Article

Article name MOLECULAR AND MORPHOLOGICAL CHARACTERISATION OF FLATWORM LARVAE PARASITISING ON FISH IN CAT TIEN NATIONAL PARK, VIETNAM
Authors

Sergey G. Sokolov, Senior Researcher of the A.N. Severtsov Institute of Ecology and Evolution of RAS (119071, Russia, Moscow, Leninsky Prospect, 33); e-mail: sokolovsg@mail.ru
Ilya I. Gordeev, Senior Researcher of the Russian Federal Research Institute of Fisheries and Oceanography; Senior Researcher of the Department of Invertebrate Zoology of the Faculty of Biology in the Lomonosov Moscow State University (V. Krasnoselskaya Street, 17, 107140, Moscow, Russia); e-mail: gordeev_ilya@bk.ru

Reference to article

Sokolov S.G., Gordeev I.I. 2020. Molecular and morphological characterisation of flatworm larvae parasitising on fish in Cat Tien National Park, Vietnam. Nature Conservation Research 5(Suppl.2): 19–30. https://dx.doi.org/10.24189/ncr.2020.039

Electronic supplement 1. List of previously published trematode sequences used in the phylogenetic analysis (Link).
Electronic supplement 2. List of previously published cestode sequences used in the phylogenetic analysis (Link).

Section Research articles
DOI https://dx.doi.org/10.24189/ncr.2020.039
Abstract

Cat Tien National Park in southern Vietnam provides a unique opportunity to study the diversity of parasites associated with animals of the plains of tropical forests and lowland river basins in Southeast Asia. In this study we provide morphological description and phylogenetic analysis based on partial sequences of the 28S rRNA gene of metacercariae belonging to five species: Clinostomum sp., Posthodiplostomum sp. 1, Posthodiplostomum sp. 2, Crassiphialinae gen. sp. 1 and Crassiphialinae gen. sp. 2, collected in four species of freshwater fish (Rasbora paviana, Trichopodus trichopterus, Anabas testudineus and Channa striata). The digenean Clinostomum sp. was found to be phylogenetically close (possibly conspecific) to metacercaria of Clinostomum sp. recorded in Australia. There are no sequences in the GenBank database identical to any Posthodiplostomum spp. nor two other crassiphialine metacercariae found by us. Phylogenetic analysis supports the sister position of Crassiphialinae gen. sp. 1 to the Uvulifer spp. + Crassiphiala spp. group of crassiphialine trematodes. At the same time, phylogenetic relationships of Crassiphialinae gen. sp. 2 were poorly resolved. We also provide morphological description, cox1 and 28S rRNA genes-based reconstruction of phylogeny for the plerocercoid of solenophorid cestode Scyphocephalus sp. ex liver of Trichopodus trichopterus. Phylogenetic analyses unite plerocercoid of Scyphocephalus sp. in one group with solenophorid Duthiersia expansa, while the type species of the genus ScyphocephalusS. bisulcatus– appears as a sister to Duthiersia fimbriata on the tree. Thus, phylogenetic data cast doubt on both the monotypy and the monophyly of the genus Scyphocephalus. This is the second record of plerocercoids of solenophorids and the first one of fish as second intermediate host of this cestode family.

Keywords

Clinostomum, Crassiphialinae, fresh waters, metacercariae, plerocercoids, Posthodiplostomum, Scyphocephalus, tropics

Artice information

Received: 21.05.2020. Revised: 11.06.2020. Accepted: 12.07.2020.

