Caracteriza??o molecular e avalia??o bioqu?mica de Trypanosoma vivax e Trypanosoma theileri no Estado do Rio de Janeiro baseado nos genes 18S rDNA e Cathepsina L-like

Detalhes bibliográficos
Autor(a) principal: Abreu, Ana Paula Martinez de
Data de Publicação: 2020
Tipo de documento: Tese
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações da UFRRJ
Texto Completo: https://tede.ufrrj.br/jspui/handle/jspui/6008
Resumo: 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
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spelling Massard, Carlos Luiz257.781.297-34https://orcid.org/0000-0002-8465-3038http://lattes.cnpq.br/7743112049924654Santos, Huarrisson Azevedo983.833.295-04https://orcid.org/0000-0002-8218-3626http://lattes.cnpq.br/3751609492049306Peixoto, Maristela Peckle110.795.51-11https://orcid.org/0000-0002-4208-1430http://lattes.cnpq.br/8817867478588076Massard, Carlos Luiz257.781.297-34https://orcid.org/0000-0002-8465-3038http://lattes.cnpq.br/7743112049924654Guedes Junior, Daniel da Silvahttp://lattes.cnpq.br/1514734234282622Roier, Erica Cristina Rochahttps://orcid.org/0000-0002-1978-9254http://lattes.cnpq.br/3538932429235543Ba?ta, Bruna de Azevedohttps://orcid.org/0000-0002-0172-556Xhttp://lattes.cnpq.br/8016012971743463Ubiali, Daniel Guimar?eshttps://orcid.org/0000-0001-8320-4567http://lattes.cnpq.br/0255736629474813051.527.387-24https://orcid.org/0000-0001-6385-3906http://lattes.cnpq.br/2317384923867126Abreu, Ana Paula Martinez de2022-09-19T13:37:20Z2020-03-11ABREU, Ana Paula Martinez de. Caracteriza??o molecular e avalia??o bioqu?mica de Trypanosoma vivax e Trypanosoma theileri no Estado do Rio de Janeiro baseado nos genes 18S rDNA e Cathepsina L-like. 2020. 113 f. Tese (Doutorado em Ci?ncias Veterin?rias) - Instituto de Veterin?ria, Universidade Federal Rural do Rio de Janeiro, Serop?dica, 2020.https://tede.ufrrj.br/jspui/handle/jspui/6008This 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 increasedEste estudo teve como objetivos realizar a caracteriza??o molecular de Trypanosoma vivax e Trypanosoma theileri atrav?s dos genes 18S e catl, bem como analisar o perf?l bioqu?mico relacionado ? infec??o para Trypanosoma vivax em bovinos do estado do Rio de Janeiro. As amostras foram coletadas, por meio de venopun??o da veia cocc?gea de 389 bovinos, sendo o sangue total coletado acondicionado em tubos contendo ?cido etileno-amino-tetrac?tico ? 10% para an?lise molecular e tubos sem anticoagulante para bioqu?mica, sendo posteriormente enviadas ao Laborat?rio de Hemoparasitos e Vetores da Universidade Federal Rural do Rio de Janeiro onde foram realizadas as an?lises. As amostras de DNA foram extra?das seguindo-se protocolo recomendado pelo fabricante (Promega?) e posteriormente armazenadas em freezer -20 ?C at? o momento das an?lises moleculares. O soro para an?lise bioqu?mica foi obtido pela centrifuga??o em centr?fuga cl?nica ? 3.000 rpm por 5 minutos e posteriormente foi enviado ao Laborat?rio de An?lises Cl?nicas do Laborat?rio de Quimioterapia Experimental em Parasitologia Veterin?ria (LQEPV/ UFRRJ), onde os n?veis s?ricos de Gama Glutamiltransferase (GGT), Prote?na S?rica Total (PTS), Albumina (ALB), Bilirrubina Total (BT) e Direta (BD), Creatinina (CR), Ur?ia (UR), Fosfatase Alcalina (FAL), Lactato Desidrogenase (LDH), Aspartato Aminotransferase (AST/TGO) e Alanina Aminotransferase (ALT/TGP) foram determinados usando o analisador bioqu?mico automatizado A15 (Biosystems?) e kits de reagentes comerciais de acordo com o equipamento utilizado. A an?lise molecular foi realizada atrav?s dos genes 18S para identifica??o de Trypanosoma sp, e catl-like (Cathepsina L-like) para identifica??o Trypanosoma vivax e T. theileri. Ap?s an?lise de frequ?ncia, algumas