Oyster hemolymph is a complex and dynamic ecosystem hosting bacteria, protists and viruses

Animal Microbiome - Tập 2 - Trang 1-16 - 2020
S. Dupont1, A. Lokmer2,3, E. Corre4, J.-C. Auguet5, B. Petton6, E. Toulza1, C. Montagnani1, G. Tanguy7, D. Pecqueur8, C. Salmeron8, L. Guillou9, C. Desnues10,11, B. La Scola12,13, J. Bou Khalil13, J. de Lorgeril1, G. Mitta1, Y. Gueguen1, J.-M. Escoubas1
1IHPE, Univ. Montpellier, CNRS, Ifremer, Univ. Montpellier, Univ. Perpignan Via Domitia, Montpellier, France
2Coastal Ecology, Wadden Sea Station Sylt, Alfred Wegener Institute - Helmholtz Centre for Polar and Marine Research, List auf Sylt, Germany
3Current affiliation UMR 7206 Eco-anthropologie et Ethnologie, CNRS – MNHN Univ. Paris Diderot Sorbonne Paris Cité, Paris, France
4Sorbonne Université, CNRS, FR2424 ABiMS (Analysis and Bioanalysis for Marine Sciences), Station Biologique de Roscoff SBR, Roscoff, France
5MARBEC, Université Montpellier, CNRS, IFREMER, IRD, CC093, place Eugène Bataillon, Montpellier, France
6Ifremer, LEMAR UMR 6539, 11 presqu’île du Vivier, Argenton-en-Landunvez, France
7Sorbonne Université, CNRS, FR2424, Genomer, Station Biologique de Roscoff SBR, Roscoff, France
8Observatoire Océanologique de Banyuls sur Mer, FR 3724, BioPIC, CNRS/SU, Banyuls-sur-Mer, France
9Sorbonne Université, CNRS, UMR7144 Adaptation et Diversité en Milieu Marin, Ecology of Marine Plankton (ECOMAP), Station Biologique de Roscoff SBR, Roscoff, France
10Aix-Marseille Université, IRD 257, Assistance-Publique des Hôpitaux de Marseille, UMR Microbes, Evolution, Phylogeny and Infections (MEPHI), IHU Méditerranée Infection, Marseille, France
11Aix-Marseille Université, Université de Toulon, CNRS, IRD, Mediterranean Institute of Oceanography, UM 110, Marseille, France
12Microbes, Evolution, Phylogeny and Infection (MEΦI), Aix-Marseille Université UM63, Institut de Recherche pour le Développement IRD 198, Assistance Publique – Hôpitaux de Marseille (AP-HM), Marseille, France
13Institut Hospitalo-Universitaire (IHU) - Méditerranée Infection, Marseille, France

Tóm tắt

The impact of the microbiota on host fitness has so far mainly been demonstrated for the bacterial microbiome. We know much less about host-associated protist and viral communities, largely due to technical issues. However, all microorganisms within a microbiome potentially interact with each other as well as with the host and the environment, therefore likely affecting the host health. We set out to explore how environmental and host factors shape the composition and diversity of bacterial, protist and viral microbial communities in the Pacific oyster hemolymph, both in health and disease. To do so, five oyster families differing in susceptibility to the Pacific oyster mortality syndrome were reared in hatchery and transplanted into a natural environment either before or during a disease outbreak. Using metabarcoding and shotgun metagenomics, we demonstrate that hemolymph can be considered as an ecological niche hosting bacterial, protist and viral communities, each of them shaped by different factors and distinct from the corresponding communities in the surrounding seawater. Overall, we found that hemolymph microbiota is more strongly shaped by the environment than by host genetic background. Co-occurrence network analyses suggest a disruption of the microbial network after transplantation into natural environment during both non-infectious and infectious periods. Whereas we could not identify a common microbial community signature for healthy animals, OsHV-1 μVar virus dominated the hemolymph virome during the disease outbreak, without significant modifications of other microbiota components. Our study shows that oyster hemolymph is a complex ecosystem containing diverse bacteria, protists and viruses, whose composition and dynamics are primarily determined by the environment. However, all of these are also shaped by oyster genetic backgrounds, indicating they indeed interact with the oyster host and are therefore not only of transient character. Although it seems that the three microbiome components respond independently to environmental conditions, better characterization of hemolymph-associated viruses could change this picture.

