Abstract

DIAZ-RIVERA, Edgar; HOLGUIN CESPEDES, Gisella  and  URREA MONTES, Daniel A.. New polymorphism in the sodium channel gene of Rhipicephalus microplus tick (Ixodida: Ixodidae) resistant to pyrethroids. Rev. biol. trop [online]. 2019, vol.67, n.4, pp.935-944. ISSN 0034-7744.  http://dx.doi.org/10.15517/rbt.v67i4.35250.

For the control of Rhipicephalus microplus, cattle tick, pyrethroid compounds are used among others. The study of the mechanisms of resistance to pyrethroids in this tick has shown the presence of mutations in the sodium channel gen, the target site of acaricide. The main objective of the present study was to detect the T2134A polymorphism in the sodium channel gene that has been related to pyrethroid resistance in carriers. To do so, 182 ticks R. microplus were collected in two farms with a history of resistance to this type of acaricide and were subjected to a dip test of females in a cypermethrin solution prepared at the recommended dose for field use, obtaining six susceptible, 92 resistants and 84 moderately resistants. DNA is extracted from all specimens using QIAamp DNA Mini Kit® and processed by conventional PCR using allele-specific primers to amplify a segment of domain IIIS6 of the sodium channel gene. The resulting amplicons were detected by electrophoresis in 2 % agarose gels stained with GelRed® and visualized with ultraviolet light. In the same way, 20 samples were sequenced of the three susceptible phenotypes, moderately resistant and resistant and the resulting sequences were aligned through the Mega 7 program and compared with the sequences stored in the GenBank®. The information of the population of R. Microplus studied did not show the expected mutation but a different mutation in the site 2134. We also observed the substitution of a thymine but by a cytosine, T2134C, which changes to a phenylalanine by a leucine, a difference from the previously identified mutation, that the change from phenylalanine to isoleucine. This shows the presence of a polymorphism that has not been reported in this species of tick and that would generate a new molecular marker of pyrethroid resistance for populations of this mite.

Keywords : ticks; chemical resistance; pesticides; mutations; polymerase chain reaction.

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