Random Amplification of Polymorphic DNA, a simple tool for the detecRandom Amplification of Polymorphic DNA, a simple tool for the detection of triclabendazole resistant strains in Fasciola hepatica

SCARCELLA S. ; LAMENZA P.; GUZMÁN M.,; MALANDRINI B.; ORTIZ OBLITAS P.; JENSEN O.; SOLANA H.

Buenos Aires

Congreso; XXIII Congreso Internacional De La Asociación Mundial Para el Avance de la Parasitología Veterinaria; 2011

Random Amplification of Polymorphic DNA, a simple tool for the detection of triclabendazole resistant strains in Fasciola hepatica   Scarcella1, S., Lamenza1, P., Guzmán2, M., Fernández1, V., Malandrini3, B., Ortiz Oblitas4 P., Jensen5, O., Solana1, H.   1 Lab. Biol. Cel. y Mol. FCV-UNCPBA – Tandil, Buenos Aires 2 Lab. Parasitologia FCV-UNCPBA – Tandil, Buenos Aires 3 Facultad de Ciencias de la Salud. Universidad Nacional de Catamarca-San Fernando del Valle de Catamarca-Catamarca 4 FCV-Universidad de Cajamarca-Cajamarca, Perú 5 Dpto. de Zoonosis - Secretaría de Salud-Chacra Nº 18 9020, Sarmiento, Provincia del Chubut, Argentina.   E-mail: silvanas@vet.unicen.edu.ar   Anthelmintic resistance in parasitic of livestock is a chronic problem in the world. Fasciolasis is a zoonotic parasitic disease caused by Fasciola hepatica. Its control is mainly based on the use of triclabendazole (TCBZ), a halogenated benzimidazole thiol derivative which shows excellent efficacy against both juvenile (immature) and adult stages.  Benzimidazoles are effective, broad-spectrum anthelmintics that bind to β-tubulins and selectively dissasembles microtubules. The intensive use of TCBZ has resulted in the development of resistant liver flukes. In H. contortus and other helminths the resistance to the benzimidazoles is caused by genetic changes in genes encoding β-tubulins. In the case of F. hepatica TCBZ resistant were not detected these genetic changes on the tubulin molecule for which the resistance obtained is from other genetic or metabolic changes. The Random Amplification of Polymorphic DNA (RAPD-PCR) technique involves the enzymatic amplification of DNA fragments, using primers of arbitrary sequence to hybridize loci randomly distributed throughout the genome. This reveals the existence of polymorphisms that are used as genetic markers and taxonomic all kinds of organisms. RAPD-PCR technique allows to comparatively evaluating the presence of genetic variations among strains of same species. The aim of the present work was to evaluate different strains of TCBZ-susceptible and TCBZ-resistant flukes of several places in Argentina and Peru using RAPD-PCR. The flukes were compared against controls pure strains TCBZ-susceptible (Cullompton strain) and flukes TCBZ-resistant (Sligo strain) both of the same geographic region. Were analyzed field strains flukes of Catamarca (northwest of Argentine), Chubut (south of Argentine) and Cajamarca (north of Peru). The primer used detected differences between Cullompton and Sligo strains and allowed us to detect probable resistant strains in the flukes field strains tested in the same geographic region. These preliminary results may be useful to better understand the mechanisms underlying the phenomenon of resistance at TCBZ in Fasciola hepatica.  Further work on this relevant area is required