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Licensed Unlicensed Requires Authentication Published by De Gruyter July 4, 2018

First molecular identification of an agent of diplostomiasis, Diplostomum pseudospathaceum (Niewiadomska 1984) in the United Kingdom and its genetic relationship with populations in Europe

Egie Elisha Enabulele, Agnes Ogheneruemu Awharitoma, Scott P. Lawton and Ruth S. Kirk
From the journal Acta Parasitologica

Abstract

Trematode genus Diplostomum comprises of parasitic species which cause diplostomiasis, the ‘white eye’ disease in fish and heavy infection can result in mortality. The increasing availability of DNA sequences of accurately identified Diplostomum species on public data base presently enables the rapid identification of species from novel sequences. We report the first molecular evidence of the occurrence of D. pseudospathaceum in the United Kingdom. Two gene regions, nuclear internal transcribed spacer cluster (ITS1-5.8S-ITS2) and mitochondrial cytochrome c oxidase subunit 1 (cox1) of cercariae from infected aquatic snails, Lymnaea stagnalis collected in several locations in Southern England were sequenced. Phylogenetic analysis based on both sequenced genes revealed that the novel sequences were D. pseudospathaceum. Molecular diversity analysis of published D. pseudospathaceum cox1 sequences from seven countries in Europe and the novel sequences from the present study revealed high diversity, but low nucleotide divergence and a lack of gene differentiation between the populations. Haplotype network analysis depicted a star-like pattern and revealed a lack of geographic structure in the population. Fixation indices confirmed gene flow between populations and we suspect high levels of dispersal facilitated by highly mobile second intermediate (fish) and definitive (piscivorous birds) host may be driving gene flow between populations. Neutrality tests and mismatch distribution indicated recent population growth/expansion for D. pseudospathaceum in Europe.

Acknowledgements

The co-operation of the Four Seasons Hotel Hampshire, Godalming Angling Club and Nature England are appreciated for allowing snail sampling in the sites visited. This research was made possible by funding from the Tertiary Education Trust Fund of Nigeria for PhD studentship to the corresponding author at Kingston University London.

References

Avise J.C. 2000. Phylogeography: The history and Formation of Species. Harvard University Press, Cambridge, MA, pp. 44710.2307/j.ctv1nzfgj7Search in Google Scholar

Behrmann-Godel J. 2013. Parasite identification, succession and infection pathways in perch fry (Perca fluviatilis): new insights through a combined morphological and genetic approach. Parasitology, 140, 509–520. 10.1017/S0031182012001989Search in Google Scholar PubMed

Blair D. 1977. A key to cercariae of British strigeoids (Digenea) for which the life-cycles are known, and notes on the characters used. Journal of Helminthology, 51, 155–166. org/10.1017/S0022149X0000743Search in Google Scholar

Brabec J., Kostadinova A., Scholz T., Littlewood D.T. 2015. Complete mitochondrial genomes and nuclear ribosomal RNA operons of two species of Diplostomum (Platyhelminthes: Trematoda): a molecular resource for taxonomy and molecular epidemiology of important fish pathogens. Parasite and Vectors, 8, 336. 10.1186/s13071-015-0949-4Search in Google Scholar PubMed PubMed Central

Chappell L.H. 1995. The biology of diplostomatid eye flukes of fishes. Journal of Helminthology, 69, 97–101. org/10.1017/S0022149X00013961Search in Google Scholar

Clement M., Posada D., Crandall K. 2000. TCS: A computer program to estimate gene genealogies. Molecular Ecology, 9, 1657–1660. 10.1046/j.1365-294x.2000.01020.xSearch in Google Scholar PubMed

Erasmus D.A. 1958. Studies on the morphology, biology, and development of a strigeid cercariae (cercaria X Baylis). Parasitology, 48, 312–335. org/10.1017/S0031182000021284Search in Google Scholar

Fu Y.X. 1997. Statistical tests of neutrality of mutations against population growth, hitchhiking and background selection. Genetics, 147, 915–92510.1093/genetics/147.2.915Search in Google Scholar PubMed PubMed Central

Georgieva S., Soldánová M., Pérez-Del-Olmo A., Dangel D.R., Sitko J., Sures B., et al. 2013. Molecular prospecting for European Diplostomum (Digenea: Diplostomidae) reveals cryptic diversity. International Journal of Parasitology, 43, 57–72. 10.1016/j.ijpara.2012.10.019Search in Google Scholar PubMed