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References

Agustí C., Aznar F.J., Olson P.D., Littlewood D.T.J., Kostadinova A., Raga J.A. 2005. Morphological and molecular characterization of tetraphyllidean merocercoids (Platyhelminthes: Cestoda) of striped dolphins (Stenella coeruleoalba) from the Western Mediterranean. Parasitology 130(4): 461–474. DOI: 10.1017/S0031182004006754
Arthur J.R., Te B.Q. 2006. Checklist of the parasites of fishes of Viet Nam. FAO Fisheries Technical Paper. No. 369/2. Rome: FAO. 133 p.
Blanc L., Maury-Lechon G., Pascal J.P. 2000. Structure, floristic composition and natural regeneration in the forests of Cat Tien National Park, Vietnam: an analysis of the successional trends. Journal of Biogeography 27(1): 141–157. DOI: 10.1046/j.1365-2699.2000.00347.x
Boone E.C., Laursen J.R., Colombo R.E., Meiners S.J., Romani M.F., Keeney D.B. 2018. Infection patterns and molecular data reveal host and tissue specificity of Posthodiplostomum species in centrarchid hosts. Parasitology 145(11): 1458–1468. DOI: 10.1017/s0031182018000306
Bush A.O., Lafferty K.D., Lotz J.M., Shostak A.W. 1997. Parasitology meets ecology on its own terms: Margolis et al. revisited. Journal of Parasitology 83(4): 575–583. DOI: 10.2307/3284227
Bykhovskaya-Pavlovskaya I.E. 1985. Parasites of fishes. Study Manual. Leningrad: Nauka. 123 p. [In Russian]
Chernov T.I., Zhelezova A.D., Tkhakakhova A.K., Bgazhba N.A., Zverev A.O. 2019. Microbiomes of Virgin Soils of Southern Vietnam Tropical Forests. Microbiology 88(4): 489–498. DOI: 10.1134/S0026261719040040
Choudhary K., Tripathi A., Pandey K.C., Agrawal N. 2017. Morphological and molecular characterisation of three Indian Neascus-type metacercariae (Digenea: Diplostomidae). Parasitology International 66(5): 652–659. DOI: 10.1016/j.parint.2017.06.007
De N.V., Le T.H. 2011. Human infections of fish-borne trematodes in Vietnam: prevalence and molecular specific identification at an endemic commune in Nam Dinh province. Experimental Parasitology 129(4): 355–361. DOI: 10.1016/j.exppara.2011.09.005
Deshcherevskaya O.A., Avilov V.K., Dinh B.D., Tran C.H., Kurbatova J.A. 2013. Modern climate of the Cat Tien National Park (Southern Vietnam): climatological data for ecological studies. Izvestiya – Atmospheric and Ocean Physics 49(8): 819–838. DOI: 10.1134/S0001433813080021
Dobson A., Lafferty K.D., Kuris A.M., Hechinger R.F., Jetz W. 2008. Homage to Linnaeus: how many parasites? How many hosts? In: J.C. Avise, S.P. Hubbell, F.J. Ayala (Eds.). In the Light of Evolution. Vol. 2: Biodiversity and Extinction. Washington: National Academies Press. P. 63–82.
Dubois G. 1968. Synopsis des Strigeidae et des Diplostomatidae (Trematoda). Mémoires de la Société Neuchâteloise des Sciences Naturelles 10: 1–258.
Faltýnková A., Georgieva S., Kostadinova A., Blasco-Costa I., Scholz T., Skírnisson K. 2014. Diplostomum von Nordmann, 1832 (Digenea: Diplostomidae) in the sub-Arctic: descriptions of the larval stages of six species discovered recently in Iceland. Systematic Parasitology 89(3): 195–213. DOI: 10.1007/s11230-014-9517-0
Felsenstein J. 1985. Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39(4): 783–791. DOI: 10.1111/j.1558-5646.1985.tb00420.x
Fischthal J.H., Kuntz R.E. 1973. Additional digenetic trematodes of birds from North Borneo (Malaysia). Proceedings of the Helminthological Society of Washington 40(2): 245–255.
Galazzo D.E., Dayanandan S., Marcogliese D.J., McLaughlin J.D. 2002. Molecular systematics of some North American species of Diplostomum (Digenea) based on rDNA-sequence data and comparisons with European congeners. Canadian Journal of Zoology 80(12): 2207–2217. DOI: 10.1139/Z02-198
