Sulphamethazole (84.2%), Ceftriaxone (78.9%), Clindamycin (68.4%) and Cefixime (65.8%). 84.2% of the isolates showed multi-drug resistance for three-to-six drug classes. All the C. lari isolates were multidrug resistant. All the three isolated species of Campylobacter were resistant to Cephalothin, and most were multidrug resistant. Isolation of Campylobacter species from faecal, carcass, knife and hand swabs revealed possible risk of contamination and exposure to Campylobacter infection of those who consume raw meat. Therefore, enactment of hygienic practices during the slaughtering process, proper handling and cooking of meat and awareness creation on jurisdictional antibiotic usage are required to avoid Campylobacter infection.

Seasonal Prevalence and Molecular Identification of Thermophilic Campylobacter from Chicken, Cattle, and Respective Drinking Water in Kajiado County, Kenya.

Thermophilic Campylobacter species are a leading cause of human gastroenteritis throughout the world and have been implicated in reproductive disorders (abortion), mastitis, enteritis, and/or diarrhoea in livestock. A cross-sectional survey was conducted in Kajiado County to determine prevalence, seasonality, and molecular detection of thermophilic Campylobacter species (with emphasis on C. jejuni, C. coli, and other thermophilic Campylobacter species) in chicken, cattle, and respective pooled drinking water. A total of 457 samples comprising 265 cattle rectal swabs, 142 chicken cloacal swabs, and 50 trough water samples were collected from 55 randomly selected smallholder farms. Individual samples were subjected to standard techniques for isolation and biochemical tests, followed by singleplex polymerase chain reaction (sPCR) assays for identification and confirmation of genus and species. Overall, thermophilic Campylobacter prevalence was 35.4% (95% confidence interval (95% CI) = 31.0–39.8), with C. jejuni dominating at 55.6% (95% CI = 47.9–63.3%) over C. coli in all sample types. The highest thermophilic Campylobacter prevalence was observed in cloacal swabs of live chicken at 44.4% (95% CI = 36.2–52.6%), followed by rectal swabs from live cattle at 30.9% (95% CI = 25.3–36.5%). Water samples from cattle drinkers/trough were found to be contaminated at 34% (95% CI = 20.9–47.1%). The isolation rate was higher in cattle under the confinement system (44.3%) (95% CI = 36.1–52.5%) than in those under the free-roaming grazing system. Thermophilic Campylobacter species were isolated in both seasons, with higher prevalence (39.8% (95% CI = 33.6–45.9)) recorded during rainy and cold season in all sample types except for water. There was significant (P < 0.05) association between season and thermophilic Campylobacter occurrence, even though there were no statistical differences in the prevalence values across the two seasons. Results of this study demonstrate that cattle, chicken, and respective drinking water harbour potentially pathogenic thermophilic campylobacters, with C. jejuni being widely distributed among farms. It is possible that seasonal variations and cattle confinement result in differences in thermophilic Campylobacter carriage. Further epidemiological and phylogenetic studies comparing distribution of thermophilic Campylobacter spp. isolates in livestock, environmental, and human samples are recommended to establish source attribution to reduce the impact of resultant diseases for the wellbeing of public and livestock.

Quantitative Assessment of First Nations Drinking Water Distribution Systems for Detection and Prevalence of Thermophilic Campylobacter Species.

Water is considered a major route for transmitting human-associated pathogens. Although microbial water quality indicators are used to test for the presence of waterborne pathogens in drinking water, the two are poorly correlated. The current study investigates the prevalence of thermophilic DNA markers specific for Campylobacter spp. (C. jejuni and C. coli) in source water and throughout the water distribution systems of two First Nations communities in Manitoba, Canada. A total of 220 water samples were collected from various points of the drinking water distribution system (DWDS) between 2016 and 2018. Target Campylobacter spp. were always (100%) detected in a home with a fiberglass (CF) cistern, as well as the community standpipe (SP). The target bacteria were also frequently detected in treated water at the Water Treatment Plant (WTP) (78%), homes with polyethylene (CP) (60%) and concrete (CC) (58%) cisterns, homes with piped (P) water (43%) and water truck (T) samples (20%), with a maximum concentration of 1.9 × 103 cells 100 mL−1 (C. jejuni) and 5.6 × 105 cells 100 mL−1 (C. coli). Similarly, target bacteria were detected in 68% of the source water samples with a maximum concentration of 4.9 × 103 cells 100 mL−1 (C. jejuni) and 8.4 × 105 cells 100 mL−1 (C. coli). Neither target Campylobacter spp. was significantly associated with free and total chlorine concentrations in water. The study results indicate that there is an immediate need to monitor Campylobacter spp. in small communities of Canada and, particularly, to improve the DWDS in First Nations communities to minimize the risk of Campylobacter infection from drinking water sources. Further research is warranted in improving/developing processes and technologies to eliminate microbial contaminants from water.

