Serological and molecular findings in diagnosis of leptospirosis serovar hardjo in a dairy bovine herd

Detalhes bibliográficos
Autor(a) principal: Hashimoto, Vanessa Yumi
Data de Publicação: 2017
Outros Autores: Chideroli, Roberta Torres, Ribeiro, Juliane, Alfieri, Amauri Alcindo, Costa, Geraldo Marcio da, Pereira, Ulisses de Padua, Freitas, Julio Cesar de
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UFLA
Texto Completo: http://repositorio.ufla.br/jspui/handle/1/30978
Resumo: The cattle are considered hosts of the Hardjo serovar, causing economic damages due to the reproductive failures like abortions and infertility. The serovar Hardjo usually remains in the reproductive tract and also in the renal tubules where it is eliminated intermittently in the urine for months. Placental remnants, the aborted fetus and contaminated urine promote the permanence of this bacterium within the herd for years. Thus, the objective of this study was to monitor for prolonged period, cows naturally infected with Leptospira ssp. through microbiological culture, serological examination and DNA detection of the pathogen in the urine. The dairy herd was composed of 50 breeding cows with a history of abortion and infertility, without leptospirosis vaccine and located in the northern region of Paraná. Blood and urine samples were collected and laboratorial diagnosis were performed five times at intervals of four months. Blood samples were collected from the all 50 animals and the serum was submitted to the microscopic agglutination test (MAT) for the detection of anti-leptospira antibodies. Of the total cows, 20 showed antibody titres ≥ 1: 100 in MAT and urine samples were collected from only those animals with higher titers to perform nested-PCR (n-PCR) and bacterial isolation per culture. In addition, two urine samples from five animals with antibody titers < 1: 100 were collected in MAT for n-PCR. Serovar Hardjo was considered the most frequent during the serological monitoring of the animals evaluated. The n-PCR technique was able to detect leptospiral DNA in the urine of animals with MAT ≥ 1: 100 antibody titers and urine from animals whose titers were < 1: 100. Sequencing of the leptospiral amplicons shared 100% nucleotide sequence identity with the Leptospira interrogans species. Positive n-PCR results from animals with titers of < 1: 100 suggest that the cut-off of MAT is could be not sufficient to detect renal carriers, so it is also important to use n-PCR as an additional diagnostic tool for identify infected animals with Hardjo serovar and whose serology was negative.
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spelling Serological and molecular findings in diagnosis of leptospirosis serovar hardjo in a dairy bovine herdEpidemiologyLeptospiraMicroscopic agglutination testRenal carriersEpidemiologiaLeptospiraPortadores renaisSoroaglutinação microscópicaThe cattle are considered hosts of the Hardjo serovar, causing economic damages due to the reproductive failures like abortions and infertility. The serovar Hardjo usually remains in the reproductive tract and also in the renal tubules where it is eliminated intermittently in the urine for months. Placental remnants, the aborted fetus and contaminated urine promote the permanence of this bacterium within the herd for years. Thus, the objective of this study was to monitor for prolonged period, cows naturally infected with Leptospira ssp. through microbiological culture, serological examination and DNA detection of the pathogen in the urine. The dairy herd was composed of 50 breeding cows with a history of abortion and infertility, without leptospirosis vaccine and located in the northern region of Paraná. Blood and urine samples were collected and laboratorial diagnosis were performed five times at intervals of four months. Blood samples were collected from the all 50 animals and the serum was submitted to the microscopic agglutination test (MAT) for the detection of anti-leptospira antibodies. Of the total cows, 20 showed antibody titres ≥ 1: 100 in MAT and urine samples were collected from only those animals with higher titers to perform nested-PCR (n-PCR) and bacterial isolation per culture. In addition, two urine samples from five animals with antibody titers < 1: 100 were collected in MAT for n-PCR. Serovar Hardjo was considered the most frequent during the serological monitoring of the animals evaluated. The n-PCR technique was able to detect leptospiral DNA in the urine of animals with MAT ≥ 1: 100 antibody titers and urine from animals whose titers were < 1: 100. Sequencing of the leptospiral amplicons shared 100% nucleotide sequence identity with the