Using cerebrospinal fluid to confirm Angiostrongylus cantonensis as the cause of canine neuroangiostrongyliasis in Australia where A. cantonensis and Angiostrongylus mackerrasae co-exist
Both Angiostrongylus cantonensis and Angiostrongylus mackerrasae have been identified along the east coast of Australia. A lack of A. mackerrasae genomic data until 2019, however, has precluded the unequivocal identification of the Angiostrongylus species responsible for neuroangiostrongyliasis in a...
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doaj-278790a26ef24ae2a98da830313d17112021-06-13T04:40:22ZengElsevierCurrent Research in Parasitology and Vector-Borne Diseases2667-114X2021-01-011100033Using cerebrospinal fluid to confirm Angiostrongylus cantonensis as the cause of canine neuroangiostrongyliasis in Australia where A. cantonensis and Angiostrongylus mackerrasae co-existJeevitheswara Thammannaya Mallaiyaraj Mahalingam0Nichola Eliza Davies Calvani1Rogan Lee2Richard Malik3Jan Šlapeta4Sydney School of Veterinary Science, Faculty of Science, University of Sydney, 2006, New South Wales, AustraliaSydney School of Veterinary Science, Faculty of Science, University of Sydney, 2006, New South Wales, Australia; Molecular Parasitology Laboratory, Centre for One Health Ryan Institute, National University of Ireland, Galway, H91 DK59, Galway, IrelandParasitology Laboratory, Centre for Infectious Diseases and Microbiology Lab Services, NSW Health Pathology, Level 3 ICPMR, Westmead Hospital, 2145, New South Wales, AustraliaCentre for Veterinary Education, University of Sydney, 2006, New South Wales, AustraliaSydney School of Veterinary Science, Faculty of Science, University of Sydney, 2006, New South Wales, Australia; Corresponding author.Both Angiostrongylus cantonensis and Angiostrongylus mackerrasae have been identified along the east coast of Australia. A lack of A. mackerrasae genomic data until 2019, however, has precluded the unequivocal identification of the Angiostrongylus species responsible for neuroangiostrongyliasis in accidental hosts such as dog and man. The availability of a whole-genome data for A. mackerrasae, including mtDNA and ITS2 rDNA, enables discrimination of A. cantonensis from A. mackerrasae. The aim of this study was to develop diagnostic PCR assays to determine the species of Angiostrongylus based on the detection of Angiostrongylus DNA sequences in the cerebrospinal fluid (CSF) of canine patients with eosinophilic meningitis. An in silico workflow utilising available cytochrome c oxidase 1 (cox1) primers streamlined the laboratory work into empirical steps, allowing optimisation and selection of a PCR assay that met the required criteria for discrimination of A. cantonensis and A. mackerrasae DNA in low-template CSF samples. The adopted cox1 qPCR assay specifically amplified and enabled the differentiation of A. cantonensis from A. mackerrasae DNA and confirmed the presence of A. cantonensis DNA in 11/50 archived CSF samples. The DNA sequences demonstrated the presence of two distinct A. cantonensis cox1 haplotypes in dogs from eastern Australia. Species identification was further confirmed via the adoption of an ITS2 rDNA assay, providing confirmation of only A. cantonensis ITS2 rDNA in the CSF samples. To our knowledge, this is the first study to unequivocally demonstrate the antemortem presence of A. cantonensis DNA in CSF from clinically affected dogs. The study confirmed the long-held assumption that A. cantonensis is the causal agent of neuroangiostrongyliasis but refutes the dogma that there was a single introduction of A. cantonensis into Australia by the demonstration of two distinct A. cantonensis cox1 haplotypes.http://www.sciencedirect.com/science/article/pii/S2667114X21000273Rat lungwormDogsHaplotypeValidationCSFMitochondrial DNA |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Jeevitheswara Thammannaya Mallaiyaraj Mahalingam Nichola Eliza Davies Calvani Rogan Lee Richard Malik Jan Šlapeta |
spellingShingle |