Tài liệu tham khảo

Rosenberg E, Koren O, Reshef L, Efrony R, Zilber-Rosenberg I. The role of microorganisms in coral health, disease and evolution. Nat Rev Microbiol. 2007;5:355–62. Rosenberg E, Zilber-Rosenberg I. The hologenome concept of evolution after 10 years. Microbiome. 2018;6:78. Chabe M, Lokmer A, Segurel L. Gut Protozoa: friends or foes of the human gut microbiota? Trends Parasitol. 2017;33:925–34. Breitbart M, Rohwer F. Method for discovering novel DNA viruses in blood using viral particle selection and shotgun sequencing. Biotechniques. 2005;39:729–36. Moustafa A, Xie C, Kirkness E, Biggs W, Wong E, Turpaz Y, Bloom K, Delwart E, Nelson KE, Venter JC, Telenti A. The blood DNA virome in 8,000 humans. PLoS Pathog. 2017;13:e1006292. Rascovan N, Duraisamy R, Desnues C. Metagenomics and the human Virome in asymptomatic individuals. Annu Rev Microbiol. 2016;70:125–41. Keen EC, Dantas G. Close encounters of three kinds: bacteriophages, commensal Bacteria, and host immunity. Trends Microbiol. 2018;26:943–54. Mirzaei MK, Maurice CF. Menage a trois in the human gut: interactions between host, bacteria and phages. Nat Rev Microbiol. 2017;7:397–408. Pfeiffer JK, Virgin HW. Transkingdom control of viral infection and immunity in the mammalian intestine. Science. 2016;351:aad5872. Hester ER, Barott KL, Nulton J, Vermeij MJ, Rohwer FL. Stable and sporadic symbiotic communities of coral and algal holobionts. ISME J. 2016;10:1157–69. Peixoto RS, Rosado PM, Leite DCD, Rosado AS, Bourne DG. Beneficial microorganisms for corals (BMC): proposed mechanisms for coral health and resilience. Front Microbiol. 2017;8:341. Bosch TC, Grasis JA, Lachnit T. Microbial ecology in Hydra: why viruses matter. J Microbiol. 2015;53:193–200. Gurung K, Wertheim B, Salles JF. The microbiome of pest insects: it is not just bacteria. Entomologia Experimentalis Et Applicata. 2019;167:156–70. Harvey E, Rose K, Eden JS, Lawrence A, Doggett SL, Holmes EC. Identification of diverse arthropod associated viruses in native Australian fleas. Virology. 2019;535:189–99. Pettersson JH, Shi M, Eden JS, Holmes EC, Hesson JC. Meta-Transcriptomic comparison of the RNA Viromes of the mosquito vectors Culex pipiens and Culex torrentium in northern Europe. Viruses. 2019;11:1033. Chiu L, Bazin T, Truchetet ME, Schaeverbeke T, Delhaes L, Pradeu T. Protective microbiota: from localized to long-reaching co-immunity. Front Immunol. 2017;8:1678. Masson F, Lemaitre B. Symbiosis: protection from within. Elife. 2017;6:e24111. Selber-Hnatiw S, Rukundo B, Ahmadi M, Akoubi H, Al-Bizri H, Aliu AF, Ambeaghen TU, Avetisyan L, Bahar I, Baird A, et al. Human gut microbiota: toward an ecology of disease. Front Microbiol. 2017;8:1265. Sweet MJ, Bulling MT. On the importance of the microbiome and Pathobiome in coral health and disease. Front Mar Sci. 2017;4(9). Vonaesch P, Anderson M, Sansonetti PJ. Pathogens, microbiome and the host: emergence of the ecological Koch's postulates. FEMS Microbiol Rev. 2018;42:273–92. Paul-Pont I, Dhand NK, Whittington RJ. Spatial distribution of mortality in Pacific oysters Crassostrea gigas: reflection on mechanisms of OsHV-1 transmission. Dis Aquat Org. 2013;105:127–38. EFSA PoAHW: Oyster mortality. EFSA J 2015, 13:4122. Barbosa Solomieu V, Renault T, Travers MA. Mass mortality in bivalves and the intricate case of the Pacific oyster, Crassostrea gigas. J Invertebr Pathol. 