The temporal pattern and relationship of Campylobacter prevalence in broiler slaughter batches and human campylobacteriosis cases in Sweden 2009–2019

Thermophilic Campylobacter species are the most common cause of bacterial gastroenteritis worldwide, and handling and consumption of broiler meat is considered a major foodborne transmission route. Both the incidence of campylobacteriosis and the prevalence of Campylobacter in broilers show seasonality but the impact of this association and broiler prevalence on human incidence is not clear. To explore this relationship we applied two approaches for analyzing time series data using different time resolutions (weekly, bi-weekly or monthly data) of human campylobacteriosis cases and prevalence of Campylobacter spp. in broiler slaughter batches in Sweden between 2009 and 2019. The decomposition of time series into seasonal (S), long-term trend (T) and residual components (STL model) showed a close overlap in seasonal patterns in terms of timing and the proportional change of peaks from normalized yearly levels. Starting 2016, when a large outbreak was reported, there was significant overlap in the trend components as well. The trend component of human cases prior to the outbreak corresponded to a linear increase of 6.5 % cases annually. In comparison, the estimated annual increase in broiler consumption was 2.7 %. An additive approach for time-series counts incorporating seasonal and epidemic (cases are a function of previous cases) components found a positive association between human cases and broiler prevalence with an optimal lag of 2 weeks, 1 bi-week, or 0 months. Considering the estimated time between slaughter and consumption, incubation time, and the time between on-set of disease and testing, a 2-week lag may be consistent with transmission via handling and consumption of fresh broiler meat. The best model included broiler prevalence as a factor in the epidemic model component, not in the seasonal component. The outcomes in terms of best model, optimal lags and significance of parameters, using weekly, bi-weekly or monthly data were, in general, in agreement but varied with data resolution when only a subset of the time series, not including any known broiler associated outbreaks, was analyzed. The optimal resolution based on the available data and conditions of the present analysis appeared to be weekly or bi-weekly data. Results suggest that broiler prevalence with a 2 week lag period can explain part of the human cases but has a smaller explanatory impact during the part of the study period not including the large known outbreaks. There is no simple relationship between broiler prevalence and human cases. Additional factors than broiler prevalence need to be evaluated in order to understand the transmission routes and epidemiology of campylobacteriosis.

Genomic diversity and antimicrobial resistance of Campylobacter spp. from humans and livestock in Nigeria

BackgroundCampylobacter spp. are zoonotic pathogens, ubiquitous and are found naturally as commensals in livestock from where they can be transmitted to humans directly or through animal products. The genomic diversity and antimicrobial resistance profile of Campylobacter was investigated with a focus on C. jejuni and C. coli in humans and livestock (poultry and cattle) from Nigeria.Methods586 human stool samples and 472 faecal samples from livestock were cultured for thermophilic Campylobacter species on modified charcoal cefoperazone deoxycholate agar (mCCDA). Culture in combination with whole genome sequencing identified and confirmed the presence of Campylobacter in humans and animals from the study area. Further analysis of the sequences was performed to determine multilocus sequence types and genetic determinants of antimicrobial resistance to fluoroquinolone, betalactam, tetracycline and macrolide classes of antimicrobials.ResultsFrom the human stool samples tested, 50 (9%) were positive of which 33 (66%) were C. jejuni, 14 (28%) were C. coli while 3 (6%) were C. hyointestinalis. In livestock, 132 (28%) were positive. Thirty one (7%) were C. jejuni while 101 (21%) were C. coli. Whole genome sequencing and MLST of the isolates revealed a total of 32 sequence types (STs) identified from 47 human isolates while 48 STs were identified in 124 isolates from livestock indicating a population which was overall, genetically diverse with a few more dominant strains. The antimicrobial resistance profiles of the isolates indicated a higher prevalence of resistance in Campylobacter isolated from livestock than in humans. Generally, resistance was greatest for betalactams (42%) closely followed by fluoroquinolones (41%), tetracyclines (15%) and lastly macrolides (2%). Multidrug resistance to three or more antimicrobials was observed in 24 (13%) isolates from humans (n = 1, 4%) and chicken (n = 23, 96%).ConclusionsThis study has further contributed information about the epidemiology, genetic diversity and antimicrobial resistance profile of thermophilic Campylobacter in Nigeria.

