Affiliation:
1. Naresuan University
2. Srinakharinwirot University
Abstract
AbstractParamphistomosis is caused by paramphistome or amphistome parasites, includingFischoederius elongatus,Gastrothylax crumenifer,Orthocoelium parvipapillatum, andParamphistomum epiclitum. The control and prevention of these parasite outbreaks are difficult because of the wide occurrence of these species. Besides, the clinical manifestations and their egg characteristics are similar to those of other intestinal flukes in the paramphistome group, leading to misdiagnosis. Here, we employed DNA barcoding using NADH dehydrogenase (ubiquinone, alpha 1) (ND1) and cytochrome c oxidase subunit I (COI), coupled with high-resolution melting analysis (Bar-HRM), for species differentiation. As a result, ParND1_3 and ParCOI4 resulted in positive amplification in the paramphistomes andFasciola gigantica, with significantly different melting curves for each species. The melting temperatures of each species obtained clearly differed. Regarding sensitivity, the limit of detection (LoD) for all species of paramphistomes was 1 pg/µl. Our findings suggest that Bar-HRM using ParND1_3 is highly suitable for the differentiation of paramphistome species. This approach can be used in parasite detection and epidemiological studies in cattle.
Publisher
Research Square Platform LLC
Reference53 articles.
1. Identification of Fasciola species isolated from Egypt based on sequence analysis of genomic (ITS1 and ITS2) and mitochondrial (NDI and COI) gene markers;Amer S;Parasitol Int,2011
2. Evaluation of formalin-ether sedimentation and trichrome staining techniques: its effectiveness in detecting Entamoeba histolytica/dispar/moshkovskii in stool samples;Anuar TS;J Microbiol Methods,2013
3. Molecular classification of rumen fluke eggs in fecal specimens from Suphanburi Province, Thailand, based on cytochrome C oxidase subunit 1;Anucherngchai S;Vet Parasitol Reg Stud Reports,2020
4. Anuracpreeda P, Chawengkirttikul R, Sobhon P (2016) Surface histology, topography, and ultrastructure of the tegument of adult Orthocoelium parvipapillatum (Stiles & Goldberger, 1910). Parasitol Res 115:2757–2769.
5. A study of Paramphistomum microbothrium in Khuzistan S.-W. Iran;Arfaa F;Ann Parasitol Hum Comp,1962