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The aim of this study was to develop a fast and usable qPCR-HRM method, which uses polymorphisms in the internal transcribed spacer 1 ( ITS-1) region to distinguish between H. HRM is a fast, simple, and cost-effective approach for genotyping and mutation scanning, and it can easily be applied to the taxonomic identification of nematodes 34, 35, 36. Differences in melting profiles are visualised as the fluorescence of saturating dye that is gradually disassociated from the dsDNA amplicons. It can detect sequence alterations, such as small deletions, insertions, or even single nucleotide polymorphisms (SNPs) in dsDNA fragments amplified by qPCR, which are subsequently denatured by increasing temperature, i.e. High Resolution Melting (HRM) analysis is a PCR-based technique available for routine diagnostic applications 31, 32, 33. Moreover, because morphological examination is time-consuming and requires a helminthologist of considerable experience, we witness a growing use of molecular tools for species identification and demand for novel approaches 28, 29, 30. Taxonomic identification based on morphological/morphometric features is possible, especially in adult male specimens, but in adult females and immature nematodes, these methods are unreliable and in the larval stages and eggs they are not applicable at all. sidemi is complicated by morphological, biochemical, and biological similarities between the two species 20, 26, 27. The impact of infection differs depending on ruminant species, age, environment, nutrition, management, the time of year, and obviously also parasitic species and its pathogenicity, therefore the proper identification of parasitic taxa is crucial both for veterinarians and producers 25, 26.Ĭorrect differentiation between H. sidemi has the potential of becoming one of the most widespread pathogenic gastrointestinal nematodes of autochthonous European ruminants.Īs noted above, GINs of domestic and wild ruminants have a negative impact on animal health, which can translate into economic losses 1, 2, 3, 4. Based on available evidence, it is to be feared that A. sidemi transmission from wildlife to livestock 23, 24, 25. In the European bison ( Bison bonasus), a new susceptible host, the intensity of infection can reach thousands of nematodes per animal, which leads to massive histopathological changes 20, 21, 22. A highly successful invasive parasite has been dynamically spreading among various species of wild ruminants and into new regions 17, 18, 19, 20.
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It is a typical parasite of Asiatic deer that was introduced into Europe probably via the sika deer ( Cervus nippon) 13, 14, 15, 16. H. contortus is currently emerging as a model organism of anthelmintic resistance in parasites, an issue that poses an increasing problem 10, 11, 12.Īshworthius sidemi is another haematophagous abomasal nematode, phylogenetically related to H. Adult nematodes attach the abomasal mucosa where they feed on host’s blood, which poses a burden on animal’s health, significant especially in sheep husbandry 8, 9. Hosts become infected after accidental ingestion of third-stage infective larvae (元) during grazing. It has a direct life cycle alternating between a parasitic and a free-living stage. 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.Īmong the GINs, trichostrongylid Haemonchus contortus belongs to the most important parasites of a wide range of small ruminants in tropical and temperate regions around the globe 5, 6, 7.
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Infection of domestic and wild ruminants by helminth parasites, especially gastrointestinal nematodes (GINs), has a considerable social and economic impact throughout the world.