Antibiotic resistance patterns of chicken and human origin Campylobacter spp. in Turkey

Campylobacteriosis is of great importance for both human and chicken populations. Unconcious and overuse of antibiotics in chickens has led to the transmission of antibiotic resistance patterns to humans. The purpose of this study was to determine the prevalence of thermophilic Campylobacter species from the cecal samples at slaughter houses, and also common antibiotic resistance patterns shared between chicken origin and human origin thermophilic Campylobacter species. Isolation and identification was performed according to EN ISO 10272-1: 2017 and Real-Time Multiplex qPCR, respectively. Antibiotic susceptibility test was performed by using the Kirby Bauer Disk Diffusion Method. Of the examined randomly collected 180 cecal samples at evisceration stage in slaughterhouses, 19 (10.5%), 17 (9.44%) and 2 (1.11%) were found to harbour Campylobacter spp., C. jejuni and C. coli, respectively. The highest resistance was determined against quinolones (86.04%) and fluoroquinolones (86.04%) among the tested 43 Campylobacter spp., comprising 19 chicken origin and 24 human origin. Except for erythromycin and gentamicin, all C. jejuni isolates from chickens and humans were found to be resistant to two or three of the antibiotics tested. The same multidrug resistance profiles observed in chicken origin C. jejuni isolates for TET/CIP/NA (70.58%) and CIP/NA (29.41%) were also determined in human origin C. jejuni isolates with the rate of 25% and 50%, respectively for each. To sum up, the same resistance patterns against common antibiotics shared in both human and chicken origin C. jejuni has pose a significant public health problem.

Isolation and characterization of thermophilic Campylobacter species from geese raised in Kars region (Turkey) using cultural, molecular and mass spectrometry methods

Background:Thermophilic Campylobacters are found in the digestive tract of wild and domestic poultry and can be transmitted to humans following their fecal discharges. Aims:This study aimed to isolate thermophilic Campylobacter by culture from cloacal swabs of geese, commonly breeding in Kars region, and to identify the isolates by PCR and mass spectrometry. Antibiotics susceptibility and resistance genes of the isolates were also analysed. Methods:The study included 400 cloacal swab samples of clinically healthy geese. The samples were cultured on mCCDA medium following the pre-enrichment in Preston broth. Identification of the isolates was performed by phenotypic methods, PCR, and MALDI-TOF MS. Antibiotic susceptibility and resistance genes of the isolates were analysed with the disc diffusion method and PCR, respectively. Results:Thermophilic Campylobacter spp. were isolated from 157 (39.3%) samples. 151 (96.2%) isolates were identified Campylobacter jejuni and 6 (3.8%) Campylobacter coli by the phenotypic tests and PCR. Among 125 isolates analysed by MALDI-TOF MS, 119 (95.2%) were identified C. jejuni and 6 (4.8%) C. coli. The isolates’ resistance to ampicillin, tetracycline, ciprofloxacin, gentamicin, and azithromycin were found 33.8%, 41.4%, 75.2%, 12.1%, and 7.6%, respectively. The distributions of blaOXA61, tetO, gyrA, and aphA-3 genes were 3.2%, 90.8%, 50.8%, and 52.7%, respectively. Conclusion:Since geese are raised in pastures in the Kars region, protecting and not polluting the existing natural environment and preventing their contact with wild birds will prevent the spread of these microorganisms.

Isolation and Characterization of Group III Campylobacter jejuni-Specific Bacteriophages From Germany and Their Suitability for Use in Food Production.

