Genetic Diversity Of Campylobacter Research Articles

Genomic Analysis Points to Multiple Genetic Mechanisms for Non-Transformable Campylobacter jejuni ST-50.

Campylobacter jejuni and Campylobacter coli are well known for their natural competence, i.e., their capacity for the uptake of naked DNA with subsequent transformation. This study identifies non-transformable C. jejuni and C. coli strains from domestic animals and employs genomic analysis to investigate the strain genotypes and their associated genetic mechanisms. The results reveal genetic associations leading to a non-transformable state, including functional DNase genes from bacteriophages and mutations within the cts-encoded DNA-uptake system, which impact the initial steps of the DNA uptake during natural transformation. Interestingly, all 38 tested C. jejuni ST-50 strains from the United States exhibit a high prevalence of non-transformability, and the strains harbor a variety of these genetic markers. This research emphasizes the role of these genetic markers in hindering the transfer of antimicrobial resistance (AMR) determinants, providing valuable insights into the genetic diversity of Campylobacter. As ST-50 is a major clone of C. jejuni globally, we additionally determined the prevalence of the genetic markers for non-transformability among C. jejuni ST-50 from different regions of the world, revealing distinct patterns of evolution and a strong selective pressure on the loss of competence in ST-50 strains, particularly in the agricultural environment in the United States. Our findings contribute to a comprehensive understanding of genetic exchange mechanisms within Campylobacter strains, and their implications for antimicrobial resistance dissemination and evolutionary pathways within specific lineages.

Genotyping and antibiotic susceptibility of Campylobacter species isolated from raw milk samples in Qazvin, Iran

ObjectiveCampylobacter species are major causes of foodborne illnesses, with unpasteurized milk being a significant carrier of these bacteria, posing a public health risk. One of the challenges in managing Campylobacter infections is the emergence and spread of antibiotic resistance. We conducted a study in Qazvin, Iran, testing 84 raw cow’s milk samples to determine the frequency of C. jejuni and C. coli using culture-based and multiplex PCR methods. Additionally, the disk diffusion and RAPD-PCR approaches were utilized to evaluate the phenotypic antibiotic resistance profile and genetic diversity of Campylobacter strains.ResultsThe findings indicated that Campylobacter spp. was present in 19.05% of the samples, with C. coli being the predominant isolate. We tested eight antibiotic agents, and the resistance levels of the isolates were as follows: erythromycin 100%, tetracycline 75%, doxycycline 56.25%, ceftriaxone 43.75%, chloramphenicol 37.5%, amoxicillin-clavulanic acid 25%, nalidixic acid 12.5%, and azithromycin 6.25%. Genetic diversity analysis categorized Campylobacter isolates into 39 clusters, indicating a wide diversity among strains. However, no significant correlation was observed between antibiotic resistance and cluster patterns. These findings underscore the role of raw milk as a reservoir for Campylobacter spp. and highlight the substantial antibiotic resistance and genetic diversity within the species population.

Highly Prevalent Multidrug-Resistant Campylobacter spp. Isolated From a Yellow-Feathered Broiler Slaughterhouse in South China.

