nach oben

Erschienen in:

2015 | Online First | Buchkapitel

Characterization of Bacterial Symbionts in Deep-Sea Fauna: Protocols for Sample Conditioning, Fluorescence In Situ Hybridization, and Image Analysis

verfasst von : Sébastien Duperron

Erschienen in: Springer Protocols Handbooks

Verlag: Springer Berlin Heidelberg

Einloggen, um Zugang zu erhalten

Aktivieren Sie unsere intelligente Suche, um passende Fachinhalte oder Patente zu finden.

search-config

KI-gestützte Suche

Aus

Abstract

Symbioses with bacteria are key adaptations allowing various groups of metazoans to reach high biomasses at deep sea reducing habitats including hydrothermal vents and cold seeps. Characterizing these associations is challenging due to the constraints associated with work on deep-sea organisms. These include limited sample availability, impact of recovery procedures and shipment on sample quality, and general lack of environmental data. In this chapter, a standard procedure for sample processing at sea which can maximize sample use back in the laboratory is presented, with example protocols for sample fixation, fluorescence in situ hybridization (FISH)-based localization of symbionts in animal tissues, and estimation of their relative abundances in the case of multiple symbioses.

McFall-Ngai M, Hadfield MG, Bosch TCG et al (2013) Animals in a bacterial world, a new imperative for the life sciences. Proc Natl Acad Sci U S A 110:3229–3236CrossRefPubMedPubMedCentral

Tunnicliffe V, Juniper SK, Sibuet M (2003) Reducing environments of the deep-sea floor. In: Tyler PA (ed) Ecosyst. Elsevier, World Deep-Sea, pp 81–110

Dubilier N, Bergin C, Lott C (2008) Symbiotic diversity in marine animals: the art of harnessing chemosynthesis. Nat Rev Microbiol 6:725–740CrossRefPubMed

Petersen JM, Dubilier N (2009) Methanotrophic symbioses in marine invertebrates. Env Microbiol Rep 1:319–335CrossRef

Zielinski FU, Pernthaler A, Duperron S et al (2009) Widespread occurrence of an intranuclear bacterial parasite in vent and seep bathymodiolin mussels. Environ Microbiol 11:1150–1167CrossRefPubMed

Duperron S (2010) The diversity of deep-sea mussels and their bacterial symbioses. In: Kiel S (ed) The vent and seep biota. Springer, Netherlands, pp 137–167

Raggi L, Schubotz F, Hinrichs K-U et al (2012) Bacterial symbionts of Bathymodiolus mussels and Escarpia tubeworms from Chapopote, an asphalt seep in the southern Gulf of Mexico. Environ Microbiol 15:1969–1987CrossRefPubMed

Rodrigues CF, Cunha MR, Genio L, Duperron S (2013) A complex picture of associations between two host mussels and symbiotic bacteria in the Northeast Atlantic. Naturwissenschaften 100:21–31CrossRefPubMed

Gros O, Liberge M, Heddi A et al (2003) Detection of the free-living forms of sulfide-oxidizing gill endosymbionts in the lucinid habitat (Thalassia testudinum environment). Appl Environ Microbiol 69:6254–6257CrossRef

10.

Harmer TL, Rotjan RD, Nussbaumer AD et al (2008) Free-living tube worm endosymbionts found at deep-sea vents. Appl Environ Microbiol 74:3895–3898CrossRefPubMedPubMedCentral

11.

Petersen JM, Wentrup C, Verna C et al (2012) Origins and evolutionary flexibility of chemosynthetic symbionts from Deep-Sea animals. Biol Bull 223:123–137CrossRefPubMed

12.

Szafranski KM, Gaudron SM, Duperron S (2014) Direct evidence for maternal inheritance of bacterial symbionts in small deep-sea clams (Bivalvia: Vesicomyidae). Naturwissenschaften 101:373–383CrossRefPubMed

13.

Vuillemin R, Le Roux D, Dorval P et al (2009) CHEMINI: A new in situ CHEmical MINIaturized analyzer. Deep Sea Res Part Oceanogr Res Pap 56:1391–1399CrossRef

14.

Wankel SD, Huang Y, Gupta M et al (2013) Characterizing the distribution of methane sources and cycling in the deep sea via in situ stable isotope analysis. Environ Sci Technol 47:1478–1486PubMed

15.

Carlier A, Ritt B, Rodrigues CF et al (2010) Heterogeneous energetic pathways and carbon sources on deep eastern Mediterranean cold seep communities. Mar Biol 157:2545–2565CrossRef

16.

Halary S, Riou V, Gaill F et al (2008) 3D FISH for the quantification of methane- and sulphur-oxidising endosymbionts in bacteriocytes of the hydrothermal vent mussel Bathymodiolus azoricus. ISME J 2:284–292CrossRefPubMed

17.

Lorion J, Halary S, do Nascimento J et al (2012) Evolutionary history of Idas sp. Med, (Bivalvia: Mytilidae), a cold seep mussel bearing multiple symbionts. Cah Biol Mar 53:77–87

18.

