Development and application of an ethically and epidemiologically advantageous assay for the multi-locus microsatellite analysis of Schistosoma mansoni | Parasitology | Cambridge Core (original) (raw)

Abstract

Non-availability of adult worms from living hosts remains a key problem in population genetic studies of schistosomes. Indirect sampling involving passage through laboratory animals presents significant ethical and practical drawbacks, and may result in sampling biases such as bottlenecking processes and/or host-induced selection pressures. The novel techniques reported here for sampling, storage and multi-locus microsatellite analysis of larval Schistosoma mansoni, allowing genotyping of up to 7 microsatellite loci from a single larva, circumvent these problems. The utility of these assays and the potential problems of laboratory passage, were evaluated using 7 S. mansoni population isolates collected from school-children in the Hoima district of Uganda, by comparing the associated field-collected miracidia with adult worms and miracidia obtained from a single generation in laboratory mice. Analyses of laboratory-passaged material erroneously indicated the presence of geographical structuring in the population, emphasizing the dangers of indirect sampling for population genetic studies. Bottlenecking and/or other sampling effects were demonstrated by reduced variability of adult worms compared to their parent field-collected larval samples. Patterns of heterozygote deficiency were apparent in the field-collected samples, which were not evident in laboratory-derived samples, potentially indicative of heterozygote advantage in establishment within laboratory hosts. Genetic distance between life-cycle stages in the majority of isolates revealed that adult worms and laboratory-passaged miracidia clustered together whilst segregating from field miracidia, thereby further highlighting the utility of this assay.

References

Blair, L.,Webster, J. P. andBarker, G. C. (2001).Isolation and characterisation of polymorphic microsatellite markers in Schistosoma mansoni from Africa.Molecular Ecology Notes 1,93–95.CrossRefGoogle Scholar

Brouwer, K. C.,Ndhlovu, P.,Munatsi, A. andShiff, C. J. (2001).Genetic diversity of a population of Schistosoma haematobium derived from schoolchildren in east central Zimbabwe.Journal of Parasitology 87,762–769.CrossRefGoogle Scholar

Brouwer, K. C.,Ndhlovu, P. D.,Wagatsuma, Y.,Munatsi, A. andShiff, C. J. (2003).Urinary tract pathology attributed to Schistosoma haematobium: does parasite genetics play a role?American Journal of Tropical Medicine and Hygiene 68,456–462.Google Scholar

Cavalli-Sforza, L. L. andEdwards, A. W. F. (1967).Phylogenetic analysis: models and estimation procedures.American Journal of Human Genetics 19,233–257.Google Scholar

Chitsulo, L.,Engels, D.,Montressor, A. andSavioli, L. (2000).The global status of schistosomiasis and its control.Acta Tropica 77,41–51.CrossRefGoogle Scholar

Cohen, L. M. andEveland, L. K. (1988).Schistosoma mansoni: characterisation of clones maintained by the microsurgical transplantation of sporocysts.Journal of Parasitology 74,963–969.CrossRefGoogle Scholar

Curtis, J. andMinchella, D. J. (2000).Schistosome population genetic structure: when clumping worms is not just splitting hairs.Parasitology Today 16,68–71.CrossRefGoogle Scholar

Curtis, J.,Sorensen, R. E. andMinchella, D. J. (2002).Schistosome genetic diversity: the implications of population structure as detected with microsatellite markers.Parasitology 125,S51–S59.CrossRefGoogle Scholar

Curtis, J.,Sorensen, R. E.,Page, L. K. andMinchella, D. J. (2001).Microsatellite loci in the human blood fluke Schistosoma mansoni and their utility for other schistosome species.Molecular Ecology Notes 1,143–145.Google Scholar

Davies, C. M.,Webster, J. P.,Kruger, O.,Munatsi, A.,Ndamba, J. andWoolhouse, M. E. J. (1999).Host-parasite population genetics: a cross-sectional comparison of Bulinus globosus and Schistosoma haematobium.Parasitology 119,295–302.CrossRefGoogle Scholar

Davies, C. M.,Webster, J. P. andWoolhouse, M. E. (2001).Trade-offs in the evolution of virulence in an indirectly transmitted macroparasite.Proceedings of the Royal Society of London, B 268,251–257.CrossRefGoogle Scholar

Durand, P.,Sire, C. andTheron, A. (2000).Isolation of microsatellite markers in the digenetic trematode Schistosoma mansoni from Guadeloupe island.Molecular Ecology 9,997–998.CrossRefGoogle Scholar

Guo, S. andThompson, E. (1992).Performing the exact test of Hardy-Weinberg proportion for multiple alleles.Biometrics 48,361–372.CrossRefGoogle Scholar

Kumar, S.,Tamura, K. andNei, M. (2004).MEGA3: integrated software for Molecular Evolutionary Genetic Analysis and sequence alignment.Briefings in Bioinformatics 5,150–163.CrossRefGoogle Scholar

Langella, O. (1999). Populations 1.2.28 (12/5/2002), CNRS UPR9034.

