Detection of anthelmintic resistance in gastrointestinal nematodes from Spanish sheep flocks comparing the egg hatch assay and the presence of resistant alleles by pyrosequencing
Trabajo presentado al: 25th International Conference of the World Association for the Advancement of Veterinary Parasitology. (Liverpool, Reino Unido, 16-20 August, 2015).
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World Association for the Advancement of Veterinary Parasitology
2015-08-16
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| Online Access: | http://hdl.handle.net/10261/133609 http://dx.doi.org/10.13039/100007652 |
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dig-igm-es-10261-1336092021-04-13T15:13:13Z Detection of anthelmintic resistance in gastrointestinal nematodes from Spanish sheep flocks comparing the egg hatch assay and the presence of resistant alleles by pyrosequencing Esteban Ballesteros, M. Calvete, C. Uriarte, J. Melville, L. Skuce, Philip González Lanza, Camino Rojo Vázquez, Francisco Antonio Martínez Valladares, María CSIC - Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA) Trabajo presentado al: 25th International Conference of the World Association for the Advancement of Veterinary Parasitology. (Liverpool, Reino Unido, 16-20 August, 2015). Study funded by Spanish lNIA(RTA2013-00064-C02-02). Anthelmintic resistance (AR) in ovine gastrointestinal nematodes (GlN) to benzimidazole (BZs) is associated with single nucleotide polymorphisms (SNPs) in the isotype 1 of B-tubulin, at codon 200 principally, but also at codons 167 and 198. ln this study we compared the levels of AR of GIN field strains by the Egg Hatch Assay (EHA) and the percentage of BZ resistance alleles obtained after pyrosequencing. Throughout the years 2012 and 2013, variation in resistance was checked by EHA in two sheep flocks (A and B). Resistance was measured by the proportion of eggs hatched at discriminating dose of 0.1 g/ml thiabendazole. DNA extraction was carríed out from a pool of larvae and eggs from faecal samples. Prior to pyrosequencing, the fragments including the SNPs198/200 were amplified for Trichostrongylus spp, Ieladorsagra circumcincta and Haemonchus conbrtus, independently for each species; a fragment for the SNP167 in T. circumcrncfa was also amplified. Flock A had a high level of hatching throughout the study, with a mean of 77.74o/o (57.21-90.64%);in flock B, the resistance level increased over time, with values of 33.25o/o in 2012 (15.81-57.460/o) and 52.02% in 2013 (46.06-55.19%). Pyrosequencing analysis detected the presence of the BZ resistance allele at codon 200 in Trichostrongylus spp in both flocks; the percentage of the resistant allele was B1.28Yo (19.53- IAAYo) in flock A and 21.37Yo (1 .57 -51 .48o/o) in B. However, for codon 198, the resistant allele in this species was at a low level in both flocks, 2.74o/o in A and 5.A4o/o in B, respectively. ln L circumcincta, the resistant allele incorporating SNP200 was not present in any flock, with a mean of 0.32%; these low levels were very similar at codons 198 and 167, with values of 0.87 and 1.61%, respectively. ln H. contorfus, pyrosequencing was only carried out in flock B, where this species was detected after morphological identification; the proportion of the BZ resistance allele was 6.330/o (0-22.77yo) at codon 200 and 1.35% (0-4.93%) at codon 198. When comparing the results of EHA and the frequency of BZ resistance alleles at codon 200 in Trichostrongylus spp, in both flocks and throughout the experiment, a significant correlation between the two techniques (¡=0.6; p<0.01) was found. These results suggest that Trichostrongylus spp is the main contributor to the development of AR in these Spanish GIN field strains. Study funded by Spanish lN|A(RTA2013-00064-C02-02) Peer Reviewed 2016-06-16T11:16:45Z 2016-06-16T11:16:45Z 2015-08-16 2016-06-16T11:16:46Z comunicación de congreso http://purl.org/coar/resource_type/c_5794 25th International Conference of the World Association for the Advancement of Veterinary Parasitology: 242 (2015) http://hdl.handle.net/10261/133609 http://dx.doi.org/10.13039/100007652 #PLACEHOLDER_PARENT_METADATA_VALUE# info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/RTA2013-00064-C02-02 http://www.waavp2015.com/ Sí none World Association for the Advancement of Veterinary Parasitology |
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Trabajo presentado al: 25th International Conference of the World Association for the Advancement of Veterinary Parasitology. (Liverpool, Reino Unido, 16-20 August, 2015). |
| author2 |
CSIC - Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA) |
| author_facet |
CSIC - Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA) Esteban Ballesteros, M. Calvete, C. Uriarte, J. Melville, L. Skuce, Philip González Lanza, Camino Rojo Vázquez, Francisco Antonio Martínez Valladares, María |
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comunicación de congreso |
| author |
Esteban Ballesteros, M. Calvete, C. Uriarte, J. Melville, L. Skuce, Philip González Lanza, Camino Rojo Vázquez, Francisco Antonio Martínez Valladares, María |
| spellingShingle |
Esteban Ballesteros, M. Calvete, C. Uriarte, J. Melville, L. Skuce, Philip González Lanza, Camino Rojo Vázquez, Francisco Antonio Martínez Valladares, María Detection of anthelmintic resistance in gastrointestinal nematodes from Spanish sheep flocks comparing the egg hatch assay and the presence of resistant alleles by pyrosequencing |
| author_sort |
Esteban Ballesteros, M. |
| title |
Detection of anthelmintic resistance in gastrointestinal nematodes from Spanish sheep flocks comparing the egg hatch assay and the presence of resistant alleles by pyrosequencing |
| title_short |
Detection of anthelmintic resistance in gastrointestinal nematodes from Spanish sheep flocks comparing the egg hatch assay and the presence of resistant alleles by pyrosequencing |
| title_full |
Detection of anthelmintic resistance in gastrointestinal nematodes from Spanish sheep flocks comparing the egg hatch assay and the presence of resistant alleles by pyrosequencing |
| title_fullStr |
Detection of anthelmintic resistance in gastrointestinal nematodes from Spanish sheep flocks comparing the egg hatch assay and the presence of resistant alleles by pyrosequencing |
| title_full_unstemmed |
Detection of anthelmintic resistance in gastrointestinal nematodes from Spanish sheep flocks comparing the egg hatch assay and the presence of resistant alleles by pyrosequencing |
| title_sort |
detection of anthelmintic resistance in gastrointestinal nematodes from spanish sheep flocks comparing the egg hatch assay and the presence of resistant alleles by pyrosequencing |
| publisher |
World Association for the Advancement of Veterinary Parasitology |
| publishDate |
2015-08-16 |
| url |
http://hdl.handle.net/10261/133609 http://dx.doi.org/10.13039/100007652 |
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