amostras de cada munic?pio foram selecionadas para an?lise filogen?tica e compara??o da identidade das sequ?ncias do presente estudo com outras sequ?ncias encontradas no GenBank. Dentre as amostras analisadas 4,6% (18/389) apresentaram-se positivas para Trypanosoma vivax em esfrega?o sangu?neo, n?o sendo visualizadas amostras positivas para Trypanosoma theileri. Para o gene 18S, 15,4% (60/389) foram positivas para Trypanosomasp, j? para o gene catl, 12,8% (50/389) foram positivos para T. vivax enquanto para T. theileri houve 3,6% (14/389) de animais positivos. A an?lise filogen?tica para T. vivax mostrou 2 clusters definidos para o gene 18S rDNA e 4 para o gene catl, j? para T. theileri, observamos 1clusters definidos para o gene 18S rDNA e 4 para o gene catl, com 15 gen?tipos distintos. O presente estudo teve por finalidade relatar a primeira caracteriza??o molecular de T. vivax e T. theileri no estado do Rio de Janeiro, bem como o primeiro relato da linhagem IB e IF de T. theileri baseado no gene catl. Al?m disso, este tamb?m foi o primeiro relato de an?lise bioqu?mica dos animais infectados naturalmente por Trypanosoma vivax no estado do Rio de Janeiro, onde foi poss?vel verificar que a LDH foi a ?nica an?lise significativamente alterada representando aproximadamente sete vezes mais chance dos bovinos serem positivos para T. vivax quando esta enzima est? aumentadaSubmitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2022-09-19T13:37:19Z No. of bitstreams: 1 2020 - Ana Paula Martinez de Abreu.pdf: 1765091 bytes, checksum: 6e5b5d16117d027a0f9f2ead2121ddf0 (MD5)Made available in DSpace on 2022-09-19T13:37:20Z (GMT). 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dc.title.por.fl_str_mv Caracteriza??o molecular e avalia??o bioqu?mica de Trypanosoma vivax e Trypanosoma theileri no Estado do Rio de Janeiro baseado nos genes 18S rDNA e Cathepsina L-like
dc.title.alternative.eng.fl_str_mv Molecular characterization and biochemical evaluation of Trypanosoma vivax and Trypanosoma theileri in the State of Rio de Janeiro based on the 18S rDNA and Cathepsin L-like gene
title Caracteriza??o molecular e avalia??o bioqu?mica de Trypanosoma vivax e Trypanosoma theileri no Estado do Rio de Janeiro baseado nos genes 18S rDNA e Cathepsina L-like
spellingShingle Caracteriza??o molecular e avalia??o bioqu?mica de Trypanosoma vivax e Trypanosoma theileri no Estado do Rio de Janeiro baseado nos genes 18S rDNA e Cathepsina L-like
Abreu, Ana Paula Martinez de
Trypanosoma vivax
Bovinos
Diagn?stico molecular
Bioqu?mica
Filogenia
Trypanosoma theileri
Trypanosoma vivax
cattle
Molecular diagnosis
PCR
Phylogeny
Trypanosoma theileri
Medicina Veterin?ria
title_short Caracteriza??o molecular e avalia??o bioqu?mica de Trypanosoma vivax e Trypanosoma theileri no Estado do Rio de Janeiro baseado nos genes 18S rDNA e Cathepsina L-like
title_full Caracteriza??o molecular e avalia??o bioqu?mica de Trypanosoma vivax e Trypanosoma theileri no Estado do Rio de Janeiro baseado nos genes 18S rDNA e Cathepsina L-like
title_fullStr Caracteriza??o molecular e avalia??o bioqu?mica de Trypanosoma vivax e Trypanosoma theileri no Estado do Rio de Janeiro baseado nos genes 18S rDNA e Cathepsina L-like
title_full_unstemmed Caracteriza??o molecular e avalia??o bioqu?mica de Trypanosoma vivax e Trypanosoma theileri no Estado do Rio de Janeiro baseado nos genes 18S rDNA e Cathepsina L-like
title_sort Caracteriza??o molecular e avalia??o bioqu?mica de Trypanosoma vivax e Trypanosoma theileri no Estado do Rio de Janeiro baseado nos genes 18S rDNA e Cathepsina L-like
author Abreu, Ana Paula Martinez de
author_facet Abreu, Ana Paula Martinez de
author_role author
dc.contributor.advisor1.fl_str_mv Massard, Carlos Luiz