Gordeev I.I., Biserova N.M., Grigorov I.V., Mustafina A.R., Golovaneva M.S. 2018. The experience of ichthyo-parasitological studies in a tropical forest. In: A.M. Orlov, I.I. Gordeev, A.A. Sergeev (Eds.): Prospects for Fisheries and Aquaculture in the Modern World. Moscow: VNIRO. P. 74. [In Russian]
Guseva E.E., Zhokhov A.E., Nguen T.H.T. 2014. Helminth fauna of freshwater fish in central and southern Vietnam. In: D.S. Pavlov, D.D. Zvorykin (Eds.): Ecology of inland waters of Vietnam. Moscow: KMK Scientific Press Ltd. P. 208–224. [In Russian]
Gupta B.K. 2016. Studies on the structure of three new species of strigeids metacercariae from Indian freshwater food fishes. Indian Journal of Pure and Applied Biosciences 4(4): 221–228. DOI: 10.18782/2320-7051.2347
Hung N.M., Dung D.T., Lan Anh, N.T., Van P.T., Thanh B.N., Van Ha N., Van Hien H., Canh L.X. 2015. Current status of fish-borne zoonotic trematode infections in Gia Vien district, Ninh Binh province, Vietnam. Parasites Vectors 8(1): 21. DOI: 10.1186/s13071-015-0643-6
Klimpel S., Kuhn T., Mьnster J., Dцrge D.D., Klapper R., Kochmann J. 2019. Parasites of marine fish and cephalopods. Springer International Publishing. 169 p.
Kumar S., Stecher G., Li M., Knyaz C., Tamura K. 2018. MEGA X: Molecular evolutionary genetics analysis across computing platforms. Molecular Biology and Evolution 35(6): 1547–1549. DOI: 10.1093/molbe v/msy09
Locke S.A., McLaughlin J.D., Marcogliese D.J. 2010. DNA barcodes show cryptic diversity and a potential physiological basis for host specificity among Diplostomoidea (Platyhelminthes: Digenea) parasitizing freshwater fishes in the St. Lawrence River, Canada. Molecular Ecology 19(13): 2813–2827. DOI: 10.1111/j.1365-294x.2010.04713.x
Locke S.A., Caffara M., Marcogliese D.J., Fioravanti M.L. 2015. A large-scale molecular survey of Clinostomum (Digenea, Clinostomidae). Zoologica Scripta 44(2): 203–217. DOI: 10.1111/zsc.12096
López-Hernández D., Locke S.A., de Melo A.L., Rabelo É.M.L., Pinto H.A. 2018. Molecular, morphological and experimental assessment of the life cycle of Posthodiplostomum nanum Dubois, 1937 (Trematoda: Diplostomidae) from Brazil, with phylogenetic evidence of the paraphyly of the genus Posthodiplostomum Dubois, 1936. Infection, Genetics and Evolution 63: 95–103. DOI: 10.1016/j.meegid.2018.05.010
Luton K., Walker D., Blair D. 1992. Comparisons of ribosomal internal transcribed spacers from two congeneric species of flukes (Platyhelminthes: Trematoda: Digenea). Molecular and Biochemical Parasitology 56(2): 323–327. DOI: 10.1016/0166-6851(92)90181-I
Lysenko V.N. 2013. Ecological-faunal analysis of parasites of carps (Cyprinidae) of the Viet Nam aquaculture. Hydrobiological Journal 49(4): 100–112. DOI: 10.1615/HydrobJ.v49.i4.90 [In Russian]
Nguyen N.T., Yen H.M. 2013. Cat Tien National Park. In: T.C.H. Sunderland, J. Sayer, M.H. Hoang (Eds.): Evidence-based conservation: lessons from the lower Mekong. Bogor: CIFOR. P. 17–28.
Nguyen T.C., Li Y.C., Makouloutou P., Jimenez L.A., Sato H. 2012. Posthodiplostomum sp. metacercariae in the trunk muscle of northern snakeheads (Channa argus) from the Fushinogawa River, Yamaguchi, Japan. Journal of Veterinary Medical Science 74(10): 1367–1372. DOI: 10.1292/jvms.12-0025
Olson P.D., Cribb T.H., Tkach V.V., Bray R.A., Littlewood D.T.J. 2003. Phylogeny and classification of the Digenea (Platyhelminthes: Trematoda). International Journal for Parasitology 33(7): 733–755. DOI: 10.1016/s0020-7519(03)00049-3
Pandey K.C., Agrawal N. 2013. Metacercarial fauna of India. Kolkata: Zoological Survey of India. 310 p.
Pandey K.C., Rajvanshi S.L. 1984. Note on metacestode of Duthiersia expansa, 1873. Current Science 53(22): 1218–1219.