Campylobacter spp. are a major cause of bacterial foodborne diarrhea worldwide. While thermophilic Campylobacter species asymptomatically colonize the intestines of chickens, most human infections in industrial countries have been attributed to consumption of chicken meat or cross-contaminated products. Bacteriophages (phages) are natural predators of bacteria and their use at different stages of the food production chain has been shown to reduce the public health burden of human campylobacteriosis. However, regarding regulatory issues, the use of lytic phages in food is still under discussion and evaluation. This study aims to identify lytic phages suitable for reducing Campylobacter bacteria along the food production chain. Therefore, four of 19 recently recovered phages were further characterized in detail for their lytic efficacy against different Campylobacter field strains and their suitability under food production settings at different temperatures and pH values. Based on the results of this study, the phages vB_CjM-LmqsCP1-4 and vB_CjM-LmqsCP1-5 appear to be promising candidates for the reduction of Campylobacter jejuni in food production settings.

Prevalence, antibiotic spectrum and genetic relatedness in thermophilic Campylobacter species from poultry production environment of Punjab

Campylobacter is responsible for human gastroenteritis worldwide. C. jejuni and C. coli are most frequently encountered in animals, birds and man, but mainly C. jejuni is particularly adapted to poultry. The disease in humans ranges from gastroenteritis to Guillain-Barre Syndrome. To know the status of Campylobacter species especially, C. jejuni in poultry environment in Punjab, 342 poultry fecal samples (228 from layers and 114 from broilers) were collected from 30 poultry farms of Punjab. Additionally, 27 fecal samples from poultry farm workers were also collected. Analysis of the samples revealed that 21 (6.14%) fecal samples (from poultry farm) and two (7.41%) from farm workers were positive for Campylobacters based on cultural and molecular detection. Out of 23 Campylobacter positive isolates, 19 (80.95%) were C. jejuni and 4 (19.04%) were C. coli. Antibiotic resistance in the isolates was low. The majority of the isolates were sensitive to macrolide and quinolone class of antibiotics which are important for the treatment of campylobacteriosis. Pulse Field Gel Electrophoresis (PFGE) showed high genetic diversity among C. jejuni, however, it was limited in C. coli isolates. Poultry production in Punjab poses a potential risk of campylobacteriosis. However, antibiotic resistance in the isolates was low.

Occurrence, Risk Factors, and Antimicrobial Susceptibility Test of Thermophilic Campylobacter Species of Bovine Carcass at Municipal Abattoir and Butcher Shops of Jimma Town, Southwest Ethiopia.

BackgroundAlthough Campylobacter jejuni (C. jejuni) and Campylobacter coli (C. coli) cause acute diarrheal diseases in people all over the world, they are most commonly seen in other mammalian species and are a seemingly healthy carrier condition. Humans in Ethiopia, on the other hand, are largely unaware of the existence of Campylobacter in food animals as potential sources of infection.ObjectiveTo determine the occurrence, risk factors, and antimicrobial susceptibility pattern of thermophilic Campylobacter species in bovine raw meat from the abattoir and butcher shops.MethodsSwab samples were collected from 177 cattle carcasses (from abattoir 93 and butcher shops 84) and cultured using standard methods. An antimicrobial susceptibility test was performed using the disk diffusion method towards eleven antimicrobial agents.ResultsThe overall prevalence of thermophilic Campylobacter species was 14 (7.9%). From these, 11 (78.6%) were identified to be C. jejuni and 3 (21.4%) were C. coli. Lack of handwashing before meat processing and after visiting the toilet, meat contact with floors, walls, or soiling during preparation, and lack of training were the most important factors independently associated with (p<0.05) the prevalence of Campylobacter species contamination. The highest level of antimicrobial resistance of the Campylobacter isolates was recorded to ampicillin (10μg) (100%), followed by amoxicillin (30μg) (78.6%) and sulphamethoxazole-trimethoprim (57.1%) while the least resisted antimicrobials were streptomycin (25μg), erythromycin (15μg), oxytetracycline (30μg) (each 28.6%), kanamycin (30μg) 14.3%, chloramphenicol (30μg) and gentamycin (10μg) (each 7.1%).ConclusionDespite the low prevalence of thermophilic Campylobacter in the current investigation, it may pose a significant public health threat. As a result, it is vital to give retailers and customers extensive education, training, and knowledge about the correct handling and cooking of animal-derived goods. Furthermore, antimicrobials should be used with caution in both veterinary and human treatment regimens as well as a wider examination of antimicrobial resistance patterns for the use of well-targeted antimicrobials.

