Heterogeneity of Campylobacter jejuni and Campylobacter coli strains from healthy sheep

The objectives of this study were to identify, at species level, thermophilic campylobacters isolated from clinically healthy sheep by a multiplex polymerase chain reaction (mPCR). The heterogeneity among Campylobacter jejuni and C. coli isolates was also investigated using a restriction fragment length polymorphism (RFLP) analysis of the flagellin (flaA) gene. Samples of intestinal contents, gall bladders and faeces were collected from 610 healthy sheep. While gall bladder samples were plated directly onto Preston agar, an enrichment stage was applied for intestinal and faecal samples. Of the 610 samples, 302 (49.5%) were positive for Campylobacter spp. Using a mPCR assay for species identification, 103 (34.1%) were positive with C. jejuni-specific primers, while 100 (33.1%) were positive with C. coli-specific primers. Additionally, 16 (11.9%) of the intestinal content samples were positive for both species by mPCR. All the isolates identified as C. jejuni and C. coli were successfully subtyped by flaA typing. Of 203 isolates tested, 48 different flaA types were found. Twenty-six flaA types were identified among C. jejuni isolates and the remaining 22 from C. coli isolates.     

Pathogenicity of Campylobacter jejuni and Campylobacter coli strains in the pregnant guinea pig model

Pathogenicity of 17 Campylobacter isolates for pregnant guinea pigs was investigated. Of 14 isolates, 12 (86%) produced rates of abortion ranging from 13% to 87%. Two isolates did not produce abortion. Reference strains of C fetus subsp venerealis produced abortion in 60% to 87% and C fetus subsp fetus produced abortion in 60% of the guinea pigs. Inoculated organisms were recovered from uterus, blood, liver, kidney, spleen, and gallbladder of the guinea pigs at rates as high as 83% for 2 ovine isolates and as low as 13% for 2 bovine and 1 human isolates. Most isolations were from the uterus. Two avian isolates were not recovered. Within the C jejuni and C coli group, the ovine and the human isolates appear to be more pathogenic. Swine, bovine, and avian isolates were less pathogenic. Seemingly, the pregnant guinea pig was a suitable and practical model for evaluating the pathogenicity of Campylobacter organisms, regardless of their host of origin.     

Fecal shedding of Campylobacter jejuni and Campylobacter coli among feral pigs in Texas

The population and range of feral pigs in the United States are rapidly expanding, yet key knowledge gaps exist regarding their role in the ecology and transmission of foodborne pathogens. Our objectives were to estimate the prevalence of Campylobacter jejuni and Campylobacter coli shedding among feral pigs throughout Texas and to identify risk factors for positive status. Faecal samples were collected from feral pigs in Texas from February 2014 through May 2015, and target organisms were detected using PCR assays. The prevalence of C. jejuni shedding was 1.6% (6/370), and the prevalence of C. coli shedding was 3.5% (13/370). C. coli shedding was significantly more common (p = .008) among female pigs than among male pigs. Feral pigs may represent a source of human campylobacteriosis.     

Risk factors for Campylobacter jejuni and Campylobacter coli in young cattle on English and Welsh farms

Campylobacter jejuni and C. coli are the most prevalent causes of bacterial diarrhoea in most of the Western World. In Great Britain, the source remains unknown for the majority of cases, though poultry is considered the main source of infection. Molecular typing methods identify cattle as a potential source of a proportion of the non-source-attributed cases, mainly through direct contact, environmental contamination or milk, but little is known about the epidemiology of Campylobacter in cattle. A cross-sectional study was undertaken on young cattle 3-17 months of age on 56 cattle farms in England and Wales to identify association between the presence of C. jejuni and C. coli and farm characteristics and management practices. Campylobacter was detected on 62.5% of the farms and the presence of dairy cows (OR: 3.7, CI(95%): 1.2; 11.7), indoor housing (OR: 4.6, CI(95%): 1.8; 12.0), private water supply (OR: 2.5, CI(95%): 1.2; 5.4), presence of horses (OR: 3.2, CI(95%):1.5; 6.9) and feeding hay (OR: 2.9, CI(95%):1.6; 5.5) were associated with detection. The model's goodness-of-fit was improved when herd size was forced in the model without being statistically significant (p=0.34).     

