Molecular and morphological variation of <i>Paragonimus westermani</i> in Vietnam with records of new second intermediate crab hosts and a new locality in a northern province

Doanh, N. Pham; Tu, A. Luu; Bui, Thi-Diem; Loan, T. Ho; Nonaka, Nariaki; Horii, Yoichiro; Blair, David; Nawa, Yukifumi

doi:10.1017/s0031182016001219

Cited by 17 publications

(17 citation statements)

References 18 publications

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“…Freshwater snails, which were molecularly identified as Triculinae gen. sp. 2 of the subfamily Triculinae Annandale, were collected in the stream where crabs were heavily infected with metacercariae of Paragonimus spp., in Huong Son Commune, Huong Hoa District, Quang Tri Province (Doanh et al 2016(Doanh et al , 2018. Snails were examined for microcercous cercariae.…”

Section: Collection and Examination Of Snailsmentioning

confidence: 99%

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Molecular identification of morphologically similar microcercous cercariae of two trematode families, Paragonimidae and Troglotrematidae, concurrently found in the same snail species of the subfamily Triculinae

Doanh¹,

Hien²,

Dung³

2020

FOLIA PARASIT

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Microcercous cercariae possess a very short tail and are produced by digenean species of several families including medically important species, such as members of the genera Paragonimus Braun, 1899, Nanophyetus Chapin, 1927 and Troglotrema Odhner, 1914. During our survey of cercariae of Paragonimus spp. in Vietnam, we found microcercous cercariae from ten (0.29%) out of 3,400 snails of Triculinae gen. sp. 2. They were morphologically and molecularly analysed for species identification. The molecular analysis, based on ITS2 sequences, revealed two distinct species: four specimens were identical to Paragonimus proliferus Hsia et Chen, 1964 (Paragonimidae Dollfus, 1939), and the other six specimens were closest to members of the family Troglotrematidae Odhner, 1914 and were temporarily named Troglotrematidae gen. sp. Morphologically, cercariae of the two species found in this study are similar to each other in their gross characteristics but can be distinguished from one another by subtle morphological details. The cercaria of P. proliferus has an I-shaped excretory bladder and does not have mucous gland cells. In contrast, that of Troglotrematidae gen. sp. has a Y-shaped excretory bladder and mucous gland cells. Besides, the redia of P. proliferus is elongate with a short intestine and contains 5-6 cercariae whereas that of Troglotrematidae gen. sp. is more round with a longer intestine and harbours 3-4 cercariae. Our results have shown the importance of the shape of the excretory bladder and the presence/absence of mucous gland cells of the cercaria as well as the shape and size of the redia, and its intestinal length as valuable taxonomic characters of intramolluscan trematode larvae. In addition, the finding of similar microcercous cercariae of different species in the same snail species suggests that careful attention to morphological details is required in the differentiation of Paragonimus cercariae and those of closely related species.

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Section: Collection and Examination Of Snailsmentioning

confidence: 99%

“…During our previous survey of snail hosts of Paragonimus spp. in a highly endemic area in central Vietnam (Doanh et al 2013(Doanh et al , 2016, we identified the cercaria of Paragonimus proliferus Hsia et Chen, 1964 in an individ-ual snail of Triculinae gen. sp. 2 (Doanh et al 2018).…”

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confidence: 91%

Molecular identification of morphologically similar microcercous cercariae of two trematode families, Paragonimidae and Troglotrematidae, concurrently found in the same snail species of the subfamily Triculinae

Doanh¹,

Hien²,

Dung³

2020

FOLIA PARASIT

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“…Different species of Paragonimus have their own predilection for infecting speci c genera of snails and crabs and hence the epidemiological prevalence of different species of Paragonimus is determined by the existence of suitable hosts. Some species of Paragonimus have been identi ed only from intermediate hosts in certain geographies which suggests infection and maintenance in non-human mammals (4,5).…”

Section: Introductionmentioning

confidence: 99%

Morphological and molecular characterization of Paragonimus species isolated from freshwater crabs in Yunnan, China.