Genomic Characterization of Fluoroquinolone-Resistant Thermophilic Campylobacter Strains Isolated from Layer Chicken Feces in Gangneung, South Korea by Whole-Genome Sequencing.

Thermophilic Campylobacter species of poultry origin have been associated with up to 80% of human campylobacteriosis cases. Layer chickens have received less attention as possible reservoirs of Campylobacter species. Initially, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of two archived Campylobacter isolates (Campylobacter jejuni strain 200605 and Campylobacter coli strain 200606) from layer chickens to five antimicrobials (ciprofloxacin, nalidixic acid, erythromycin, tetracycline, and gentamicin) were determined using broth microdilution while the presence of selected antimicrobial resistance genes was performed by polymerase chain reaction (PCR) using specific primers. Whole-genome sequencing (WGS) was performed by the Illumina HiSeq X platform. The analysis involved antimicrobial resistance genes, virulome, multilocus sequence typing (MLST), and phylogeny. Both isolates were phenotypically resistant to ciprofloxacin (MIC: 32 vs. 32 µg/mL), nalidixic acid (MIC: 128 vs. 64 µg/mL), and tetracycline (MIC: 64 vs. 64 µg/mL), but sensitive to erythromycin (MIC: 1 vs. 2 µg/mL) and gentamicin (MIC: 0.25 vs. 1 µg/mL) for C. jejuni strain 200605 and C. coli strain 200606, respectively. WGS confirmed C257T mutation in the gyrA gene and the presence of cmeABC complex conferring resistance to FQs in both strains. Both strains also exhibited tet(O) genes associated with tetracycline resistance. Various virulence genes associated with motility, chemotaxis, and capsule formation were found in both isolates. However, the analysis of virulence genes showed that C. jejuni strain 200605 is more virulent than C. coli strain 200606. The MLST showed that C. jejuni strain 200605 belongs to sequence type ST-5229 while C. coli strain 200606 belongs to ST-5935, and both STs are less common. The phylogenetic analysis clustered C. jejuni strain 200605 along with other strains reported in Korea (CP028933 from chicken and CP014344 from human) while C. coli strain 200606 formed a separate cluster with C. coli (CP007181) from turkey. The WGS confirmed FQ-resistance in both strains and showed potential virulence of both strains. Further studies are recommended to understand the reasons behind the regional distribution (Korea, China, and Vietnam) of such rare STs.

Antimicrobial Resistance, FlaA Sequencing, and Phylogenetic Analysis of Campylobacter Isolates from Broiler Chicken Flocks in Greece.

Human campylobacteriosis caused by thermophilic Campylobacter species is the most commonly reported foodborne zoonosis. Consumption of contaminated poultry meat is regarded as the main source of human infection. This study was undertaken to determine the antimicrobial susceptibility and the molecular epidemiology of 205 Campylobacter isolates derived from Greek flocks slaughtered in three different slaughterhouses over a 14-month period. A total of 98.5% of the isolates were resistant to at least one antimicrobial agent. In terms of multidrug resistance, 11.7% of isolates were resistant to three or more groups of antimicrobials. Extremely high resistance to fluoroquinolones (89%), very high resistance to tetracycline (69%), and low resistance to macrolides (7%) were detected. FlaA sequencing was performed for the subtyping of 64 C. jejuni and 58 C. coli isolates. No prevalence of a specific flaA type was observed, indicating the genetic diversity of the isolates, while some flaA types were found to share similar antimicrobial resistance patterns. Phylogenetic trees were constructed using the neighbor-joining method. Seven clusters of the C. jejuni phylogenetic tree and three clusters of the C. coli tree were considered significant with bootstrap values >75%. Some isolates clustered together were originated from the same or adjacent farms, indicating transmission via personnel or shared equipment. These results are important and help further the understanding of the molecular epidemiology and antimicrobial resistance of Campylobacter spp. derived from poultry in Greece.

