Raman microspectrometry sulfur detection and characterization in the marine ectosymbiotic nematode Eubostrichus dianae (Desmodoridae, Stilbonematidae).
BACKGROUND
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 (transmission electron microscopy) analyses, extracellular bacteria surrounding Eubostrichus dianae possess these spherical periplasmic granules. Few investigative techniques can be used to identify elemental sulfur, S(8), such as EDXS (energy dispersive X-ray spectroscopy) and EELS (electron energy loss spectroscopy), which are 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.
Himmel
David
D
GTSI - Université des Antilles et de la Guyane, UFR des Sciences Exactes et Naturelles, Département de Physique. B.P. 592. 97159 Pointe-à-Pitre Cedex, Guadeloupe, France. dhimmel@univ-ag.fr
Maurin
Leslie Candice
LC
Gros
Olivier
O
Mansot
Jean-Louis
JL
D000818:Animals; D001419:Bacteria / Q000737:chemistry; D049872:Bivalvia / Q000382:microbiology; D008853:Microscopy; D009348:Nematoda / Q000382:microbiology*; D013059:Spectrum Analysis, Raman / Q000379:methods*; D013455:Sulfur / Q000032:analysis*; D013559:Symbiosis
D013455:Sulfur