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

Autor(es)

Amarante, M. R. V. ^[1] ; Bassetto, C. C. ^[1] ; Neves, J. H. ^[1] ; Amarante, A. F. T. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Estadual Paulista, UNESP, Inst Biociencias, Dept Parasitol, BR-18618970 Botucatu, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: JOURNAL OF HELMINTHOLOGY; v. 88, n. 4, p. 447-452, DEC 2014.
• Citações Web of Science: 2

Resumo

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. (AU)

• Processo FAPESP: 10/18678-5 - Proteção de ovinos e bovinos contra haemoncose após imunização com antígenos oriundos da membrana intestinal de Haemonchus contortus.
• Beneficiário: César Cristiano Bassetto
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 07/07182-6 - Identificacao de nematodeos parasitas de bovinos e ovinos com a utilizacao de marcadores moleculares.
• Beneficiário: Mônica Regina Vendrame Amarante
• Modalidade de apoio: Auxílio à Pesquisa - Regular