Stewart FJ, Young CR, Cavanaugh CM (2008) Lateral symbiont acquisition in a maternally transmitted chemosynthetic clam endosymbiosis. Mol Biol Evol 25:673–683CrossRefPubMed

19.

Decker C, Olu K, Arnaud-Haond S, Duperron S (2013) Physical proximity may promote lateral acquisition of bacterial symbionts in vesicomyid clams. PLoS One 8(7):e64830CrossRefPubMedPubMedCentral

20.

Shillito B, Jollivet D, Sarradin PM et al (2001) Temperature resistance of Hesiolyra bergi, a polychaetous annelid living on vent smoker walls. Mar Ecol Prog Ser 216:141–149CrossRef

21.

Shillito B, Gaill F, Ravaux J (2014) The ipocamp pressure incubator for deep-sea fauna. J Mar Sci Technol Taiwan 22:97–102

22.

Pradillon F, Shillito B, Chervin JC et al (2004) Pressure vessels for in vivo studies of deep-sea fauna. High Press Res 24:237–246CrossRef

23.

Shillito B, Hamel G, Duchi C et al (2008) Live capture of megafauna from 2300 m depth, using a newly designed Pressurized Recovery Device. Deep-Sea Res Part -Oceanogr Res Pap 55:881–889CrossRef

24.

Mullineaux LS, Mills SW, Sweetman AK et al (2005) Vertical, lateral and temporal structure in larval distributions at hydrothermal vents. Mar Ecol Prog Ser 293:1–16CrossRef

25.

Beaulieu SE, Mullineaux LS, Adams DK, Mills SW (2009) Comparison of a sediment trap and plankton pump for time-series sampling of larvae near deep-sea hydrothermal vents. Limnol Oceanogr Methods 7:235–248CrossRef

26.

Gaudron SM, Pradillon F, Pailleret M et al (2010) Colonization of organic substrates deployed in deep-sea reducing habitats by symbiotic species and associated fauna. Mar Env Res 70:1–12CrossRef

27.

Gros O, Maurin LC (2008) Easy flat embedding of oriented samples in hydrophilic resin (LR White) under controlled atmosphere: application allowing both nucleic acid hybridizations (CARD-FISH) and ultrastructural observations. Acta Histochem 110:427–431CrossRefPubMed

28.

Verna C, Ramette A, Wiklund H et al (2010) High symbiont diversity in the bone-eating worm Osedax mucofloris from shallow whale-falls in the North Atlantic. Environ Microbiol 12:2355–2370CrossRefPubMed

29.

Duperron S, De Beer D, Zbinden M et al (2009) Molecular characterization of bacteria associated with the trophosome and the tube of Lamellibrachia sp., a siboglinid annelid from cold seeps in the eastern Mediterranean. FEMS Microbiol Ecol 69:395–409CrossRefPubMed

30.

Zimmermann J, Lott C, Weber M et al (2014) Dual symbiosis with co-occurring sulfur-oxidizing symbionts in vestimentiferan tubeworms from a Mediterranean hydrothermal vent. Environ Microbiol 16:3638–3656CrossRefPubMed

31.

Lösekann T, Robador A, Niemann H et al (2008) Endosymbioses between bacteria and deep-sea siboglinid tubeworms from an Arctic Cold Seep (Haakon Mosby Mud Volcano, Barents Sea). Environ Microbiol 10:3237–3254CrossRefPubMed

32.

Nussbaumer AD, Fisher CR, Bright M (2006) Horizontal endosymbiont transmission in hydrothermal vent tubeworms. Nature 441:345–348CrossRefPubMed

33.

Goffredi SK, Orphan VJ, Rouse GW et al (2005) Evolutionary innovation: a bone-eating marine symbiosis. Environ Microbiol 7:1369–1378CrossRefPubMed

34.

Polz MF, Cavanaugh CM (1995) Unique dominance of one bacterial phylotype at a Mid-Atlantic Ridge hydrothermal vent site. Proc Nat Acad Sci USA 92:7232–7236CrossRefPubMedPubMedCentral

35.

Petersen JM, Ramette A, Lott C et al (2010) Dual symbiosis of the vent shrimp Rimicaris exoculata with filamentous gamma- and epsilonproteobacteria at four Mid-Atlantic Ridge hydrothermal vent fields. Environ Microbiol 12:2204–2218PubMed

36.

Duperron S, Pottier M-A, Leger N et al (2013) A tale of two chitons: is habitat specialisation linked to distinct associated bacterial communities? FEMS Microbiol Ecol 83:552–567CrossRefPubMed

37.

Rodrigues CF, Duperron S (2011) Distinct symbiont lineages in three thyasirid species (Bivalvia: Thyasiridae) form the eastern Atlantic and Mediterranean Sea. Naturwissenschaften 98:281–287CrossRefPubMed

38.

Distel DL, Beaudoin DJ, Morrill W (2002) Coexistence of multiple proteobacterial endosymbionts in the gills of the wood-boring bivalve Lyrodus pedicellatus (Bivalvia: Teredinidae). Appl Environ Microbiol 2002:6292–6299CrossRef

39.

