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Genotypic variation in the parasitic dinoflagellate Hematodinium perezi along the Delmarva Peninsula, Virginia

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Abstract

Hematodinium perezi (genotype III) is a parasitic dinoflagellate that infects blue crabs along the eastern seaboard and Gulf of Mexico, USA. In order to examine the intra-specific genetic variation of this parasite, eleven microsatellite markers from H. perezi (III) were amplified from 227 infected blue crabs collected during 2008–2009 from six sites in Virginia. Simultaneous infections with multiple genetic types in a single-host individual were common and observed in 42 % of the samples. The remaining 58 % of samples had a single allele per locus at all eight polymorphic loci suggesting that the life history stages of the parasite in the host hemolymph are likely haploid. The composition and distribution of multi-locus genotypes (MLG) from samples with infections of a single genetic type indicated high genotypic variation along the Delmarva Peninsula, Virginia, with no evidence of population structure. The lack of linkage disequilibrium combined with the large number of unique MLGs (84 %) is strong evidence for recombination in the life cycle, but the sexual stages remain undetermined. This is the first evidence of ploidy level, infections by multiple genetic types in an individual host animal, high levels of genotypic variation, and sexual reproduction for any species of Hematodinium.

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References

  • Ajzenberg D, Banuls AL, Tibayrenc M, Darde ML (2002) Microsatellite analysis of Toxoplasma gondii shows considerable polymorphism structured into two main clonal groups. Int J Parasitol 32:27–38

    Article  CAS  Google Scholar 

  • Alpermann TJ, Beszteri B, John U, Tillmann U, Cembella AD (2009) Implications of life-history transitions on the population genetic structure of the toxigenic marine dinoflagellate Alexandrium tamarense. Mol Ecol 18:2122–2133

    Article  Google Scholar 

  • Alpermann TJ, Tillmann U, Beszteri B, Cembella AD, John U (2010) Phenotypic variation and genotypic diversity in a planktonic population of the toxigenic marine dinoflagellate Alexandrium tamarense (Dinophyceae). J Phycol 46:18–32

    Article  CAS  Google Scholar 

  • Anderson TJC, Haubold B, Williams JT, Estrada-Franco JG, Richardson L, Mollinedo R, Bockarie M, Mokili J, Mharakurwa S, French N, Whitworth J, Velez ID, Brockman AH, Nosten F, Ferreira MU, Day KP (2000) Microsatellite markers reveal a spectrum of population structures in the malaria parasite Plasmodium falciparum. Mol Biol Evol 17:1467–1482

    Article  CAS  Google Scholar 

  • Audemard C, Reece KS, Burreson EM (2004) Real-time PCR for detection and quantification of the protistan parasite Perkinsus marinus in environmental waters. Appl Environ Microbiol 70:6611–6618

    Article  CAS  Google Scholar 

  • Babiker HA, Walliker D (1997) Current views on the population structure of Plasmodium falciparum: implications for control. Parasitol Today 13:262–267

    Article  CAS  Google Scholar 

  • Balmer O, Stearns SC, Schotzau A, Brun R (2009) Intraspecific competition between co-infecting parasite strains enhances host survival in African trypanosomes. Ecology 90:3367–3378

    Article  Google Scholar 

  • Belkhir K, Borsa P, Chikhi L, Raufaste N, Catch F (1996–2002) GENETIX 4.04, software under Windows TM for the genetics of the populations. Laboratory Genome, Populations, Interactions, CNRS UMR 5000, University of Montpellier II, Montpellier, France. http://kimura.univ-montp2.fr/genetix/. Accessed 6 May 2009

  • Brown SP, Hochberg ME, Grenfell BT (2002) Does multiple infection select for raised virulence? Trends Microbiol 10:401–405

    Article  CAS  Google Scholar 

  • Colwell RK (2011) EstimateS, Version 8.2: statistical estimation of species richness and shared species from samples. http://viceroy.eeb.uconn.edu/EstimateS/

  • Doerder FP, Arslanyolu M, Saad Y, Kaczmarek M, Mendoza M, Mita B (1996) Ecological genetics of Tetrahymena thermophila: mating types, i-antigens, multiple alleles, and epistasis. J Eukaryot Microbiol 43:95–100