2015;131:2–10. Martenot C, Oden E, Travaille E, Malas JP, Houssin M. Detection of different variants of Ostreid Herpesvirus 1 in the Pacific oyster, Crassostrea gigas between 2008 and 2010. Virus Res. 2011;160:25–31. Renault T, Moreau P, Faury N, Pepin JF, Segarra A, Webb S. Analysis of clinical Ostreid Herpesvirus 1 (Malacoherpesviridae) specimens by sequencing amplified fragments from three virus genome areas. J Virol. 2012;86:5942–7. Segarra A, Pepin JF, Arzul I, Morga B, Faury N, Renault T. Detection and description of a particular Ostreid herpesvirus 1 genotype associated with massive mortality outbreaks of Pacific oysters, Crassostrea gigas, in France in 2008. Virus Res. 2010;153:92–9. Abbadi M, Zamperin G, Gastaldelli M, Pascoli F, Rosani U, Milani A, Schivo A, Rossetti E, Turolla E, Gennari L, et al. Identification of a newly described OsHV-1 microvar from the North Adriatic Sea (Italy). J Gen Virol. 2018;99:693–703. Burioli EA, Prearo M, Riina MV, Bona MC, Fioravanti ML, Arcangeli G, Houssin M. Ostreid herpesvirus type 1 genomic diversity in wild populations of Pacific oyster Crassostrea gigas from Italian coasts. J Invertebr Pathol. 2016;137:71–83. Burioli EAV, Prearo M, Houssin M. Complete genome sequence of Ostreid herpesvirus type 1 microVar isolated during mortality events in the Pacific oyster Crassostrea gigas in France and Ireland. Virology. 2017;509:239–51. Lynch SA, Carlsson J, Reilly AO, Cotter E, Culloty SC. A previously undescribed ostreid herpes virus 1 (OsHV-1) genotype detected in the pacific oyster, Crassostrea gigas, in Ireland. Parasitology. 2012;139:1526–32. Peeler EJ, Reese RA, Cheslett DL, Geoghegan F, Power A, Thrush MA. Investigation of mortality in Pacific oysters associated with Ostreid herpesvirus-1 mu Var in the Republic of Ireland in 2009. Preventive Veterinary Med. 2012;105:136–43. Lemire A, Goudenege D, Versigny T, Petton B, Calteau A, Labreuche Y, Le Roux F. Populations, not clones, are the unit of vibrio pathogenesis in naturally infected oysters. ISME J. 2014;9:1523–31. Bruto M, James A, Petton B, Labreuche Y, Chenivesse S, Alunno-Bruscia M, Polz MF, Le Roux F. Vibrio crassostreae, a benign oyster colonizer turned into a pathogen after plasmid acquisition. ISME J. 2017;11:1043–52. Faury N, Saulnier D, Thompson FL, Gay M, Swings J, Le Roux F. Vibrio crassostreae sp. nov., isolated from the haemolymph of oysters (Crassostrea gigas). Int J Syst Evol Microbiol. 2004;54:2137–40. de Lorgeril J, Lucasson A, Petton B, Toulza E, Montagnani C, Clerissi C, Vidal-Dupiol J, Chaparro C, Galinier R, Escoubas J-M, et al. Immune-suppression by OsHV-1 viral infection causes fatal bacteraemia in Pacific oysters. Nat Commun. 2018;9:4215. Li Z, Nicolae V, Akileh R, Liu T. A brief review of oyster-associated microbiota. Microbiol Res J Int. 2017;20:1–14. Pierce ML, Ward JE. Microbial ecology of the Bivalvia, with an emphasis on the family Ostreidae. J Shellfish Res. 2018;37:793–806. Lokmer A, Kuenzel S, Baines J, Wegner KM. The role of tissue-specific microbiota in initial establishment success of Pacific oysters. Environ Microbiol. 2016;18:970–87. Garnier M, Labreuche Y, Garcia C, Robert M, Nicolas JL. Evidence for the involvement of pathogenic bacteria in summer mortalities of the Pacific oyster Crassostrea gigas. Microb Ecol. 2007;53:187–96. Olafsen JA, Fletcher TC, Grant PT. Agglutinin activity in Pacific oyster (Crassostrea gigas) hemolymph following in vivo Vibrio anguillarum challenge. Dev Comp Immunol. 