Thioautotrophic bacterial endosymbionts are degraded by enzymatic digestion during starvation: Case study of two lucinids Codakia orbicularis and C. orbiculata

The Caribbean bivalves Codakia orbicularis (Linné, 1758) and C. orbiculata (Montagu, 1808) live in seagrass beds of Thalassia testudinum and harbor intracellular sulfur‐oxidizing gamma‐proteobacteria. These bacterial symbionts fix CO2 via the Calvin Benson cycle and provide organic compounds to the bivalve. During experimentally induced starvation, no reduced sulfur compounds and no organic particle food are available; the symbionts could be considered as the sole nutrient source of the host bivalve. A previous study has shown that the intracellular bacterial population decreased considerably during starvation and that bacterial endosymbionts were not released by the bivalves. In this study, the activity of two lysosomal marker enzymes (acid phosphatase and arylsulfatase) was detected using cytochemical experiments coupled with energy‐dispersive X‐ray transmission electron microscopy during sulfide and organic particle starvation. The degradation of bacterial endosymbionts began after 2 weeks of starvation in C. orbiculata and after 3 weeks in C. orbicularis. Degradation processes seem to be continuous over several months and could be responsible for the disappearance of the bacterial endosymbionts within the gills during starvation. These data suggest that the host use symbionts as a nutrient source to survive a hunger crisis. The carbon transfer from the symbionts to the host could be flexible and could consist in transfer of organic matter, “milking,” under normal feeding conditions and digestion of the symbionts under starved conditions. Microsc. Res. Tech. 78:173–179, 2015. © 2014 Wiley Periodicals, Inc.

[1]  O. Gros,et al.  Cell proliferation and apoptosis in gill filaments of the lucinid Codakia orbiculata (Montagu, 1808) (Mollusca: Bivalvia) during bacterial decolonization and recolonization , 2012, Microscopy research and technique.

[2]  N. Dubilier,et al.  Plasticity of symbiont acquisition throughout the life cycle of the shallow-water tropical lucinid Codakia orbiculata (Mollusca: Bivalvia). , 2012, Environmental microbiology.

[3]  M. Troussellier,et al.  Effects of Long-Term Starvation on a Host Bivalve (Codakia orbicularis, Lucinidae) and Its Symbiont Population , 2009, Applied and Environmental Microbiology.

[4]  O. Gros,et al.  Lack of endosymbiont release by two Lucinidae (Bivalvia) of the genus Codakia: consequences for symbiotic relationships. , 2009, FEMS microbiology ecology.

[5]  O. Gros,et al.  In situ characterization of sulphur in gill-endosymbionts of the shallow water lucinid Codakia orbicularis (Linné, 1758) by high-pressure cryofixation and EFTEM microanalysis , 2008 .

[6]  A. Lobo-da-Cunha,et al.  Cytoenzymatic investigation of intracellular digestion in the symbiont-bearing hydrothermal bivalve Bathymodiolus azoricus , 2008 .

[7]  P. Berg,et al.  Lucinid clam influence on the biogeochemistry of the seagrassThalassia testudinum sediments , 2007 .

[8]  M. Troussellier,et al.  Characterization of the Population of the Sulfur-Oxidizing Symbiont of Codakia orbicularis (Bivalvia, Lucinidae) by Single-Cell Analyses , 2007, Applied and Environmental Microbiology.

[9]  C. Fisher,et al.  The color of the trophosome: elemental sulfur distribution in the endosymbionts of Riftia pachyptila (Vestimentifera; Siboglinidae) , 2005 .

[10]  S. Dufour,et al.  Anatomical and experimental evidence for particulate feeding in Lucinoma aequizonata and Parvilucina tenuisculpta (Bivalvia: Lucinidae) from the Santa Barbara Basin , 2004 .

[11]  A. Heddi,et al.  Detection of the Free-Living Forms of Sulfide-Oxidizing Gill Endosymbionts in the Lucinid Habitat (Thalassia testudinum Environment) , 2003, Applied and Environmental Microbiology.

[12]  O. Gros,et al.  Lysosomes and sulfide-oxidizing bodies in the bacteriocytes of Lucinapectinata, a cytochemical and microanalysis approach , 2001 .

[13]  C. Fernandez,et al.  Bacterial symbiosis in Loripes lucinalis (Mollusca: Bivalvia) with comments on reproductive strategy , 2001, Journal of the Marine Biological Association of the United Kingdom.

[14]  Cindy Lee Van Dover,et al.  The Ecology of Deep-Sea Hydrothermal Vents , 2000 .

[15]  Carlos M. Duarte,et al.  Seagrass Biomass And Production: A Reassessment , 1999 .