    Article  CAS  Google Scholar 

  • Eaton WD, Love DC, Botelho C, Meyers TR, Imamura K, Koeneman T (1991) Preliminary results on the seasonality and life cycle of the parasitic dinoflagellate causing Bitter Crab disease in Alaskan Tanner crabs (Chionoecetes bairdi). J Invertebr Pathol 57:426–434

    Article  CAS  Google Scholar 

  • Ellegren H (2004) Microsatellites: simple sequences with complex evolution. Nat Rev Genet 5:435–445

    Article  CAS  Google Scholar 

  • Excoffier LGL, Schneider S (2005) Arlequin ver. 3.0: an integrated software package for population genetics data analysis. Evol Bioinform Online 1:47–50

    CAS  Google Scholar 

  • Frischer ME, Lee RF, Sheppard MA, Mauer A, Rambow F, Neumann M, Brofft JE, Wizenmann T, Danforth JM (2006) Evidence for a free-living stage of the blue crab parasitic dinoflagellate, Hematodinium sp. Harmful Algae 5:548–557

    Article  Google Scholar 

  • Giraud T, Enjalbert J, Fournier E, Delmotte F, Dutech C (2008a) Population genetics of fungal diseases of plants. Parasite 15:449–454

    Article  CAS  Google Scholar 

  • Giraud T, Yockteng R, López-Villavicencio M, Refrégier G, Hood ME (2008b) Mating system of the Anther Smut Fungus Microbotryum violaceum: selfing under heterothallism. Eukaryot Cell 7:765–775

    Article  CAS  Google Scholar 

  • Gruebl T, Frischer ME, Sheppard M, Neumann M, Maurer AN, Lee RF (2002) Development of an 18 s rRNA gene-targeted PCR-based diagnostic for the blue crab parasite Hematodinium sp. Dis Aquat Org 49:61–70

    Article  CAS  Google Scholar 

  • Hanif A, Bowers HA, Mcintosh D, Dyson WE, Pitula JS, Jagus R, Schott E (2011) Using PCR methodology to search for an environmental presence of Hematodinium sp., a lethal parasite of the blue crab Callinectes sapidus. Abstract. National Shellfisheries Association, Baltimore

    Google Scholar 

  • Howells EJ, van Oppen MJH, Willis BL (2009) High genetic differentiation and cross-shelf patterns of genetic diversity among Great Barrier Reef populations of Symbiodinium. Coral Reefs 28:215–225

    Article  Google Scholar 

  • Kirk NL, Andras JP, Harvell CD, Santos SR, Coffroth MA (2009) Population structure of Symbiodinium sp. associated with the common sea fan, Gorgonia ventalina, in the Florida Keys across distance, depth, and time. Mar Biol 156:1609–1623

    Article  Google Scholar 

  • Kumar S, Tamura K, Nei M (2004) MEGA3: integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform 5:150–163

    Article  CAS  Google Scholar 

  • Li YC, Korol AB, Fahima T, Beiles A, Nevo E (2002) Microsatellites: genomic distribution, putative functions and mutational mechanisms: a review. Mol Ecol 11:2453–2454

    Article  CAS  Google Scholar 

  • Li C, Shields JD, Miller TL, Small HJ, Pagenkopp KM, Reece KS (2010) Detection and quantification of the free-living stage of the parasitic dinoflagellate Hematodinium sp. in laboratory and environmental samples. Harmful Algae 9:515–521

    Article  CAS  Google Scholar 

  • Li C, Miller TL, Small HJ, Shields JD (2011) In vitro culture and developmental cycle of the parasitic dinoflagellate Hematodinium sp. from the blue crab Callinectes sapdius. Parasitol 138:1924–1934

    Article  Google Scholar 

  • Ligges U, Mächler M (2003) Scatterplot3d- an R package for visualizing multivariate data. J Stat Softw 8:1–20

    Google Scholar 

  • Lowe CD, Montagnes DJS, Martin LE, Watts PC (2010) High genetic diversity and fine-scale spatial structure in the marine flagellate Oxyrrhis marina (Dinophyceae) uncovered by microsatellite loci. PLoS ONE. doi:10.1371/journal.pone.0015557