Campylobacter jejuni and Campylobacter coli inoculation of neonatal calves

Three groups of neonatal calves were inoculated orally with pathogenic strains of Campylobacter jejuni or C coli. The calves developed a mild, self-limiting enteritis characterized by thick mucoid feces. Bacteremia and fecal shedding of Campylobacter were sporadic in all inoculated calves. Two groups of calves were killed 1 to 3 weeks after inoculation to study the pathogenesis of infection. Postmortem culture of tissues revealed C jejuni or C coli most frequently in the ileum, cecum, colon, and blood. Clinical or pathologic differences between C jejuni-inoculated and C coli-inoculated calves were minimal.     

Occurrence of virulence genes among Campylobacter jejuni and Campylobacter coli isolates from domestic animals and children

The presence of the flaA, cadF, cdtB and iam genes of Campylobacter spp. was determined with the PCR method. The materials to investigate were 56 C. jejuni and 23 C. coli strains isolated from clinical samples (children and domestic animals). It was found that all of the Campylobacter spp. isolates from children with diarrhoea and domestic animals had cadF gene, responsible for adherence. The flaA gene was present in all Campylobacter spp. isolates derived from children and cats. Occurrence of flaA gene was confirmed in 100% of C. jejuni strains obtained from dogs. The high prevalence of the cdtB gene associated with toxin production was observed in this study (100%-Campylobacter spp. isolates obtained from dogs and cats, 97.9%-Campylobacter spp. isolates from children). The isolates showed a wide variation for the presence of iam gene. The lowest prevalence (23.5%) was detected in Campylobacter spp. obtained from dogs. The highest rates of iam detection (91.6%) were revealed in C. coli isolates from children.     

Detection and genotypic differentiation of Campylobacter jejuni and Campylobacter coli strains from laying hens by multiplex PCR and fla-typing

In total, 26 Campylobacter (C.) strains, isolated from liver, spleen, caecal or jejunal content of laying hens from different flocks were examined. In these flocks a drop in egg production, an increasing mortality and livers with whitish-grey lesions as post-mortem finding were observed. Suspected Campylobacter colonies were differentiated using a modified m-PCR in 13 Campylobacter jejuni and 13 Campylobacter coli strains. All isolates were characterised by typing of the flaA and flaB gene each with two restriction enzymes. To compare the four different profiles for all strains an artificial “fla-type” was generated. Different and identical fla-types of C. jejuni and C. coli were recovered from both intestinal and extra-intestinal organs of the laying hens and even from individual birds. One significant observation is that some fla-types of C. jejuni or C. coli were detected in intestinal and systemic sites but not all fla-types of both species appeared to be equally able to invade internal organs.     

Detection and genotypic differentiation of Campylobacter jejuni and Campylobacter coli strains from laying hens by multiplex PCR and fla-typing

In total, 26 Campylobacter (C.) strains, isolated from liver, spleen, caecal or jejunal content of laying hens from different flocks were examined. In these flocks a drop in egg production, an increasing mortality and livers with whitish-grey lesions as post-mortem finding were observed. Suspected Campylobacter colonies were differentiated using a modified m-PCR in 13 Campylobacter jejuni and 13 Campylobacter coli strains. All isolates were characterised by typing of the flaA and flaB gene each with two restriction enzymes. To compare the four different profiles for all strains an artificial “fla-type” was generated. Different and identical fla-types of C. jejuni and C. coli were recovered from both intestinal and extra-intestinal organs of the laying hens and even from individual birds. One significant observation is that some fla-types of C. jejuni or C. coli were detected in intestinal and systemic sites but not all fla-types of both species appeared to be equally able to invade internal organs.     

Campylobacter jejuni abortions in two beef cattle herds in Saskatchewan

Abortions, accompanied by placental retention and weight loss, occurred during February and March in 19% of 120 and 10% of 108 beef cows and heifers on two neighboring ranches in southern Saskatchewan. A diagnosis of Campylobacter jejuni abortion was made based on lesions of necrotizing and suppurative placentitis and fetal bronchopneumonia in association with the culture of large numbers of C. jejuni from placentas and fetal tissues.

Campylobacter jejuni was isolated with variable frequency from fecal samples of aborting and healthy cows, and scouring and healthy calves. Campylobacter jejuni serotype 2 (Lior) was isolated from fetal tissues and feces of a scouring calf, whereas C. jejuni serotypes 1, 4, 5 and 99 were isolated from feces of in-contact cattle. We hypothesized that the source and mode of transmission of C. jejuni was fecal contamination of water supplies and feeding grounds by carrier cows or wildlife.

