It is generally assumed that hypersaline environments with sodium chloride concentrations close to saturation are dominated by halophilic members of the domain Archaea, while Bacteria are not considered to be relevant in this kind of environment. Here, we report the high abundance and growth of a new group of hitherto-uncultured Bacteria in crystallizer ponds (salinity, from 30 to 37%) from multipond solar salterns. In the present study, these Bacteria constituted from 5 to 25% of the total prokaryotic community and were affiliated with the Cytophaga-Flavobacterium-Bacteroides phylum. Growth was demonstrated in saturated NaCl. A provisional classification of this new bacterial group as "Candidatus Salinibacter gen. nov." is proposed. The perception that Archaea are the only ecologically relevant prokaryotes in hypersaline aquatic environments should be revised.Hypersaline environments, such as the crystallizer ponds (i.e., ponds where sodium chloride precipitates) of multipond solar salterns, have been shown to harbor a very low prokaryotic diversity, to the point of having been described as "almost monospecific cultures of halophilic archaea" (9). The use of molecular techniques has also revealed a very low diversity, although the most abundant haloarchaeon (5) corresponding to the unique archaeal 16S ribosomal DNA (rDNA) sequences most frequently recovered from several crystallizer ponds by PCR-based methods did not correspond to any previously described microorganism (5,6,26). This apparent discrepancy between molecular and culture-based data has been found in other environments (4).When fluorescence in situ hybridization (FISH) was used to analyze the prokaryotic community inhabiting crystallizer ponds (around 37% salinity) of a marine solar saltern located in Alicante, Spain (5), Bacteria in high numbers (around 3 ϫ 10 6 /ml) were unexpectedly found. The cells, which accounted for 18% of total cell counts, contained large numbers of ribosomes, as indicated by the intense and uniform FISH signals obtained with the 16S rRNA-targeted oligonucleotide probe EUB338 (3), which is specific for members of the Bacteria domain. The finding of abundant Bacteria with high cellular rRNA content in such a hypersaline environment was unexpected in light of previous reports (20-23) suggesting that almost all the active biomass was of archaeal origin.Solar salterns consist of a series of shallow ponds connected in a sequence of increasingly saline brines. Crystallizers are the last ponds and have a salinity above 30% (6). Therefore, the presence in these ponds of Bacteria could be due either to their import from previous ponds with lower salinity or to their active growth in the crystallizers. The key point that would determine the relevance of finding Bacteria in the crystallizers was, then, whether they were in fact extreme halophiles forming part of the autochthonous microbiota of these ponds.In order to characterize this bacterial community and assess this point, we used the rRNA approach (4, 19), which allows direct, cu...