Please cite this article in press as: Furtado, L.F.V., et al., First identification of the F200Y SNP in the -tubulin gene linked to benzimidazole resistance in Ancylostoma caninum. Vet. Parasitol. (2014), http://dx.doi.org/10.1016/j.vetpar.2014.10.021 ARTICLE IN PRESS G Model VETPAR-7426; No. of Pages 4 Veterinary Parasitology xxx (2014) xxx–xxx Contents lists available at ScienceDirect Veterinary Parasitology jo u r nal homep age: www.elsevier.com/locate/vetpar Short Communication First identification of the F200Y SNP in the -tubulin gene linked to benzimidazole resistance in Ancylostoma caninum Luis Fernando Viana Furtado a , Ana Cristina Passos de Paiva Bello a , Hudson Andrade dos Santos a , Maria Raquel Santos Carvalho b , Élida Mara Leite Rabelo a,∗ a Universidade Federal de Minas Gerais, Instituto de Ciências Biológicas, Departamento de Parasitologia, Belo Horizonte, Minas Gerais, Brazil b Universidade Federal de Minas Gerais, Instituto de Ciências Biológicas, Departamento de Biologia Geral, Belo Horizonte, Minas Gerais, Brazil a r t i c l e i n f o Article history: Received 25 June 2014 Received in revised form 2 October 2014 Accepted 12 October 2014 Keywords: Ancylostoma caninum Drug resistance Benzimidazoles -Tubulin ARMS-PCR a b s t r a c t Single nucleotide polymorphisms (SNPs) in the -tubulin isotype 1 gene in codons 167, 198 and 200 have been associated with benzimidazole resistance in some nematodes, although no polymorphisms in this gene have been described in the hookworm Ancylostoma can- inum to date. This study aimed to screen for SNPs in the -tubulin isotype 1 gene at codons 198 and 200 associated with benzimidazole resistance in A. caninum worms recovered from naturally infected dogs from two Brazilian states (Minas Gerais and Piauí). To perform this analysis, a molecular technique based on the amplification refractory mutation system (ARMS-PCR) was standardized. A total of 110 individual worms recovered from 9 dogs in Piauí and 124 individual worms recovered from 10 dogs in Minas Gerais were analyzed using this methodology. The presence of a SNP at position 200 was observed at a low fre- quency (0.8%), which was only detected in the worm population from Minas Gerais State. A total of 75 randomly selected samples were sequenced to validate this technique, and the presence of the SNP was confirmed. Furthermore, SNP at position 198 were evaluated by sequencing, and none were detected in this region. This is the first report of the presence of a SNP in the -tubulin isotype 1 gene associated with benzimidazole resistance in an A. caninum population. © 2014 Elsevier B.V. All rights reserved. 1. Introduction The hookworm Ancylostoma caninum is globally dis- tributed and typically affects canines, to which it can be transmitted preferentially orally but also by cutaneous penetration (Hotez et al., 2004). Although this parasite generally does not infect the intestines of humans, some reports have described A. caninum-associated eosinophilic ∗ Corresponding author. Tel.: +55 31 34092851; fax: +55 31 34092970. E-mail address: elidam.rabelo@gmail.com (É.M.L. Rabelo). enteritis in humans (Prociv and Croese, 1996). Addition- ally, this nematode is a relevant zoonotic agent, causing cutaneous larva migrans in humans (Leeming, 1966). Given the high morbidity resulting from infection by hookworms, strategies are often implemented to control for the diseases caused by these parasites in humans and animals (Hotez et al., 2004). These strategies include a mass drug administration (MDA) program, which can select for strains resistant to treatment. Benzimidazoles are the anti- helminthics of choice for MDA (Prichard et al., 2012). These drugs block the polymerization of parasite microtubules by binding to the -tubulin protein (Lacey, 1988). Single http://dx.doi.org/10.1016/j.vetpar.2014.10.021 0304-4017/© 2014 Elsevier B.V. All rights reserved.