The purpose of this study was to investigate the prevalence, antimicrobial resistance, virulence genes, and genetic diversity of Campylobacter spp. along the yellow-feathered broiler slaughtering line in Southern China from December 2018 to June 2019. A total of 157 Campylobacter spp. isolates were identified from 1,102 samples (including 53.6% (75/140) of live chicken anal swab samples, 27.5% (44/160) of defeathering samples, 18.1% (29/160) of evisceration samples, 2.1% (3/140) of washing samples, 1.4% (2/140) of chilling samples, and 1.1% (4/362) of environmental samples). The prevalence of Campylobacter spp. was 14.2%, including 43.9% Campylobacter jejuni, 53.5% Campylobacter coli, and 2.5% other Campylobacter species. The highest antimicrobial resistance rate was found to be against sulfamethoxazole (138/157, 87.9%), and 90.4% (142/157) of the isolates were multidrug resistant (MDR). Examination of resistance-related genes revealed the double base mutated Thr-86-Ile, which informed ACA-TTA, with an Arg-79-Lys substitution in gyrA. Eleven virulence-associated genes (cadF, cdtA, cdtB, ciaB, flaA, imaA, dnaJ, plaA, virB11, racR, and cdtC) were also detected by a polymerase chain reaction (PCR) analysis, and cadF (81.5%) was the most prevalent. Based on an analysis of pulsed-field gel electrophoresis (PFGE) results, we found that Campylobacter spp. could be cross-contaminated throughout the entire slaughtering line. These results show that it is imperative to study the Campylobacter spp. from the yellow-feathered broiler along the slaughtering line in China to develop preventative and treatment measures for the poultry industry, as well as food safety and public health.

Campylobacter jejuni Strain Dynamics in a Raccoon (Procyon lotor) Population in Southern Ontario, Canada: High Prevalence and Rapid Subtype Turnover.

Free-ranging wildlife are increasingly recognized as potential reservoirs of disease-causing Campylobacter species such as C. jejuni and C. coli. Raccoons (Procyon lotor), which live at the interface of rural, urban, and more natural environments, are ideal subjects for exploring the potential role that wildlife play in the epidemiology of campylobacteriosis. We studied the prevalence and genetic diversity of Campylobacter from live-captured raccoons on five swine farms and five conservation areas in southwest Ontario. From 2011 to 2013, we collected fecal swabs (n = 1,096) from raccoons, and (n = 50) manure pit samples from the swine farm environment. We subtyped the resulting Campylobacter isolates (n = 581) using Comparative Genomic Fingerprinting (CGF) and 114 distinct subtypes were observed, including 96 and 18 subtypes among raccoon and manure pit isolates, respectively. Campylobacter prevalence in raccoons was 46.3%, with 98.7% of isolates recovered identified as C. jejuni. Novel raccoon-specific CGF subtypes (n = 40/96) accounted for 24.6% (n = 143/581) of Campylobacter isolates collected in this study. Our results also show that C. jejuni is readily acquired and lost in this wild raccoon population and that a high Campylobacter prevalence is observed despite transient carriage typically lasting 30 days or fewer. Moreover, although raccoons appeared to be colonized by species-adapted subtypes, they also harbored agriculture-associated genotypes that accounted for the majority of isolates observed (66.4%) and that are strongly associated with human infections. This suggests that raccoons may act as vectors in the transmission of clinically-relevant C. jejuni subtypes at the interface of rural, urban, and more natural environments.

Prevalence and genetic diversity of Campylobacter spp. in the production chain of broiler chickens in Lebanon and its association with the intestinal protozoan Blastocystis sp

Campylobacter jejuni is recognized as the most common foodborne pathogen associated with human gastroenteritis worldwide. Broilers are frequently infected by the bacteria and are considered the main source of exposure to humans. However, despite its public health impact, no recent data are currently available in Lebanon about Campylobacter spp. in poultry and human population. Therefore, this study aimed to determine the prevalence and genetic diversity of Campylobacter spp. in 227 ceca and on 227 carcasses of broiler chickens collected in Lebanese slaughterhouses. Overall, the prevalence of Campylobacter was shown to reach 67.0% in ceca and 17.2% on carcasses of Lebanese poultry. The only 2 Campylobacter species identified were C. jejuni and C. coli, with a slightly higher prevalence of C. coli in ceca and of C. jejuni on carcasses. A high level of genetic diversity was reported among the 51 C. jejuni isolates selected, since 25 distinct profiles were identified according to the comparative genomic fingerprinting typing method based on a subset of 40 genes using the 90% similarity threshold. Predominant clusters observed in Lebanese poultry isolates were also frequently found among French human clinical cases, highlighting that broiler chickens represent a potential reservoir for human campylobacteriosis. In addition, a significantly higher prevalence of Campylobacter spp. was found in slaughterhouse workers than in a cohort of hospitalized patients with no contact with poultry, confirming that contaminated broiler chickens in slaughterhouse appeared to be a non-negligible source of Campylobacter spp. transmission. Interestingly, a significant association between Campylobacter spp. and Blastocystis sp. has been observed. This correlation suggested that the presence of Campylobacter spp. would be favored when Blastocystis sp. is present and, similarly, the absence of one would favor the absence of the other. This is the first large-scale investigation focusing on the impact of Campylobacter spp. in broiler chickens in Lebanon and confirmed the need to implement prevention and control measures in the poultry production to reduce the burden of campylobacteriosis in the human population.