Haarder S., Jørgensen K., Kania P.W., Skovgaard A., Buchmann K. 2013. Occurrence of Diplostomum pseudospathaceum Niewiadomska, 1984 and D. mergi Dubois, 1932 (Digenea: Diplostomidae) in Danish freshwater snails: ecological and molecular data. Folia Parasitology, 60, 177–180. 10.14411/fp.2013.020Search in Google Scholar PubMed

Hall T.A. 1999. BioEdit: a user friendly biological sequence alignment program editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series, 41, 95–98Search in Google Scholar

Harpending H.C. 1994. Signature of ancient population growth in a low-resolution mitochondrial DNA mismatch distribution. Human Biology, 66, 591–600Search in Google Scholar

Harrison R.G. 1989. Animal mitochondrial DNA as a genetic marker in population and evolutionary biology. Trends in Ecology and Evolution, 4, 6–11. 10.1016/0169-5347(89)90006-2Search in Google Scholar

Hudson R.R., Slatkin M., Maddison W.P. 1992. Estimation of levels of gene flow from DNA sequence data. Genetics 132 (2), 583 – 58910.1093/genetics/132.2.583Search in Google Scholar

Horák P., Kolářová L., Mikeš L. 2014. Schistosomatoidea and Diplostomoidea. Advances in Experimental Medicine and Biology, 766, 331–364. 10.1007/978-1-4939-0915-5_10Search in Google Scholar

Kudlai O., Oros M., Kostadinova A., Georgieva S. 2017. Exploring the diversity of Diplostomum (Digenea: Diplostomidae) in fishes from the River Danube using mitochondrial DNA barcodes. Parasite and Vectors, 10, 592. 10.1186/s13071-017-2518-5Search in Google Scholar

Librado P., Rozas J. 2009. DnaSP v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics, 25(11), 1451–1452. 10.1093/bioinformatics/btp187Search in Google Scholar

Leigh J.W., Bryant D. 2015. POPART: full-feature software for haplotype network construction. Methods in Ecology and Evolution, 6, 1110–1116. 10.1111/2041-210X.12410Search in Google Scholar

Locke S.A., Al-Nasiri F.S., Caffara M., Drago F., Kalbe M., Lapierre A.R., et al. 2015. Diversity, specificity and speciation in larval Diplostomidae (Platyhelminthes: Digenea) in the eyes of freshwater fish, as revealed by DNA barcodes. International Journal of Parasitology, 45, 841–55. 10.1016/j.ij-para.2015.07.001Search in Google Scholar

Louhi K.R., Karvonen A., Rellstab C., Jokela J. 2010. Is the population genetic structure of complex life cycle parasites determined by the geographic range of the most motile host? Infections Genetics and Evolution, 10, 1271–1277. 10.1016/j.meegid.2010.08.013Search in Google Scholar

Mckeown C.A., Irwin S.W. 1995. The life cycle stages of three Diplostomum species maintained in the laboratory. International Journal of Parasitology, 25, 897–906. org/10.1016/0020-7519(94)00217-CSearch in Google Scholar

Moritz C., Dowling T.E., Brown W.M. 1987. Evolution of animal mitochondrial DNA: relevance for population biology and systematics. Annual Review of Ecology and Systematics, 18, 269–292. 10.1146/annurev.es.18.110187.001413Search in Google Scholar

Morley N.J., Lewis J.W. 2007. Anthtropogenic pressure on a molluscan-trematode community over a long-term period in the Basingstoke Canal, UK, and its implications for ecosystem health. EcoHealth, 3, 269–28010.1007/s10393-006-0058-0Search in Google Scholar

Moszczynska A., Locke S.A., Mclaughlin J.D., Marcogliese D.J., Crease T.J. 2009. Development of primers for the mitochondrial cytochrome c oxidase I gene in digenetic trematodes (Platyhelminthes) illustrates the challenge of barcoding parasitic helminths. Molecular Ecology Resources, 9, 75–82. 10.1111/j.1755-0998.2009.02634.xSearch in Google Scholar

Nei M. 1973. Analysis of gene diversity in subdivided populations. Proceedings of the National Academy of Sciences, 70, 3321–332310.1073/pnas.70.12.3321Search in Google Scholar

Niewiadomska K. 1984. Present status of Diplostomum spathaceum (Rudolphi, 1819) and differentiation of Diplostomum pseudospathaceum nom. nov. (Trematoda: Diplostomatidae). Systematic Parasitology, 6, 81–86. org/10.1007/BF02185515Search in Google Scholar

