Ocorrência e caracterização molecular de Giardia spp. em amostras fecais de cães naturalmente infectados em Santa Maria, RS
Autor(a) principal: | |
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Data de Publicação: | 2019 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Manancial - Repositório Digital da UFSM |
dARK ID: | ark:/26339/001300000qvmh |
Texto Completo: | http://repositorio.ufsm.br/handle/1/16544 |
Resumo: | Giardiasis is an important and prevalent zoonosis in dogs and humans, being caused by protozoa of the genus Giardia. The close relationship between pets and humans has many benefits, both physical, emotional and social. However, the presence of these animals without proper care may pose a risk to human health. As dogs have an important role in the cycle and transmission of Giardia duodenalis, the objective of this study was to verify the occurrence of the parasite in dogs in the city of Santa Maria, Rio Grande do Sul State (RS). Additionally, in this research, the objective was to carry out a molecular characterization and phylogenetic analyses of Giardia duodenalis. In this study, 230 samples of dog feces were collected from April to October 2018. Of these, 10 were from the routine of coproparasitological examinations performed at the Laboratory of Parasitary Diseases (LADOPAR) of the Federal University of Santa Maria (UFSM), 74 were collected in 3 commercial kennels and 166 were collected at the UFSM Veterinary University Hospital (HVU), all located in the city of Santa Maria, Rio Grande do Sul State, Brazil. All samples were submitted to the Faust coproparasitological technique, after which the total DNA extraction was carried out for the subsequent molecular analysis through the nested PCR technique. Afterwards, the positive samples in the nested PCR were sent for DNA sequencing. The sequences obtained from this study were compared with the sequences for the β-giardin (bg) gene deposited in GenBank and phylogenetic analyses were elaborated using the Neighbor-Joining method using bootstrap of 1000 replicates, where the evolutionary distances were calculated by the Jukes- Cantor method, and Mega X software was used to verify the relationship between assemblages or genotypes found in isolates. From the faecal samples analysed, there were no differences between the variables: diagnostic techniques, local, sex and age of the animals (p<0.05). In the Faust technique, 5.6% (13/230) of the samples were positive for the presence of Giardia spp. In the nested PCR, 4.3% (10/230) of the analysed samples were positive for the amplification of protozoan DNA. There was no significant difference in the results found (p>0.05), and there was no difference in the sensitivity of the tests used. The assemblages found in the positive samples were C and D, which are specific and frequently reported in dogs. In this way, we observed that Giardia duodenalis occurs in the dog populations evaluated in this study. Due to the proximity of dogs and their tutors, there may be the possibility of coinfection with other assemblages, thus increasing the risk of transmission of this zoonosis. |
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Ocorrência e caracterização molecular de Giardia spp. em amostras fecais de cães naturalmente infectados em Santa Maria, RSOccurrence and molecular characterization of Giardia spp. in fecal samples of naturally infected dogs in Santa Maria, RSProtozoárioGiardíaseNested PCRFilogeniaCãesProtozoaGiardiasisNested PCRPhylogenyDogsCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIAGiardiasis is an important and prevalent zoonosis in dogs and humans, being caused by protozoa of the genus Giardia. The close relationship between pets and humans has many benefits, both physical, emotional and social. However, the presence of these animals without proper care may pose a risk to human health. As dogs have an important role in the cycle and transmission of Giardia duodenalis, the objective of this study was to verify the occurrence of the parasite in dogs in the city of Santa Maria, Rio Grande do Sul State (RS). Additionally, in this research, the objective was to carry out a molecular characterization and phylogenetic analyses of Giardia duodenalis. In this study, 230 samples of dog feces were collected from April to October 2018. Of these, 10 were from the routine of coproparasitological examinations performed at the Laboratory of Parasitary Diseases (LADOPAR) of the Federal University of Santa Maria (UFSM), 74 were collected in 3 commercial kennels and 166 were collected at the UFSM Veterinary University Hospital (HVU), all located in the city of Santa Maria, Rio Grande do Sul State, Brazil. All samples were submitted to the Faust coproparasitological technique, after which the total DNA extraction was carried out for the subsequent molecular analysis through the nested PCR technique. Afterwards, the positive samples in the nested PCR were sent for DNA sequencing. The sequences obtained from this study were compared with the sequences for the β-giardin (bg) gene deposited in GenBank and phylogenetic analyses were elaborated using the Neighbor-Joining method using bootstrap of 1000 replicates, where the evolutionary distances were calculated by the Jukes- Cantor method, and Mega X software was used to