Loverde, P. T.,Dewald, J.,Minchella, D. J.,Bosshardt, S. C. andDamian, R. T. (1985).Evidence for host-induced selection in Schistosoma mansoni.Journal of Parasitology 71,297–301.CrossRefGoogle Scholar

Manning, S. D.,Woolhouse, M. E. J. andNdamba, J. (1995).Geographic compatibility of the freshwater snail Bulinus globusus and schistosomes from the Zimbabwe Highveld.International Journal for Parasitology 25,37–42.CrossRefGoogle Scholar

Morand, S.,Manning, S. D. andWoolhouse, M. E. J. (1996).Parasite-host coevolution and geographic patterns of parasite infectivity and host susceptibility.Proceedings of the Royal Society of London, B 263,119–128.CrossRefGoogle Scholar

Nelson, G. S. andSaoud, M. F. A. (1968).A comparison of the pathogenicity of two geographical strains of Schistosoma mansoni in rhesus monkeys.Journal of Helminthology 17,339–362.CrossRefGoogle Scholar

Prugnolle, F.,De Meeus, T.,Durand, P.,Sire, C. andTheron, A. (2002).Sex-specific genetic structure in Schistosoma mansoni: evolutionary and epidemiological implications.Molecular Ecology 11,1231–1238.CrossRefGoogle Scholar

Prugnolle, F.,Choisy, M.,Theron, A.,Durand, P. andDe Meeus, T. (2004).Sex-specific correlation between heterozygosity and clone size in the trematode Schistosoma mansoni.Molecular Ecology 13,2859–2864.CrossRefGoogle Scholar

Rodrigues, N. B.,Filho, P. C.,Souza, C. P.,Passos, L. K. J.,Dias-Neto, E. andRomanha, A. J. (2002).Population structure of Schistosoma mansoni assessed by DNA microsatelites.International Journal for Parasitology 32,843–851.CrossRefGoogle Scholar

Rollinson, D. andJohnston, D. A. (1996).Schistosomiasis: a persistent parasitic disease.Interdisciplinary Science Reviews 21,140–154.CrossRefGoogle Scholar

Rosenberg N. A.,Pritchard, J. K.,Weber, J. L.,Cann, H. M.,Kidd, K. K.,Zhivotovsky, L. A. andFeldman, M. W. (2002).Genetic structure of human populations.Science 298,2381–2383.CrossRefGoogle Scholar

Schneider, S.,Roesli, D. andExcoffier, L. (2000).Arlequin ver 2.000: A Software for Population Genetic Data Analysis. Genetics and Biometry Laboratory, University of Geneva,Switzerland.

Shrivastava, J.,Barker, G. C.,Johansen, M. V.,Xiaonong, Z.,Aligui, G. D.,Mcgarvey, S. T. andWebster, J. P. (2003).Isolation and characterisation of polymorphic DNA microsatellite markers from Schistosoma japonicum.Molecular Ecology Notes 3,406–408.CrossRefGoogle Scholar

Shrivastava, J.,Gower, C. M.,Balolong, E. J.,Wang, T. P.,Qian, B. Z. andWebster, J. P. (2005).Population genetics of multihost parasites – the case for natural sampling of Schistosoma japonicum larval stages.Parasitology 31,617–626.CrossRefGoogle Scholar

Sire, C.,Durand, P.,Pointier, J. P. andTheron, A. (1999).Genetic diversity and recruitment pattern of Schistosoma mansoni in a Biomphalaria glabrata snail population: a field study using random-amplified polymorphic DNA markers.Journal of Parasitology 85,436–441.CrossRefGoogle Scholar

Sire, C.,Durand, P.,Pointier, J. P. andTheron, A. (2001).Genetic diversity of Schistosoma mansoni within and among individual hosts (Rattus rattus): infrapopulation differentiation at microspatial scale.International Journal for Parasitology 31,1609–1616.CrossRefGoogle Scholar

Smithers, S. R. andTerry, R. J. (1965).The infection of laboratory hosts with cercariae of Schistosoma mansoni and the recovery of adult worms.Parasitology 55,695–700.CrossRefGoogle Scholar

Stohler, R. A.,Curtis, J. andMinchella, D. J. (2004).A comparison of microsatellite polymorphism and heterozygosity among field and laboratory populations of Schistosoma mansoni.International Journal for Parasitology 34,595–610.CrossRefGoogle Scholar

Thiongo, F. W.,Madsen, H.,Ouma, J. H.,Andreassen, J. andChristensen, N. O. (1997).Host-parasite relationships in infections with two Kenyan isolates of Schistosoma mansoni in NMRI mice.Journal of Parasitology 83,330–332.CrossRefGoogle Scholar

Thompson, J. N. (1994).The Coevolutionary Process.Chicago University Press,Chicago.CrossRef

Webster, J. P. andDavies, C. M. (2001).Coevolution and compatibility in the snail-schistosome system.Parasitology 123,S41–S56.CrossRefGoogle Scholar

Webster, J. P.,Gower, C. M. andBlair, L. (2004).Do hosts and parasites coevolve?: empirical support from the Schistosoma system.American Naturalist 164,S33–S51.CrossRefGoogle Scholar

Wolfensohn, S. andLloyd, M. (1999).Handbook of Laboratory Animal Management and Welfare.Blackwell Science,Oxford.