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• F. gigantica was the main liver fluke in Cambodia as identified by mPCR- PEPCK gene. • All flukes were classified into F. gigantica haplogroup C based on ND1 gene. • Nucleotide diversity of ND1 of Cambodian flukes was higher than that from Thailand. • Nucleotide diversity of Cambodian population was higher than that from Thailand. • Population genetics suggest that F. gigantica spread from Cambodia to other areas. Fasciola gigantica and hybrid Fasciola are distributed throughout Asia. Herein, we investigated the species of the Fasciola fluke distributed in three hotspots of fascioliasis in Cambodia. A total of 92 flukes collected from 21 slaughtered cattle from Kandal (44), Battambang (41), and Kratie (7) Provinces were identified as F. gigantica using multiplex PCR for a nuclear phosphoenolpyruvate carboxykinase (PEPCK) gene. The overall prevalence of F. gigantica infestation was 7.14% (21/294). Phylogenetic as well as population genetics analyses were performed using the mitochondrial NADH dehydrogenase subunit 1 (ND1). The 19 ND1 haplotypes were identified from Cambodian F. gigantica (haplotype diversity, 0.83). All of the haplotypes were classified into F. gigantica haplogroup C, which includes ND1 haplotypes detected from Thailand, Vietnam, Indonesia, Myanmar, and China. Among haplogroup C, novel and unique haplotypes of Cambodia were found in the Battambang and Kandal Provinces, and the nucleotide diversity of the Cambodian population (0.00532) was the highest. Pairwise fixation indices among the F. gigantica populations from these countries indicated that the Cambodian and Thailand populations were related to each other. The highest genetic diversity in the Cambodian population suggests that F. gigantica in Cambodia may be the ancestor of the populations in Southeast Asian countries. Most likely, livestock movement, including Zebu cattle, played an important role in the transmission of F. gigantica. In this study, the hybrid Fasciola flukes that are commonly found in neighboring countries, were not found in Cambodia. Further comprehensive investigations of Fasciola prevalence should be conducted by analyzing a wider range of hosts throughout Cambodia to reach a more solid conclusion about the absence of hybrid flukes. [ABSTRACT FROM AUTHOR]

Fasciola gigantica and hybrid Fasciola flukes, responsible for the disease fasciolosis, are found in Southeast Asian countries. In the present study, we performed molecular species identification of Fasciola flukes distributed in Terengganu, Malaysia using multiplex PCR for phosphoenolpyruvate carboxykinase (pepck) and PCR-restriction fragment length polymorphism (RFLP) for DNA polymerase delta (pold). Simultaneously, phylogenetic analysis based on mitochondrial NADH dehydrogenase subunit 1 (nad1) was performed for the first time on Malaysian Fasciola flukes to infer the dispersal direction among neighboring countries. A total of 40 flukes used in this study were identified as F. gigantica. Eight nad1 haplotypes were identified in the F. gigantica population of Terengganu. Median-joining network analysis revealed that the Malaysian population was related to those obtained from bordering countries such as Thailand and Indonesia. However, genetic differentiation was detected using population genetics analyses. Nevertheless, the nucleotide diversity (π) value suggested that F. gigantica with the predominant haplotypes was introduced into Malaysia from Thailand and Indonesia. The dispersal direction suggested by population genetics in the present study may not be fully reliable since Fasciola flukes were collected from a single location in one state of Malaysia. Further studies analyzing more samples from many locations are required to validate the dispersal direction proposed herein. [Display omitted] • Molecular species identification of Fasciola flukes in Terengganu was performed. • They were identified as F. gigantica using nuclear markers, pepck and pold. • Eight mitochondrial nad1 haplotypes were identified in Terengganu. • Population genetics analyses were conducted to infer the expansion route. • Malaysian F. gigantica was speculated to be introduced from Thailand and Indonesia. [ABSTRACT FROM AUTHOR]

Fasciola gigantica and aspermic (hybrid) Fasciola flukes are thought to be distributed in Southeast Asian countries. The objectives of this study were to investigate the distribution of these flukes from unidentified ruminants in western Java, Indonesia, and to determine their distribution history into the area. Sixty Fasciola flukes from western Java were identified as F. gigantica based on the nucleotide sequences of the nuclear phosphoenolpyruvate carboxykinase (pepck) and DNA polymerase delta (pold) genes. The flukes were then analyzed phylogenetically based on the nucleotide sequence of the mitochondrial NADH dehydrogenase subunit 1 (nad1) gene, together with Fasciola flukes from other Asian countries. All but one F. gigantica fluke were classified in F. gigantica haplogroup C, which mainly contains nad1 haplotypes detected in flukes from Thailand, Vietnam, and China. A population genetic analysis suggested that haplogroup C spread from Thailand to the neighboring countries including Indonesia together with domestic ruminants, such as the swamp buffalo, Bubalus bubalis. The swamp buffalo is one of the important definitive hosts of Fasciola flukes in Indonesia, and is considered to have been domesticated in the north of Thailand. The remaining one fluke displayed a novel nad1 haplotype that has never been detected in the reference countries. Therefore, the origin of the fluke could not be established. No hybrid Fasciola flukes were detected in this study, in contrast to neighboring Asian countries., (Copyright © 2016. Published by Elsevier Ireland Ltd.)