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Species-specific PCR for the identification of Cooperia curticei (Nematoda: Trichostrongylidae) in sheep

Published online by Cambridge University Press:  31 May 2013

M.R.V. Amarante ,
C.C. Bassetto ,
J.H. Neves  and
A.F.T. Amarante
M.R.V. Amarante*
Affiliation:
Departamento de Parasitologia, Instituto de Biociências, Unesp-Universidade Estadual Paulista, Botucatu, SP, BrazilCEP 18618-970
C.C. Bassetto
Affiliation:
Departamento de Parasitologia, Instituto de Biociências, Unesp-Universidade Estadual Paulista, Botucatu, SP, BrazilCEP 18618-970
J.H. Neves
Affiliation:
Departamento de Parasitologia, Instituto de Biociências, Unesp-Universidade Estadual Paulista, Botucatu, SP, BrazilCEP 18618-970
A.F.T. Amarante
Affiliation:
Departamento de Parasitologia, Instituto de Biociências, Unesp-Universidade Estadual Paulista, Botucatu, SP, BrazilCEP 18618-970
*
* E-mail: monicarv@ibb.unesp.br
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Abstract

Agricultural ruminants usually harbour mixed infections of gastrointestinal nematodes. A specific diagnosis is important because distinct species can differ significantly in their fecundity and pathogenicity. Haemonchus spp. and Cooperia spp. are the most important gastrointestinal nematodes infecting ruminants in subtropical/tropical environments. In Brazil, C. punctata is more adapted to cattle than sheep. Additionally, C. spatulata appears to be more adapted to cattle, whereas C. curticei is more adapted to sheep. However, infection of sheep with C. punctata is common when cattle and sheep share the same pasture. Although morphological analyses have been widely used to identify nematodes, molecular methods can overcome technical limitations and help improve species-specific diagnoses. Genetic markers in the first and second internal transcribed spacers (ITS-1 and ITS-2, respectively) of nuclear ribosomal DNA (rDNA) have been used successfully to detect helminths. In the present study, the ITS-1 region was analysed and used to design a species-specific oligonucleotide primer pair to identify C. curticei. The polymerase chain reaction (PCR) product was sequenced and showed 97% similarity to C. oncophora partial ITS-1 clones and 99% similarity to the C. curticei sequence JF680982. The specificity of this primer pair was corroborated by the analysis of 17 species of helminths, including C. curticei, C. punctata and C. spatulata. Species-specific diagnosis, which has implications for rapid and reliable identification, can support studies on the biology, ecology and epidemiology of trichostrongylid nematodes in a particular geographical location.

Type
Research Papers
Information
Journal of Helminthology , Volume 88 , Issue 4 , December 2014 , pp. 447 - 452
Copyright
Copyright © Cambridge University Press 2013 

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