Molecular biology, genetics and biotechnology Azorean wild rabbits as reservoirs of antimicrobial resistant Escherichia coli Catarina Marinho a, b, c, d , Gilberto Igrejas a, b , Alexandre Gonçalves a, b, c, d , Nuno Silva c , Tiago Santos a, b, c, d , Ricardo Monteiro a, b, c, d , David Gonçalves e, f , Tiago Rodrigues e , Patrícia Poeta c, d, * a Institute for Biotechnology and Bioengineering, Centre of Genomics and Biotechnology, University of Tr as-os-Montes and Alto Douro, Apartado 1013, 5001-801 Vila Real, Portugal b Department of Genetics and Biotechnology, University of Tr as-os-Montes and Alto Douro, Apartado 1013, 5001-801 Vila Real, Portugal c Centre of Studies of Animal and Veterinary Sciences, Apartado 1013, 5001-801 Vila Real, Portugal d Department of Veterinary Sciences, University of Tr as-os-Montes and Alto Douro, Apartado 1013, 5001-801 Vila Real, Portugal e CIBIO, Research Centre in Biodiversity and Genetic Resources of the University of Porto, R. Padre Armando Quintas, 4485-661 Vair~ ao, Portugal f Biology Department, Sciences Faculty, University of Porto, Rua Campo Alegre s/n, 4169-007 Porto, Portugal article info Article history: Received 29 April 2014 Received in revised form 8 September 2014 Accepted 12 September 2014 Available online 20 September 2014 Keywords: Antibiotic resistance Escherichia coli Wild rabbit abstract Antibiotic resistance in bacteria is an increasing problem that is not only constrained to the clinical setting but also to other environments that can lodge antibiotic resistant bacteria and therefore they may serve as reservoirs of genetic determinants of antibiotic resistance. One hundred and thirty-six faecal samples from European wild rabbits (Oryctolagus cuniculus algirus) were collected on S~ ao Jorge Island in Azores Archipelago, and analysed for Escherichia coli isolates. Seventy-seven isolates (56.6%) were recovered and studied for antimicrobial resistance, one isolate per positive sample. Thirteen (16.9%), 19 (24.7%), 25 (32.4%) and 20 (26%) isolates were ascribed to A, B1, B2 and D phylogenetic groups, respectively, by specific primer polymerase chain reaction. Different E. coli iso- lates were found to be resistant to ampicillin (16.9%), tetracycline (1.3%), streptomycin (42.9%), sul- famethoxazoleetrimethoprim (1.3%), amikacin (1.3%), tobramycin (2.6%) and nalidixic acid (1.3%). Additionally, the bla TEM , tetA, strA/strB, aadA, sul1, intI, intI2 and qacED þ sul1 genes were found in most resistant isolates. This study showed that E. coli from the intestinal tract of wild rabbits from Azores Archipelago are resistant to widely prescribed antibiotics in medicine and they constitute a reservoir of antimicrobial resistant genes, which may play a significant role in the spread of antimi- crobial resistance. Therefore, antibiotic resistant E. coli from Azorean wild rabbits may represent an ecological and public health problem. © 2014 Elsevier Ltd. All rights reserved. 1. Introduction Antibiotic resistant bacteria from different habitats have been increasing, mainly as a result of the intensive use of antibacterial drugs in human activities, veterinary medicine, and bacterial ge- netic transfer that allow antibiotic resistant strains to emerge and spread through wildlife sources [1,2]. Commensal bacteria from the intestinal microflora constitute a reservoir of resistance genes for facultative and obligate pathogenic bacteria [3,4]. Dissemina- tion of resistant bacteria to antibiotics commonly used in the treatment of human and animal infections is an emergent public health problem. Escherichia coli colonize the gastrointestinal tract of many animals and are also commonly found in soil, plants, vegetables and water [5]. Also, E. coli are expert in horizontal genes transfer even to phylogenetically distant bacteria. This microor- ganism is considered an important “indicator bacteria” in order to track the development of antibiotic resistance in different eco- systems [3]. The European wild rabbit (Oryctolagus cuniculus) is endemic in the Iberian Peninsula and native in Azorean Archi- pelago. These animals represent an essential species in Iberian ecosystems where it is hunted and eaten by carnivorous animals and humans [6]. Interaction between wild rabbits and the * Corresponding author. Department of Veterinary Sciences, University of Tr as- os-Montes and Alto Douro, Vila Real, Portugal. Tel.: þ351 259350466; fax: þ351 259350629. E-mail address: ppoeta@utad.pt (P. Poeta). Contents lists available at ScienceDirect Anaerobe journal homepage: www.elsevier.com/locate/anaerobe http://dx.doi.org/10.1016/j.anaerobe.2014.09.009 1075-9964/© 2014 Elsevier Ltd. All rights reserved. Anaerobe 30 (2014) 116e119