Genetic diversity and population structure of the helminths intermediate hostAnentome helena(von dem Busch, 1847) in northern Thailand

Abstract

Objective: To investigate the genetic diversity and population structure of the helminth intermediate host, Anentome (A.) helena, populations from northern Thailand and reveal the effect of parasitic infection on snail host populations.

Methods: A. helena was collected throughout river drainages from northern Thailand and investigated for larval trematode infection. Seventy-three A. helena specimens were used to examine and re-construct the molecular phylogenetics (neighbour-joining, maximum likelihood, and Bayesian inference methods), genetic diversity, and population structure based on the concatenated datasets of COI and 16S rRNA (1 178 bp).

Results: The infection of zoogonid cercariae, heterophyid metacercariae, and echinostome metacercariae in the snail was found from 2 localities, including Mae Sa and Mae Pu Kha populations, with a total prevalence of 6.7% and 16.3%, respectively. The phylogenetic tree and haplotype network revealed three main clades and three haplogroups for each region (COI and 16S rRNA). High levels of genetic diversity in terms of haplotype diversity were found in parasitic infected populations.

Conclusions: This is the first time that Echinostoma mekongi has been discovered from A. helena in northern Thailand. The results also showed that parasitic infection can affect the population genetic structure of the snail intermediate host, A. helena. Moreover, habitat and historical demographic changes could be other important factors that can affect the degree of population genetic structure in A. helena. Understanding the population structure of intermediate host species can inform parasite transmission dynamics and is useful for the creation of effective public health management strategies.

Keywords

Cercariae / Metacercariae / Phylogeny / Parasite / Host / Freshwater snail

References

[1] Altaf J, Qureshi NA, Javed M, Siddiqui I. Genetic diversity analysis of snail species (Mollusca: Gastropoda) using molecular marker technology. J Biodivers Environ Sci 2017; 10(1): 26-35.

[2] Wiroonpan P, Chontananarth T, Chai JY, Purivirojkul W. High diversity of trematode metacercariae that parasitize freshwater gastropods in Bangkok, Thailand, and their infective situations, morphologies and phylogenetic relationships. Parasitology 2022; 149(7): 913-933.

[3] Rabone M, Wiethase JH, Allan F, Gouvras AN, Pennance T, Hamidou AA, et al. Freshwater snails of biomedical importance in the Niger River Valley: Evidence of temporal and spatial patterns in abundance, distribution and infection with Schistosoma spp. Parasit Vectors 2019; 12(1): 498. doi: 10.1186/s13071-019-3745-8.

[4] Chantima K, Rika C. Snail-borne zoonotic trematodes in edible viviparid snails obtained from wet markets in Northern Thailand. J Helminthol 2020; 94(e198): 1-6.

[5] Chomchoei N, Backeljau T, Segers B, Wongsawad C, Butboonchoo P, Nantarat N. Morphological and molecular characterization of larval trematodes infecting the assassin snail genus Anentome in Thailand. J Helminthol 2022; 96(e52): 1-11.

[6] Gordy MA, Kish L, Tarrabain M, Hanington PC. A comprehensive survey of larval digenean trematodes and their snail hosts in central Alberta, Canada. Parasitol Res 2016; 115(10): 3867-3880.

[7] Abe EM, Guan W, Guo YH, Kassegne K, Qin ZQ, Xu J, et al. Differentiating snail intermediate hosts of Schistosoma spp. using molecular approaches: Fundamental to successful integrated control mechanism in Africa. Infect Dis Poverty 2018; 7(29): 1-13.

[8] Standley CJ, Goodacre SL, Wade CM, Stothard JR. The population genetic structure of Biomphalaria choanomphala in Lake Victoria, East Africa: Implications for schistosomiasis transmission. Parasit Vectors 2014; 7(524): 1-10.

[9] Caldeira RL, Vidigal THDA, Paulinelli ST, Simpson AJG, Carvalho OS. Molecular identification of similar species of the genus Biomphalaria (Mollusca: Planorbidae) determined by a polymerase chain reaction-restriction fragment length polymorphism. Mem Inst Oswaldo Cruz 1998; 93(1994): 219-225.

[10] Webster JP, Davies CM, Hoffman JI, Ndamba J, Noble LR, Woolhouse MEJ. Population genetics of the schistosome intermediate host Biomphalaria pfeifferi in the Zimbabwean highveld: Implications for co-evolutionary theory. Ann Trop Med Parasitol 2001; 95(2): 203-214.

