Urania basin in the deep Mediterranean Sea houses a lake that is >100 m deep, devoid of oxygen, 6 times more saline than seawater, and has very high levels of methane and particularly sulfide (up to 16 mM), making it among the most sulfidic water bodies on Earth. Along the depth profile there are 2 chemoclines, a steep one with the overlying oxic seawater, and another between anoxic brines of different density, where gradients of salinity, electron donors and acceptors occur. To identify and differentiate the microbes and processes contributing to the turnover of organic matter and sulfide along the water column, these chemoclines were sampled at a high resolution. Bacterial cell numbers increased up to a hundredfold in the chemoclines as a consequence of elevated nutrient availability, with higher numbers in the upper interface where redox gradient was steeper. Bacterial and archaeal communities, analyzed by DNA fingerprinting, 16S rRNA gene libraries, activity measurements, and cultivation, were highly stratified and metabolically more active along the chemoclines compared with seawater or the uniformly hypersaline brines. Detailed analysis of 16S rRNA gene sequences revealed that in both chemoclines ␦-and -Proteobacteria, predominantly sulfate reducers and sulfur oxidizers, respectively, were the dominant bacteria. In the deepest layers of the basin MSBL1, putatively responsible for methanogenesis, dominated among archaea. The data suggest that the complex microbial community is adapted to the basin's extreme chemistry, and the elevated biomass is driven largely by sulfur cycling and methanogenesis.deep anoxic hypersaline lake ͉ element cycling ͉ geosphere-biosphere interaction ͉ Mediterranean Sea ͉ microbial diversity T he Urania basin is one of the deep-sea hypersaline anoxic basins (DHABs) located in the eastern Mediterranean Sea. DHABs are far below the photic zone (3,200-3,600 m deep) and contain brines, the origin of which has been attributed to dissolution of 5.9-to 5.3-million-year-old Messinian evaporites (1). Urania is less saline than the other Mediterranean DHABs, with NaCl concentrations 5.4-7 times higher than normal seawater, but has higher concentrations of methane (5.56 mM) and exceptionally high levels of sulfide (up to 16 mM), making Urania basin among the most sulfidic marine water bodies on Earth (2-4).Interfaces are considered to be hot spots for biological activity (2, 5), and environmental gradients represent an important part of the biosphere that must be accounted for in models of global biogeochemical cycles, especially in otherwise oligotrophic environments like the Eastern Mediterranean (6).In the present study, we discovered 2 different environmental chemoclines within the Urania basin. We finely dissected the gradients and compared the oxic/anoxic upper interface of Urania basin with those found in chemically different DHABs. We concluded that the lower overall salinity but higher sulfide and methane concentrations in Urania DHAB are the primary factors determining the observed ...