Molecular characterization of Dirofilaria spp. circulating in Portugal
Abstract 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 D...
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doaj-87471024bc424a42a15c14f34cf604082020-11-25T00:20:25ZengBMCParasites & Vectors1756-33052017-05-011011810.1186/s13071-017-2180-yMolecular characterization of Dirofilaria spp. circulating in PortugalCátia Ferreira0Ana Afonso1Manuela Calado2Isabel Maurício3Ana Margarida Alho4José Meireles5Luís Madeira de Carvalho6Silvana Belo7Global Health and Tropical Medicine, GHTM, Instituto de Higiene e Medicina Tropical, IHMT, Universidade Nova de Lisboa, UNLGlobal Health and Tropical Medicine, GHTM, Instituto de Higiene e Medicina Tropical, IHMT, Universidade Nova de Lisboa, UNLGlobal Health and Tropical Medicine, GHTM, Instituto de Higiene e Medicina Tropical, IHMT, Universidade Nova de Lisboa, UNLGlobal Health and Tropical Medicine, GHTM, Instituto de Higiene e Medicina Tropical, IHMT, Universidade Nova de Lisboa, UNLCIISA, Faculdade de Medicina Veterinária, Universidade de Lisboa (ULisboa)CIISA, Faculdade de Medicina Veterinária, Universidade de Lisboa (ULisboa)CIISA, Faculdade de Medicina Veterinária, Universidade de Lisboa (ULisboa)Global Health and Tropical Medicine, GHTM, Instituto de Higiene e Medicina Tropical, IHMT, Universidade Nova de Lisboa, UNLAbstract 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.http://link.springer.com/article/10.1186/s13071-017-2180-yDirofilariaPCRInternal transcribed spacerDogPortugal |
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DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Cátia Ferreira Ana Afonso Manuela Calado Isabel Maurício Ana Margarida Alho José Meireles Luís Madeira de Carvalho Silvana Belo |
spellingShingle |
Cátia Ferreira Ana Afonso Manuela Calado Isabel Maurício Ana Margarida Alho José Meireles Luís Madeira de Carvalho Silvana Belo Molecular characterization of Dirofilaria spp. circulating in Portugal Parasites & Vectors Dirofilaria PCR Internal transcribed spacer Dog Portugal |
author_facet |
Cátia Ferreira Ana Afonso Manuela Calado Isabel Maurício Ana Margarida Alho José Meireles Luís Madeira de Carvalho Silvana Belo |
author_sort |
Cátia Ferreira |
title |
Molecular characterization of Dirofilaria spp. circulating in Portugal |
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 |
publisher |
BMC |
series |
Parasites & Vectors |
issn |
1756-3305 |
publishDate |
2017-05-01 |
description |
Abstract 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. |
topic |
Dirofilaria PCR Internal transcribed spacer Dog Portugal |
url |
http://link.springer.com/article/10.1186/s13071-017-2180-y |
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