[11] Webster JP, Woolhouse MEJ. Cost of resistance: Relationship between reduced fertility and increased resistance in a snail schistosome host-parasite system. Proc R Soc B Biol Sci 1999; 266(1417): 391-396.

[12] Brandt RAM. The non-marine aquatic Mollusca of Thailand. Arch für Molluskenkd 1974; 105: 1-423.

[13] Chantima K, Chai JY, Wongsawad C. Echinostoma revolutum: Freshwater snails as the second intermediate hosts in Chiang Mai, Thailand. Korean J Parasitol 2013; 51(2): 183-189.

[14] Chomchoei N, Wongsawad C, Nantarat N. Investigation of cryptic diversity and occurrence of echinostome metacercariae infection in Anentome helena (von dem Busch, 1847). Asian Pac J Trop Med 2018; 11(10): 590-596.

[15] Mkize LS, Mukaratirwa S, Zishiri OT. Population genetic structure of the freshwater snail, Bulinus globosus, (Gastropoda: Planorbidae) from selected habitats of KwaZulu-Natal, South Africa. Acta Trop 2016; 161: 91-99.

[16] Lounnas M, Vázquez AA, Alda P, Sartori K, Pointier JP, David P, et al. Isolation, characterization and population-genetic analysis of microsatellite loci in the freshwater snail Galba cubensis (Lymnaeidae). J Molluscan Stud 2017; 83(1): 63-68.

[17] Galindo LA, Puillandre N, Utge J. The phylogeny and systematics of the Nassariidae revisited (Gastropoda, Buccinoidea). Mol Phylogenet Evol 2016; 99: 337-353.

[18] Strong EE, Galindo LA, Kantor YI. Quid est Clea helena? Evidence for a previously unrecognized radiation of assassin snails (Gastropoda: Buccinoidea: Nassariidae). PeerJ 2017; 2017(8): 1-41.

[19] Sanches Palasio RG, Zanotti-Magalhães EM, Tuan R. Genetic diversity of the freshwater snail Biomphalaria tenagophila (d’Orbigny, 1835) (Gastropoda: Hygrophila: Planorbidae) across two coastal areas of southeast Brazil. Folia Malacol 2018; 26(4): 221-229.

[20] Tantrawatpan C, Saijuntha W, Bunchom N, Suksavate W, Pilap W, Walalite T, et al. Genetic structure and geographical variation of Bithynia siamensis goniomphalos sensu lato (Gastropoda: Bithyniidae), the snail intermediate host of Opisthorchis viverrini sensu lato (Digenea: Opisthorchiidae) in the Lower Mekong Basin revealed by mitochondrial DNA sequences. Int J Parasitol 2020; 50(1): 55-62.

[21] Bunchom N, Saijuntha W, Pilap W, Suksavate W, Vaisusuk K, Suganuma N, et al. Genetic variation of a freshwater snail Hydrobioides nassa (Gastropoda: Bithyniidae) in Thailand examined by mitochondrial DNA sequences. Hydrobiologia 2019;2. doi: 10.1007/s10750-019-04013-2.

[22] Olivier L, Schneiderman M. A method for estimating the density of aquatic snail populations. Exp Parasitol 1956; 5(2): 109-117.

[23] United States Agency for International Development. Resources and environmental problems. Draft Environmental Report on Thailand 1979. [Online]. Available from: https://pdf.usaid.gov/pdf_docs/PNAAG972.pdf. [Accessed on 4 July 2023].

[24] Caron Y, Rondelaud D, Losson B. The detection and quantification of a digenean infection in the snail host with special emphasis on Fasciola sp. Parasitol Res 2008; 103(4): 735-744.

[25] Frandsen F, Christensen NO. An introductory guide to the identification of cercariae from African freshwater snails with special reference to cercariae of trematode species of medical and veterinary importance. Acta Trop 1984; 41(2): 181-202.

[26] Kanev I, Fried B, Radev V. Collar spine models in the genus Echinostoma (Trematoda: Echinostomatidae). Parasitol Res 2009; 105(4): 921-927.

[27] Schell SC. How to know the trematodes. Iowa: W.C. Brown Company; 1970.

[28] Bowles J, Blair D, McManus DP. Genetic variants within the genus Echinococcus identified by mitochondrial DNA sequencing. Mol Biochem Parasitol 1992; 54(2): 165-173.

