Emerging pathogen Arcobacter spp. in acute gastroenteritis: molecular identification, antibiotic susceptibilities and genotyping of the isolated arcobacters.

The aims of this study were as follows: (i) to isolate Arcobacter spp. from the stool samples of patients with gastroenteritis; (ii) to identify them with molecular methods; (iii) to genotype them using enterobacterial repetitive intergenic consensus (ERIC)-PCR; and (iv) to determine their antibiotic susceptibilities. For the study, a total of 3287 diarrhoeal stool samples submitted to the Microbiology Laboratory of the Kayseri Training and Research Hospital, Kayseri, Turkey, between 2010 and 2011 were analysed. Campylobacter blood-free selective medium supplemented with cefoperazone, amphotericin B and teicoplanin was used for isolation. Medium inoculated with stool samples was incubated microaerobically at 37 °C for 72-96 h. Phenotypic tests, a genus-specific PCR and a multiplex PCR were used to identify the arcobacters, whilst ERIC-PCR was used for genotyping and the antibiotic susceptibilities of the isolates were detected by E-test. Arcobacter spp. were isolated from nine of the 3287 samples. These nine isolates were identified as Arcobacter butzleri and all showed different ERIC-PCR profiles. All nine isolates were resistant to ampicillin and susceptible to gentamicin, tetracycline, erythromycin and ciprofloxacin. As far as is known, this is the first study in which A. butzleri has been isolated from human acute gastrointestinal infections in Turkey. According to these results, it is recommended that, when investigating the aetiology of infections of the digestive system in humans, Arcobacter spp. be considered for inclusion. The results of this study should contribute to our knowledge related to A. butzleri infections in humans.

Seroepidemiological investigation of brucellosis in sheep abortions in Kars, Turkey.

This study was undertaken to investigate the seroprevalence of brucellosis in unvaccinated sheep from the flocks having previous abortion cases in Kars and around, Turkey and to compare the efficacy of each serological test used. Four hundred serum samples collected from 16 different flocks of sheep having a history of abortions in Kars and its surrounding area in Turkey were examined for the presence of antibodies raised against Brucella using Rose Bengal Plate Test (RBPT), Serum Agglutination Test (SAT), Rivanol Agglutination Test (RAT) and Complement Fixation Test (CFT). All animals were unvaccinated against Brucella. Of the serum samples tested, 147 (%36.7), 142 (%35.5), 139 (%34.75) and 135 (%33.75) were found positive by SAT, RAT, RBPT and CFT, respectively. No statistically significant difference was found between the serological tests used (p > 0.05). It is concluded from this study that brucellosis continues to be an important problem for ovine abortions and poses a risk both for human and other animals in this area. Therefore, adequate intervention measures should be implemented to control and eradicate brucellosis. In addition, if conventional serological tests are used at least two tests, RPBT for screening and CFT for the confirmation of the positive samples are preferable, should be used in parallel for detection of brucellosis effectively.

Isolation of various Arcobacter species from domestic geese (Anser anser).

In this study, the prevalence and distribution of various Arcobacter spp. were investigated in samples taken from the cloacae of healthy domestic geese raised in Turkey. A membrane filtration technique with a non-selective blood agar was employed after enrichment in Arcobacter enrichment broth (AEB) to isolate a wide range of Arcobacter spp. In addition, the isolates were characterized phenotypically and identified at species level using a multiplex-PCR assay. A total of 90 cloacal swab samples taken from geese, collected on three farms (18, 25, 47 samples, respectively), were examined. Of the samples examined, 16 (18%) were found positive for Arcobacter. One Arcobacter species was isolated from each bird. Of the 16 Arcobacter isolates, 7 (44%), 7 (44%) and 2 (12.5%) were identified by m-PCR as A. cryaerophilus, A. skirrowii and A. butzleri, respectively. The present study indicates that domestic geese can harbour a variety of Arcobacter spp. in their cloacae. The presence of Arcobacter in geese may be of significance as reservoirs in their dissemination. Detailed research is needed for better understanding of the epidemiology and zoonotic potential of this emerging pathogen.

Detection and diversity of various Arcobacter species in Danish poultry.

The prevalence and diversity of different Arcobacter spp. in various poultry species in Denmark were investigated using cultural and multiplex PCR methods. A pool of three fresh droppings obtained at the production site from 70 broiler chicken flocks aged 4-5 weeks was examined. In addition, pools of 10 cloacal swabs taken at the abattoir prior to stunning from each of 15, and 37 duck and turkey flocks, respectively, were analyzed. Thirty fresh broiler chicken carcasses and 29 cloacal swabs from the respective viscera were also examined at the abattoir. Finally, 10 caecal and 10 cloacal swabs from ducks at the abattoir were analyzed individually. In total, 85 Arcobacter isolates were obtained. Of these 45, 20 and 7 were identified as Arcobacter butzleri, Arcobacter cryaerophilus and Arcobacter skirrowii, respectively, using a multiplex PCR. Interestingly, some chicken isolates of A. butzleri showed urease activity, and 6 out of seven A. skirrowi isolates were unable to hydrolyse indoxyl acetate. All chicken carcasses examined were found positive for A. butzleri and/or A. cryaerophilus, whereas 21 (72%) of the 29 chicken cloacal swabs were positive for either A. butzleri (13) or A. cryaerophilus (9). Three (4.3%) out of 70 chicken flocks analyzed were positive only for A. cryaerophilus. Of the ten ducks examined individually, 7 carried A. skirrowii and/or A. cryaerophilus in their cloacae. None of the respective caecal samples were positive. Of the remaining 15 duck flocks, 11 (73%) were positive for A. cryaerophilus (7), A. butzleri (2) or A. skirrowii (2). Four (11%) of the 37 turkey flocks analyzed harboured either A. butzleri or A. cryaerophilus. The carriage rate of Arcobacter was higher in live ducks than those of live broiler chickens and turkeys in the present study. In addition, chicken carcasses slaughtered in Denmark were found to be contaminated with Arcobacter. The presence of Arcobacter spp. both on chicken carcasses and in poultry intestine may be of significance to human health.

The prevalence of Arcobacter spp. on chicken carcasses sold in retail markets in Turkey, and identification of the isolates using SDS-PAGE.

In this study, the prevalence of Arcobacter spp. on chicken carcasses sold in various retail markets in Turkey was investigated. The isolates were characterized and identified using various phenotypic and molecular tests. The membrane filtration technique employing 0.45-microm pore size membrane filters laid onto a nonselective blood agar was used after enrichment in Oxoid Arcobacter Enrichment Broth (AEB) to examine a total of 75 chicken carcasses (44 fresh and 31 frozen). Species level identification was performed using SDS-PAGE of whole-cell proteins and a recently developed multiplex-PCR assay. All isolates were identified as Arcobacter butzleri. Of the 44 fresh chicken carcasses examined, 42 (95%) were positive for A. butzleri. A. butzleri was also recovered from seven (23%) of the 31 frozen carcasses examined.