[16]  O. Gros,et al.  Gill filament differentiation and experimental colonization by symbiotic bacteria in aposymbiotic juveniles of Codakia orbicularis (Bivalvia: Lucinidae) , 1998 .

[17]  Megan E. Streams,et al.  Methanotrophic symbiont location and fate of carbon incorporated from methane in a hydrocarbon seep mussel , 1997 .

[18]  O. Gros,et al.  Environmental transmission of a sulfur-oxidizing bacterial gill endosymbiont in the tropical lucinid bivalve Codakia orbicularis , 1996, Applied and environmental microbiology.

[19]  D. Prieur,et al.  Phylogenetic characterization of sulfur-oxidizing bacterial endosymbionts in three tropical Lucinidae by 16S rDNA sequence analysis , 1996 .

[20]  R. Narbaitz Cytochemical staining methods for electron microscopy, by P. R. Lewis and D. P. Knight. vol. 14 in “practical methods in electron microscopy,” A. M. glauert, ed., elsevier, Amsterdam, London, New york, Tokyo, 1992, 321 pp., $59 (paperback); $192 (hardcover) , 1995 .

[21]  A. Boetius,et al.  Digestive enzymes in marine invertebrates from hydrothermal vents and other reducing environments , 1995 .

[22]  L. Frenkiel,et al.  Gill ultrastructure and symbiotic bacteria in Codakia orbicularis (Bivalvia, Lucinidae) , 1995, Zoomorphology.

[23]  M. Diouris,et al.  Chemoautotrophic symbionts and translocation of fixed carbon from bacteria to host tissues in the littoral bivalve Loripes lucinalis (Lucinidae) , 1989 .

[24]  M. Moore,et al.  The ultrastructural localization of lysosomal acid hydrolases in developing oocytes of the common marine musselMytilus edulis , 1985, The Histochemical Journal.

[25]  B. Patel,et al.  Effect of environmental parameters on lysosomal marker enzymes in the tropical blood clam Anadara granosa , 1985 .

[26]  H. Kunze,et al.  A combined assay of three lysosomal marker enzymes: Acid phosphatase, β-d-glucuronidase, and β-N-acetyl-d-hexosaminidase , 1984 .

[27]  H. Felbeck Sulfide oxidation and carbon fixation by the gutless clamSolemya reidi: an animal-bacteria symbiosis , 1983, Journal of comparative physiology.

[28]  J. Ott,et al.  New Mouth less Interstitial Worms from the Sulfide System: Symbiosis with Prokaryotes , 1982 .

[29]  B. Jørgensen Mineralization of organic matter in the sea bed—the role of sulphate reduction , 1982, Nature.

[30]  J. Waterbury,et al.  Prokaryotic Cells in the Hydrothermal Vent Tube Worm Riftia pachyptila Jones: Possible Chemoautotrophic Symbionts. , 1981, Science.

[31]  H. Felbeck Chemoautotrophic Potential of the Hydrothermal Vent Tube Worm, Riftia pachyptila Jones (Vestimentifera). , 1981, Science.

[32]  D. Chandramohan,et al.  Arylsulfatase activity in marine sediments , 1974 .

[33]  G. Glenner,et al.  Improvements in the method for the electron microscopic localization of arylsulphatase activity , 1966, Histochemie.

[34]  P. J. Anderson,et al.  HISTOCHEMICAL METHODS FOR ACID PHOSPHATASE USING HEXAZONIUM PARAROSANILIN AS COUPLER , 1962 .

[35]  F. J. Reithel,et al.  Acid phosphatases of Escherichia coli. , 1960, Archives of biochemistry and biophysics.

[36]  F. Stewart,et al.  Symbiosis of thioautotrophic bacteria with Riftia pachyptila. , 2006, Progress in molecular and subcellular biology.

[37]  D. Dorman,et al.  Cytochrome oxidase inhibition induced by acute hydrogen sulfide inhalation: correlation with tissue sulfide concentrations in the rat brain, liver, lung, and nasal epithelium. , 2002, Toxicological sciences : an official journal of the Society of Toxicology.

[38]  E. Glover,et al.  The lucinid bivalve genus Cardiolucina (Mollusca, Bivalvia, Lucinidae): systematics, anatomy and relationships , 1997 .

[39]  J. Michalski,et al.  Lysosomic and lysozyme activities in the gill of bivalves from deep hydrothermal vents , 1994 .

[40]  D. Knight,et al.  Cytochemical staining methods for electron microscopy , 1992 .

[41]  H. Felbeck,et al.  Significance of the occurrence of chemoautotrophic bacterial endosymbionts in lucinid clams from Bermuda , 1985 .