    Google Scholar 

  • Magalon H, Baudry E, Huste A, Adjeroud M, Veuille M (2006) High genetic diversity of the symbiotic dinoflagellates in the coral Pocillopora meandrina from the South Pacific. Mar Biol 148:913–922

    Article  Google Scholar 

  • Masseret E, Grzebyk D, Nagai S, Genovesi B, Lasserre B, Laabir M, Collos Y, Vaquer A, Berrebi P (2009) Unexpected genetic diversity among and within populations of the toxic dinoflagellate Alexandrium catenella as revealed by nuclear microsatellite markers. Appl Environ Microbiol 75:2037–2045

    Article  CAS  Google Scholar 

  • Maynard Smith J, Smith NH, O’Rourke M, Spratt BG (1993) How clonal are bacteria? Proc Natl Acad Sci 90:4384–4388

    Article  Google Scholar 

  • McCauley LAR, Erdner DL, Nagai S, Richlen ML, Anderson DM (2009) Biogeographic analysis of the globally distributed harmful algal bloom species Alexandrium minutum (Dinophyceae) based on rRNA gene sequences and microsatellite markers. J Phycol 45:454–463

    Article  CAS  Google Scholar 

  • Medlin L, Elwood HJ, Stickel S, Sogin ML (1988) The characterization of enzymatically amplified eukaryotic 16S-like rRNA- coding regions. Gene 71:491–499

    Article  CAS  Google Scholar 

  • Meirmans PG, Van Tienderen PH (2004) GENOTYPE and GENODIVE: two programs for the analysis of genetic diversity of asexual organisms. Mol Ecol Notes 4:792–794

    Article  Google Scholar 

  • Messick GA (1994) Hematodinium perezi infections in adult and juvenile blue crabs Callinectes sapidus from coastal bays of Maryland and Virginia, USA. Dis Aquat Org 19:77–82

    Article  Google Scholar 

  • Messick GA, Shields JD (2000) Epizootiology of the parasitic dinoflagellate Hematodinium sp. in the American blue crab Callinectes sapidus. Dis Aquat Org 43:139–152

    Article  CAS  Google Scholar 

  • Mosquera J, Adler FR (1998) Evolution of virulence: a unified framework for coinfection and superinfection. J Theor Biol 195:293–313

    Article  CAS  Google Scholar 

  • Moss JA, Xiao J, Dungan CF, Reece KS (2008) Description of Perkinsus beihaiensis n. sp., a new Perkinsus sp. parasite in oysters of southern China. J Eukaryot Microbiol 55:117–130

    Article  CAS  Google Scholar 

  • Nagai S, Lian C, Yamaguchi S, Hamaguchi M, Matsuyama Y, Itakura S, Shimada H, Kaga S, Yamauchi H, Sonda Y, Nishikawa T, Kim CH, Hogetsu T (2007) Microsatellite markers reveal population genetic structure of the toxic dinoflagellate Alexandrium tamarense (Dinophyceae) in Japanese coastal waters. J Phycol 43:43–54

    Article  CAS  Google Scholar 

  • Nagai S, Nishitani G, Sakamoto S, Sugaya T, Lee CK, Kim CH, Itakura S, Yamaguchi M (2009) Genetic structuring and transfer of marine dinoflagellate Cochlodinium polykrikoides in Japanese and Korean coastal waters revealed by microsatellites. Mol Ecol 18:2337–2352

    Article  CAS  Google Scholar 

  • Nei M (1978) Estimation of average heterozygosity and genetic distance from a small number of individuals. Genet 89:583–590

    CAS  Google Scholar 

  • Newman MW, Johnson CA (1975) A disease of blue crabs (Callinectes sapidus) caused by a parasitic dinoflagellate, Hematodinium sp. J Parasitol 63:554–557

    Article  Google Scholar 

  • Oliveira RP, Broude NE, Macedo AM, Cantor CR, Smith CL, Pena SDJ (1998) Probing the genetic population structure of Trypanosoma cruzi with polymorphic microsatellites. Proc Natl Acad Sci 95:3776–3780

