Miranda-Miranda E, Liébano-Hernández E, López-Arellano ME, Mendoza-de-Gives P, Cossío-Bayúgar R

Language: Spanish
References: 22
Page: 6-12
PDF size: 102.05 Kb.

ABSTRACT

Anthelmintic resistance detection on the gastroenteric parasitic nematode Heamonchus contortus, is achieved by a complex bioassay which requires reference strains of nematodes as well as field isolated worms, cultured and exposed to different anthelmintic products. In order to assess an alternate diagnostics procedure, enzymatic assays were performed on SDS polyacrylamide gels, using larval nematodes extracts from three reference strains with variable levels of resistance and susceptibility to bencimidazole and ivermectine products. Zymograms were designed for detection of esterase (ES), phosphodiesterase (PDE), and glutathione-S transferase (GST). Zymograms were processed as digital images obtaining their respective densitograms and molecular weight (MW) in thousands of Daltons (kDa). Two ES were located with a MW of 17 and 60 kDa shared among all strains used, as well as a 24 kDa ES found only in both resistant strains. Several PDE were detected in all strains with a MW of 160, 130 and 70 kDa, as well as an isozyme of 41 kDa found only on the ivermectine resistant strain which also showed PDE enzymatic activity up to 94% above the sensitive strain basal activity. GST assay showed activity of 49 kDa band not detectable on the sensitive strain, as well as 24 and 6 kDa bands shared among all strains. We concluded that these zymograms assays as well as the data obtained from them may be used as anthelmintic resistance markers on field nematodes.

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