[29] Johnson M, Zaretskaya I, Raytselis Y, Merezhuk Y, McGinnis S, Madden TL. NCBI BLAST: A better web interface. Nucleic Acids Res 2008; 36(Web Server issue): W5-W9.

[30] Kessing B, Croom H, Martin A, McIntosh C, McMillan W, Palumbi S. The simple fool’s guide to PCR. Hawaii, Honolulu: Department of Zoology, University of Hawaii; 1989.

[31] Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R. DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Mol Mar Biol Biotechnol 1994; 3(5): 294-299.

[32] Thompson JD, Higgins DG, Gibson TJ. CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 1994; 22(22): 4673-4680.

[33] Kumar S, Stecher G, Tamura K. MEGA7: Molecular evolutionary genetics analysis version 7. 0 for bigger datasets. Mol Biol Evol 2016; 33(7): 1870-1874.

[34] Saitou N, Nei M. The neighbor-joining method: A new method for reconstructing phylogenetic trees. Mol Biol Evol 1987; 4(4): 406-425.

[35] Sullivan J. Maximum-likelihood methods for phylogeny estimation. Mol Evol Prod Biochem Data 2005; 395: 757-779.

[36] Guindon S, Delsuc F, Dufayard JF, Gascuel O. Estimating maximum likelihood phylogenies with PhyML. Methods Mol Biol 2009; 537: 113-137.

[37] Ronquist F, Teslenko M, van der Mark P, Ayres D, Darling A, Höhna S, et al. MrBayes 3. 2: Efficient Bayesian phylogenetic inference and model choice across a large model space. Syst Biol 2012; 61(3): 539-542.

[38] Rambaut A. FigTree v1. 4.3 2010. [Online]. Available from: http://tree.bio.ed.ac. [Accessed on 29 July 2023].

[39] Leigh JW, Bryant D. POPART: Full-feature software for haplotype network construction. Methods Ecol Evol 2015; 6(9): 1110-1116.

[40] Rozas J. morePhyML: Improving the phylogenetic tree space exploration with PhyML 3. In: Posada D, editor. Methods in molecular biology. Totowa, NJ: Humana Press; 2009, p. 337-350.

[41] IBM Corp. IBM SPSS statistics for windows, version 20. 0 2013. [Online]. Available from: https://www.ibm.com/support/pages/downloading-ibm-spss-statistics-20. [Accessed on 29 September 2023].

[42] Excoffier L, Lischer HEL. Arlequin suite ver 3. 5: A new series of programs to perform population genetics analyses under Linux and Windows. Mol Ecol Resour 2010; 10: 564-567.

[43] Chantima K, Suk-Ueng K, Kampan M. Freshwater snail diversity in Mae Lao agricultural basin (Chiang Rai, Thailand) with a focus on larval trematode infections. Korean J Parasitol 2018; 56(3): 247-257.

[44] Yutemsuk N, Krailas D, Anancharoenkit C, Phanpeng L, Dechruksa W. Trematode infections of freshwater snails genus Clea A. Adams, 1855 in the reservoir of lower northeast Thailand. Jt Int Trop Med Meet 2017; 6: 7-16.

[45] Thaenkham U, Phuphisut O, Nuamtanong S, Yoonuan T, Sa-Nguankiat S, Vonghachack Y, et al. Genetic differences among Haplorchis taichui populations in Indochina revealed by mitochondrial COX1 sequences. J Helminthol 2017; 91(5): 597-604.

[46] Butboonchoo P, Wongsawad C, Wongsawad P, Chai JY. Morphology and molecular identification of Echinostoma revolutum and Echinostoma macrorchis in freshwater snails and experimental hamsters in upper northern Thailand. Korean J Parasitol 2020; 58(5): 499-511.

[47] Ewers WH. The influence of the density of snails on the incidence of larval trematodes. Parasitology 1964; 54(3): 579-583.

[48] Mereta ST, Abaya SW, Tulu FD, Takele K, Ahmednur M, Melka GA, et al. Effects of land-use and environmental factors on snail distribution and trematode infection in Ethiopia. Trop Med Infect Dis 2023; 8(3): 1-13.

[49] Chai JY. Human intestinal flukes: From discovery to treatment and control. Dordrecht: Springer; 2019, p. 1-549.

[50] Wardle WJ. A new zoogonid cercaria (Trematoda: Digenea) from the Florida horse conch, Pleuroploca gigantea in the Northwestern Gulf of Mexico. J Helminthol Soc Washingt 1993; 60(2): 216-218.