Leptospira interrogans species. Positive n-PCR results from animals with titers of < 1: 100 suggest that the cut-off of MAT is could be not sufficient to detect renal carriers, so it is also important to use n-PCR as an additional diagnostic tool for identify infected animals with Hardjo serovar and whose serology was negative.Os bovinos são considerados hospedeiros do sorovar Hardjo, causando prejuízos econômicos devido aos problemas reprodutivos como abortos e infertilidade. O sorovar Hardjo costuma permanecer no trato reprodutivo e também nos túbulos renais onde é eliminado de forma intermitente na urina por meses. Restos placentários, o feto abortado e a urina contaminada favorecem a permanência dessa bactéria dentro do rebanho por anos. Assim, o objetivo deste estudo foi monitorar por período prolongado, vacas naturalmente infectadas por Leptospira ssp. através de cultura microbiológica, exame sorológico e detecção de DNA do patógeno na urina. O rebanho leiteiro estudado era constituído por 50 vacas reprodutoras com histórico de abortos e infertilidade, sem uso de vacina contra a leptospirose e localizada na região Norte do Paraná. As amostras de sangue e urina foram coletadas e submetidas a análises laboratoriais em cinco vezes em intervalos espaçados de quatro meses. As amostras de sangue foram coletadas dos 50 animais e o soro foi submetido ao teste de soroaglutinação microscópica (SAM) para detecção de anticorpos anti-leptospira. Do total vacas, 20 demonstraram títulos de anticorpos ≥ 1: 100 na SAM e foram coletadas amostras de urina apenas destes animais com maiores título para realizar a nested-PCR (n-PCR) e isolamento bacteriano por cultura. Adicionalmente, foram coletadas duas amostras de urina de cinco animais com títulos de anticorpos < 1: 100 na SAM para n-PCR. O Sorovar Hardjo foi considerado como sendo o mais frequente durante o monitoramento sorológico dos animais avaliados. A técnica de n-PCR foi capaz de detectar DNA leptospiral na urina de animais com títulos de anticorpos ≥ 1: 100 e na urina de animais cujos títulos eram < 1:100. O sequenciamento dos amplicons leptospíricos compartilhou 100% de identidade da sequência de nucleotídeos com a espécie Leptospira interrogans. Os resultados positivos da n-PCR dos animais com títulos < 1: 100 sugerem que a diminuição do ponto de corte da SAM pode não ser suficiente para detectar os portadores renais, por isso também é importante usar a n-PCR como uma ferramenta diagnóstica adicional para identificar os animais infectados com serovar Hardjo e cuja sorologia foi negativa.Universidade Estadual de Londrina2018-10-09T13:46:48Z2018-10-09T13:46:48Z2017info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfHASHIMOTO, V. Y. et al. Serological and molecular findings in diagnosis of leptospirosis serovar hardjo in a dairy bovine herd. Semina: Ciências Agrárias, Londrina, v. 38, n. 5, 2017.http://repositorio.ufla.br/jspui/handle/1/30978Semina: Ciências Agráriasreponame:Repositório Institucional da UFLAinstname:Universidade Federal de Lavras (UFLA)instacron:UFLAAttribution-NonCommercial 4.0 Internationalhttp://creativecommons.org/licenses/by-nc/4.0/info:eu-repo/semantics/openAccessHashimoto, Vanessa YumiChideroli, Roberta TorresRibeiro, JulianeAlfieri, Amauri AlcindoCosta, Geraldo Marcio daPereira, Ulisses de PaduaFreitas, Julio Cesar deeng2018-10-09T13:46:49Zoai:localhost:1/30978Repositório InstitucionalPUBhttp://repositorio.ufla.br/oai/requestnivaldo@ufla.br || repositorio.biblioteca@ufla.bropendoar:2018-10-09T13:46:49Repositório Institucional da UFLA - Universidade Federal de Lavras (UFLA)false
dc.title.none.fl_str_mv Serological and molecular findings in diagnosis of leptospirosis serovar hardjo in a dairy bovine herd
title Serological and molecular findings in diagnosis of leptospirosis serovar hardjo in a dairy bovine herd
spellingShingle Serological and molecular findings in diagnosis of leptospirosis serovar hardjo in a dairy bovine herd
Hashimoto, Vanessa Yumi
Epidemiology
Leptospira
Microscopic agglutination test
Renal carriers
Epidemiologia
Leptospira
Portadores renais
Soroaglutinação microscópica
title_short Serological and molecular findings in diagnosis of leptospirosis serovar hardjo in a dairy bovine herd
title_full Serological and molecular findings in diagnosis of leptospirosis serovar hardjo in a dairy bovine herd
title_fullStr Serological and molecular findings in diagnosis of leptospirosis serovar hardjo in a dairy bovine herd
title_full_unstemmed Serological and molecular findings in diagnosis of leptospirosis serovar hardjo in a dairy bovine herd
title_sort Serological and molecular findings in diagnosis of leptospirosis serovar hardjo in a dairy bovine herd