Molecular epidemiology of Campylobacter isolates from broiler slaughterhouses in Tripoli, North of Lebanon

ABSTRACT 1. The real burden of Campylobacter spp. in Lebanon is still unknown. The aims of this study were to unravel the epidemiology of Campylobacter spp. in broilers at slaughterhouses in Tripoli, North of Lebanon and to characterise their antibiotic resistance profiles. 2. From May to November 2015, sampling was performed through five repeated surveys from 15 slaughterhouses that sold chicken directly to Lebanese customers. Isolates were subjected to pulsed field gel electrophoresis (PFGE) and flaA-restriction fragment length polymorphism (flaA-RFLP). 3. All investigated slaughterhouses were found to be positive for Campylobacter spp. Campylobacter coli was the predominant species (38 isolates) followed by C. jejuni (eight isolates). A noticeable level of resistance was detected among isolates against ciprofloxacin (97% of C. coli and 87.5% of C. jejuni), amoxicillin (89% of C. coli and 75% of C. jejuni), gentamicin (79% of C. coli and 50% of C. jejuni), and co-amoxiclav (24% of C. coli and 25% of C. jejuni). Erythromycin and ertapenem resistance were observed only in C. coli with the following percentages 74% and 13% respectively, but not in C. jejuni. PFGE and flaA-RFLP using DdeI as restriction enzyme divided the strains into 27 and 25 types respectively. 4. The high observed genetic diversity of Campylobacter spp. revealed the complexity of the spread of this genus in broilers. This study highlighted the pressing need to monitor antibiotic resistance and to ensure food safety from ‘farm to fork’ in Lebanon.

Genetic Diversity of Campylobacter jejuni and Campylobacter coli Isolates from Conventional Broiler Flocks and the Impacts of Sampling Strategy and Laboratory Method.

The genetic diversity of Campylobacter jejuni and Campylobacter coli isolates from commercial broiler farms was examined by multilocus sequence typing (MLST), with an assessment of the impact of the sample type and laboratory method on the genotypes of Campylobacter isolated. A total of 645 C. jejuni and 106 C. coli isolates were obtained from 32 flocks and 17 farms, with 47 sequence types (STs) identified. The Campylobacter jejuni isolates obtained by different sampling approaches and laboratory methods were very similar, with the same STs identified at similar frequencies, and had no major effect on the genetic profile of Campylobacter population in broiler flocks at the farm level. For C. coli, the results were more equivocal. While some STs were widely distributed within and among farms and flocks, analysis of molecular variance (AMOVA) revealed a high degree of genetic diversity among farms for C. jejuni, where farm effects accounted for 70.5% of variance, and among flocks from the same farm (9.9% of variance for C. jejuni and 64.1% for C. coli). These results show the complexity of the population structure of Campylobacter in broiler production and that commercial broiler farms provide an ecological niche for a wide diversity of genotypes. The genetic diversity of C. jejuni isolates among broiler farms should be taken into account when designing studies to understand Campylobacter populations in broiler production and the impact of interventions. We provide evidence that supports synthesis of studies on C. jejuni populations even when laboratory and sampling methods are not identical.