1992;16:123–38. Wendling CC, Batista FM, Wegner KM. Persistence, seasonal dynamics and pathogenic potential of Vibrio communities from pacific oyster hemolymph. PLoS One. 2014;9:e94256. Bachere E, Rosa RD, Schmitt P, Poirier AC, Merou N, Charriere GM, Destoumieux-Garzon D. The new insights into the oyster antimicrobial defense: cellular, molecular and genetic view. Fish Shellfish Immunol. 2015;46(1):50–64. Desriac F, Le Chevalier P, Brillet B, Leguerinel I, Thuillier B, Paillard C, Fleury Y. Exploring the hologenome concept in marine bivalvia: haemolymph microbiota as a pertinent source of probiotics for aquaculture. FEMS Microbiol Lett. 2014;350:107–16. Lokmer A, Wegner KM. Hemolymph microbiome of Pacific oysters in response to temperature, temperature stress and infection. ISME J. 2015;9:670–82. Lokmer A, Goedknegt MA, Thieltges DW, Fiorentino D, Kuenzel S, Baines JF, Wegner KM. Spatial and temporal dynamics of Pacific oyster Hemolymph microbiota across multiple scales. Front Microbiol. 2016;7:1367. Wegner KM, Volkenborn N, Peter H, Eiler A. Disturbance induced decoupling between host genetics and composition of the associated microbiome. BMC Microbiol. 2013;13:252. Dégremont L, Nourry M, Maurouard E. Mass selection for survival and resistance to OsHV-1 infection in Crassostrea gigas spat in field conditions: response to selection after four generations. Aquaculture. 2015;446:111–21. Petton B, Bruto M, James A, Labreuche Y, Alunno-Bruscia M, Le Roux F. Crassostrea gigas mortality in France: the usual suspect, a herpes virus, may not be the killer in this polymicrobial opportunistic disease. Front Microbiol. 2015;6:686. John SG, Mendez CB, Deng L, Poulos B, Kauffman AK, Kern S, Brum J, Polz MF, Boyle EA, Sullivan MB. A simple and efficient method for concentration of ocean viruses by chemical flocculation. Environ Microbiol Rep. 2011;3:195–202. Herlemann DPR, Labrenz M, Jurgens K, Bertilsson S, Waniek JJ, Andersson AF. Transitions in bacterial communities along the 2000 km salinity gradient of the Baltic Sea. ISME J. 2011;5:1571–9. Medlin L, Elwood HJ, Stickel S, Sogin ML. The characterization of enzymatically amplified eukaryotic 16s-like Rrna-coding regions. Gene. 1988;71:491–9. Guillou L, Bachar D, Audic S, Bass D, Berney C, Bittner L, Boutte C, Burgaud G, de Vargas C, Decelle J, et al. The Protist ribosomal reference database (PR2): a catalog of unicellular eukaryote small sub-unit rRNA sequences with curated taxonomy. Nucleic Acids Res. 2013;41:D597–604. Escudie F, Auer L, Bernard M, Mariadassou M, Cauquil L, Vidal K, Maman S, Hernandez-Raquet G, Combes S, Pascal G. FROGS: find, rapidly, OTUs with galaxy solution. Bioinformatics. 2018;34:1287–94. Mahe F, Rognes T, Quince C, de Vargas C, Dunthorn M. Swarm: robust and fast clustering method for amplicon-based studies. PeerJ. 2014;2:e593. Bolger AM, Lohse M, Usadel B. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics. 2014;30:2114–20. Kopylova E, Noe L, Touzet H. SortMeRNA: fast and accurate filtering of ribosomal RNAs in metatranscriptomic data. Bioinformatics. 