In situ characterization of sulphur in gill-endosymbionts of the shallow water lucinidCodakia orbicularis(Linné, 1758) by high-pressure cryofixation and EFTEM microanalysis

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In this study, Codakia orbicularis gill-tissues were cryo-fixed by using high-pressure freezing, a freeze substitution process and finally by cryo-embedding in Lowicryl. Ultrathin sections were then used for an EFTEM microanalysis. Results show that intracellular bacterial symbionts contain elemental sulphur in periplasmic vesicles as indicated by conventional TEM. When sulphur is temporarily depleted in the environment, such structures may act as energy sources for bacterial metabolism. Moreover, sulphate was detected in the cytoplasm of the bacterial symbionts, suggesting the oxidation of elemental sulphur, located in periplasmic granules, to sulphate (the final step in sulphur oxidation) by these chemoautotrophic bacteria. To assess the effects of host starvation on the bacterial sulphur content, adult individuals of C. orbicularis were maintained in starvation for 6 weeks in sterile artificial seawater depleted in sulphur. During starvation, both (1) the number of bacteria inside the bacteriocytes and (2) the number of periplasmic granules per prokaryotic cell decreased. The content of the remaining periplasmic granules had been modified to sulphate. This observation suggests that bacterial gill-endosymbionts used the elemental sulphur in their periplasmic granules as stored substrate for oxidation in order to produce energy in case of sulphur depletion.

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