Is species identification of Echinostoma revolutum using mitochondrial DNA barcoding feasible with high-resolution melting analysis?
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The taxonomic evaluation of Echinostoma species is controversial. Echinostoma species are recognized as complex, leading to problems associated with accurate identification of these species. The aim of this study was to test the feasibility of using DNA barcoding of cytochrome c oxidase subunit I (COI) and NADH dehydrogenase subunit 1 (ND1) conjugated with high-resolution melting (HRM) analysis to identify Echinostoma revolutum. HRM using COI and ND1 was unable to differentiate between species in the “revolutum complex” but did distinguish between two isolates of 37-collar-spined echinostome species, including E. revolutum (Asian lineage) and Echinostoma sp. A from different genera, e.g., Hypoderaeum conoideum, Haplorchoides mehrai, Fasciola gigantica, and Thapariella anastomusa, based on the Tm values derived from HRM analysis. Through phylogenetic analysis, a new clade of the cryptic species known as Echinostoma sp. A was identified. In addition, we found that the E. revolutum clade of ND1 phylogeny obtained from the Thailand strain was from a different lineage than the Eurasian lineage. These findings reveal the complexity of the clade, which is composed of 37-collar-spined echinostome species found in Southeast Asia. Taken together, the systematic aspects of the complex revolutum group are in need of extensive investigation by integrating morphological, biological, and molecular features in order to clarify them, particularly in Southeast Asia.
KeywordsComplex species Cercaria Intermediate host Metacercaria Snail Southeast Asia
Finally, we would like to thank Mr. Simon McIver and Dr. Russell Kirk Hollis for their assistance in editing and proofreading this manuscript.
Thapana Chontananarth (TC) contributed, designed, and participated in all part of study, collected and identified the samples. Kittisak Buddhachat (KB) conducted the experiment in the laboratory and analyzed the data. KB and TC were working together for writing the manuscript. All authors have approved this manuscript.
This research was supported by Srinakharinwirot University (Research Grant No. 309/2559 and 043/2560).
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