dc.contributor.advisor1ID.fl_str_mv 257.781.297-34
https://orcid.org/0000-0002-8465-3038
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/7743112049924654
dc.contributor.advisor-co1.fl_str_mv Santos, Huarrisson Azevedo
dc.contributor.advisor-co1ID.fl_str_mv 983.833.295-04
https://orcid.org/0000-0002-8218-3626
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/3751609492049306
dc.contributor.advisor-co2.fl_str_mv Peixoto, Maristela Peckle
dc.contributor.advisor-co2ID.fl_str_mv 110.795.51-11
https://orcid.org/0000-0002-4208-1430
dc.contributor.advisor-co2Lattes.fl_str_mv http://lattes.cnpq.br/8817867478588076
dc.contributor.referee1.fl_str_mv Massard, Carlos Luiz
dc.contributor.referee1ID.fl_str_mv 257.781.297-34
https://orcid.org/0000-0002-8465-3038
dc.contributor.referee1Lattes.fl_str_mv http://lattes.cnpq.br/7743112049924654
dc.contributor.referee2.fl_str_mv Guedes Junior, Daniel da Silva
dc.contributor.referee2Lattes.fl_str_mv http://lattes.cnpq.br/1514734234282622
dc.contributor.referee3.fl_str_mv Roier, Erica Cristina Rocha
dc.contributor.referee3ID.fl_str_mv https://orcid.org/0000-0002-1978-9254
dc.contributor.referee3Lattes.fl_str_mv http://lattes.cnpq.br/3538932429235543
dc.contributor.referee4.fl_str_mv Ba?ta, Bruna de Azevedo
dc.contributor.referee4ID.fl_str_mv https://orcid.org/0000-0002-0172-556X
dc.contributor.referee4Lattes.fl_str_mv http://lattes.cnpq.br/8016012971743463
dc.contributor.referee5.fl_str_mv Ubiali, Daniel Guimar?es
dc.contributor.referee5ID.fl_str_mv https://orcid.org/0000-0001-8320-4567
dc.contributor.referee5Lattes.fl_str_mv http://lattes.cnpq.br/0255736629474813
dc.contributor.authorID.fl_str_mv 051.527.387-24
https://orcid.org/0000-0001-6385-3906
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/2317384923867126
dc.contributor.author.fl_str_mv Abreu, Ana Paula Martinez de
contributor_str_mv Massard, Carlos Luiz
Santos, Huarrisson Azevedo
Peixoto, Maristela Peckle
Massard, Carlos Luiz
Guedes Junior, Daniel da Silva
Roier, Erica Cristina Rocha
Ba?ta, Bruna de Azevedo
Ubiali, Daniel Guimar?es
dc.subject.por.fl_str_mv Trypanosoma vivax
Bovinos
Diagn?stico molecular
Bioqu?mica
Filogenia
Trypanosoma theileri
topic Trypanosoma vivax
Bovinos
Diagn?stico molecular
Bioqu?mica
Filogenia
Trypanosoma theileri
Trypanosoma vivax
cattle
Molecular diagnosis
PCR
Phylogeny
Trypanosoma theileri
Medicina Veterin?ria
dc.subject.eng.fl_str_mv Trypanosoma vivax
cattle
Molecular diagnosis
PCR
Phylogeny
Trypanosoma theileri
dc.subject.cnpq.fl_str_mv Medicina Veterin?ria
description 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
publishDate 2020
dc.date.issued.fl_str_mv 2020-03-11
dc.date.accessioned.fl_str_mv 2022-09-19T13:37:20Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
status_str publishedVersion
dc.identifier.citation.fl_str_mv ABREU, Ana Paula Martinez de. Caracteriza??o molecular e avalia??o bioqu?mica de Trypanosoma vivax e Trypanosoma theileri no Estado do Rio de Janeiro baseado nos genes 18S rDNA e Cathepsina L-like. 2020. 113 f. Tese (Doutorado em Ci?ncias Veterin?rias) - Instituto de Veterin?ria, Universidade Federal Rural do Rio de Janeiro, Serop?dica, 2020.
dc.identifier.uri.fl_str_mv https://tede.ufrrj.br/jspui/handle/jspui/6008
identifier_str_mv ABREU, Ana Paula Martinez de. Caracteriza??o molecular e avalia??o bioqu?mica de Trypanosoma vivax e Trypanosoma theileri no Estado do Rio de Janeiro baseado nos genes 18S rDNA e Cathepsina L-like. 2020. 113 f. Tese (Doutorado em Ci?ncias Veterin?rias) - Instituto de Veterin?ria, Universidade Federal Rural do Rio de Janeiro, Serop?dica, 2020.
url https://tede.ufrrj.br/jspui/handle/jspui/6008
dc.language.iso.fl_str_mv por
language por
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