    Article  CAS  Google Scholar 

  • Pagenkopp Lohan KM, Reece KS, Miller TL, Wheeler KN, Small HJ, Shields JD (2012a) The role of alternate hosts in the ecology and life history of Hematodinium sp., a parasitic dinoflagellate of the blue crab (Callinectes sapidus). J Parasitol 98:73–84

    Article  Google Scholar 

  • Pagenkopp Lohan KM, McDowell JR, Shields JD, Xiao J, Miller TL, Reece KS (2012b) Isolation and characterization of microsatellite loci for the parasitic dinoflagellate, Hematodinium perezi genotype III, a parasite of Callinectes sapidus. Mol Ecol Resour 12:570–572

    Article  Google Scholar 

  • Pagenkopp Lohan KM, Shields JD, Place AR, Small HJ, Reece KS (2013) Conservation in the first internal transcribed spacer (ITS1) region of Hematodinium perezi genotype III from Callinectes sapidus. Dis Aquat Org 13:65–76

    Article  Google Scholar 

  • Pitula JS, Dyson WD, Bakht HB, Njoku I, Chen F (2012) Temporal distribution of genetically homogenous ‘free-living’ Hematodinium sp. in a Delmarva coastal ecosystem. Aquat Biosyst 8:16

    Article  Google Scholar 

  • R Core Team (2012) R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna. ISBN: 3-900051-07-0. http://www.R-project.org/

  • Raymond M, Rousset F (1995) GENEPOP (version 2.1): population genetics software for exact test and ecumenicism. J Hered 86:248–249

    Google Scholar 

  • Razakandrainibe FG, Durand P, Koella JC, Meeus TD, Rousset F, Ayala FJ, Renaud F (2002) “Clonal” population structure of the malaria agent Plasmodium falciparum in high-infection regions. Proc Natl Acad Sci 102:17388–17393

    Article  Google Scholar 

  • Reece KS, Bushek D, Hudson KL, Graves JE (2001) Geographic distribution of Perkinsus marinus genetic strains along the Atlantic and Gulf coasts of the USA. Mar Biol 139:1047–1055

    Article  Google Scholar 

  • Rice WR (1989) Analyzing tables of statistical tests. Evolution 43:223–225

    Article  Google Scholar 

  • Santos SR, Coffroth MA (2003) Molecular genetic evidence that dinoflagellates belonging to the genus Symbiodinium Freudenthal are haploid. Biol Bull 204:10–20

    Article  CAS  Google Scholar 

  • Santos SR, Gutierrez-Rodriguez C, Lasker HR, Coffroth MA (2003) Symbiodinium sp. associations in the gorgonian Pseudopterogorgia elisabethae in the Bahamas: high levels of genetic variability and population structure in symbiotic dinoflagellates. Mar Biol 143:111–120

    Article  Google Scholar 

  • Shields JD, Squyars CM (2000) Mortality and hematology of blue crabs, Callinectes sapidus, experimentally infected with the parasitic dinoflagellate Hematodinium perezi. Fish Bull 98:139–152

    Google Scholar 

  • Small HJ, Shields JD, Hudson KL, Reece KS (2007) Molecular detection of Hematodinium sp. infecting the blue crab, Callinectes sapidus. J Shellfish Res 26:131–139

    Article  Google Scholar 

  • Small HJ, Shields J, Reece K, Bateman K, Stentiford G (2012) Morphological and molecular characterization of Hematodinium perezi (Dinophyceae: Syndiniales), a dinoflagellate parasite of the harbour crab, Liocarcinus depurator. J Eukaryot Microbiol 59:54–66

    Article  CAS  Google Scholar 

  • Stentiford GD, Shields JD (2005) A review of the parasitic dinoflagellates Hematodinium species and Hematodinium-like infections in marine crustaceans. Dis Aquat Org 66:47–70

    Article  Google Scholar 

  • Taylor F (1990) Phylum Dinoflagellata. In: Margulis L, Corliss J, Melkonian M, Chapman D (eds) Handbook of protoctista. Jones and Bartlett Publishers, Sudbury, pp 419–437