Comparative analysis of antimicrobial resistance and genetic diversity of Campylobacter from broilers slaughtered in Poland

In the current study, the relationship of Campylobacter jejuni and Campylobacter coli strains isolated at slaughter was investigated using comparative analysis of antimicrobial resistance (AMR), virulence gene (VG) and PFGE profiling. A total of 254 Campylobacter isolates from poultry caeca and corresponding carcasses, including 139 C. jejuni and 115 C. coli strains were tested. The most prevalent resistance profiles observed in C. jejuni were ciprofloxacin, nalidixic acid and tetracycline (46 out of 139, 33.1% isolates) as well as ciprofloxacin, nalidixic acid, tetracycline and streptomycin among C. coli strains (34 out of 115, 29.6%). Multi-resistance was found more frequently among C. coli than C. jejuni (P<0.05). The presence of 11 virulence genes exhibited 19 different VG profiles in Campylobacter isolates tested. All Campylobacter strains were classified into 154 different PFGE types. Among them, 56 profiles (28 C. jejuni and 28 C. coli) were common for at least two isolates including 9 clusters covering from 4 to 9 strains. Campylobacter composite types generated by a combination of 154 PFGE types, 10 AMR profiles and 19 VG patterns divided 178 distinct types with 95% similarity. The majority of the composite profiles (76 for C. jejuni and 58 for C. coli; 75.3% in total) included only one bacterial isolate. Furthermore, 11 pairs of C. jejuni and 12 pairs of C. coli from caeca and the corresponding carcasses isolated from the same places possessed the identical PFGE, AMR and VG patterns.This study demonstrated that C. jejuni and C. coli isolated from poultry in Poland showed to have a high genetic diversity and a weak clonal population structure. However, the composite analysis revealed a strong evidence for cross-contamination of chicken carcasses during the slaughter process. Additionally, our results confirm that Campylobacter may easily contaminate poultry carcasses at slaughter process and spread around country. More than half of Campylobacter strains tested (50.4%) were resistant to at least two classes of antimicrobials, i.e. quinolones and tetracyclines, which may cause a public health risk.

Prevalence, quantitative load and genetic diversity of Campylobacter spp. in dairy cattle herds in Lithuania.

BackgroundCampylobacteriosis is a zoonotic disease, and animals such as poultry, pigs and cattle may act as reservoirs for Campylobacter spp. Cattle shed Campylobacter spp. into the environment and they can act as a reservoir for human infection directly via contact with cattle or their faeces or indirectly by consumption of contaminated food. The aim of this study was to determine the prevalence, the quantitative load and the genetic strain diversity of Campylobacter spp. in dairy cattle of different age groups.ResultsFaecal samples of 200 dairy cattle from three farms in the central part of Lithuania were collected and examined for Campylobacter. Cattle herds of all three farms were Campylobacter spp. positive, with a prevalence ranging from 75% (farm I), 77.5% (farm II) to 83.3% (farm III). Overall, the highest prevalence was detected in calves (86.5%) and heifers (86.2%). In contrast, the lowest Campylobacter prevalence was detectable in dairy cows (60.6%). C. jejuni, C. coli, C. lari and C. fetus subsp. fetus were identified in faecal samples of dairy cattle. C. upsaliensis was not detectable in any sample. The high counts of Campylobacter spp. were observed in faecal material of dairy cattle (average 4.5 log10 cfu/g). The highest numbers of Campylobacter spp. were found in faecal samples from calves (average 5.3 log10 cfu/g), whereas, faecal samples from cows harboured the lowest number of Campylobacter spp. (average 3.7 log10 cfu/g). Genotyping by flaA PCR-RFLP analysis of selected C. jejuni isolates showed that some genotypes were present in all farms and all age groups. However, farm or age specific genotypes were also identified.ConclusionsFuture studies are needed to investigate risk factors related to the degree of colonisation in cattle. Based on that, possible measures to reduce the colonisation and subsequent shedding of Campylobacter in cattle could be established. It is important to further investigate the epidemiology of Campylobacter in the cattle population in order to assess associated risks to public health.