Antimicrobial Resistance Profiles, Virulence Genes, and Genetic Diversity of Thermophilic Campylobacter Species Isolated From a Layer Poultry Farm in Korea.

Thermophilic Campylobacter species are among the major etiologies of bacterial enteritis globally. This study aimed at assessing the antimicrobial resistance (AMR) profiles, virulence genes, and genetic diversity of thermophilic Campylobacter species isolated from a layer poultry farm in South Korea. One hundred fifty-three chicken feces were collected from two layer poultry farms in Gangneung, South Korea. The Campylobacter species were isolated by cultural techniques, while PCR and sequencing were used for species confirmation. Antimicrobial susceptibility testing for six antimicrobials [ciprofloxacin (CIP), nalidixic acid (NAL), sitafloxacin (SIT), erythromycin (ERY), tetracycline (TET), and gentamicin (GEN)] was carried out by broth microdilution. Three AMR and nine virulence genes were screened by PCR. Genotyping was performed by flaA-restriction fragment length polymorphism (RFLP) and multilocus sequence typing (MLST). Of the 153 samples, Campylobacter spp. were detected in 55 (35.9%), with Campylobacter jejuni and Campylobacter coli being 49 (89.1%) and six (10.9%), respectively. High-level resistance was observed for CIP (100%), NAL (100%), and TET (C. jejuni, 93.9%; C. coli: 83.3%). No resistance was observed for SIT. The missense mutation (C257T) in gyrA gene was confirmed by sequencing, while the tet(O) gene was similar to known sequences in GenBank. The rate of multidrug-resistant (MDR) strains was 8.2%, and they all belonged to C. jejuni. All Campylobacter isolates possessed five virulence genes (cdtB, cstII, flaA, cadF, and dnaJ), but none possessed ggt, while the rates for other genes (csrA, ciaB, and pldA) ranged between 33.3 and 95.9%. The flaA-RFLP yielded 26 flaA types (C. jejuni: 21 and C. coli: five), while the MLST showed 10 sequence types (STs) for C. jejuni and three STs for C. coli, with CC-607 (STs 3611) and CC-460 (ST-460) being predominant. Among the 10 STs of C. jejuni, three were newly assigned. The findings of this study highlight the increased resistance to quinolones and TET, the virulence potential, and the diverse genotypes among Campylobacter strains isolated from the layer poultry farm.

Antimicrobial Susceptibility and Genomic Analysis of Aliarcobacter cibarius and Aliarcobacter thereius, Two Rarely Detected Aliarcobacter Species.

Aliarcobacter cibarius and Aliarcobacter thereius are two rarely detected Aliarcobacter species. In the study, we analyzed the antimicrobial susceptibility and provide detailed insights into the genotype and phylogeny of both species using whole-genome sequencing. Thermophilic Campylobacter species are the most common bacterial foodborne pathogens causing gastroenteritis in humans worldwide. The genus Aliarcobacter is part of the Campylobacteraceae family and includes the species Aliarcobacter butzleri, Aliarcobacter cryaerophilus, Aliarcobacter skirrowii, and the rarely described Aliarcobacter cibarius, Aliarcobacter faecis, Aliarcobacter lanthieri, Aliarcobacter thereius, and Acrobarter trophiarum. Aliarcobacter are emergent enteropathogens and potential zoonotic agents. Here, we generated, analyzed, and characterized whole-genome sequences of Aliarcobacter cibarius and Aliarcobacter thereius. They were isolated from water poultry farms in Germany, cultured and identified by MALDI-TOF MS. With PCR the identity was verified. Antibiotic susceptibility testing was carried out with erythromycin, ciprofloxacin, doxycycline, tetracycline, gentamicin, streptomycin, ampicillin, and cefotaxime using the gradient strip method (E-test). Whole-genome sequences were generated including those of reference strains. Complete genomes for six selected strains are reported. These provide detailed insights into the genotype. With these, we predicted in silico known AMR genes, virulence-associated genes, and plasmid replicons. Phenotypic analysis of resistance showed differences between the presence of resistance genes and the prediction of phenotypic resistance profiles. In Aliarcobacter butzleri, the nucleotide sequence of the gyrA gene (DQ464331) can show a signature mutation resulting in an amino acid change T85>I. Acrobarter cibarius and Acrobarter thereius showed the same gene as assessed by similarity annotation of the mutations 254C>G. Most of the isolates were found to be sensitive to ciprofloxacin. The ciprofloxacin-resistant Aliarcobacter thereius isolate was associated with the amino acid change T85>I. But this was not predicted with antibiotic resistance databases, before. Ultimately, a phylogenetic analysis was done to facilitate in future outbreak analysis.

