Caracterização molecular e avaliação bioquímica de Trypanosoma vivax e Trypanosoma theileri no Estado do Rio de Janeiro baseado nos genes 18S rDNA e Cathepsina L-like

Abstract

This study aimed to perform the molecular characterization of Trypanosoma vivax and Trypanosoma theileri through the 18S and catl genes, as well as to analyze the biochemical profile related to infection for Trypanosoma vivax in cattle from the state of Rio de Janeiro. The samples were collected by means of venocuncture of the coccygeal vein of 389 cattle, and the whole blood collected was placed in tubes containing 10% ethylene-amino-tetracetic acid for molecular analysis and tubes without anticoagulant for biochemistry, being subsequently sent to the Hemoparasites and vectors from the Federal Rural University of Rio de Janeiro where the analyzes were performed. The DNA samples were extracted following the protocol recommended by the manufacturer (Promega®) and subsequently stored in a -20 ° C freezer until the moment of molecular analysis. The serum for biochemical analysis was obtained by centrifugation in a clinical centrifuge at 3,000 rpm for 5 minutes and was later sent to the Clinical Analysis Laboratory of the Experimental Chemotherapy Laboratory in Veterinary Parasitology (LQEPV / UFRRJ), where the serum levels of Glutamyltransferase (GGTT) ), Total Serum Protein (PTS), Albumin (ALB), Total Bilirubin (BT) and Direct (BD), Creatinine (CR), Urea (UR), Alkaline Phosphatase (FAL), Lactate Dehydrogenase (LDH), Aspartate Aminotransferase (AST / TGO) and Alanine Aminotransferase (ALT / TGP) were determined using the automated biochemical analyzer A15 (Biosystems®) and commercial reagent kits according to the equipment used. Molecular analysis was performed using the 18S genes to identify Trypanosoma sp, and catl-like to identify Trypanosoma vivax and T. theileri. After frequency analysis, some samples from each municipality were selected for phylogenetic analysis and comparison of the identity of the sequences of the present study with other sequences found in GenBank. Among the analyzed samples 4.6% (18/389) were positive for Trypanosoma vivax in a blood smear, with no positive samples for Trypanosoma theileri being seen. For the 18S gene, 15.4% (60/389) were positive for Trypanosoma sp, while for the catl gene, 12.8% (50/389) were positive for T. vivax while for T. theileri there was 3.6 % (14/389) of positive animals. Phylogenetic analysis for T. vivax showed 2 clusters defined for the 18S rDNA gene and 4 for the catl gene, whereas for T. theileri, we observed 1 clusters defined for the 18S rDNA gene and 4 for the catl gene, with 15 distinct genotypes. This study aimed to report the first molecular characterization of T. vivax and T. theileri in the state of Rio de Janeiro, as well as the first report of the IB and IF lineage of T. theileri based on the catl gene. In addition, this was also the first report of biochemical analysis of animals naturally infected with Trypanosoma vivax in the state of Rio de Janeiro, where it was possible to verify that LDH was the only significantly altered analysis representing approximately seven times more chance of cattle being positive for T. vivax when this enzyme is increased