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Seventy-four Gram-negative, motile, slightly curved rod-shaped, microaerophilic, oxidase-positive and catalase-negative isolates, recovered from fecal samples of the Anatolian ground squirrel (Spermophilus xanthoprymnus) in Kayseri, Turkey, were subjected to a polyphasic taxonomic study. Results of a genus-specific PCR indicated that all isolates belonged to the genus Campylobacter. 16S rRNA gene sequence analyses revealed the closest match as Campylobacter curvus DSM 6644 T with identity levels of 96.41-96.70%. Based on the 16S rRNA gene phylogeny of the 74 isolates, six isolates (faydin-G24, faydin-G52, faydin-G105, faydin-G114, faydin-G129 and faydin-G140 T ) were chosen as representatives for further characterization. The overall genome relatedness indices for the strain faydin-G140 T , compared to the most closely related type strain C. curvus ATCC 35224 T , were calculated as 15.2%, 72.5%, and 83.7% for digital DNA-DNA hybridization (dDDH), and average nucleotide identity (ANIb and ANIm), respectively. The G+C content and genome size of the strains ranged between 35.2-35.4 mol% and 1.7-1.8 Mb, respectively. Based on data obtained from the polyphasic taxonomy approach, including phenotypic characterization as well as genomic and chemotaxonomic analyses, these strains are concluded to represent a novel species, for which the name Campylobacter anatolicus sp. nov. is proposed with faydin-G140 T as the type strain (=DSM 112311 T  = LMG 32238 T )., (Copyright © 2021 Elsevier GmbH. All rights reserved.)

Aims: Campylobacter sp. are important causes of reproductive disease in ruminants worldwide. Although healthy bulls are well-known carriers for infection of cows, the role of rams as a potential source for infecting ewes is unclear. This study aimed to determine prevalence, species distribution, genetic diversity and antimicrobial susceptibility profiles of Campylobacter sp. isolated from the preputial cavity of healthy rams., Methods and Results: The material of this prospective study comprised 191 swab samples taken from the preputial cavity of healthy rams. Enrichment and membrane filtration were employed for the isolation of Campylobacter. Presumptive isolates were confirmed as Campylobacter by phenotypic and molecular tests. 16S rRNA gene sequence analysis was used for the definitive identification of the isolates at species level, and genotyping was performed using pulsed-field gel electrophoresis (PFGE). The susceptibility of the Campylobacter sp. isolates to various antibiotics was determined by the disk diffusion test. In all, 27 of the 191 (14·13%) swab samples were found to be positive for Campylobacter sp. (28 isolates were recovered in total). Per phenotypic and genotypic analyses, one isolate was identified as Campylobacter mucosalis and the remaining 27 isolates were identified as Campylobacter sputorum bv. faecalis. The PFGE analysis of the C. sputorum biovar faecalis isolates produced 17 clusters and 24 different pulsotypes, indicating high genetic heterogeneity. All 28 isolates were found to be susceptible to all of the antibiotics tested., Conclusions: Healthy rams may be an important reservoir of different Campylobacter species in the preputium., Significance and Impact of the Study: This study demonstrated for the first time that healthy rams can carry different Campylobacter sp. including genetically diverse C. sputorum bv. faecalis and C. mucosalis in the preputial cavity. Further investigation on the potential implication of this finding on sheep reproductive health (e.g. infectious infertility, and abortion) and overall epidemiology of Campylobacter may be warranted., (© 2020 The Society for Applied Microbiology.)

The aims of this study were to detect the frequency of isolation of thermophilic Campylobacter spp. from acute gastroenteritis cases by phenotypic and molecular methods and to evaluate the antibiotic susceptibilities of the isolates. A total of 3287 stool samples obtained from diarrheal patients who were admitted to Kayseri Training and Research Hospital, Kayseri, Turkey, between March 2010 - March 2011 and sent to the microbiology laboratory, were included in the study. Cefoperazone, amphotericin B and teicoplanin (CAT) supplemented Campylobacter Blood-free Selective Medium (modified CCDA-Preston, Oxoid CM739, UK) was used for the isolation of Campylobacter spp. The media inoculated with stool samples were incubated at 42°C microaerobically for 72 to 96 hours. The genus and species level identifications of the thermophilic Campylobacter spp. isolates defined by phenotypic tests were carried out with multiplex polymerase chain reaction (mPCR). Antibiotic susceptibilities of the isolates were detected by disc diffusion method and the results were evaluated according to the CLSI guidelines. The thermophilic Campylobacter spp. were isolated from 5.4% (179/3287) of the patients' stool samples. Of Campylobacter positive cases 71% (127/179) were children and 58% (104/179) were male. The prevalence rate was estimated as 7.5% (127/1683) for children and 3.2% (52/1604) for adults. Of the isolates, 146 (82%) were identified as C.jejuni, 24 (13%) were C.coli, 6 (3%) were C.lari and 3 (2%) were C.upsaliensis with phenotypic tests. By using mPCR, 152 (85%) and 27 (15%) of 179 isolates were identified as C.jejuni and C.coli, respectively. Three of the six isolates identified as C.lari by the phenotypic methods were identified as C.jejuni and the remaining three as C.coli by mPCR. Phenotypically identified three C.upsaliensis isolates were shown to be C.jejuni (n= 2) and C.coli (n= 1). On the other hand one C.coli isolate was found to be C.jejuni by mPCR. The rates of resistance of the isolates were 92.6% for trimethoprim-sulfamethoxazole, 79.5% for nalidixic acid, 75.6% for levofloxacin, 73.9% for ciprofloxacin, 40.3% for ampicillin, 35% for cefotaxime, 33.4% for piperacillin-tazobactam, 24% for tetracycline, 14.6% for clindamycin, 11.2% for amikacin and 6.3% for erythromycin. During the 13 months study period, the highest isolation rates were detected between March-June (mean rate 66%). Our data concerning the prevalence and antibiotic resistance rates revealed the significance of campylobacters in gastroenteritis cases. Therefore, specific microbiological isolation and identification methods should be applied in routine microbiology laboratories to investigate the presence of campylobacters in gastroenteritis etiology. Besides, determination of the antibiotic susceptibilities of the isolates on routine basis should be encouraged to help to guide the antimicrobial treatment approaches in case of gastroenteritis. The results of this study also indicated that phenotypic tests were adequate for the identification of campylobacters at the genus-level, however, for accurate identification at the species level and for reliable epidemiological data molecular analysis might be added to the detailed identification procedures.