Fish-borne trematode metacercariae detected in fish commonly used for raw consumption in Ninh Binh Province, Vietnam.

Raw or undercooked fish dishes are the major sources of human infection of fishborne trematodes (FBT) and the situation of metacercarial infection in fish greatly affect the prevalence in humans, especially those fish that are commonly used for raw consumption. To investigate the situation of infection with metacercaria of FBT in fish often used to prepare raw fish dishes by local people to assess the risk of infection to humans in Ninh Binh province, Vietnam. 345 fish belonging to five species of freshwater and one species of brackish water fish were collected from fishermen or small-scale fish dealers in Kim Son and Yen Khanh districts, Ninh Binh province between May 2017 and May 2018. Metacercaria of FBT was discovered by pepsin and hydrochloric acid digestion techniques and identified by the morphological and molecular analysis. Among examined fish, 44.06% infected with FBT metacercaria and the highest prevalence was in Cyprinus carpio (86.54%), Ctenopharyngodon idellus (78.43%) and Hypophthalmichthys molitrix (66.67%) while Konosirus punctatus - the brackish water fish - were free from infection. Three species of FBT were found; namely Haplorchis pumilio (accounting for 99.84% of collected metacercariae), Haplorchis taichui and Clonorchis sinensis. The average density was 1.06 metacercariae per gram of freshwater fish and the highest number was of C. idellus (6.38 cysts/gram) followed by Cirrhinus molitorella and C. carpio. Results of the study show the high prevalence of infection of FBT metacercariae among freshwater fish often used to prepare raw fish dishes in Ninh Binh province. These findings suggest the need for greater awareness of the risk from raw fish dishes among public health authorities and people.

Fish-borne trematode metacercariae detected in fish commonly used for raw consumption in Ninh Binh Province, Vietnam

@#Raw or undercooked fish dishes are the major sources of human infection of fishborne trematodes (FBT) and the situation of metacercarial infection in fish greatly affect the prevalence in humans, especially those fish that are commonly used for raw consumption. To investigate the situation of infection with metacercaria of FBT in fish often used to prepare raw fish dishes by local people to assess the risk of infection to humans in Ninh Binh province, Vietnam. 345 fish belonging to five species of freshwater and one species of brackish water fish were collected from fishermen or small-scale fish dealers in Kim Son and Yen Khanh districts, Ninh Binh province between May 2017 and May 2018. Metacercaria of FBT was discovered by pepsin and hydrochloric acid digestion techniques and identified by the morphological and molecular analysis. Among examined fish, 44.06% infected with FBT metacercaria and the highest prevalence was in Cyprinus carpio (86.54%), Ctenopharyngodon idellus (78.43%) and Hypophthalmichthys molitrix (66.67%) while Konosirus punctatus – the brackish water fish – were free from infection. Three species of FBT were found; namely Haplorchis pumilio (accounting for 99.84% of collected metacercariae), Haplorchis taichui and Clonorchis sinensis. The average density was 1.06 metacercariae per gram of freshwater fish and the highest number was of C. idellus (6.38 cysts/gram) followed by Cirrhinus molitorella and C. carpio. Results of the study show the high prevalence of infection of FBT metacercariae among freshwater fish often used to prepare raw fish dishes in Ninh Binh province. These findings suggest the need for greater awareness of the risk from raw fish dishes among public health authorities and people.

Genetic differences among Haplorchis taichui populations in Indochina revealed by mitochondrial COX1 sequences.

Haplorchis taichui is an intestinal heterophyid fluke that is pathogenic to humans. It is widely distributed in Asia, with a particularly high prevalence in Indochina. Previous work revealed that the lack of gene flow between three distinct populations of Vietnamese H. taichui can be attributed to their geographic isolation with no interconnected river basins. To test the hypothesis that interconnected river basins allow gene flow between otherwise isolated populations of H. taichui, as previously demonstrated for another trematode, Opisthorchis viverrini, we compared the genetic structures of seven populations of H. taichui from various localities in the lower Mekong Basin, in Thailand and Laos, with those in Vietnam, using the mitochondrial cytochrome c oxidase subunit 1 (COX1) gene. To determine the gene flow between these H. taichui populations, we calculated their phylogenetic relationships, genetic distances and haplotype diversity. Each population showed very low nucleotide diversity at this locus. However, high levels of genetic differentiation between the populations indicated very little gene flow. A phylogenetic analysis divided the populations into four clusters that correlated with the country of origin. The negligible gene flow between the Thai and Laos populations, despite sharing the Mekong Basin, caused us to reject our hypothesis. Our data suggest that the distribution of H. taichui populations was incidentally associated with national borders.

Genetic differences among Vietnamese Haplorchis taichui populations using the COI genetic marker.

Adults of the fish-borne intestinal trematode species Haplorchis taichui were collected from humans in three provinces of Vietnam: Ha Giang, Thanh Hoa and Quang Tri. Genetic analysis revealed three groups of the parasite from clustering dendrograms, correlating with the localities in which they were collected. Measurements of evolutionary divergence over sequence pairs were greater between the different populations than within them, which indicated that the three populations were genetically different. The significance (Fst= 0.73; P value < 0.05) of the genetic variation of the three studied populations implied that genetic separation of the populations had already occurred, which may have been caused by a low gene flow among the different H. taichui populations. Factors contributing to the low gene flow may include isolation resulting from the intermediate-host fish rarely being sold outside of the rural commune where they are raised and the enclosed aquacultural areas themselves.