verify the relationship between assemblages or genotypes found in isolates. From the faecal samples analysed, there were no differences between the variables: diagnostic techniques, local, sex and age of the animals (p<0.05). In the Faust technique, 5.6% (13/230) of the samples were positive for the presence of Giardia spp. In the nested PCR, 4.3% (10/230) of the analysed samples were positive for the amplification of protozoan DNA. There was no significant difference in the results found (p>0.05), and there was no difference in the sensitivity of the tests used. The assemblages found in the positive samples were C and D, which are specific and frequently reported in dogs. In this way, we observed that Giardia duodenalis occurs in the dog populations evaluated in this study. Due to the proximity of dogs and their tutors, there may be the possibility of coinfection with other assemblages, thus increasing the risk of transmission of this zoonosis.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESA giardíase é uma zoonose importante e prevalente em cães e humanos, sendo causada por protozoários do gênero Giardia. A estreita relação entre animais de estimação e seres humanos traz muitos benefícios, tanto físicos, emocionais e sociais. No entanto, a presença desses animais sem o devido cuidado pode representar um risco para a saúde humana. Como os cães possuem um papel importante no ciclo e transmissão de Giardia duodenalis, o objetivo desse estudo foi verificar a ocorrência do parasito em cães na cidade de Santa Maria, Estado do Rio Grande do Sul (RS). Adicionalmente, nesta pesquisa, objetivou-se realizar uma caracterização molecular e análises filogenéticas de Giardia duodenalis. Neste estudo, foram coletadas 230 amostras de fezes de cães, obtidas de abril a outubro de 2018. Destas, 10 eram provenientes da rotina de exames coproparasitológicos realizados no Laboratório de Doenças Parasitárias (LADOPAR) da Universidade Federal de Santa Maria (UFSM), 74 foram coletadas em 3 canis de criação comercial e 166 foram coletados no Hospital Universitário Veterinário (HVU) da UFSM, todos localizados na cidade de Santa Maria, no Rio Grande do Sul, Brasil. Todas as amostras foram submetidas à técnica coproparasitológica de Faust, e, após, foi realizada a extração do DNA total para a posterior análise molecular através da técnica de nested PCR. Posteriormente, as amostras positivas no nested PCR foram enviadas para o sequenciamento de DNA. As sequências obtidas desse estudo foram comparadas com as sequências para o gene da β-giardina (bg) depositadas no GenBank e foram elaboradas análises filogenéticas através do método Neighbor - Joining utilizando bootstrap de 1000 replicatas, onde as distâncias evolutivas foram calculadas pelo método de Jukes – Cantor, e, o software Mega X foi utilizado para verificar a relação entre as assemblages ou genótipos encontrados nos isolados. Das amostras de fezes analisadas, não houve diferença entre as variáveis: técnicas de diagnóstico, locais de coleta, sexo e idade dos animais (p<0,05). Na técnica de Faust, 5,6% (13/230) das amostras foram positivas para a presença de Giardia spp. No nested PCR, 4,3% (10/230) das amostras analisadas foram positivas para a amplificação do DNA do protozoário. Não houve diferença significativa nos resultados encontrados (p>0,05), e não houve diferença na sensibilidade dos testes utilizados. As assemblages encontradas nas amostras positivas foram C e D, que são específicas e frequentemente reportadas nos cães. Desta forma, observamos que Giardia duodenalis ocorre nas populações de cães avaliadas nesse estudo. Devido à proximidade dos cães e seus tutores, pode existir a possibilidade de coinfecções com outras assemblages, aumentando assim o risco de transmissão dessa zoonose.Universidade Federal de Santa MariaBrasilMedicina VeterináriaUFSMPrograma de Pós-Graduação em Medicina VeterináriaCentro de Ciências RuraisSangioni, Luis Antoniohttp://lattes.cnpq.br/8056805667740451Botton, Sônia de Avilahttp://lattes.cnpq.br/0814772095155945Pereira, Daniela Isabel Brayerhttp://lattes.cnpq.br/3382450720179401Osmari, Vanessa2019-05-13T16:18:13Z2019-05-13T16:18:13Z2019-03-08info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://repositorio.ufsm.br/handle/1/16544ark:/26339/001300000qvmhporAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2019-05-14T06:01:57Zoai:repositorio.ufsm.br:1/16544Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2019-05-14T06:01:57Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false |
dc.title.none.fl_str_mv |
Ocorrência e caracterização molecular de Giardia spp. em amostras fecais de cães naturalmente infectados em Santa Maria, RS Occurrence and molecular characterization of Giardia spp. in fecal samples of naturally infected dogs in Santa Maria, RS |
title |
Ocorrência e caracterização molecular de Giardia spp. em amostras fecais de cães naturalmente infectados em Santa Maria, RS |
spellingShingle |
Ocorrência e caracterização molecular de Giardia spp. em amostras fecais de cães naturalmente infectados em Santa Maria, RS Osmari, Vanessa Protozoário Giardíase Nested PCR Filogenia Cães Protozoa Giardiasis Nested PCR Phylogeny Dogs CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
title_short |
