Article

Nadezhda Yu. Kirillova, Senior Researcher of the Laboratory of Populational Ecology, Samara Federal Research Scientific Center of RAS, Institute of Ecology of Volga River Basin of RAS (445003, Russia, Samara region, Togliatti, Komzina str., 10); e-mail: parasitolog@yandex.ru
Alexander A. Kirillov, Senior Researcher of the Laboratory of Populational Ecology, Samara Federal Research Scientific Center of RAS, Institute of Ecology of Volga River Basin of RAS (445003, Russia, Samara region, Togliatti, Komzina str., 10); e-mail: parasitolog@yandex.ru
Sergei V. Shchenkov, Department of Invertebrate Zoology, Faculty of Biology, Saint-Petersburg State University (199034, Russia, St. Petersburg, Universitetskaya emb., 7/9); e-mail: sergei.shchenkov@gmail.com
Igor V. Chikhlyaev, Senior Researcher of the Laboratory of Populational Ecology, Samara Federal Research Scientific Center of RAS, Institute of Ecology of Volga River Basin of RAS (445003, Russia, Samara region, Togliatti, Komzina str., 10); e-mail: diplodiscus@mail.ru

Kirillova N.Yu., Kirillov A.A., Shchenkov S.V., Chikhlyaev I.V. 2020. Oswaldocruzia filiformis sensu lato (Nematoda: Molineidae) from amphibians and reptiles in European Russia: morphological and molecular data. Nature Conservation Research 5(Suppl.2): 41–56. https://dx.doi.org/10.24189/ncr.2020.041

Nematodes of the genus Oswaldocruzia parasitise the small intestine of amphibians and reptiles. Their biodiversity remains unknown. We studied Oswaldocruzia nematodes from nine species of amphibians and reptiles (Pelophylax ridibundus, Rana arvalis, Rana temporaria, Bufo bufo, Lacerta agilis, Zootoca vivipara, Anguis fragilis, Natrix natrix, Vipera berus) at six localities in European Russia in 2018–2019. To identify their nematode species, we analysed the morphological characters traditionally used in the taxonomy of nematodes of this genus together with new molecular phylogenetic data. The results of partial sequencing and molecular phylogenetic analysis of CoxI mtDNA gene showed that all Oswaldocruzia specimens in this study belonged to the same species. We observed a broad morphological variability of nematodes both from different host species and from the same host individual. Morphological variation in nematodes from various host species could be host-induced, while that in nematodes from the same host individuals could be due to phenotypic plasticity of the species. Molecular data indicate that only one species of the genus Oswaldocruzia, O. filiformis s.l., which has a broad morphological variability, parasitises amphibians and reptiles in European Russia. The results of our study highlight the necessity of studying the diversity of morphologically similar Oswaldocruzia spp. from the Western Palearctic by molecular genetic methods.

CoxI mtDNA, molecular phylogenetic analysis, morphological variability, trichostrongylids, Western Palearctic

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

Anderson R.C. 2000. Nematode Parasites of Vertebrates: Their Development and Transmission. 2^nd ed. Wallingford: CABI Publishing. 650 p.
Amin O.M. 1975. Variability in Acanthocephalus parksidei Amin, 1974 (Acanthocephala: Echinorhynchidae). Journal of Parasitology 61(2): 307–317. DOI: 10.2307/3279011
Anikieva L.V. 2004. Variability and phenotypic structure of Proteocephalus torulosus (Cestoda: Proteocephalidae) – a parasite of cyprinid fishes. Parazitologiya 38(2): 171–179. [In Russian]
Anikieva L.V. 2005. Phenotypic variability of a perch parasite – cestode Proteocephalus percae (Muller, 1780) (Proteocephalidea) in different parts of the species range. Parazitologiya 39(5): 386–396. [In Russian]
Anikieva L.V. 2008. Populational variability of Proteocephalus longicollis (Cestoda: Proteocephalidae) in the vendace Coregonus albula from lakes of Karelia. Parazitologiya 42(1): 3–12. [In Russian]
Baker M.R. 1981. On three Oswaldocruzia spp. (Trichostrongyloidea: Molineidea) in Amphibians from Africa. Canadian Journal of Zoology 59(2): 246–251. DOI: 10.1139/z81-039
Ben Slimane B., Durette-Desset M.C. 1993. Quatre nouvelles espéces du genre Oswaldocruzia Travassos, 1917 (Nematoda: Trichostrongyloidea) parasites d'Amphibiens d' Equateur. Revue Suisse de Zoologie 100: 113–136.