2012;28:3211–7. Nurk S, Meleshko D, Korobeynikov A. Pevzner PA: metaSPAdes: a new versatile metagenomic assembler. Genome Res. 2017;5:824–34. Bolduc B, Youens-Clark K, Roux S, Hurwitz BL, Sullivan MB. iVirus: facilitating new insights in viral ecology with software and community data sets imbedded in a cyberinfrastructure. ISME J. 2017;11:7–14. Roux S, Enault F, Hurwitz BL, Sullivan MB. VirSorter: mining viral signal from microbial genomic data. PeerJ. 2015;3:e985. Arndt D, Grant JR, Marcu A, Sajed T, Pon A, Liang YJ, Wishart DS. PHASTER: a better, faster version of the PHAST phage search tool. Nucleic Acids Res. 2016;44:W16–21. Bettarel Y, Halary S, Auguet JC, Mai TC, Van Bui N, Bouvier T, Got P, Bouvier C, Monteil-Bouchard S, Christelle D. Corallivory and the microbial debacle in two branching scleractinians. ISME J. 2018;4:1109–26. Hyatt D, et al. Prodigal: prokaryotic gene recognition and translation initiation site identification. BMC Bioinformatics. 2010;11:119. Mojica KDA, Evans C, Brussaard CPD. Flow cytometric enumeration of marine viral populations at low abundances. Aquat Microb Ecol. 2014;71:203–9. Caporaso JG, Kuczynski J, Stombaugh J, Bittinger K, Bushman FD, Costello EK, Fierer N, Pena AG, Goodrich JK, Gordon JI, et al. QIIME allows analysis of high-throughput community sequencing data. Nat Methods. 2010;7:335–6. R Core Team. R: a language and environment for statistical computing. Vienna: R Foundation for statistical Computing; 2017. Schloss PD, Westcott SL, Ryabin T, Hall JR, Hartmann M, Hollister EB, Lesniewski RA, Oakley BB, Parks DH, Robinson CJ, et al. Introducing mothur: open-source, platform-independent, community-supported software for describing and comparing microbial communities. Appl Environ Microbiol. 2009;75:7537–41. Pinheiro J, Bates D, DebRoy S, Sarkar D, R Development Core Team: nlme: Linear and Nonlinear Mixed Effects Models. R package version 3.1–131. Availlabe at: https://CRAN.R-project.org/package=nlme edition; 2017. Gastwirth JL, Gel YR, Wallace Hui WL, Lyubchich V, Miao W, Noguchi K. lawstat: Tools for Biostatistics, Public Policy, and Law. R package version 3.1 edition; 2017. Anderson MJ. A new method for non-parametric multivariate analysis of variance. Austral Ecol. 2001;26:32–46. Oksanen J, Blanchet FG, Kindt R, Legendre P, Minchin PR, O'Hara RB, Simpson GL, Solymos P, Henry M, Stevens H, Wagner H: vegan: Community Ecology Package. R package version 2.4–3. Available at: https://cran.r-project.org edition; 2017. De Caceres M, Legendre P. Associations between species and groups of sites: indices and statistical inference. Ecology. 2009;90:3566–74. Gotelli NJ, McCabe DJ. Species co-occurrence: a meta-analysis of J. M Diamond’s assembly rules model. Ecology. 2002;83:2091–6. Stone L, Roberts A. The checkerboard score and species distributions. Oecologia. 1990;85:74–9. Jeanbille M, Gury J, Duran R, Tronczynski J, Agogue H, Ben Said O, Ghiglione JF, Auguet JC. Response of Core microbial consortia to chronic hydrocarbon contaminations in coastal sediment habitats. Front Microbiol. 2016;7:1637. Newman MEJ. The structure and function of complex networks. SIAM Rev. 2003;45:167–256. Csardi G, Nepusz T. The igraph software package for complex network research. Inter J Complex Syst. 2005. p. 1695. Langfelder P, Horvath S. WGCNA: an R package for weighted correlation network analysis. Bmc Bioinformatics. 