Ocorrência e caracterização molecular de Giardia spp. em amostras fecais de cães naturalmente infectados em Santa Maria, RS |
title_full |
Ocorrência e caracterização molecular de Giardia spp. em amostras fecais de cães naturalmente infectados em Santa Maria, RS |
title_fullStr |
Ocorrência e caracterização molecular de Giardia spp. em amostras fecais de cães naturalmente infectados em Santa Maria, RS |
title_full_unstemmed |
Ocorrência e caracterização molecular de Giardia spp. em amostras fecais de cães naturalmente infectados em Santa Maria, RS |
title_sort |
Ocorrência e caracterização molecular de Giardia spp. em amostras fecais de cães naturalmente infectados em Santa Maria, RS |
author |
Osmari, Vanessa |
author_facet |
Osmari, Vanessa |
author_role |
author |
dc.contributor.none.fl_str_mv |
Sangioni, Luis Antonio http://lattes.cnpq.br/8056805667740451 Botton, Sônia de Avila http://lattes.cnpq.br/0814772095155945 Pereira, Daniela Isabel Brayer http://lattes.cnpq.br/3382450720179401 |
dc.contributor.author.fl_str_mv |
Osmari, Vanessa |
dc.subject.por.fl_str_mv |
Protozoário Giardíase Nested PCR Filogenia Cães Protozoa Giardiasis Nested PCR Phylogeny Dogs CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
topic |
Protozoário Giardíase Nested PCR Filogenia Cães Protozoa Giardiasis Nested PCR Phylogeny Dogs CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
description |
Giardiasis is an important and prevalent zoonosis in dogs and humans, being caused by protozoa of the genus Giardia. The close relationship between pets and humans has many benefits, both physical, emotional and social. However, the presence of these animals without proper care may pose a risk to human health. As dogs have an important role in the cycle and transmission of Giardia duodenalis, the objective of this study was to verify the occurrence of the parasite in dogs in the city of Santa Maria, Rio Grande do Sul State (RS). Additionally, in this research, the objective was to carry out a molecular characterization and phylogenetic analyses of Giardia duodenalis. In this study, 230 samples of dog feces were collected from April to October 2018. Of these, 10 were from the routine of coproparasitological examinations performed at the Laboratory of Parasitary Diseases (LADOPAR) of the Federal University of Santa Maria (UFSM), 74 were collected in 3 commercial kennels and 166 were collected at the UFSM Veterinary University Hospital (HVU), all located in the city of Santa Maria, Rio Grande do Sul State, Brazil. All samples were submitted to the Faust coproparasitological technique, after which the total DNA extraction was carried out for the subsequent molecular analysis through the nested PCR technique. Afterwards, the positive samples in the nested PCR were sent for DNA sequencing. The sequences obtained from this study were compared with the sequences for the β-giardin (bg) gene deposited in GenBank and phylogenetic analyses were elaborated using the Neighbor-Joining method using bootstrap of 1000 replicates, where the evolutionary distances were calculated by the Jukes- Cantor method, and Mega X software was used to verify the relationship between assemblages or genotypes found in isolates. From the faecal samples analysed, there were no differences between the variables: diagnostic techniques, local, sex and age of the animals (p<0.05). In the Faust technique, 5.6% (13/230) of the samples were positive for the presence of Giardia spp. In the nested PCR, 4.3% (10/230) of the analysed samples were positive for the amplification of protozoan DNA. There was no significant difference in the results found (p>0.05), and there was no difference in the sensitivity of the tests used. The assemblages found in the positive samples were C and D, which are specific and frequently reported in dogs. In this way, we observed that Giardia duodenalis occurs in the dog populations evaluated in this study. Due to the proximity of dogs and their tutors, there may be the possibility of coinfection with other assemblages, thus increasing the risk of transmission of this zoonosis. |
publishDate |
2019 |
dc.date.none.fl_str_mv |
2019-05-13T16:18:13Z 2019-05-13T16:18:13Z 2019-03-08 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://repositorio.ufsm.br/handle/1/16544 |
dc.identifier.dark.fl_str_mv |
ark:/26339/001300000qvmh |
url |
http://repositorio.ufsm.br/handle/1/16544 |
identifier_str_mv |
ark:/26339/001300000qvmh |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Brasil Medicina Veterinária UFSM Programa de Pós-Graduação em Medicina Veterinária Centro de Ciências Rurais |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Brasil Medicina Veterinária UFSM Programa de Pós-Graduação em Medicina Veterinária Centro de Ciências Rurais |
dc.source.none.fl_str_mv |
reponame:Manancial - Repositório Digital da UFSM instname:Universidade Federal de Santa Maria (UFSM) instacron:UFSM |
instname_str |
Universidade Federal de Santa Maria (UFSM) |
instacron_str |
UFSM |
institution |
UFSM |
reponame_str |
Manancial - Repositório Digital da UFSM |
collection |
Manancial - Repositório Digital da UFSM |
repository.name.fl_str_mv |
Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM) |
repository.mail.fl_str_mv |
atendimento.sib@ufsm.br||tedebc@gmail.com |
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1822612325435179008 |