Ben Slimane B., Durette-Desset M.C. 1996a. Four New Species of Oswaldocruzia (Nematoda: Trichostrongylina, Molineoidea) Parasitizing Amphibians and Lizards from Ecuador. Memórias do Instituto Oswaldo Cruz 91(3): 317–328. DOI: 10.1590/S0074-02761996000300012
Ben Slimane B., Durette-Desset M.C. 1996b. New Oswaldocruzia (Nematoda, Trichostrongylina, Molineoidea) parasites of Amphibians from French Guyana and Ecuador. Miscellánia Zoológica 19(1): 55–66.
Ben Slimane B., Durette-Desset M.C. 1997. Révision du genre Oswaldocruzia (Nematoda, Trichostrongylina, Molineoidea) en zone néarctique avec description de cinq nouvelles espéces. Zoosystema 19(1): 61–79.
Ben Slimane B., Durette-Desset M.C., Chabaud A.G. 1993. Oswaldocruzia (Trichostrongyloidea) Parasites d'amphibiens des collections du Muséum de Paris. Annales De Parasitologie Humaine et Comparée 68(2): 88–100. DOI: 10.1051/parasite/199368288
Ben Slimane B., Chabaud A.G., Durette-Desset M.C. 1996a. Les Nématodes Trichostrongylina parasites d'Amphibiens et de Reptiles: problémes taxonomiques, phylétiques et biogéographiques. Systematic Parasitology 35: 179–206.
Ben Slimane B., Guerrero R., Durette-Desset M.C. 1996b. Oswaldocruzia venezuelensis sp. n. (Nematoda: Trichostrongylina, Molineoidea), a parasite of Bufo marinus from Venezuela. Folia Parasitologica 43(4): 297–300.
Ben Slimane B., Lluch J., Durette-Desset M.C. 1995. Two new species of the genus Oswaldocruzia Travassos, 1917 (Nematode: Trichostrongylina: Molineidae) parasitizing Spanish amphibians. Research and Review in Parasitology 55(4): 209–215.
Bertman M., Okulewicz A. 1987. Blindworm – Anguis fragilis L. and the grass snake – Natrix natrix (L.) new hosts of Oswladocruzia filiformis (Goeze, 1782) Travassos 1917 (Nematoda) in Poland. Wiadomosti Parazytologichni 33: 209–212.
Bjelić-Čabrilo O., Popović E., Paunović A. 2009. Helminthofauna of Pelophylax kl. esculentus (Linne, 1758) from Petrovaradinski Rit Marsh (Serbia). Helminthologia 46(2): 107–111. DOI: 10.2478/s11687-009-0021-z
Bowles J., Blair D., McManus D.P. 1992. Genetic variants within the genus Echinococcus identified by mitochondrial DNA sequencing. Molecular and Biochemical Parasitology 54(2): 165–173. DOI: 10.1016/0166-6851(92)90109-w
Bursey C.R., Goldberg S.R. 2011. New species of Oswaldocruzia (Nematoda: Molineidae) and other helminths in Bolitoglossa subpalmata (Caudata: Plethodontidae) from Costa Rica. Journal of Parasitology 97(2): 286–292. DOI: 10.1645/GE-2622.1
Bursey C.R., Goldberg S.R., Telford S.R. 2007. Gastrointestinal Helminths of 14 Species of Lizards from Panama with Descriptions of Five New Species. Comparative Parasitology 74(1): 108–140. DOI: 10.1654/4228.1
Catalano S., Lejeune M., van Paridon B., Pagan C.A., Wasmuth J.D., Tizzani P., Duignan P.J., Nadler S.A. 2015. Morphological variability and molecular identification of Uncinaria spp. (Nematoda: Ancylostomatidae) from grizzly and black bears: new species or phenotypic plasticity? Journal of Parasitology 101(2): 182–192. DOI: 10.1645/14-621.1
Chikhlyaev I.V., Ruchin A.B. 2014. The helminth fauna study of European common brown frog (Rana temporaria Linnaeus, 1758) in the Volga basin. Acta Parasitologica 59(3): 459–471. DOI: 10.2478/s11686-014-0268-5
Chikhlyaev I.V., Ruchin A.B., Fayzulin A.I. 2016. The Helminth Fauna Study of European Common Toad in the Volga Basin. Nature Environment and Pollution Technology 15(3): 1103–1109.