    Google Scholar 

  • Thompson PC, Rosenthal BM, Hare MP (2011) An evolutionary legacy of sex and clonal reproduction in the protistan oyster parasite Perkinsus marinus. Infect Genet Evol 11:598–609

    Article  Google Scholar 

  • Thornhill DJ, Xiang Y, Fitt WK, Santos SR (2009) Reef endemism, host specificity, and temporal stability in populations of symbiotic dinoflagellates from two ecologically dominant Caribbean corals. PLoS ONE. doi:10.1371/journal.pone.0006262

    Google Scholar 

  • Tibayrenc M, Ayala FJ (2002) The clonal theory of parasitic protozoa: 12 years on. Trends Parasitol 18:405–410

    Article  CAS  Google Scholar 

  • Tibayrenc M, Kjellberg F, Ayala FJ (1990) A clonal theor of parasitic protozoa: the population structures of Entamoeba, Giardia, Leishmania, Naegleria, Plasmodium, Trichomonas, and Trypanosoma and their medical and taxonomical consequences. Proc Natl Acad Sci 87:2414–2418

    Article  CAS  Google Scholar 

  • Van Baalen M, Sabelis MW (1995) The dynamics of multiple infection and the evolution of virulence. Am Nat 146:881–910

    Article  Google Scholar 

  • Vardo-Zalik AM, Schall JJ (2008) Clonal diversity within infections and the virulence of a malaria parasite, Plasmodium mexicanum. Parasitology 135:1363–1372

    Article  CAS  Google Scholar 

  • Vardo-Zalik AM, Ford AF, Schall JJ (2009) Detecting number of clones, and their relative abundance, of a malaria parasite (Plasmodium mexicanum) infecting its vertebrate host. Parasitol Res 105:209–215

    Article  Google Scholar 

  • Vilas R, Cao A, Pardo BG, Fernández S, Villalba A, Martínez P (2011) Very low microsatellite polymorphism and large heterozygote deficits suggest founder effects and cryptic structure in the parasite Perkinsus olseni. Infect Genet Evol 11:904–911

    Article  Google Scholar 

  • Wheeler K, Shields JD, Taylor DM (2007) Pathology of Hematodinium infections in snow crabs (Chionoecetes opilio) from Newfoundland, Canada. J Invertebr Pathol 95:93–100

    Article  Google Scholar 

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Acknowledgments

We are grateful to those who helped with the sample collection including Dr. Caiwen Li, Dr. Tom Dolan, Kersten Wheeler, Anna Coffey, Peter Coffey, Christopher Magel, Kelly Delaney, Bhae-Jin Peemoeller, and David Gibbs. Drs. Hamish Small and Jessica Moss Small helped with sample collection and discussion. We thank Sean Fate, Alan Birch, and Edward Smith of the VIMS Eastern Shore Laboratory for their able boat handling and critical help in the field. Gail Scott, Alanna MacIntyre, and Annie Dershem assisted with the molecular laboratory work. Drs. David Gauthier, Allen Place, Emmett Duffy, and three anonymous reviewers offered comments on early versions of this manuscript. This project was funded by Evolution and Ecology of Infectious Diseases Program Grant, National Science Foundation (OCE BE-UF #0723662). This manuscript is contribution number 3315 of the Virginia Institute of Marine Science, College of William & Mary.

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  1. Katrina M. Pagenkopp Lohan

    Present address: Smithsonian Conservation Biology Institute, Center for Conservation and Evolutionary Genetics, National Zoological Park, Washington, DC, 20008, USA

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  1. The Virginia Institute of Marine Science, The College of William and Mary, PO Box 1346, Gloucester Point, VA, 23062, USA

    Katrina M. Pagenkopp Lohan, Jan R. McDowell, Jeffrey D. Shields & Kimberly S. Reece

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Pagenkopp Lohan, K.M., McDowell, J.R., Shields, J.D. et al. Genotypic variation in the parasitic dinoflagellate Hematodinium perezi along the Delmarva Peninsula, Virginia. Mar Biol 161, 261–273 (2014). https://doi.org/10.1007/s00227-013-2332-3

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