Diagnóstico morfológico e molecular de Haemonchus spp. em bovinos e ovinos

• Autor(a) principal: Silva, Maria Regina Lucas da [UNESP]
• Data de Publicação: 2014
• Tipo de documento: Dissertação
• Idioma: por
• Título da fonte: Repositório Institucional da UNESP
• Texto Completo: http://hdl.handle.net/11449/111041
• topic: Haemonchus; Bovino - Infecções; Ovino - Doenças; Sistema gastrointestinal - Doenças - Diagnóstico; Diagnóstico molecular; Molecular diagnosis

Nematodes gastrointestinal infection causes considerable losses in the livestock industry. Haemonchus spp. is major nematode that affects cattle and sheep mainly in tropical and subtropical region. Therefore, the aim of the present study was to assess diagnosis methodologies based in morphological and molecular analysis of infective larvae and adults of Haemonchus spp. Twenty-three calves and twenty lambs naturally infected with gastrointestinal nematodes were utilized. Before necropsy of animals, faecal samples were collated for preparing faecal cultures and obtaining infective larvae that were morphometrically analyzed. After the euthanasia of animals the abomasums were removed and frozen for posterior recovered Haemonchus specimens that were stored in alcohol 70º. In the case sheep, the parasites remained stored in alcohol 70º for more than one year before be analyzed, while the parasites of cattle were stored for only four months. The morphological diagnosis of males Haemonchus spp. was released by synlophe analysis and spicule measurements. The same parasites used in the morphological analysis were submitted in the molecular analysis, using PCR, with primers 52/159, 75/86 and HplacBotu. The “gold standard” method used to identify Haemonchus similis was spicules analysis and to identify Haemonchus placei was PCR with the primers HplacBotu. All Haemonchus specimens recovered from sheep were assumed to be Haemonchus contortus based on host specificity, and because the molecular analysis did not show any evidence of other Haemonchus species. The results presented statistic differences in the sheath tail length of infective larvae of three species. There was overlapping of measurements was only 1.26% between H.contortus and H. placei. In the espicule analysis also were observed same overlapping of measurements that caused misidentification of H. placei and H. contortus. In relation the synlophe most samples ...