Chikhlyaev I.V., Kirillova N.Yu., Kirillov A.A. 2018. Overwiew of helminths of amphibians (Amphibia) from Samara region. Proceedings of Samara Scientific Centre RAS 20(5–3): 385–400. [In Russian]
Chikhlyaev I.V., Ruchin A.B., Fayzulin A.I. 2019. Parasitic nematodes of Pool frog (Pelophylax lessonae) in the Volga basin. Journal MVZ Cordoba 24(3): 7314–7321. DOI: 10.21897/rmvz.1501
Durette-Desset M.C. 1985. Trichostrongyloid Nematodes and their Vertebrate Hosts: Reconstruction of the Phylogeny of a Parasitic Group. Advances in Parasitology 24: 239–306.
Durette-Desset M.C., Chabaud A.G. 1981. Nouvel essai de classification des Nématodes Trichostrongyloidea. Annales De Parasitologie Humaine et Comparée 56: 297–312.
Durette-Desset M.C., Batcharov A., Ben Slimane B., Chabaud A. 1993. Some Oswaldocruzia (Nematoda: Trichostrongyloidea) parasites of Amphibia in Bulgaria. Redescription of Oswaldocruzia bialata (Molin, 1860). Helminthologia 30(1/2): 99–104.
Durette-Desset M.C., Alves dos Anjos L., Vrcibradic D. 2006. Three new species of the genus Oswaldocruzia Travassos, 1917 (Nematoda, Trichostrongylina, Molineoidea) parasites of Enyalius spp. (Iguanidae) from Brazil. Parasite 13(2): 115–125. DOI: 10.1051/parasite/2006132115
Edgar R.C. 2004. MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Research 32(5): 1792–1797. DOI: 10.1093/nar/gkh340
Galli P., Crosa G., Gentilli A., Santagostino M. 2001. New geographical records of parasitic nematodes from Bufo bufo in Italy. Parassitologia 43(4): 147–149.
Gouy M., Guindon S., Gascuel O. 2010. SeaView version 4: a multiplatform graphical user interface for sequence alignment and phylogenetic tree building. Molecular Biology and Evolution 27(2): 221–224. DOI: 10.1093/molbev/msp259
Guerrero R. 2013. Two new species of Oswaldocruzia (Nematoda: Trichostrongylina: Molineoidea) parasites of the cane toad Rhinella marina (Amphibia: Anura) from Peru. Acta Parasitologica 58(1): 30–36. DOI: 10.2478/s11686-013-0103-4
Herczeg D., Voros L., Vegvari Z., Kuzmin Yu., Brooks D.R. 2016. Helminth Parasites of the Pelophylax esculentus Complex (Anura: Ranidae) in Hortobagy National Park (Hungary). Comparative Parasitology 83(1): 36–48. DOI: 10.1654/1525-2647-83.1.36
Hendrikx W.M.L., van Moppes M.C. 1983. Oswaldocruzia filiformis (Nematoda: Trichostrongylidae): Morphology of developmental stages, parasitic development and some pathological aspects of the infection in amphibians. Zeitschrift fur Parasitenkunde 69(4): 523–537. DOI: 10.1007/BF00927709
Hu W., Yu X.G., Wu S., Tan L.P., Song M.R., Abdulahi A.Y., Wang Z., Jiang B., Li G.Q. 2016. Levels of Ancylostoma infections and phylogenetic analysis of cox 1 gene of A. ceylanicum in stray cat faecal samples from Guangzhou, China. Journal of Helminthology 90(4): 392–397. DOI: 10.1017/S0022149X15000413
Kirillov A.A. 2000. Helminth fauna of reptiles from Samara region. Proceedings of Samara Scientific Centre RAS 2(2): 324–329. [In Russian]
Kirillov A.A. 2010. Helminths of reptiles from the Volga region peculiar to other animals. Vestnik of Samara University 80(6): 196–205. [In Russian]
Kirillov A.A., Kirillova N.Yu. 2010. Morphological variability of Plagiorchis elegans (Rudolphi, 1802) (Trematoda: Plagiorchiidae) from type lizard in Middle Volga region. Proceedings of Samara Scientific Centre RAS 12(1): 138–141. [In Russian]
Kirillov A.A., Kirillova N.Yu. 2015. Variability and determining factors of the body size structure of the infrapopulation of Cosmocerca ornata (Nematoda: Cosmocercidae) in marsh frogs. Parazitologiya 49(2): 104–118. [In Russian]
Kirillov A.A., Kirillova N.Yu. 2018. Overview of helminths in reptiles of the National Park «Samarskaya Luka» (Russia). Nature Conservation Research 3(Suppl.1): 73–82. DOI: 10.24189/ncr.2018.049 [In Russian]
Kirillov A.A., Kirillova N.Yu., Ruchin A.B. 2019. Overview of the Helminth Fauna of Slow Worms of Genus Anguis (Reptilia, Anguidae) in the Western Palearctic. Entomology and Applied Science Letters 6(1): 51–65.
