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High-throughput multiplex qPCRs for the surveillance of zoonotic species of canine hookworms.

ABSTRACT: The canine hookworms Ancylostoma braziliense, Ancylostoma ceylanicum, Ancylostoma caninum and Uncinaria stenocephala are not only capable of producing morbidity and mortality in dogs but are also neglected tropical zoonoses. Each hookworm species differs considerably in its geographical distribution, life cycle, biology, pathogenic impacts on both canine and human hosts, zoonotic potential, and response to treatment with anthelminthics. Here we describe the development and validation of two Taq-Man based multiplex PCR assays capable of detecting and differentiating all four canine hookworm species in faeces of naturally infected dogs. The analytical sensitivity of both assays was assessed using 10-fold serial dilutions of synthetic gene block fragments containing individual sequence targets of each hookworm species. The sensitivity of the assays and ability to detect mixed species infections were compared to a conventional PCR-Restriction Fragment Length Polymorphism based-approach when applied to laboratory and field samples from endemic areas. The qPCRs detected at least one species of hookworms in 82.4% of PCR-RFLP-negative but microscopy-positive samples. The qPCRs detected an additional 68% mixed infections with different species of canine hookworms, and additional single species infection with A. caninum (47%), U. stenocephala (33%) and A. ceylanicum (0.02%) that were missed by PCR-RFLP. These multiplex qPCR assays will assist field based epidemiological surveillance studies towards an accurate and sensitive monitoring of canine hookworm infections in dogs, to inform their species-specific zoonotic risks to populations living in endemic areas, globally.

SUBMITTER: Massetti L

PROVIDER: S-EPMC7316352 | biostudies-literature | 2020 Jun

REPOSITORIES: biostudies-literature

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High-throughput multiplex qPCRs for the surveillance of zoonotic species of canine hookworms.

Massetti Luca L Colella Vito V Zendejas Patsy A PA Ng-Nguyen Dinh D Harriott Lana L Marwedel Lara L Wiethoelter Anke A Traub Rebecca J RJ

PLoS neglected tropical diseases 20200615 6

The canine hookworms Ancylostoma braziliense, Ancylostoma ceylanicum, Ancylostoma caninum and Uncinaria stenocephala are not only capable of producing morbidity and mortality in dogs but are also neglected tropical zoonoses. Each hookworm species differs considerably in its geographical distribution, life cycle, biology, pathogenic impacts on both canine and human hosts, zoonotic potential, and response to treatment with anthelminthics. Here we describe the development and validation of two Taq- ...[more]

PMID: 32542036

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Project description:BACKGROUND: Canine hookworm infection is endemic in Southeast Asian countries with a prevalence ranging from 70% to 100%, with zoonotic transmission representing a potentially significant public health concern. However, there are limited data available on the prevalence of canine hookworms in Malaysia. This study was conducted to determine the prevalence of hookworm and Ancylostoma species among dogs in Malaysia. METHODS: Faecal samples were collected from 221 dogs living in urban areas, rural areas and animal shelters in Selangor. Faecal samples were processed using the formal-ether concentration technique followed by wet mount preparation and iodine staining for the detection of hookworm eggs. Samples positive for hookworm eggs were examined using PCR, targeting ITS2 and 28?s rRNA region, and subsequently sequenced in both directions. The sequences were phylogenetically analysed using MrBayes for Bayesian Inference. RESULTS: The overall prevalence of hookworm among dogs was 48% (95%CI; 41.41-54.95). Rural stray dogs had the highest prevalence 71.4% (95%CI; 61.13-81.49) followed by urban stray dogs, recording 48% (95%CI; 34.15-61.85) and lastly dogs in shelters with 28.7% (95%CI; 19.56-37.84). Logistic regression identified rural stray dogs as a high risk group (OR?=?4.55, 95%; 2.50-8.31) and keeping dogs in shelters as a protective factor (OR?=?0.24, 95%; 0.14-0.43). Molecular methods identified both Ancylostoma ceylanicum and Ancylostoma caninum with A. ceylanicum being predominant among urban stray dogs. Rural dogs had a higher prevalence of A. caninum than A. ceylanicum, while both species showed equal distribution among dogs in shelters. Phylogenetic analysis placed A. ceylanicum isolated from dogs in one group with A. ceylanicum human isolates. CONCLUSION: This study indicates that dogs have the potential to act as reservoir hosts of human hookworm infection in Malaysia. This finding necessitates the inclusion of dogs in any interventions to combat hookworm in the country.

Dataset Information

High-throughput multiplex qPCRs for the surveillance of zoonotic species of canine hookworms.

Publications

High-throughput multiplex qPCRs for the surveillance of zoonotic species of canine hookworms.

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