[51] Barnett LJ, Miller TL, Cribb TH. A review of the currently recognised zoogonid cercariae, including the identification and emergence ecology of Cercaria capricornia XI (Digenea: Zoogonidae) from Nassarius olivaceus (Gastropoda: Nassariidae) in Central Queensland, Australia. Folia Parasitol (Praha) 2014; 61(4): 322-330.

[52] Gilardoni C, Etchegoin J, Cribb T, Pina S, Rodrigues P, Diez ME, et al. Cryptic speciation of the zoogonid digenean Diphterostomum flavum n. sp. demonstrated by morphological and molecular data. Parasite 2020; 27(44): 1-11.

[53] Barnett LJ, Miller TL. Host specificity and ecology of digenean parasites of nassariid gastropods in Central Queensland, Australia, with comments on host-parasite associations of the Nassariidae. Yale J Biol Med 2018; 91(4): 361-374.

[54] Cho J, Jung BK, Chang T, Sohn WM, Sinuon M, Chai JY. Echinostoma mekongi n. sp. (Digenea: Echinostomatidae) from riparian people along the Mekong River in Cambodia. Korean J Parasitol 2020; 58(4): 431-443.

[55] Pantoja C, Faltýnková A, O’Dwyer K, Jouet D, Skírnisson K, Kudlai O. Diversity of echinostomes (Digenea: Echinostomatidae) in their snail hosts at high latitudes. Parasite 2021; 28(59): 1-31.

[56] Zimmermann MR, Luth KE, Esch GW. Microhabitat differences surrounding a pond affects the distribution of trematode parasites among a pulmonate snail community. Helminthologia 2014; 51(4): 301-308.

[57] Wood S, Ziegler A. Floodplain sediment from a 100-year-recurrence flood in 2005 of the Ping River in northern Thailand. Hydrol Earth Syst Sci 2008; 12: 959-973.

[58] Bogan A, Hanneman EH. A carnivorous aquatic gastropod in the pet trade in North America: The next threat to freshwater gastropods. Ellipsaria 2013; 15(2): 18-19.

[59] Ng TH, Junn Kitt F, Tan SK, Chan MKK, Yeo DCJ. First non-native establishment of the carnivorous assassin snail, Anentome helena (von dem Busch in Philippi, 1847). BioInvasions Rec 2016; 5: 143-148.

[60] Mendez-Harclerode FM, Strauss RE, Fulhorst CF, Milazzo ML, Ruthven III DC, Bradley RD. Molecular evidence for high levels of intrapopulation genetic diversity in woodrats (Neotoma micropus). J Mammal 2007; 88(2): 360-370.

[61] Feng Y, Li Q, Kong L, Zheng X. DNA barcoding and phylogenetic analysis of Pectinidae (Mollusca: Bivalvia) based on mitochondrial COI and 16S rRNA genes. Mol Biol Rep 2011; 38(1): 291-299.

[62] Webster JP, Shrivastava J, Johnson PJ, Blair L. Is host-schistosome coevolution going anywhere? BMC Evol Biol 2007; 7: 1-11.

[63] Webster JP, Davies CM, Ndamba J, Noble LR, Jones CS, Woolhouse MEJ. Spatio-temporal genetic variability in the schistosome intermediate host Biomphalaria pfeifferi. Ann Trop Med Parasitol 2001; 95(5): 515-527.

[64] Viard F, Bremond P, Labbo R, Delay B, Jarne P. Microsatellites and the genetics of highly selfing populations in the freshwater snail Bulinus truncatus. Genetics 1996; 142(4): 1237-1247.

[65] Viard F, Justy F, Jarne P. Population dynamics inferred from temporal variation at microsatellite loci in the selfing snail Bulinus truncatus. Genetics 1997; 146(3): 973-982.

[66] Zein-Eddine R, Djuikwo-Teukeng FF, Dar Y, Dreyfuss G, Van den Broeck F. Population genetics of the Schistosoma snail host Bulinus truncatus in Egypt. Acta Trop 2017; 172: 36-43.

[67] Djuikwo-Teukeng FF, Da Silva A, Njiokou F, Kamgang B, Ekobo AS, Dreyfuss G. Significant population genetic structure of the Cameroonian fresh water snail, Bulinus globosus, (Gastropoda: Planorbidae) revealed by nuclear microsatellite loci analysis. Acta Trop 2014; 137: 111-117.