Shu¹,

Shude²,

Tian³

et al. 2020

Preprint

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Background Paragonimus species are highly prevalent in ethnic minority areas of Yunnan, China. The objective of the study is to isolate and identify Paragonimus from natural habitat and compare the phylogenetic diversity of Paragonimus in Yunnan province, China. Methods Metacercariae of Paragonimus was isolated from crabs and morphologic identification was performed by microscopy. Metacercariae were injected into experimental Paragonimus free Sprague Dawley rats. After 112 days, adult worms and eggs were isolated from multiple organs. Morphologic identification confirmed the initial identification. DNA was extracted from the adult worms and molecular characterization was performed by amplification and sequencing of CO1 and ITS2 regions followed by phylogenetic analysis. Results Out of 447 crabs captured, 186 crabs were found to be infected. A total of 5 occurrences of Paragonimus was observed from naturally infected crabs. Paragonimus microrchis (2), P,heterotremus (1), P.proliferus (1) and P.skrjabini (1) were isolated and identified. A total of 32 sequences downloaded from national center for biotechnology information and 5 sequences generated in the study was used for phylogenetic analysis. Phylogenetic analysis revealed robustness of the double loci method as against the single locus method with either CO1 or ITS2 alone. Considerable phylogenetic diversity depending on the geographic location was observed. Conclusion Paragonimus species isolated from Yunnan province, China were phylogenetically diverse and the analysis revealed the clustering of multiple species of Paragonimus isolated from different geographic locations.

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“…The cercaria was, therefore, identified as P. heterotremus without molecular evidence [ 6 ]. However, subsequent surveys have revealed the presence of six additional Paragonimus species, and metacercariae of two to four species has been found in the same area [ 5 , 8 ]. Moreover, the taxonomy of the “ Oncomelania ” snails in Vietnam has been revised and remains uncertain [ 9 – 11 ].…”

Section: Introductionmentioning

confidence: 99%

“…The aim of this study was, therefore, to investigate snail hosts of Paragonimus spp. in Yen Bai and Quang Tri Provinces, where Paragonimus metacercariae are highly prevalent with the dominance of P. heterotremus and P. westermani , respectively [ 8 ]. In this survey, we found cercariae of three Paragonimus species, each in a different snail species.…”

Section: Introductionmentioning

confidence: 99%

First intermediate hosts of Paragonimus spp. in Vietnam and identification of intramolluscan stages of different Paragonimus species

Doanh

Hien

et al. 2018

Parasites Vectors

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BackgroundMembers of the genus Paragonimus require at least three hosts in their life-cycles. The obligatory first intermediate hosts are freshwater snails. In Vietnam, although seven Paragonimus species have been recorded, the natural first intermediate hosts of almost all species have not been confirmed. The aim of this study was, therefore, to investigate snail hosts of Paragonimus species in Vietnam, and to identify Paragonimus species at intramolluscan stages.MethodsFreshwater snails were collected from streams in Yen Bai and Quang Tri Provinces, where high prevalences of Paragonimus metacercariae in crab hosts have been reported. Snails were morphologically identified and then examined individually for Paragonimus cercariae using shedding and crushing methods. Chaetomicrocercous cercariae, the morphological class to which Paragonimus cercariae belong, were collected for morphological description and molecular species identification by analyses of ITS2 sequences. The infected snail species were identified based on analyses of nucleotide sequences of the cox1 gene.ResultsThree snail species were found to be infected with Paragonimus cercariae at low infection rates, ranging between 0.07–1.0%. The molecular analyses identified them as Sulcospira quangtriensis and 2 species of subfamily Triculinae. In a phylogenetic tree, these two triculine snails were related to the genera Gammatricula and Tricula with low posterior probabilities. Thus we named them as Triculinae sp. 1 and Triculinae sp. 2. Cercariae from the three snail species, Sulcospira quangtriensis, Triculinae sp. 1 and Triculinae sp. 2, were molecularly identified as Paragonimus westermani, P. heterotremus and P. proliferus, respectively. The cercariae of the three species are morphologically similar to each other, but their daughter rediae can be distinguished by the length of the intestine and the number of cercariae per redia. The rediae of P. westermani have a long intestine and each contain 6–8 cercariae. In contrast, those of P. heterotremus and P. proliferus have a short intestine and each redia contain 10–12 and 5–6 cercariae, respectively.ConclusionsThree snail species, Sulcospira quangtriensis, Triculinae sp. 1 and Triculinae sp. 2, serve as the first intermediate hosts of P. westermani, P. heterotremus and P. proliferus, respectively, in Vietnam. The length of the intestine of rediae and the number of cercariae per redia are valuable characteristics for distinguishing between larvae of these Paragonimus species.

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Molecular and morphological variation of Paragonimus westermani in Vietnam with records of new second intermediate crab hosts and a new locality in a northern province

Cited by 17 publications

References 18 publications

Molecular identification of morphologically similar microcercous cercariae of two trematode families, Paragonimidae and Troglotrematidae, concurrently found in the same snail species of the subfamily Triculinae

Molecular identification of morphologically similar microcercous cercariae of two trematode families, Paragonimidae and Troglotrematidae, concurrently found in the same snail species of the subfamily Triculinae

Morphological and molecular characterization of Paragonimus species isolated from freshwater crabs in Yunnan, China.

First intermediate hosts of Paragonimus spp. in Vietnam and identification of intramolluscan stages of different Paragonimus species

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