     

Campylobacter jejuni, Campylobacter coli, and cytolethal distending toxin genes in laying hens

As no data are available on the prevalence of cytolethal distending toxin (cdt) genes carried by Campylobacter spp. in laying hens, this study was conducted with the aim to evaluate the prevalence of both Campylobacter spp. and cdt genes in 1680 laying hens from four different farms. The samples were analyzed by culture methods and by polymerase chain reaction. Campylobacter spp. were isolated from 1097/1680 cloacal swabs. Among the isolates, 913 were identified as Campylobacter jejuni whereas 345 were identified as Campylobacter coli. All isolates carried cdt genes. The results presented here confirm the very common occurrence of C. jejuni and C. coli in laying hens and underline that the cdt genes may also be frequently present in both C. jejuni and C. coli isolates from laying hens.     

Higher resistance of Campylobacter coli compared to Campylobacter jejuni at chicken slaughterhouse

In order to compare the prevalence of Campylobacter coli and Campylobacter jejuni during the processing of broilers at slaughterhouse a total of 848 samples were analyzed during 2012 in southern Spain. Four hundred and seventy six samples were collected from cloaca, carcass surfaces and quartered carcasses. Moreover, 372 environmental swabs from equipment and scalding water were collected. Minimum inhibitory concentration (MIC) to ciprofloxacin, erythromycin, streptomycin, tetracycline and gentamicin was determined for isolates from chicken meat. The general prevalence of Campylobacter was 68.8% (40.2% of C. coli and 28.5% of C. jejuni). The relative prevalence of C. coli increased from loading dock area (41.5%) to packing area (64.6%). In contrast, the relative prevalence of C. jejuni decreased from 58.5% to 35.4%. These differences between species from initial to final area were significant (p = 0.02). The highest antimicrobial resistance for C. jejuni and C. coli was detected to tetracycline (100%) and ciprofloxacin (100%), respectively. Campylobacter coli showed an antimicrobial resistance significantly higher than C. jejuni to streptomycin (p = 0.002) and erythromycin (p < 0.0001).     

Fla-DGGE analysis of Campylobacter jejuni and Campylobacter coli in cecal samples of broilers without cultivation

In a commercial broiler flock during rearing multiple genotypes of Campylobacter jejuni may be present as well as in gastrointestinal tracts of individual birds. The aim of this study was to optimize and apply a denaturing gradient gel electrophoresis assay of the flagellin gene (fla-DGGE) for analysis of C. jejuni and Campylobacter coli in cecal samples of broilers without prior cultivation. One C. coli and 21 C. jejuni strains isolated from broiler flocks, of which 14 typed as unique by restriction fragment length polymorphism of flaA and two undefined strains, were clustered into 9 groups when applying fla-DGGE. Spiking of cecal samples revealed that fla-DGGE is able to detect at least 4.55-5.96logCFUCampylobacter/mlcecal material. The presence of 3 strains spiked in cecal material was demonstrated by fla-DGGE as the corresponding bands were visible on the DGGE gel. Naturally contaminated cecal samples were shown to contain different types of C. jejuni and C. coli. Fla-DGGE has some potential as a cultivation-independent fast primary subtyping method for C. jejuni and C. coli in cecal samples of broilers.     

鸡源空肠弯曲菌和结肠弯曲菌的临床分离及多重PCR鉴定

弯曲菌(Campylobacter)尤其是空肠弯曲菌(C.jejuni)是重要的人畜共患病病原菌,在人可引起急性胃肠炎,感染严重者伴有心内膜炎、肺炎、败血症、血栓静脉炎、脑膜炎、多发性关节炎以及格林-巴利综合征等疾病[1-3].家禽、家畜、宠物等动物是弯曲菌的常见宿主,因此研究动物源弯曲菌具有重要的公共卫生意义.     

Serotypes of Campylobacter jejuni and C coli isolated from dogs

The serotypes of Campylobacter jejuni and C coli isolated from 56 dogs were established by the Penner serotyping scheme. A total of 37 C jejuni and 19 C coli were typed. Only two of the C coli strains were typable by the Penner method compared to 29 of 37 C jejuni strains. Pen 2 and 4 were the most predominant serotypes, constituting 41-4 per cent of the typable C jejuni strains. All but one of the C jejuni strains belonging to serotypes pen 1 and 2 were isolated from dogs with diarrhoea.