2008;9:559. Vezzulli L, Stagnaro L, Grande C, Tassistro G, Canesi L, Pruzzo C. Comparative 16SrDNA gene-based microbiota profiles of the Pacific oyster (Crassostrea gigas) and the Mediterranean mussel (Mytilus galloprovincialis) from a shellfish farm (Ligurian Sea, Italy). Microb Ecol. 2018;75:495–504. Faust K, Raes J. Microbial interactions: from networks to models. Nat Rev Microbiol. 2012;10:538–50. Sun MY, Dafforn KA, Johnston EL, Brown MV. Core sediment bacteria drive community response to anthropogenic contamination over multiple environmental gradients. Environ Microbiol. 2013;15:2517–31. Deng Y, Jiang YH, Yang Y, He Z, Luo F, Zhou J. Molecular ecological network analyses. BMC Bioinformatics. 2012;13:113. Zhou JZ, Deng Y, Luo F, He ZL, Tu QC, Zhi XY. Functional molecular ecological networks. Mbio. 2010;1:e00169–10. Montoya JM, Pimm SL, Sole RV. Ecological networks and their fragility. Nature. 2006;442:259–64. Steele JA, Countway PD, Xia L, Vigil PD, Beman JM, Kim DY, Chow CET, Sachdeva R, Jones AC, Schwalbach MS, et al. Marine bacterial, archaeal and protistan association networks reveal ecological linkages. ISME J. 2011;5:1414–25. Paine RT. A note on trophic complexity and community stability. Am Nat. 1969;103:91–3. Berry D, Widder S. Deciphering microbial interactions and detecting keystone species with co-occurrence networks. Front Microbiol. 2014;5:219. Sole RV, Montoya JM. Complexity and fragility in ecological networks. Proc Royal Soc B-Biological Sci. 2001;268:2039–45. Prol-Garcia M, Pintado J. Effectiveness of probiotic Phaeobacter Bacteria grown in biofilters against Vibrio anguillarum infections in the rearing of turbot (Psetta maxima) larvae. Mar Biotechnol. 2013;15:726–38. Karim M, Zhao WJ, Rowley D, Nelson D, Gomez-Chiarri M. Probiotic strains for shellfish aquaculture: protection of eastern oyster, Crassostrea Virginica, larvae and juveniles against bacterial challenge. J Shellfish Res. 2013;32:401–8. Azema P, Lamy JB, Boudry P, Renault T, Travers MA, Degremont L. Genetic parameters of resistance to Vibrio aestuarianus, and OsHV-1 infections in the Pacific oyster, Crassostrea gigas, at three different life stages. Genet Sel Evol. 2017;49:23. Gutierrez AP, Bearf TP, Hooper C, Stenton CA, Sanders MB, Paley RK, Rastas P, Bryrom M, Matika O, Houston RD. A genome-wide association study for host resistance to Ostreid Herpesvirus in Pacific oysters (Crassostrea gigas). bioRxiv. 2017;8(4):1273–80. https://doi.org/10.1534/g3.118.200113. Martenot C, Gervais O, Chollet B, Houssin M, Renault T. Haemocytes collected from experimentally infected Pacific oysters, Crassostrea gigas: detection of ostreid herpesvirus 1 DNA, RNA, and proteins in relation with inhibition of apoptosis. PLoS One. 2017;12:e0177448. Morga B, Faury N, Guesdon S, Chollet B, Renault T. Haemocytes from Crassostrea gigas and OsHV-1: a promising in vitro system to study host/virus interactions. J Invertebr Pathol. 2017;150:45–53. King WL, Jenkins C, Go J, Siboni N, Seymour JR, Labbate M. Characterisation of the Pacific oyster microbiome during a summer mortality event. Microb Ecol. 2018;77:502–12. Theis KR, Dheilly NM, Klassen JL, Brucker RM, Baines JF, Bosch TCG, Cryan JF, Gilbert SF, Goodnight CJ, Lloyd EA, et al. Getting the Hologenome Concept Right: an Eco-Evolutionary Framework for Hosts and Their Microbiomes. mSystems. 2016;1(2):e00028–16. Krishnamurthy SR, Wang D. Origins and challenges of viral dark matter. Virus Res. 2017;239:136–42.