A random selection of ampr isolates all showed β-lactamase activity, but when tested by bla TEM PCR, only 4 out of 144 isolates were positive. This indicates a low level of bla TEM alleles. The four isolates were all identified as E. coli, and the bla TEM alleles were inserted in a Tn3 transposon which is found

in a wide variety of bacteria. The presence of bla TEM alleles has previously been reported in wild animals in Portugal, where they detected the alleles in E. coli isolated from faeces from deer, fox, owl, and birds of prey [38]. Others have identified bla TEM in faecal E. coli isolates from pigs, dogs, and cats [17, 39]. The bla TEM PCR on total DNA extracted was negative for the two rectal swabs, and two of KU-57788 in vivo the three faecal samples were bla TEM PCR positive (Fig. 3). Previous studies on Arctic soil samples suggest that the detection limit for total DNA extracted was < 21 bla TEM alleles (pUC18) per PCR sample [15]. The diversity analysis of polar bear faeces showed a dominance of clostridiales in which there has been no reports of β-lactamase production. This is consistent with the low levels

of bla TEM alleles detected in the samples. Conclusions This study showed that the bacterial diversity in faeces from polar bears in their natural environment in the pristine Svalbard area were low, all obtained clones affiliated to Firmicutes. As with any PCR-based method, 16S rRNA gene clone libraries are biased [40] and the gastrointestinal microbiota of more polar bears should be studied to give a more complete MAPK inhibitor picture of the microbial diversity. buy VS-4718 Furthermore, only low levels of bla TEM alleles were detected in contrast to their increasing prevalence in some clinical and commensal bacterial populations. Methods Sampling Ten samples from ten polar bears were collected on two occasions. Faeces were sampled from five individuals March 30th-April

12th 2004 and from five individuals March 30th-April 9th 2006 (Table 5). Sampling occurred on both occasions at the coast or the surrounding sea ice at Spitsbergen and Liothyronine Sodium Nordaustlandet in Svalbard, Norway (Fig. 1). Bears were caught by remote injection of a dart (Palmer Cap-Chur Equipment) containing the drug Zoletil® (Virbac, Carros Cedex, France) fired from a helicopter [41]. Animal handling methods were approved by the National Animal Research Authority (Norwegian Animal Health Authority, P.O. Box 8147 Dep., N-0033 Oslo, Norway). The sex, reproductive status, and a series of standardized morphometric measurements were collected from each bear (Table 5). In 2004, the samples were collected by swabbing rectum and the samples were kept frozen in LB-broth (Luria Broth, Fluka BioChemica) with 20% glycerol. In 2006, faeces was collected with a sterile glove and kept in sterilized plastic bags. The amount of sample ranged from 0.2 g to 2 g.