Molecular characterization of Dirofilaria spp. circulating in Portugal

Detalhes bibliográficos
Autor(a) principal: Ferreira, Cátia
Data de Publicação: 2017
Outros Autores: Afonso, Ana, Calado, Manuela, Maurício, Isabel, Alho, Ana Margarida, Meireles, José, Madeira de Carvalho, Luís, Belo, Silvana
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: https://doi.org/10.1186/s13071-017-2180-y
Resumo: Background: Dirofilariosis is a potentially zoonotic parasitic disease, mainly transmitted by mosquito vectors in many parts of the world. Data concerning the canine Dirofilaria species currently circulating in Portugal is scarce. Thereby, a large-scale study was conducted to determine the Dirofilaria spp. present in Portugal, based on a molecular approach, and also to optimize a reliable and highly sensitive species-specific polymerase chain reaction (PCR) assay that could be used for the simultaneous detection and differentiation of Dirofilaria immitis, Dirofilaria repens, and other concurrent filarial species in animal reservoirs. Methods: Blood samples were collected from three districts of Portugal (Coimbra, Santarém and Setúbal) between 2011 and 2013. Samples were tested using rapid immunomigration tests (Witness® Dirofilaria), modified Knott’s technique and acid phosphatase histochemical staining. In addition, molecular analysis was performed by amplification of the internal transcribed spacer (ITS) region using two different PCR protocols, specific for molecular screening of canine filarial species. Results: Of the 878 dogs sampled, 8.8% (n = 77) were positive for D. immitis circulating antigen and 13.1% (n = 115) positive for microfilariae by the modified Knott’s technique. Of the 134 samples tested by acid phosphatase histochemical staining, 100 (74.6%) were positive for D. immitis. Overall, 13.7% (n = 120) were positive by PCR for D. immitis by ITS2, of which 9.3% (67/720) were also positive by ITS1. ITS2 PCR was the most sensitive and specific method, capable of detecting mixed D. immitis and A. reconditum infections. Heterozygosity, in the form of double peaks, was detected by sequencing of both ITS regions. No D. repens was detected by any of the diagnostic methods. Conclusions: The present study confirmed D. immitis as the dominant species of the genus Dirofilaria infecting Portuguese dogs, based on sequencing of ITS1 and ITS2 PCR fragments. Additionally, ITS2 PCR was the most adequate method for diagnosis and prevalence estimation.
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spelling Molecular characterization of Dirofilaria spp. circulating in PortugalDirofilariaDogInternal transcribed spacerPCRPortugalParasitologyInfectious DiseasesSDG 3 - Good Health and Well-beingBackground: Dirofilariosis is a potentially zoonotic parasitic disease, mainly transmitted by mosquito vectors in many parts of the world. Data concerning the canine Dirofilaria species currently circulating in Portugal is scarce. Thereby, a large-scale study was conducted to determine the Dirofilaria spp. present in Portugal, based on a molecular approach, and also to optimize a reliable and highly sensitive species-specific polymerase chain reaction (PCR) assay that could be used for the simultaneous detection and differentiation of Dirofilaria immitis, Dirofilaria repens, and other concurrent filarial species in animal reservoirs. Methods: Blood samples were collected from three districts of Portugal (Coimbra, Santarém and Setúbal) between 2011 and 2013. Samples were tested using rapid immunomigration tests (Witness® Dirofilaria), modified Knott’s technique and acid phosphatase histochemical staining. In addition, molecular analysis was performed by amplification of the internal transcribed spacer (ITS) region using two different PCR protocols, specific for molecular screening of canine filarial species. Results: Of the 878 dogs sampled, 8.8% (n = 77) were positive for D. immitis circulating antigen and 13.1% (n = 115) positive for microfilariae by the modified Knott’s technique. Of the 134 samples tested by acid phosphatase histochemical staining, 100 (74.6%) were positive for D. immitis. Overall, 13.7% (n = 120) were positive by PCR for D. immitis by ITS2, of which 9.3% (67/720) were also positive by ITS1. ITS2 PCR was the most sensitive and specific method, capable of detecting mixed D. immitis and A. reconditum infections. Heterozygosity, in the form of double peaks, was detected by sequencing of both ITS regions. No D. repens was detected by any of the diagnostic methods. Conclusions: The present study confirmed D. immitis as the dominant species of the genus Dirofilaria infecting Portuguese dogs, based on sequencing of ITS1 and ITS2 PCR fragments. Additionally, ITS2 PCR was the most adequate method for diagnosis and prevalence estimation.Instituto de Higiene e Medicina Tropical (IHMT)Global Health and Tropical Medicine (GHTM)Vector borne diseases and pathogens (VBD)RUNFerreira, CátiaAfonso, AnaCalado, ManuelaMaurício, IsabelAlho, Ana MargaridaMeireles, JoséMadeira de Carvalho, LuísBelo, Silvana2018-05-10T22:16:40Z2017-05-192017-05-19T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article8application/pdfhttps://doi.org/10.1186/s13071-017-2180-yengPURE: 3213600http://www.scopus.com/inward/record.url?scp=85019868830&partnerID=8YFLogxKhttps://doi.org/10.1186/s13071-017-2180-yinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-05-22T17:32:30Zoai:run.unl.pt:10362/36481Portal AgregadorONGhttps://www.rcaap.pt/oai/openairemluisa.alvim@gmail.comopendoar:71602024-05-22T17:32:30Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Molecular characterization of Dirofilaria spp. circulating in Portugal
