Detection of Dirofilaria immitis via integrated serological and molecular analyses in coyotes from Texas, United States

Wild canids serve as reservoir for various vector-borne pathogens of veterinary and medical importance, including the canine heartworm, Dirofilaria immitis. In North and Central America, coyotes (Canis latrans) may be a relevant reservoir host for heartworm transmission. The objective of this study...

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Main Authors: Caroline Sobotyk, Nathalia Nguyen, Veronica Negrón, Amanda Varner, Meriam N. Saleh, Clayton Hilton, John M. Tomeček, Maria D. Esteve -Gasent, Guilherme G. Verocai
Format: Article
Language:English
Published: Elsevier 2022-08-01
Series:International Journal for Parasitology: Parasites and Wildlife
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2213224422000311
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author Caroline Sobotyk
Nathalia Nguyen
Veronica Negrón
Amanda Varner
Meriam N. Saleh
Clayton Hilton
John M. Tomeček
Maria D. Esteve -Gasent
Guilherme G. Verocai
author_facet Caroline Sobotyk
Nathalia Nguyen
Veronica Negrón
Amanda Varner
Meriam N. Saleh
Clayton Hilton
John M. Tomeček
Maria D. Esteve -Gasent
Guilherme G. Verocai
author_sort Caroline Sobotyk
collection DOAJ
description Wild canids serve as reservoir for various vector-borne pathogens of veterinary and medical importance, including the canine heartworm, Dirofilaria immitis. In North and Central America, coyotes (Canis latrans) may be a relevant reservoir host for heartworm transmission. The objective of this study was to determine the occurrence of D. immitis in coyotes across Texas using integrated antigen detection test and molecular assays. Matching whole blood and serum samples were collected from 122 coyotes from different locations across the state of Texas, United States, encompassing nine counties. Collections occurred from February to April 2016, and December 2016. Samples were assessed serologically using a commercial microtiter plate ELISA (DiroCHEK®), and molecularly by conventional PCR targeting the cytochrome oxidase c subunit 1 (cox1) and NADH dehydrogenase subunit 5 (nad5) of the mitochondrial DNA, and via a TaqMan© probe-based real-time PCR protocol, also targeting a fragment of the cox1 gene. Overall, 12 (9.83%) samples tested positive when serological and molecular results were combined. Seven of 122 samples (5.73%) were antigen-positive, 8 (6.55%) were qPCR-positive, and 4 (3.27%) were positive using conventional PCR. Of 12 positive samples, 4 tested antigen-positive by DiroCHEK® but were negative in all molecular tests, another 4 tested positive by at least one of the molecular assays but tested negative by DiroCHEK®, and 3 samples tested positive by both antigen test and at least one of the molecular assays. Two samples (16.67%) tested positive on both the antigen test and both conventional PCR and qPCR. Our study confirmed the presence of D. immitis infection in coyotes from southern and northern Texas. The combination of serologic and molecular diagnostic tests was proven synergistic for the identification of D. immitis infections, including occult dirofilariosis, and revealed a more accurate picture of heartworm occurrence in the sampled coyotes.
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spelling doaj.art-8109799133e74507a845aa59d458f42d2022-12-21T22:00:08ZengElsevierInternational Journal for Parasitology: Parasites and Wildlife2213-22442022-08-01182024Detection of Dirofilaria immitis via integrated serological and molecular analyses in coyotes from Texas, United StatesCaroline Sobotyk0Nathalia Nguyen1Veronica Negrón2Amanda Varner3Meriam N. Saleh4Clayton Hilton5John M. Tomeček6Maria D. Esteve -Gasent7Guilherme G. Verocai8Department of Veterinary Pathobiology, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, 77843, USADepartment of Veterinary Pathobiology, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, 77843, USADepartment of Veterinary Pathobiology, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, 77843, USADepartment of Veterinary Pathobiology, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, 77843, USADepartment of Veterinary Pathobiology, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, 77843, USADepartment of Animal Science and Veterinary Technology, Caesar Kleberg Wildlife Research Institute, Texas A&M University-Kingsville, Kingsville, TX, 78363, USADepartment of Wildlife and Fisheries Sciences Texas A&M University, College Station, TX, 77843, USADepartment of Veterinary Pathobiology, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, 77843, USADepartment of Veterinary Pathobiology, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, 77843, USA; Corresponding author. Department of Pathobiology, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, 77843, USA.Wild canids serve as reservoir for various vector-borne pathogens of veterinary and medical importance, including the canine heartworm, Dirofilaria immitis. In North and Central America, coyotes (Canis latrans) may be a relevant reservoir host for heartworm transmission. The objective of this study was to determine the occurrence of D. immitis in coyotes across Texas using integrated antigen detection test and molecular assays. Matching whole blood and serum samples were collected from 122 coyotes from different locations across the state of Texas, United States, encompassing nine counties. Collections occurred from February to April 2016, and December 2016. Samples were assessed serologically using a commercial microtiter plate ELISA (DiroCHEK®), and molecularly by conventional PCR targeting the cytochrome oxidase c subunit 1 (cox1) and NADH dehydrogenase subunit 5 (nad5) of the mitochondrial DNA, and via a TaqMan© probe-based real-time PCR protocol, also targeting a fragment of the cox1 gene. Overall, 12 (9.83%) samples tested positive when serological and molecular results were combined. Seven of 122 samples (5.73%) were antigen-positive, 8 (6.55%) were qPCR-positive, and 4 (3.27%) were positive using conventional PCR. Of 12 positive samples, 4 tested antigen-positive by DiroCHEK® but were negative in all molecular tests, another 4 tested positive by at least one of the molecular assays but tested negative by DiroCHEK®, and 3 samples tested positive by both antigen test and at least one of the molecular assays. Two samples (16.67%) tested positive on both the antigen test and both conventional PCR and qPCR. Our study confirmed the presence of D. immitis infection in coyotes from southern and northern Texas. The combination of serologic and molecular diagnostic tests was proven synergistic for the identification of D. immitis infections, including occult dirofilariosis, and revealed a more accurate picture of heartworm occurrence in the sampled coyotes.http://www.sciencedirect.com/science/article/pii/S2213224422000311HeartwormImmunodiagnosticsReal-time PCRVector-borne diseaseWildlife reservoir
spellingShingle Caroline Sobotyk
Nathalia Nguyen
Veronica Negrón
Amanda Varner
Meriam N. Saleh
Clayton Hilton
John M. Tomeček
Maria D. Esteve -Gasent
Guilherme G. Verocai
Detection of Dirofilaria immitis via integrated serological and molecular analyses in coyotes from Texas, United States
International Journal for Parasitology: Parasites and Wildlife
Heartworm
Immunodiagnostics
Real-time PCR
Vector-borne disease
Wildlife reservoir
title Detection of Dirofilaria immitis via integrated serological and molecular analyses in coyotes from Texas, United States
title_full Detection of Dirofilaria immitis via integrated serological and molecular analyses in coyotes from Texas, United States
title_fullStr Detection of Dirofilaria immitis via integrated serological and molecular analyses in coyotes from Texas, United States
title_full_unstemmed Detection of Dirofilaria immitis via integrated serological and molecular analyses in coyotes from Texas, United States
title_short Detection of Dirofilaria immitis via integrated serological and molecular analyses in coyotes from Texas, United States
title_sort detection of dirofilaria immitis via integrated serological and molecular analyses in coyotes from texas united states
topic Heartworm
Immunodiagnostics
Real-time PCR
Vector-borne disease
Wildlife reservoir
url http://www.sciencedirect.com/science/article/pii/S2213224422000311
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