Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
The mangrove swamp environment constitutes a sulphide rich habitat harbouring some thioautotrophic organisms. The ciliate Zoothamnium niveum and the nematode Eubostrichus dianae, both known to live associated with bacterial sulphide-oxidizing ectosymbionts, were analysed as positive controls by Raman microspectrometry. The detection of the 3 Raman bands characteristic of elemental sulphur (S8) allows us to define a positive model of sulphide-oxidizing symbiotic invertebrates and by extrapolation, of thioautotrophic organisms. A fast screening using this tool was carried out on eukaryotic organisms such as hydrozoan, nematodes, annelids, copepods, and ciliate (Pseudovorticella sp.) and on free-living filamentous bacteria found on decomposing leaves in order to detect thioautotrophic organisms. The Raman microspectrometry permits us: (i) to reveal thioautotrophic metabolism of free-living bacteria (Beggiatoa sp.) and even for Archaea and (ii) to detect sulphide-oxidizing endosymbiotic and ectosymbiotic bacteria associated with the Bivalve Lucina pectinata and Pseudovorticella sp., respectively. Raman microspectrometry represents a fast, easy and non destructive technique which can be used on living organisms without constraints of sample size. The Raman analysis can also be completed by ultrastructural analysis (SEM, TEM) on the same sample.
ISSN: 0141-1136

hal-00715329
https://hal.univ-antilles.fr/hal-00715329
DOI : 10.1016/j.marenvres.2010.02.001

Éditeur(s) : HAL CCSD Wiley
Résumé : International audience
Background information. Marine nematodes belonging to the Stilbonematidae (Desmodoridae) family are described as living in obligatory association with sulfur-oxidizing chemoautotrophic ectosymbionts. The symbiotic bacteria carrying out this chemosynthesis should contain elemental sulfur in periplasmic granules as sulfur granules of chemoautotrophic endosymbionts described in various marine invertebrates. Results. Based on TEM analyses, extracellular bacteria surrounding Eubostrichus dianae possess these spherical periplasmic granules. Few investigative techniques can be used to identify elemental sulfur S8 such as EDXS (energy dispersive X-ray spectroscopy) and EELS (electron energy loss spectroscopy) associated with cryo-fixation of the sample to avoid sulfur loss. These techniques are time consuming, expensive and require technical skills. Raman microspectrometry applied to the analysis of E. dianae allowed us to detect elemental sulfur S8 and confirmed the location of these sulfur clusters in the bacterial coat. In the same way, Raman spectrometry was positively applied to the endosymbiotic bivalve Codakia orbicularis, suggesting that this technique can be used to characterize sulfur in ecto- as well as in endo-symbiotic sulfur-oxidizing bacteria. Conclusions. As Raman spectrometry can be used on living organisms (without preliminary fixation) without sample damage and preserving the molecular structure of the sulfur (denatured during chemical fixation) it represents a very well-adapted investigative tool for biologists. This technique therefore permits us to detect quickly and easily (in a few seconds and on entire living animals) the presence of sulfur compounds in the symbiotic nematode.
ISSN: 0248-4900

hal-00753017
https://hal.univ-antilles.fr/hal-00753017

Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
Ciliates represent a diversified group of protists known to establish symbioses with prokaryotic microorganisms. They are mainly phagotrophs and symbiotic relationships with bacteria can give them an important advantage in chemosynthetic environments. The aim of this study is to describe the thiotrophic association that occurs between the peritrich ciliate Pseudovorticella sp. and potential sulfur-oxidizing bacteria. Investigations at microscopic scale (LM, SEM, TEM) showed ectosymbiotic bacteria covering the surface of the body of Pseudovorticella sp. According to 16S rDNA phylogenetic analysis, these ectosymbiotic bacteria belong to-proteobacteria and are phylogenetically close to the symbiont of the recently described Zoothamnium ignavum, which inhabits shallow-water wood falls. FISH experiments, using symbiont specific probes, clearly indicate that these ectosymbiotic bacteria are also ingested into food vacuoles. Electron lucent granules observed in TEM in the cytoplasm of the ectosymbiotic bacteria have been identified as sulfur granules by Raman microspectrometry analyses. Raman microspec-trometry analyses confirmed the thiotrophic nature of this relationship already suggested by the results obtained by TEM and phylogeny. A complete sulfur map was then performed to investigate the sulfur distribution in the zooid. Results show that the relationship between this protist and its bacterial partner is a thiotrophic ectosymbiosis.
ISSN: 0932-4739

hal-01655933
http://hal.upmc.fr/hal-01655933
http://hal.upmc.fr/hal-01655933/document
http://hal.upmc.fr/hal-01655933/file/GRIMONPREZ_Adrien_sans%20marque.pdf
DOI : 10.1016/j.ejop.2017.11.002