title Molecular characterization of Dirofilaria spp. circulating in Portugal
spellingShingle Molecular characterization of Dirofilaria spp. circulating in Portugal
Ferreira, Cátia
Dirofilaria
Dog
Internal transcribed spacer
PCR
Portugal
Parasitology
Infectious Diseases
SDG 3 - Good Health and Well-being
title_short Molecular characterization of Dirofilaria spp. circulating in Portugal
title_full Molecular characterization of Dirofilaria spp. circulating in Portugal
title_fullStr Molecular characterization of Dirofilaria spp. circulating in Portugal
title_full_unstemmed Molecular characterization of Dirofilaria spp. circulating in Portugal
title_sort Molecular characterization of Dirofilaria spp. circulating in Portugal
author Ferreira, Cátia
author_facet Ferreira, Cátia
Afonso, Ana
Calado, Manuela
Maurício, Isabel
Alho, Ana Margarida
Meireles, José
Madeira de Carvalho, Luís
Belo, Silvana
author_role author
author2 Afonso, Ana
Calado, Manuela
Maurício, Isabel
Alho, Ana Margarida
Meireles, José
Madeira de Carvalho, Luís
Belo, Silvana
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Instituto de Higiene e Medicina Tropical (IHMT)
Global Health and Tropical Medicine (GHTM)
Vector borne diseases and pathogens (VBD)
RUN
dc.contributor.author.fl_str_mv Ferreira, Cátia
Afonso, Ana
Calado, Manuela
Maurício, Isabel
Alho, Ana Margarida
Meireles, José
Madeira de Carvalho, Luís
Belo, Silvana
dc.subject.por.fl_str_mv Dirofilaria
Dog
Internal transcribed spacer
PCR
Portugal
Parasitology
Infectious Diseases
SDG 3 - Good Health and Well-being
topic Dirofilaria
Dog
Internal transcribed spacer
PCR
Portugal
Parasitology
Infectious Diseases
SDG 3 - Good Health and Well-being
description Background: Dirofilariosis is a potentially zoonotic parasitic disease, mainly transmitted by mosquito vectors in many parts of the world. Data concerning the canine Dirofilaria species currently circulating in Portugal is scarce. Thereby, a large-scale study was conducted to determine the Dirofilaria spp. present in Portugal, based on a molecular approach, and also to optimize a reliable and highly sensitive species-specific polymerase chain reaction (PCR) assay that could be used for the simultaneous detection and differentiation of Dirofilaria immitis, Dirofilaria repens, and other concurrent filarial species in animal reservoirs. Methods: Blood samples were collected from three districts of Portugal (Coimbra, Santarém and Setúbal) between 2011 and 2013. Samples were tested using rapid immunomigration tests (Witness® Dirofilaria), modified Knott’s technique and acid phosphatase histochemical staining. In addition, molecular analysis was performed by amplification of the internal transcribed spacer (ITS) region using two different PCR protocols, specific for molecular screening of canine filarial species. Results: Of the 878 dogs sampled, 8.8% (n = 77) were positive for D. immitis circulating antigen and 13.1% (n = 115) positive for microfilariae by the modified Knott’s technique. Of the 134 samples tested by acid phosphatase histochemical staining, 100 (74.6%) were positive for D. immitis. Overall, 13.7% (n = 120) were positive by PCR for D. immitis by ITS2, of which 9.3% (67/720) were also positive by ITS1. ITS2 PCR was the most sensitive and specific method, capable of detecting mixed D. immitis and A. reconditum infections. Heterozygosity, in the form of double peaks, was detected by sequencing of both ITS regions. No D. repens was detected by any of the diagnostic methods. Conclusions: The present study confirmed D. immitis as the dominant species of the genus Dirofilaria infecting Portuguese dogs, based on sequencing of ITS1 and ITS2 PCR fragments. Additionally, ITS2 PCR was the most adequate method for diagnosis and prevalence estimation.
publishDate 2017
dc.date.none.fl_str_mv 2017-05-19
2017-05-19T00:00:00Z
2018-05-10T22:16:40Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
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dc.identifier.uri.fl_str_mv https://doi.org/10.1186/s13071-017-2180-y
url https://doi.org/10.1186/s13071-017-2180-y
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv PURE: 3213600
http://www.scopus.com/inward/record.url?scp=85019868830&partnerID=8YFLogxK
https://doi.org/10.1186/s13071-017-2180-y
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
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dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
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reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
repository.mail.fl_str_mv mluisa.alvim@gmail.com
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