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Thirty fecal samples from wild specimens of Iberian lynx were collected and analyzed for Enterococcus spp. (27 isolates) and Escherichia coli (18 isolates) recovery. The 45 isolates obtained were tested for antimicrobial resistance, molecular mechanisms of resistance, and presence of virulence genes. Among the enterococci, Enterococcus faecium and Enterococcus hirae were the most prevalent species (11 isolates each), followed by Enterococcus faecalis (5 isolates). High percentages of resistance to tetracycline and erythromycin (33% and 30%, respectively) were detected among enterococcal isolates. The tet(M) and/or tet(L), erm(B), aac(6')-Ie-aph(2″)-Ia, ant(6)-Ia, or aph(3')-IIIa genes were detected among resistant enterococci. Virulence genes were detected in one E. faecalis isolate (cpd, cylB, and cylL) and one E. hirae isolate (cylL). High percentages of resistance were detected in E. coli isolates to tetracycline (33%), streptomycin (28%), nalidixic acid (28%), and sulfamethoxazole-trimethoprim (SXT, 22%). Additionally, the blaTEM, tet(A), aadA, cmlA, and different combinations of sul genes were detected among most ampicillin, tetracycline, streptomycin, chloramphenicol and SXT-resistant isolates, respectively. Two isolates contained a class 1 integron with the gene cassette arrays dfrA1 + aadA1 and dfrA12 + aadA2. The E. coli isolates were ascribed to phylo-groups A (n=5); B1 (n=4); B2 (n=6), and D (n=3), with the virulence gene fimA present in all E. coli isolates. This study found resistance genes in wild specimens of Iberian lynx. Thus, it is important to notice that multiresistant bacteria have reached species as rare and completely non-synanthropic as the Iberian lynx. Furthermore, the susceptibility of this endangered species to bacterial infection may be affected by the presence of these virulence and resistance genes., (Copyright © 2013 Elsevier B.V. All rights reserved.)

The aim of this study was to perform the molecular characterization of vancomycin resistant enterococci (VRE) within the faecal flora of Iberian wolf and Iberian lynx. The association with other resistance genes and the detection of virulence genes were also analysed. From 2008 to 2010, 365 faecal samples from Iberian wolf and Iberian lynx were collected and tested for VRE recovery. Mechanisms of resistance to vancomycin and other antibiotics, as well as genes encoding virulence factors were detected through PCR. Multilocus Sequence Typing (MLST) was performed for Enterococcus faecium strains. VRE were recovered in 8 of the 365 analysed samples. The vanA gene was identified in two E. faecium isolates recovered from Iberian wolf faecal samples and the remaining six showed intrinsic resistance (3 vanC1-E. gallinarum and 3 vanC2-E. casseliflavus, from Iberian wolf and Iberian lynx faecal samples, respectively). One vanA-containing isolate showed tetracycline and erythromycin resistance [with erm(B) and tet(L) genes] and the other one also exhibited ampicillin and kanamycin resistance [with erm(B), tet(M) and aph(3')-III genes]. One of the vanA-isolates revealed a new sequence type named ST573 and the other one belonged to the CC17 clonal complex (ST18). The hyl gene was detected in one E. casseliflavus and three E. gallinarum but not among vanA-positive isolates, and the occurrence of cylA and cylL genes was confirmed in two E. casseliflavus isolates. A low prevalence of VRE has been detected in faecal samples of Iberian wolf and Iberian lynx and strains with an acquired mechanism of resistance to vancomycin have not been detected among Iberian lynx., (Copyright © 2011 Elsevier B.V. All rights reserved.)

A total of 44 Escherichia coli and 64 enterococci recovered from 77 intestinal samples of wild European rabbits in Portugal were analyzed for resistance to antimicrobial agents. Resistance in E. coli isolates was observed for ampicillin, tetracycline, sulfamethoxazole/trimethoprim, streptomycin, gentamicin, tobramycin, nalidixic acid, ciprofloxacin and chloramphenicol. None of the E. coli isolates produced extended-spectrum beta-lactamases (ESBLs). The bla(TEM), aadA, aac(3)-II, tet(A) and/or tet(B), and the catA genes were demonstrated in all ampicillin, streptomycin, gentamicin, tetracycline, and chloramphenicol-resistant isolates respectively, and the sul1 and/or sul2 and/or sul3 genes in 4 of 5 sulfamethoxazole/trimethoprim resistant isolates. Of the enterococcal isolates, Enterococcus faecalis was the most prevalent detected species (39 isolates), followed by E. faecium (21 isolates) and E. hirae (4 isolates). More than one-fourth (29.7%) of the isolates were resistant to tetracycline; 20.3% were resistant to erythromycin, 14.1% were resistant to ciprofloxacin and 10.9% were resistant to high-level-kanamycin. Lower level of resistance (<10%) was detected for ampicillin, quinupristin/dalfopristin and high-level-gentamicin, -streptomycin. No vancomycin-resistance was detected in the enterococci isolates. Resistance genes detected included aac(6')-aph(2''), ant(6)-Ia, tet(M) and/or tet(L) in all gentamicin, streptomycin and tetracycline-resistant isolates respectively. The aph(3')-IIIa gene was detected in 6 of 7 kanamycin-resistant isolates, the erm(B) gene in 11 of 13 erythromycin-resistant isolates and the vat(D) gene in the quinupristin/dalfopristin-resistant E. faecium isolate. This survey showed that faecal bacteria such as E. coli and enterococci of wild rabbits could be a reservoir of antimicrobial resistance genes., (Copyright 2010 Elsevier B.V. All rights reserved.)