The Prevalence and Genetic Diversity of Campylobacter spp. in Domestic ‘Backyard’ Poultry in Canterbury, New Zealand

Campylobacteriosis is the most commonly notified illness in New Zealand. Whilst the importance of commercial poultry in campylobacteriosis is well established, little is known about the possible role of chickens kept at home as a direct animal/faecal contact or consumption exposure pathway. The aim of this study was to determine the prevalence and genetic diversity of Campylobacter spp. in domestic backyard chicken flocks in the Canterbury region of New Zealand. Poultry faecal samples were collected from 35 domestic 'backyard' poultry flocks from urban and rural properties around the Canterbury Region of New Zealand. A total of 291 samples were collected and tested for the presence of thermotolerant Campylobacter spp. and positive isolates were analysed using pulsed-field gel electrophoresis (PFGE) using both SmaI and KpnI enzymes. There was a high prevalence of Campylobacter spp. with 86% of flocks testing positive. Campylobacter jejuni alone, Campylobacter coli alone and both C. jejuni and C. coli were detected in 20 (57%), 2 (6%) and 8 (23%) of the flocks respectively. SmaI/KpnI PFGE analysis identified 50 different genotypes across the 35 flocks. Genotype diversity richness was highest on the lifestyle block and farm properties with 43 different genotypes isolated, whilst urban properties displayed the least richness with 12 genotypes isolated. Rural flocks tended to have more different genotypes in a given flock than urban flocks. Comparison of the genotypes with the PulseNet Aotearoa Campylobacter database showed that 28 of the genotypes had previously been isolated from human cases of campylobacteriosis. Many of these were also indistinguishable from Campylobacter spp. previously isolated from retail chicken. Therefore, contact with backyard poultry or their faecal material is a potential additional infection pathway outside of exposure to the established pathways associated with the consumption of Campylobacter-contaminated commercial meat or foods cross-contaminated from contaminated poultry.

Prevalence and Characterization ofCampylobacterspp. Isolated from Domestic and Imported Poultry Meat in Korea, 2004–2008

Campylobacteriosis in humans is primarily caused by handling or consuming contaminated poultry or their products. The aims of this study were to estimate the prevalence of Campylobacter spp. in domestic and imported poultry meat in Korea and to further characterize the obtained isolates. From 2004 to 2008, a total of 475 domestic and 867 imported raw poultry meat samples were examined for the presence of Campylobacter spp. Among 475 domestic poultry meat samples, Campylobacter jejuni and Campylobacter coli were isolated from 219 (46.1%) and 156 (32.8%), respectively. Relative prevalence of C. jejuni and C. coli was higher in meat from Brazil (39/78, 50.0% and 7/78, 8.9%) and France (32/96, 33.3% and 8/96, 8.3%), whereas lower in meat from Denmark (72/516, 14.0% and 12/516, 2.3%) and Thailand (5/39, 12.8% and 3/39, 7.6%). The resistance to ampicillin and tetracycline was highly prevalent in Campylobacter spp. from most countries investigated, whereas lower in meat from Denmark. On the other hand, the prevalence of erythromycin and gentamicin resistance was less than 10% in most countries. The resistance rate to nalidixic acid, ciprofloxacin, and enrofloxacin ranged from 11.9% to 87.5%. The use of fla-polymerase chain reaction–restriction fragment length polymorphism for epidemiological analysis found that some pattern types were considerably more frequent and distinct in meat from each country. In conclusion, we report the presence of high contamination in domestic and imported poultry meat in Korea and the antimicrobial and genetic diversity of Campylobacter spp. between each country.

