Molecular confirmation of trematodes in the snail intermediate hosts from Ratchaburi Province, Thailand

Abstract

Objective: To analyze the prevalence of trematode infections in the snails intermediate host Tarebia granifera and Melanoides tuberculata collected from Ratchaburi Province by a molecular approach in order to construct a dendrogram to identify species at the cercarial stage. Methods: Specimens were collected from nine districts located in Ratchaburi Province using the stratified sampling method. The samples were examined for cercarial infection using the crushing method. All specimens were amplified by region of internal transcribed spacer 2 (ITS2) based on the PCR technique, and the sequence data were aligned in order to construct the dendrogram by maximum likelihood with 10 000 bootstrap replicates. Results: Five cercarial types were observed comprising parapleurolophocercous cercaria, xiphidiocercaria, megarulous cercaria, furcocercous cercaria and transversotrema cercaria. The overall prevalence of cercarial infection was found to be 7.92% (63 / 795). The parapleurolophocercous cercaria was found in the highest level of prevalence followed by xiphidiocercaria, megarulous cercaria, furcocercous cercaria and transversotrema cercaria, respectively. The parapleurolophocercous cercaria samples were identified and separated into Haplorchis taichui and Haplorchis pumilio, while xiphidiocercaria were identified as Lecithodendriidae and megarulous cercaria were identified as Philophthalmus. The dendrogram separated the cercariae into five groups, which were comprised of parapleurolophocercous cercaria, xiphidiocercaria, megarulous cercaria, furcocercous cercaria and transversotrema cercaria using the sequence of Angiostrongylus cantonensis as an out-group. Conclusions: This is the first study to report on the prevalence of cercarial infection in Ratchaburi Province, Thailand. The high prevalence of cercariae was revealed by the infection in Tarebia granifera and Melanoides tuberculata. Thus, the sequence data of ITS2 can be used to investigate the dendrogram of trematodes and can help to separate the collected samples at the species level. This information can be used to create the prevention program for parasite infection from intermediate hosts in the future. © 2017 by the Asian Pacific Journal of Tropical Disease.