MOLECULAR CHARACTERIZATION OF CAMPYLOBACTER SPECIES FROM TURKEYS FLOCKS IN DELTA GOVERNMENTS

Two hundred and forty samples were collected from turkey farms and various markets in Delta Governorates for Campylobacter spp. isolation. Multiplex PCR assay was used to identify 2 Thermophilic Campylobacter species (C. jejuni and C. coli). The prevalence of Campylobacter in turkeys(22.5%) observed in this study; C. jejuni was 15.4% and C. coli 7%. The high incidence of Campylobacter in liver was (30%) followed by skin (26.7%), cloacal swabs (21.3%) and finally intestinal content (16.7%). A food borne disease infected with campylobacteriosis resulting from consuming or handling of contaminated poultry meat. The high incidence of Campylobacter due to contamination to intestinal tract during evisceration can lead to direct contamination of the carcasses or indirectly through the hands of the processors, material or instrument used in processing. The amplification of the DNA belonging to Campylobacter genus specific (C. jejuni hipO) was (19%); (13.8%) C. jejuni at 323 bp and (5.8%) C. coli at 126 bp, while for C. jejuni 23S rRNA at 650 bp. PCR analysis for detection of Campylobacter can be utilized as a simple, rapid and sensitive tool to discriminate stains recovered from different sources, especially when used as profile analysis for a control strategy of C. jejuni in turkey farms to reduce the risk of human exposure to Campylobacter and is an important food safety issue. Multiplex PCR was found to be more reliable than the conventional cultural methods in species level for identification of Campylobacter isolates. Further epidemiological studies at different geographical areas, carried to discover the prevalence, magnitude and importance of Campylobacter infection in turkey farms.

Prevalence, Risk Factors, and Antimicrobial Resistance Profiles of Thermophilic Campylobacter Species in Humans and Animals in Sub-Saharan Africa: A Systematic Review.

Thermophilic Campylobacter species are clinically important aetiologies of gastroenteritis in humans throughout the world. The colonization of different animal reservoirs by Campylobacter poses an important risk for humans through shedding of the pathogen in livestock waste and contamination of water sources, environment, and food. A review of published articles was conducted to obtain information on the prevalence and antimicrobial resistance (AMR) profiles of thermophilic Campylobacter species in humans and animals in sub-Saharan Africa (SSA). Electronic databases, namely, PubMed, Google Scholar, Research4life-HINARI Health, and Researchgate.net, were searched using the following search terms “thermophilic Campylobacter,” “Campylobacter jejuni,” “Campylobacter coli,” “diarrhea/diarrhoea,” “antimicrobial resistance,” “antibiotic resistance,” “humans,” “animals,” “Sub-Saharan Africa,” and “a specific country name.” Initially, a total of 614 articles were identified, and the lists of references were screened in which 22 more articles were identified. After screening, 33 articles on humans and 34 on animals and animal products were included in this review. In humans, Nigeria reported the highest prevalence (62.7%), followed by Malawi (21%) and South Africa (20.3%). For Campylobacter infections in under-five children, Kenya reported 16.4%, followed by Rwanda (15.5%) and Ethiopia (14.5%). The country-level mean prevalence in all ages and under-five children was 18.6% and 9.4%, respectively. The prevalence ranged from 1.7%–62.7% in humans and 1.2%–80% in animals. The most reported species were C. jejuni and C. coli. The AMR to commonly used antimicrobials ranged from 0–100% in both humans and animals. Poultry consumption and drinking surface water were the main risk factors for campylobacteriosis. The present review provides evidence of thermophilic Campylobacter occurrence in humans and animals and high levels of AMR in SSA, emphasizing the need for strengthening both national and regional multisectoral antimicrobial resistance standard surveillance protocols to curb both the campylobacteriosis burden and increase of antimicrobial resistance in the region.