Small-subunit (16S) ribosomal DNA clone libraries were constructed using DNA isolated from the anoxic sediments underlying the cyanobacterial mats from two sampling stations of different salinity (Station A, 150-200 per thousand salinity; Station B, 250-320 per thousand salinity) located in the Mediterranean salterns of Salin-de-Giraud (France). Previous studies have shown that the mats at these two sites differ greatly in physicochemical and microbial composition. Sequence analysis of the clone libraries indicated that prokaryotic diversity was high in the sediments from both stations, in both the Bacteria and Archaea domains. Clones related to the alpha- and delta-Proteobacteria (phylum Proteobacteria), the strictly anaerobic fermentative bacteria (phylum Firmicutes), and the Cytophaga-Flavobacterium-Bacteroides (CFB) group (phylum Bacteroidetes) were found in the libraries from both sediments and accounted for the majority of Bacteria domain clones. The results indicated that the populations of delta-Proteobacteria (principally sulfate-reducing bacteria) were significantly different in the two sediments. In addition, several clones from Station A were related either to the gamma-Proteobacteria (phylum Proteobacteria) or to the Spirochaeta, whereas the library from Station B contained several clones related to the uncultured, deep-branching 'KTK group' of Bacteria. Among the Archaea domain clones, all from Station B and the majority from Station A were related to the order Halobacteriales (phylum Euryarchaeota, class Halobacteria). In addition, 12% of the Archaea domain clones from Station A were related to the Methanococci group (phylum Euryarchaeota, class Methanobacteria) and 32% to the phylum Crenarchaeota. This study represents the first molecular analysis of the diversity of halophilic prokaryotes present in these sediments and the results are discussed in the light of our current knowledge of the microbial ecology of these hypersaline ecosystems.

A novel halophilic fermentative bacterium has been isolated from the black sediment below a gypsum crust and a microbial m a t in hypersaline ponds of Mediterranean salterns. Morphologically, physiologically and genetically this organism belongs to the genus Haloanaerobacter. Haloanaerobacter strain SG 3903T (T = type strain) is composed of non-sporulating long flexible rods with peritrichous flagella, able to grow in the salinity range of 5-30% NaCI, with an optimum a t 14-15%. The strain grows by fermenting carbohydrates or by using the Stickland reaction with either serine or H, as electron donors and glycine-betaine as acceptor, which is reduced to trimethylamine. The two species described so far in the genus Haloanaerobacter are not capable of Stickland reaction with glycine-betaine + serine; however, Haloanaerobacter chitinovorans can use glycine-betaine with H, as electron donor. Strain SG 3903T thus represents the first described strain in the genus Haloanaerobacter capable of the Stickland reaction with two amino acids. Although strain SG 3903T showed 67 O/ O DNA-DNA relatedness to H. chitinovorans, it is physiologically sufficiently different from the two described species to be considered as a new species which has been named Haloanaerobacter salinarius sp. nov.

Three new spirilloid phototrophic purple nonsulfur bacteria were isolated in pure culture from three different environments: strain CE2105 from a brackish lagoon in the Arcachon Bay (Atlantic coast, France), strain SE3104 from a saline sulfur spring in the Pyrenees (Navarra, Spain), and strain AT2115 a microbial mat (Tetiaroa Atoll, Society Islands). Single cells of the three strains were spiral-shaped and highly motile. Their intracellular photosynthetic membranes were of the vesicular type. Bacteriochlorophyll a and carotenoids of the normal spirilloxanthin series were present as photosynthetic pigments. Optimal growth occurred under photoheterotrophic conditions and in the presence of 0.5-4% w/v NaCl. These features are similar to those described for Roseospira mediosalina. Comparative sequence analysis of their 16S rRNA genes placed these strains within the alpha-subclass of Proteobacteria, in a cluster together with Roseospira mediosalina and Rhodospira trueperi. They form a closely related group of slightly to moderately halophilic spiral-shaped purple nonsulfur bacteria.However, the three new isolates exhibited some differences in their physiology and genetic characteristics. Consequently, we propose that they are members of three new species within the genus Roseospira, Roseospira marina sp. nov., Roseospira navarrensis sp. nov., and Roseospira thiosulfatophila sp. nov., with strains CE2105, SE3104, and AT2115 as the type strains, respectively. As a consequence, an emended description of the genus Roseospira is also given.

Microbial mats developing in the hypersaline lagoons of a commercial saltern in the Salin-de-Giraud (Rhô ne delta) were found to contain a red layer fully dominated by spirilloid phototrophic purple bacteria underlying a cyanobacterial layer. From this layer four strains of spirilloid purple bacteria were isolated, all of which were extremely halophilic. All strains were isolated by using the same medium under halophilic photolithoheterotrophic conditions. One of them, strain SG 3105 was a purple non-sulfur bacterial strain closely related to Rhodovibrio sodomensis with a 16S rDNA sequence similarity of 98?8 %. The three other isolated strains, SG 3301 T , SG 3302 and SG 3304, were purple sulfur bacteria and were found to be very similar. The cells were motile by a polar tuft of flagella. Photosynthetic intracytoplasmic membranes of the lamellar stack type contained BChl a and spirilloxanthin as the major carotenoid. Phototrophic growth with sulfide as electron donor was poor; globules of elemental sulfur were present outside the cells. In the presence of sulfide and CO 2 good growth occurred with organic substrates. Optimum growth occurred in the presence of 9-12 % (w/v) NaCl at neutral pH (optimal pH 6?8-7) and at 30-35˚C. The DNA base composition of strains SG 3301 T and SG 3304 were 74?5 and 74?1 mol% G+C, respectively.According to the 16S rDNA sequences, strains SG 3301 T and SG 3304 belonged to the genus Halorhodospira, but they were sufficiently separated morphologically, physiologically and genetically from other recognized Halorhodospira species to be described as a new species of the genus. They are, therefore, described as Halorhodospira neutriphila sp. nov. with strain SG 3301 T as the type strain (=DSM 15116 T ).