Patarwut L., Chontananarth T., Chai J.Y., Purivirojkul W. 2020. Infections of Digenetic Trematode Metacercariae in Wrestling Halfbeak, Dermogenys pusilla from Bangkok Metropolitan Region in Thailand. Korean Journal of Parasitology 58(1): 27–35. DOI: 10.3347/kjp.2020.58.1.27
Pérez-Ponce de León G., Garcia-Varela M., Pinacho-Pinacho C.D., Sereno-Uribe A.L., Poulin R. 2016. Species delimitation in trematodes using DNA sequences: Middle-American Clinostomum as a case study. Parasitology 143(13): 1773–1789. DOI: 10.1017/S0031182016001517
Posada D. 2008. jModelTest: phylogenetic model averaging. Molecular Biology and Evolution 25(7): 1253–1256. DOI: 10.1093/molbev/msn083
Poulin R. 2010. Latitudinal gradients in parasite diversity: bridging the gap between temperate and tropical areas. Neotropical Helminthology 4(2): 169–177.
Riggenbach E. 1899. Scyphocephalus bisulcatus n. g. n. sp., ein neuer Reptiliencestode. Zoologischer Jahrbücher. Abtheilung für Systematik, Geographie und Biologie der Thiere 12: 145–154.
Rim H.J., Sohn W.M., Yong T.S., Eom K.S., Chai J.Y., Min D.Y., Lee S.H., Hoang E.H., Phommasack B., Insisengmay S. 2008. Fishborne trematode metacercariae detected in freshwater fish from Vientiane Municipality and Savannakhet Province, Lao PDR. Korean Journal of Parasitology 46(4): 253–260. DOI: 10.3347/kjp.2008.46.4.253
Ronquist F., Huelsenbeck J.P. 2003 MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19(12): 1572–1574. DOI: 10.1093/bioinformatics/btg180
Schmidt G.D., Kuntz R.E. 1974. Tapeworms from Philippine reptiles, with two new species of Proteocephalata. Proceedings of the Helminthological Society of Washington 41(2): 195–199.
Sokolov S.G., Gordeev I.I. 2019. Asaccotrema vietnamiense n. gen.; n. sp. (Trematoda: Monorchioidea), a new aberrant representative of lissorchiid trematodes from the sidestripe rasbora, Rasbora paviana Tirant (Actinopterygii: Cyprinidae), Vietnam. Zootaxa 4674(4): 451–462. DOI: 10.11646/zootaxa.4674.4.4
Tkach V.V., Littlewood D.T.J., Olson P.D., Kinsella J.M., Swiderski Z. 2003. Molecular phylogenetic analysis of the Microphalloidea Ward, 1901 (Trematoda: Digenea). Systematic Parasitology 56(1): 1–15. DOI: 10.1023/A:1025546001611
Tordoff A.W., Tran Quoc Bao, Nguyen Duc Tu, Le Manh Hung (Eds.). 2004. Sourcebook of existing and proposed protected areas in Vietnam. Second edition. Hanoi: BirdLife International in Indochina and the Ministry of Agriculture and Rural Development. 268 p.
Truett G.E. 2006. Preparation of genomic DNA from animal tissues. In: J. Kieleczawa (Ed.): The DNA Book: Protocols and Procedures for the Modern Molecular Biology. Udbury, Massachusetts: Jones & Bartlett Publisher. P. 33–46.
Vlnová I. 2014. Revision of tapeworms of family Diphyllobothriidae (Eucestoda: Solenophoridae) from the monitor lizards. Mgr. Thesis. České Budějovice, Czech Republic: University of South Bohemia. 61 p.
Waeschenbach A., Brabec J., Scholz T., Littlewood D.T.J., Kuchta R. 2017. The catholic taste of broad tapeworms – multiple routes to human infection. International Journal for Parasitology 47(13): 831–843. DOI: 10.1016/j.ijpara.2017.06.004
Waeschenbach A., Littlewood D.T.J. 2017. A molecular framework for the Cestoda. In: J.N. Caira, K. Jensen (Eds.): Planetary Biodiversity Inventory (2008–2017): Tapeworms from the vertebrate bowels of the Earth. Special Publication No. 25. Lawrence: University of Kansas. P. 431–451.
Zhang G., Chen J., Yang Y., Liu N., Jiang W., Gu S., Wang X., Wang Z. 2014. Utility of DNA barcoding in distinguishing species of the family Taeniidae. Journal of Parasitology 100(4): 542–546. DOI: 10.1645/13-224.1