Wentrup C, Wendeberg A, Schimak M et al (2014) Forever competent: deep-sea bivalves are colonized by their chemosynthetic symbionts throughout their lifetime. Environ Microbiol 16:3699–3713CrossRefPubMed

40.

Duperron S, Rodrigues CF, Leger N et al (2012) Diversity of symbioses between chemosynthetic bacteria and metazoans at the Guiness cold seep site (Gulf of Guinea, West Africa). MicrobiologyOpen 1:467–480CrossRefPubMedPubMedCentral

41.

Duperron S, Nadalig T, Caprais JC et al (2005) Dual symbiosis in a Bathymodiolus mussel from a methane seep on the Gabon continental margin (South East Atlantic): 16S rRNA phylogeny and distribution of the symbionts in the gills. Appl Environ Microbiol 71:1694–1700CrossRefPubMedPubMedCentral

42.

Duperron S, Sibuet M, MacGregor BJ et al (2007) Diversity, relative abundance, and metabolic potential of bacterial endosymbionts in three Bathymodiolus mussels (Bivalvia: Mytilidae) from cold seeps in the Gulf of Mexico. Env Microbiol 9:1423–1438CrossRef

43.

Distel DL, Cavanaugh CM (1994) Independent phylogenetic origins of methanotrophic and chemoautotrophic bacterial endosymbioses in marine bivalves. J Bacteriol 176:1932–1938CrossRefPubMedPubMedCentral

44.

Distel DL, Lee HKW, Cavanaugh CM (1995) Intracellular coexistence of methano- and thioautotrophic bacteria in a hydrothermal vent mussel. Proc Natl Acad Sci USA 92:9598–9602CrossRefPubMedPubMedCentral

45.

Duperron S, Bergin C, Zielinski F et al (2006) A dual symbiosis shared by two mussel species, Bathymodiolus azoricus and B. puteoserpentis (Bivalvia: Mytilidae), from hydrothermal vents along the northern Mid-Atlantic Ridge. Environ Microbiol 8:1441–1447CrossRefPubMed

46.

Duperron S, Halary S, Lorion J et al (2008) Unexpected co-occurrence of 6 bacterial symbionts in the gill of the cold seep mussel Idas sp. (Bivalvia: Mytilidae). Environ Microbiol 10:433–445CrossRefPubMed

47.

Urakawa H, Dubilier N, Fujiwara Y et al (2005) Hydrothermal vent gastropods from the same family (Provannidae) harbour ε- and γ-proteobacterial endosymbionts. Environ Microbiol 7:750–754CrossRefPubMed

48.

Bates AE, Harmer TL, Roeselers G, Cavanaugh CM (2011) Phylogenetic characterization of episymbiotic bacteria hosted by a hydrothermal vent limpet (Lepetodrilidae, Vetigastropoda). Biol Bull 220:118–127CrossRefPubMed

49.

Cavanaugh CM, Levering PR, Maki JS et al (1987) Symbiosis of methylotrophic bacteria and deep-sea mussels. Nature 325:346–347CrossRef

50.

Fiala-Médioni A, McKiness ZP, Dando P et al (2002) Ultrastructural, biochemical and immunological characterisation of two populations of the Mytilid mussel Bathymodiolus azoricus from the Mid Atlantic Ridge: evidence for a dual symbiosis. Mar Biol 141:1035–1043CrossRef

51.

Lorion J, Buge B, Cruaud C, Samadi S (2010) New insights into diversity and evolution of deep-sea Mytilidae (Mollusca: Bivalvia). Mol Phyl Evol 57:71–83CrossRef

52.

Pernthaler A, Pernthaler J, Amann R (2002) Fluorescence in situ hybridization and catalysed reporter deposition for the identification of marine Bacteria. Appl Environ Microbiol 68:3094–3101CrossRefPubMedPubMedCentral

53.

Stoecker K, Dorninger C, Daims H, Wagner M (2010) Double labeling of oligonucleotide probes for fluorescence in situ hybridization (DOPE-FISH) improves signal intensity and increases rRNA accessibility. Appl Environ Microbiol 76:922–926CrossRefPubMed

54.

Pond DW, Bell MV, Dixon DR et al (1998) Stable-carbon-isotope composition of fatty acids in hydrothermal vent mussels containing methanotrophic and thiotrophic bacterial endosymbionts. Appl Environ Microbiol 64:370–375PubMedPubMedCentral

55.

Guezi H, Boutet I, Andersen AC et al (2014) Comparative analysis of symbiont ratios and gene expression in natural populations of two Bathymodiolus mussel species. Symbiosis 63:19–29CrossRef

Titel: Characterization of Bacterial Symbionts in Deep-Sea Fauna: Protocols for Sample Conditioning, Fluorescence In Situ Hybridization, and Image Analysis
verfasst von: Sébastien Duperron
Verlag: Springer Berlin Heidelberg
DOI: https://doi.org/10.1007/8623_2015_73

Springer Professional

Abstract

Bitte loggen Sie sich ein, um Zugang zu Ihrer Lizenz zu erhalten.