Comparison of Campylobacter populations isolated from a free-range broiler flock before and after slaughter

Relatively little is known about the Campylobacter genotypes colonizing extensively reared broiler flocks and their survival through the slaughter process, despite the increasing demand for free-range and organic products by the consumer. Campylobacter isolates from a free-range boiler flock, sampled before and after slaughter, were genotyped by MLST (multilocus sequence typing) and sequence analysis of the flaA short variable region (SVR). The Campylobacter genotypes isolated before and after slaughter were diverse, with up to five sequence types (STs) (seven-locus allelic profiles resulting from MLST) identified per live bird, up to eight STs identified per carcass and 31 STs identified in all. The majority (72.0%) of isolates sampled from carcasses post-slaughter were indistinguishable from those isolated from the live flock before slaughter by ST and flaA SVR type, however, sampling ‘on-farm’ failed to capture all of the diversity seen post-slaughter. There were statistically significant increases in the genetic diversity of Campylobacter ( p = 0.005) and the proportion of C. coli ( p = 0.002), with some evidence for differential survival of genotypes contaminating the end product. C. coli genotypes isolated after slaughter were more similar to those from free-range and organic meat products sampled nationally, than from the live flock sampled previously. This study demonstrated the utility of MLST in detecting genetic diversity before and after the slaughter process.

Occurrence and genotypes of Campylobacter in broiler flocks, other farm animals, and the environment during several rearing periods on selected poultry farms

On 15 Swiss poultry farms, broiler flocks, other farm animals, and the environment were examined during consecutive rearing periods to investigate the occurrence and genetic diversity of Campylobacter. Of the 5154 collected samples, 311 (6%) from 14 farms were Campylobacter positive by culture. Amongst the positive samples, 228 tested positive for Campylobacter jejuni and 92 for Campylobacter coli. Positive samples originated from broilers, the broiler houses, cattle, pigs, bantams, laying hens, a horse, and a mouse. Feed, litter, flies, and the supply air to the broiler house tested negative. By flagellin gene typing ( fla-RFLP) and pulsed-field gel electrophoresis (PFGE), 917 Campylobacter isolates were genotyped. Additionally, amplified fragment length polymorphism (AFLP) analysis was performed on 15 assorted strains. On eight farms, matching genotypes were isolated from broiler flocks and other farm animals: Certain genotypes from cattle (farms H, K, L, and M), pigs (farms D and P), or laying hens (farm L) were subsequently found in the broiler flocks, whereas other genotypes initially present in the broiler flocks turned up in cattle (farms A, D, and O). These results emphasize the importance of other farm animals on poultry farms for broiler flock colonization. Indications of persistent contamination of the broiler house were evident on four farms (C, D, I, and L) where matching genotypes were detected in consecutive broiler flocks, but not concurrently in other samples. By fla-RFLP, PFGE, and confirmed by AFLP, some genotypes proofed to be identical across different farms.

Campylobacterspp. in Dogs and Cats in Switzerland: Risk Factor Analysis and Molecular Characterization with AFLP

A 1-year cross-sectional study was carried out to determine the prevalence, risk factors for carriage, and genetic diversity of Campylobacter spp. in healthy dogs and cats in Switzerland. Veterinary practitioners collected samples from 1268 animals (all ages) presented for vaccination. The prevalence of Campylobacter spp. in 634 dogs and 596 cats that were eligible for the study was 41.2% (confidence interval 95%: 37.3-45.1%) and 41.9% (CI 95%: 37.9-46%), respectively. Risk factors identified for carriage of Campylobacter jejuni were found to be different from risk factors for C. upsaliensis/C. helveticus. Young animals (< or =3 years) had significantly higher odds of carrying C. upsaliensis/C. helveticus than older animals (OR 1.8-3.3), whereas for C. jejuni carriage, the age was not a risk factor. Amplified fragment length polymorphism (AFLP) genotyping revealed heterogeneity among C. jejuni strains and was found to clearly separate C. helveticus from C. upsaliensis. It was shown that cats more often carry C. helveticus with an estimated prevalence of 28.2%, whereas dogs mainly are carrying C. upsaliensis.