Species-specific PCR for the identification of Cooperia curticei (Nematoda: Trichostrongylidae) in sheep. Issue 4 (December 2014)

• Record Type: Journal Article
• Authors: Amarante, M.R.V.; Bassetto, C.C.; Neves, J.H.; Amarante, A.F.T.
• Is Part Of: Journal of helminthology. Volume 88:Issue 4(2014)
• Page Start: 447
• Page End: 452
• Publication Date: 2014-12
• Subjects: Helminthology -- Periodicals; 592.3
• DOI: 10.1017/S0022149X13000412
• Languages: English
• ISSNs: 0022-149X

Abstract

<abstract abstract-type="normal"> <title>Abstract</title> <p>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. <italic>Haemonchus</italic> spp. and <italic>Cooperia</italic> spp. are the most important gastrointestinal nematodes infecting ruminants in subtropical/tropical environments. In Brazil, <italic>C. punctata</italic> is more adapted to cattle than sheep. Additionally, <italic>C. spatulata</italic> appears to be more adapted to cattle, whereas <italic>C. curticei</italic> is more adapted to sheep. However, infection of sheep with <italic>C. punctata</italic> 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 <italic>C. curticei.</italic> The polymerase chain reaction (PCR) product was sequenced and showed 97% similarity to <italic>C. oncophora</italic> partial ITS-1 clones and 99% similarity to the <italic>C. curticei</italic> sequence JF680982. The specificity of this primer pair was corroborated by the analysis of 17 species of helminths, including <italic>C. curticei, C. punctata</italic> and <italic>C. spatulata.</italic> 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.</p> </abstract>