Kirillov A.A., Ruchin A.B., Fayzulin A.I., Chikhlyaev I.V. 2015. Helminths of reptiles from Mordovia: advance information. Proceedings of the Mordovia State Nature Reserve 14: 243–255. [In Russian]
Kirillova N.Yu., Kirillov A.A., Evlanov I.A. 2012. Variability of the dimensional structure in the hemipopulation of Thominx neopulchra (Nematoda: Capillariidae) females from bats of the genus Myotis (Chiroptera: Vespertilionidae). Parazitologiya 46(1): 11–15. [In Russian]
Machado-Silva J.R., Galvao C., Presgrave O.A.F., Rey L., Gomes D.C. 1994. Host-induced morphological changes of Schistosoma mansoni Sambon, 1907 male worms. Memorias do Instituto Oswaldo Cruz 89(3): 411–416.
Miller M.A., Pfeiffer W., Schwartz T. 2010. Creating the CIPRES Science Gateway for inference of large phylogenetic trees. Proceedings of the Gateway Computing Environments Workshop (GCE). New Orleans, LA: IEEE. P. 1–8. DOI: 10.1109/GCE.2010.5676129
Murvanidze L., Nikolaishvili K., Lomidze Ts. 2008. The annotated list of amphibians of Georgia. Proceedings of the Institute of Zoology 23: 43–49.
Nadler S.A., Lyons E.T., Pagan C., Hyman D., Lewis E.E., Beckman K., Bell C.M., Castinel A., DeLong R.L., Duignan P.J., Farinpour C., Huntington K.B., Kuiken T., Morgades D., Naem S., Norman R., Parker C., Ramos R., Spraker T.R., Beron-Vera B. 2013. Molecular systematics of pinniped hookworms (Nematoda: Uncinaria): species delimitation, host associations and host-induced morphometric variation. International Journal for Parasitology 43(14): 1119–1132. DOI: 10.1016/j.ijpara.2013.08.006
Novokhatskaya O. 2008. On the occurrence of the nematode Oswaldocruzia filiformis (Strongylida: Molineidae) in Karelia. Parazitologiya 42(3): 204–209. [In Russian]
Okulewicz A., Hildebrand J., Łysowski R., Buńkowska K., Perec-Matysiak A. 2014. Helminth Communities of Green and Brown Frogs from Poland (Lower Silesia Region). Journal of Herpetology 48(1): 34–37. DOI: 10.1670/12-108
Popiołek M., Witkowski A., Kotusz J., Kusznierz J., Baldy K. 2004. Intestinal parasites of brown trout (Salmo trutta fario L.) from streams of the Stołowe Mountains National Park. Parki Narodowe i Rezerwaty Przyrody 23: 121–127.
