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

Abstract

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 6C for 72-96 h. Phenotypic tests, a genusspecific 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. INTRODUCTIONThe genus Arcobacter is a member of the family Campylobacteraceae. Currently, the genus Arcobacter has a total of 15 recognized species: Arcobacter butzleri, Arcobacter cryaerophilus, Arcobacter skirrowii, Arcobacter nitrofigilis, Arcobacter cibarius, Arcobacter halophilus, Arcobacter mytili, Arcobacter thereius, Arcobacter marinus, Arcobacter trophiarum, Arcobacter defluvii, Arcobacter molluscorum, Arcobacter ellisi, Arcobacter bivalviorum and Arcobacter venerupis Figueras et al., 2011; Levican et al., 2012). Three species, namely A. butzleri, A. cryaerophilus and A. skirrowii, have been associated with gastrointestinal infections (Burnens et al., 1992;Jiang et al., 2010;Vandamme et al., 1992a) and extra-intestinal invasive diseases (Lau et al., 2002;On et al., 1995;Yan et al., 2000). In addition, asymptomatic arcobacter carriage in type 2 diabetic patients has been reported (Fera et al., 2010). Because of its isolation from many human disease cases, A. butzleri is considered to be the most important species of the genus. Some studies have reported the isolation and molecular analyses of arcobacters from miscellaneous sources in Turkey (Atabay et al., 2003(Atabay et al., , 2008Aydin et al., 2007;Ertas et al., 2010). However, in human medicine, no studies have been conducted on arcobacters, except for one case report (Kayman et al., 2010).The current study was undertake...

Comparison of Conventional PCR, Multiplex PCR, and Loop-Mediated Isothermal Amplification Assays for Rapid Detection of Arcobacter Species

“…Bacterial culture is not practical for the rapid detection of Arcobacter species because of their slow growth in Campylobacter blood-free selective medium; therefore, conventional PCR, multiplex PCR, and enterobacterial repetitive intergenic consensus (ERIC)-PCR were used in previous studies for the rapid detection of Arcobacter spp. (23,35,36). In this study, five C. jejuni strains and one C. coli strain were detected as false positives by multiplex PCR described previously (26).…”

“…In severe chronic human diseases caused by Arcobacter spp., the rapid detection and treatment of Arcobacter infection are required (35). Nine A. butzleri isolates were cultured from diarrheal stool samples in Turkish hospitals, although A. butzleri was first isolated from the blood of a Korean patient with liver cirrhosis (36)(37)(38). Bacterial culture is not practical for the rapid detection of Arcobacter species because of their slow growth in Campylobacter blood-free selective medium; therefore, conventional PCR, multiplex PCR, and enterobacterial repetitive intergenic consensus (ERIC)-PCR were used in previous studies for the rapid detection of Arcobacter spp.…”

Bacteremia Caused by Arcobacter butzleri in an Immunocompromised Host

“…In Turkey, A. butzleri was identified as a cause of gastroenteritis, at a rate of 0.3% in 3,297 stools examined. In that study, one third of the patients with A. butzleri infections presented with diarrhea, and all of them presented with abdominal pain and nausea (18). Arcobacter butzleri has also been detected in travelers' diarrhea at a rate of 8% (16/201 stool specimens) in U.S. and European travelers returning from Mexico, Guatemala, and India (19).…”

“…A case of A. butzleri bacteremia was described in a 60-year-old male in Taiwan with liver cirrhosis, fever, and esophageal variceal bleeding (20). In Hong Kong, a 69-year-old female with appendicitis and local perforation developed A. butzleri bacteremia (18). In 1995, a case report from London described a neonate with bacteremia, suggesting possible vertical transmission of this organism (21).…”

Comparative Detection and Quantification of Arcobacter butzleri in Stools from Diarrheic and Nondiarrheic People in Southwestern Alberta, Canada

“…Most methods for the isolation of A. butzleri from microbiologically complex matrices rely on selective enrichments and/or antibiotics to inhibit the growth of nontarget microorganisms (8,9). In addition, the incubation temperature and atmosphere utilized for isolation have been inconsistent; temperatures vary from 25°C (10) to 37°C (11), and atmospheres range from aerobic (9,12) to microaerobic (5 to 6% O 2 , 6 to 10% CO 2 , 0 to 7% H 2 , and 79 to 85% N 2 ) and anaerobic (10,(13)(14)(15). Accumulated evidence indicates that no single medium, temperature, or atmosphere will isolate all strains of A. butzleri.…”