author Hashimoto, Vanessa Yumi
author_facet Hashimoto, Vanessa Yumi
Chideroli, Roberta Torres
Ribeiro, Juliane
Alfieri, Amauri Alcindo
Costa, Geraldo Marcio da
Pereira, Ulisses de Padua
Freitas, Julio Cesar de
author_role author
author2 Chideroli, Roberta Torres
Ribeiro, Juliane
Alfieri, Amauri Alcindo
Costa, Geraldo Marcio da
Pereira, Ulisses de Padua
Freitas, Julio Cesar de
author2_role author
author
author
author
author
author
dc.contributor.author.fl_str_mv Hashimoto, Vanessa Yumi
Chideroli, Roberta Torres
Ribeiro, Juliane
Alfieri, Amauri Alcindo
Costa, Geraldo Marcio da
Pereira, Ulisses de Padua
Freitas, Julio Cesar de
dc.subject.por.fl_str_mv Epidemiology
Leptospira
Microscopic agglutination test
Renal carriers
Epidemiologia
Leptospira
Portadores renais
Soroaglutinação microscópica
topic Epidemiology
Leptospira
Microscopic agglutination test
Renal carriers
Epidemiologia
Leptospira
Portadores renais
Soroaglutinação microscópica
description The cattle are considered hosts of the Hardjo serovar, causing economic damages due to the reproductive failures like abortions and infertility. The serovar Hardjo usually remains in the reproductive tract and also in the renal tubules where it is eliminated intermittently in the urine for months. Placental remnants, the aborted fetus and contaminated urine promote the permanence of this bacterium within the herd for years. Thus, the objective of this study was to monitor for prolonged period, cows naturally infected with Leptospira ssp. through microbiological culture, serological examination and DNA detection of the pathogen in the urine. The dairy herd was composed of 50 breeding cows with a history of abortion and infertility, without leptospirosis vaccine and located in the northern region of Paraná. Blood and urine samples were collected and laboratorial diagnosis were performed five times at intervals of four months. Blood samples were collected from the all 50 animals and the serum was submitted to the microscopic agglutination test (MAT) for the detection of anti-leptospira antibodies. Of the total cows, 20 showed antibody titres ≥ 1: 100 in MAT and urine samples were collected from only those animals with higher titers to perform nested-PCR (n-PCR) and bacterial isolation per culture. In addition, two urine samples from five animals with antibody titers < 1: 100 were collected in MAT for n-PCR. Serovar Hardjo was considered the most frequent during the serological monitoring of the animals evaluated. The n-PCR technique was able to detect leptospiral DNA in the urine of animals with MAT ≥ 1: 100 antibody titers and urine from animals whose titers were < 1: 100. Sequencing of the leptospiral amplicons shared 100% nucleotide sequence identity with the Leptospira interrogans species. Positive n-PCR results from animals with titers of < 1: 100 suggest that the cut-off of MAT is could be not sufficient to detect renal carriers, so it is also important to use n-PCR as an additional diagnostic tool for identify infected animals with Hardjo serovar and whose serology was negative.
publishDate 2017
dc.date.none.fl_str_mv 2017
2018-10-09T13:46:48Z
2018-10-09T13:46:48Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv HASHIMOTO, V. Y. et al. Serological and molecular findings in diagnosis of leptospirosis serovar hardjo in a dairy bovine herd. Semina: Ciências Agrárias, Londrina, v. 38, n. 5, 2017.
http://repositorio.ufla.br/jspui/handle/1/30978
identifier_str_mv HASHIMOTO, V. Y. et al. Serological and molecular findings in diagnosis of leptospirosis serovar hardjo in a dairy bovine herd. Semina: Ciências Agrárias, Londrina, v. 38, n. 5, 2017.
url http://repositorio.ufla.br/jspui/handle/1/30978
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv Attribution-NonCommercial 4.0 International
http://creativecommons.org/licenses/by-nc/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Attribution-NonCommercial 4.0 International
http://creativecommons.org/licenses/by-nc/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Estadual de Londrina
publisher.none.fl_str_mv Universidade Estadual de Londrina
dc.source.none.fl_str_mv Semina: Ciências Agrárias
reponame:Repositório Institucional da UFLA
instname:Universidade Federal de Lavras (UFLA)
instacron:UFLA
instname_str Universidade Federal de Lavras (UFLA)
instacron_str UFLA
institution UFLA
reponame_str Repositório Institucional da UFLA
collection Repositório Institucional da UFLA
repository.name.fl_str_mv Repositório Institucional da UFLA - Universidade Federal de Lavras (UFLA)
repository.mail.fl_str_mv nivaldo@ufla.br || repositorio.biblioteca@ufla.br
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