Jeevitheswara Thammannaya Mallaiyaraj Mahalingam Nichola Eliza Davies Calvani Rogan Lee Richard Malik Jan Šlapeta Using cerebrospinal fluid to confirm Angiostrongylus cantonensis as the cause of canine neuroangiostrongyliasis in Australia where A. cantonensis and Angiostrongylus mackerrasae co-exist Current Research in Parasitology and Vector-Borne Diseases Rat lungworm Dogs Haplotype Validation CSF Mitochondrial DNA |
author_facet |
Jeevitheswara Thammannaya Mallaiyaraj Mahalingam Nichola Eliza Davies Calvani Rogan Lee Richard Malik Jan Šlapeta |
author_sort |
Jeevitheswara Thammannaya Mallaiyaraj Mahalingam |
title |
Using cerebrospinal fluid to confirm Angiostrongylus cantonensis as the cause of canine neuroangiostrongyliasis in Australia where A. cantonensis and Angiostrongylus mackerrasae co-exist |
title_short |
Using cerebrospinal fluid to confirm Angiostrongylus cantonensis as the cause of canine neuroangiostrongyliasis in Australia where A. cantonensis and Angiostrongylus mackerrasae co-exist |
title_full |
Using cerebrospinal fluid to confirm Angiostrongylus cantonensis as the cause of canine neuroangiostrongyliasis in Australia where A. cantonensis and Angiostrongylus mackerrasae co-exist |
title_fullStr |
Using cerebrospinal fluid to confirm Angiostrongylus cantonensis as the cause of canine neuroangiostrongyliasis in Australia where A. cantonensis and Angiostrongylus mackerrasae co-exist |
title_full_unstemmed |
Using cerebrospinal fluid to confirm Angiostrongylus cantonensis as the cause of canine neuroangiostrongyliasis in Australia where A. cantonensis and Angiostrongylus mackerrasae co-exist |
title_sort |
using cerebrospinal fluid to confirm angiostrongylus cantonensis as the cause of canine neuroangiostrongyliasis in australia where a. cantonensis and angiostrongylus mackerrasae co-exist |
publisher |
Elsevier |
series |
Current Research in Parasitology and Vector-Borne Diseases |
issn |
2667-114X |
publishDate |
2021-01-01 |
description |
Both Angiostrongylus cantonensis and Angiostrongylus mackerrasae have been identified along the east coast of Australia. A lack of A. mackerrasae genomic data until 2019, however, has precluded the unequivocal identification of the Angiostrongylus species responsible for neuroangiostrongyliasis in accidental hosts such as dog and man. The availability of a whole-genome data for A. mackerrasae, including mtDNA and ITS2 rDNA, enables discrimination of A. cantonensis from A. mackerrasae. The aim of this study was to develop diagnostic PCR assays to determine the species of Angiostrongylus based on the detection of Angiostrongylus DNA sequences in the cerebrospinal fluid (CSF) of canine patients with eosinophilic meningitis. An in silico workflow utilising available cytochrome c oxidase 1 (cox1) primers streamlined the laboratory work into empirical steps, allowing optimisation and selection of a PCR assay that met the required criteria for discrimination of A. cantonensis and A. mackerrasae DNA in low-template CSF samples. The adopted cox1 qPCR assay specifically amplified and enabled the differentiation of A. cantonensis from A. mackerrasae DNA and confirmed the presence of A. cantonensis DNA in 11/50 archived CSF samples. The DNA sequences demonstrated the presence of two distinct A. cantonensis cox1 haplotypes in dogs from eastern Australia. Species identification was further confirmed via the adoption of an ITS2 rDNA assay, providing confirmation of only A. cantonensis ITS2 rDNA in the CSF samples. To our knowledge, this is the first study to unequivocally demonstrate the antemortem presence of A. cantonensis DNA in CSF from clinically affected dogs. The study confirmed the long-held assumption that A. cantonensis is the causal agent of neuroangiostrongyliasis but refutes the dogma that there was a single introduction of A. cantonensis into Australia by the demonstration of two distinct A. cantonensis cox1 haplotypes. |
topic |
Rat lungworm Dogs Haplotype Validation CSF Mitochondrial DNA |
url |
http://www.sciencedirect.com/science/article/pii/S2667114X21000273 |
work_keys_str_mv |
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