The use of high resolution melting analysis of ITS-1 for rapid differentiation of parasitic nematodes Haemonchus contortus and Ashworthius sidemi

Lucie Skorpikova,Jan Magdalek,Jaroslav Vadlejch,Nikol Reslova,Martin Kasny

doi:10.1038/s41598-020-73037-9

Lucie Skorpikova, Jan Magdalek + Show 3 more

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https://doi.org/10.1038/s41598-020-73037-9

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Abstract

Among gastrointestinal nematodes, haematophagous strongylids Haemonchus contortus and Ashworthius sidemi belong to the most pathogenic parasites of both domestic and wild ruminants. Correct identification of parasitic taxa is of crucial importance in many areas of parasite research, including monitoring of occurrence, epidemiological studies, or testing of effectiveness of therapy. In this study, we identified H. contortus and A. sidemi in a broad range of ruminant hosts that occur in the Czech Republic using morphological/morphometric and molecular approaches. As an advanced molecular method, we employed qPCR followed by High Resolution Melting analysis, specifically targeting the internal transcribed spacer 1 (ITS-1) sequence to distinguish the two nematode species. We demonstrate that High Resolution Melting curves allow for taxonomic affiliation, making it a convenient, rapid, and reliable identification tool.

Highlights

IntroductionHaematophagous strongylids Haemonchus contortus and Ashworthius sidemi belong to the most pathogenic parasites of both domestic and wild ruminants
Among gastrointestinal nematodes, haematophagous strongylids Haemonchus contortus and Ashworthius sidemi belong to the most pathogenic parasites of both domestic and wild ruminants
The aim of this study was to develop a fast and usable qPCR-High Resolution Melting (HRM) method, which uses polymorphisms in the internal transcribed spacer 1 (ITS-1) region to distinguish between H. contortus and A. sidemi

Summary

Introduction

Haematophagous strongylids Haemonchus contortus and Ashworthius sidemi belong to the most pathogenic parasites of both domestic and wild ruminants. Infection of domestic and wild ruminants by helminth parasites, especially gastrointestinal nematodes (GINs), has a considerable social and economic impact throughout the world These infections can lead to significant economic losses both in livestock industry and wildlife ranching due to decreased productivity or even animal death[1,2,3,4]. H. contortus is currently emerging as a model organism of anthelmintic resistance in parasites, an issue that poses an increasing problem[10,11,12] Ashworthius sidemi is another haematophagous abomasal nematode, phylogenetically related to H. contortus. The impact of infection differs depending on ruminant species, age, environment, nutrition, management, the time of year, and obviously parasitic species and its pathogenicity, the proper identification of parasitic taxa is crucial both for veterinarians and p roducers[25,26]

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