Popiołek M., Rozenblut-Kościsty B., Kot M., Nosal W., Ogielska M. 2011. Endoparasitic helminths of water frog complex in Poland: do differences exist between the parental species Pelophylax ridibundus and Pelophylax lessonae, and their natural hybrid Pelophylax esculentus? Helminthologia 48(2): 108–115. DOI: 10.2478/s11687-011-0020-8
Rambaut A., Drummond A.J., Xie D., Baele G., Suchard M.A. 2018. Posterior summarization in Bayesian phylogenetics using Tracer 1.7. Systematic Biology 67(5): 901–904. DOI: 10.1093/sysbio/syy032
Roytman V.A., Kazakov B.E. 1977. Some aspects of study of morphological variability in helminths (on example of trematodes of genus Azygia). Cestodes and Trematodes: Proceedings of Helminthological Laboratory of AS USSR 27: 110–128. [In Russian]
Ryzhikov K.M., Sharpilo V.P., Shevchenko N.N. 1980. Helminths of the amphibians of the fauna of the USSR. Moscow: Nauka. 276 p. [In Russian]
Sanchis V., Roing J., Carretero M., Roca V., Llorente G. 2000. Host-parasites relationships of Zootoca vivipara (Sauria: Lacertidae) in the Pyrenees (North Spain). Folia Parasitologica 47(2): 118–122. DOI: 10.14411/fp.2000.024
Schotthoefer A.M., Bolek M.G., Cole R.A., Beasley V.R. 2009. Parasites of the Mink Frog (Rana septentrionalis) from Minnesota, U.S.A. Comparative Parasitology 76(2): 240–246. DOI: 10.1654/4353.1
Sharpilo V.P. 1976. Parasitic worms of reptiles of the fauna of the USSR. Kiev: Naukova Dumka. 287 p. [In Russian]
Sinsch U., Heneberg P., Těšínsky M., Balczun C., Scheid P. 2019. Helminth endoparasites of the smooth newt Lissotriton vulgaris: linking morphological identification and molecular data. Journal of Helminthology 93(3): 332–341. DOI: 10.1017/S0022149X18000184
Skrjabin K.I., Shikhobalova N.P., Schulz R.S. 1954. Trichostrongylids of animals and man. Essentials of nematodology. Vol. 3. Moscow: AS USSR. 683 p. [In Russian]
Svitin R. 2015. New data on the morphology and distribution of Oswaldocruzia skrjabini (Nematoda, Molineidae). Vestnik Zoologii 49(5): 447–452.
Svitin R. 2016. Nematodes of the Genus Oswaldocruzia Travassos, 1917 of Western Palaearctic. PhD Thesis. Kyiv: Institute of Zoology. [In Ukrainian]
Svitin R. 2017. Two new species of Oswaldocruzia (Nematoda, Molineidae) parasitising lizards in Ukraine. Zootaxa 4263(2): 358–368. DOI: 10.11646/zootaxa.4263.2.9
Svitin R., Kuzmin Yu. 2012. Oswaldocruzia duboisi (Nematoda, Molineidae): morphology, hosts and distribution in Ukraine. Vestnik Zoologii 46(3): 195–203.
Svitin R., Gorobchishin V. 2015. Postcyclic parasitism of the nematodes of genus Osvaldoruzia Travassos, 1917 in some reptilian species on the territory of Ukraine. Proceedings of the Kiev National University 1(18): 68–70. [In Ukrainian]
Tarasovskaya N.E. 2009. Intraspecific interactions of the nematode Oswaldocruzia filiformis from Rana arvalis and their influence on the helminth body sizes. Gylym zhene bilim 3(16): 97–104.
Tarasovskaya N.E. 2011. The use of morphometric analysis in the study of interspecific interactions of lung helminths of Rana arvalis in Pavlodar region. KazNU Bulletin. Biological series 1(47): 81–85. [In Russian]
Tarasovskaya N.E., Pashkevich V.I. 2011. To the helminthofauna of Rana arvalis in Akmola region. Biological Sciences of Kazakhstan 1: 87–94. [In Russian]
Travassos L. 1937. Sur les espéces européennes du genre Oswaldocruzia. In: Proceedings on helminhology published in commemoration of the 30-year anniversary of K.I. Skrjabin and of 15^th anniversary of the All-Union Institute of Helminthology 1: 725–733.
Viney M., Diaz A. 2012. Phenotypic plasticity in nematodes. Evolutionary and ecological significance. Worm 1(2): 98–106. DOI: 10.4161/worm.21086
Zhao L., Liu G.-H., Zhao G.-H., Cai J.-Z., Zhu X.-Q., Qian A.-D. 2013. Genetic differences between Chabertia ovina and C. erschowi revealed by sequence analysis of four mitochondrial genes. Mitochondrial DNA 26(2): 167–170. DOI: 10.3109/19401736.2013.843089