Niewiadomska K. 1986. Verification of the life-cycles of Diplostomum spathaceum (Rudolphi, 1819) and D. pseudospathaceum Niewiadomska, 1984 (Trematoda, Diplostomidae). Systematic Parasitology, 8, 23–31. org/10.1007/BF00010306Search in Google Scholar

Niewiadomska K. 1996. The genus Diplostomum – taxonomy, morphology and biology. Acta Parasitologica, 41, 55–66Search in Google Scholar

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 of Parasitology, 33, 733–755. org/10.1016/S0020-7519(03)00049-3Search in Google Scholar

Pérez-Del-Olmo A., Georgieva S., Pula H.J., Kostadinova A. 2014. Molecular and morphological evidence for three species of Diplostomum (Digenea: Diplostomidae), parasites of fishes and fish-eating birds in Spain. Parasite and Vectors, 7, 502. 10.1186/s13071-014-0502-xSearch in Google Scholar

Posada D., Crandall K.A. 2001. Intraspecific gene genealogies: trees grafting into networks. Trends in Ecology and Evolution, 16, 37–45. org/10.1016/S0169-5347(00)02026-7Search in Google Scholar

RamõÂrez-Soriano A., Ramos-Onsins S.E., Rozas J., Calafell F., Navarro A. 2008. Statistical power analysis of neutrality tests under demographic expansions, contractions and bottlenecks with recombination. Genetics, 179, 555–567. 10.1534/genetics.107.083006Search in Google Scholar PubMed PubMed Central

Ramos-Onsins S.E., Rozas J. 2002. Statistical properties of new neutrality test against population growth. Molecular Biology and Evolution, 19, 2092–2100. org/10.1093/oxfordjournals.molbev.a004034Search in Google Scholar

Rees G. 1955. The adult and Diplostomulum stage (Diplostomulum phoxini (Faust)) of Diplostomum pelmatoides Dubois and an experimental demonstration of part of the life cycle. Parasitology, 45, 295–312. org/10.1017/S0031182000027670Search in Google Scholar

Rogers A.R., Harpending H. 1992. Population growth makes waves in the distribution of pairwise genetic differences. Molecular Biology and Evolution, 9, 552–569. org/10.1093/oxfordjournals.molbev.a040727Search in Google Scholar

Selbach C., Soldánová M., Georgieva S., Kostadinova A., Sures B. 2015. Integrative taxonomic approach to the cryptic diversity of Diplostomum spp. in lymnaeid snails from Europe with a focus on the ‘Diplostomum mergi’ species complex. Parasite and Vectors, 8, 300. 10.1186/s13071-015-0904-4Search in Google Scholar PubMed PubMed Central

Slatkin M., Hudson R. R. 1991. Pairwise comparison of mitochondrial DNA sequences in stable and exponentially growing populations. Genetics, 129, 555–56210.1093/genetics/129.2.555Search in Google Scholar PubMed PubMed Central

Tajima F. 1993. Statistical analysis of DNA polymorphism. The Japanese Journal of Genetics, 68, 567–595. org/10.1266/jjg.68.567Search in Google Scholar

Tamura K., Stecher G., Peterson D., Filipski A., Kumar S. 2013. MEGA6: molecular evolutionary genetics analysis version 6.0. Molecular Biology and Evolution, 30, 2725–2729. 10.1093/molbev/mst197Search in Google Scholar PubMed PubMed Central

Templeton A.R., Crandall K. A., Sing C.F. 1992. A cladistic analysis of phenotypic associations with haplotypes inferred from restriction endonuclease mapping and DNA sequences data.III. Cladogram estimation. Genetics, 134, 659–66910.1093/genetics/134.2.659Search in Google Scholar PubMed PubMed Central

Wang C.R., Li L., Ni H.B., Zhai Y.Q., Chen A.H., Chen J., et al. 2009. Orientobilharzia turkestanicum is a member of Schistosoma genus based on phylogenetic analysis using ribosomal DNA sequences. Experimental Parasitology, 121, 193–197. 10.1016/j.exppara.2008.10.012Search in Google Scholar PubMed

Williams M.O. 1966. On some larval trematodes from Lymnaea peregra (Muller) in Scotland. Journal of Helminthology, 40, 245–252. org/10.1017/S0022149X00034234Search in Google Scholar

Received: 2017-10-30
Revised: 2018-02-20
Accepted: 2018-03-02
Published Online: 2018-07-04
Published in Print: 2018-09-25

© 2018 W. Stefański Institute of Parasitology, PAS