Diversity of extremely halophilic bacteria

In this review, the history of the classification of the family Halobacteriaceae, the extremely halophilic aerobic Archaea, is reviewed with some emphasis on the recently described new genera Halobaculum, Halorubrum, Natrialba, Natronomonas, and "Haloterrigena." Speculation is made about the evolutionary relationship between members of the Halobacteriaceae and the extremely halophilic, anaerobic methanogens of the genera Methanohalobium and Methanohalophilus. Efforts to find missing links between the two groups are also reviewed. Received: January 22, 1998 / Accepted: February 16, 1998     

Denitrification by extremely halophilic bacteria

Abstract Extremely halophilic bacteria were isolated from widely separated sites by anaerobic enrichment in the presence of nitrate. The anaerobic growth of several of these isolates was accompanied by the production of nitrite, nitrous oxide, and dinitrogen. These results are a direct confirmation of the existence of extremely halophilic denitrifying bacteria, and suggest that such bacteria may be common inhabitants of hypersaline environments.     

Substrate uptake in extremely halophilic microbial communities revealed by microautoradiography and fluorescence in situ hybridization

The combination of fluorescence in situ hybridization and microautoradiography (FISH-MAR approach) was applied to brine samples of a solar saltern crystallizer pond from Mallorca (Spain) where the simultaneous occurrence of Salinibacter spp. and the conspicuous square Archaea had been detected. Radioactively labeled bicarbonate, acetate, glycerol, and an amino acid mixture were tested as substrates for the microbial populations inhabiting such brines. The results indicated that hitherto uncultured 'square Archaea' do actively incorporate amino acids and acetate. However, Salinibacter spp. only showed amino acid incorporation in pure culture, but no evidence of such activity in their natural environment could be demonstrated. No glycerol incorporation was observed for any component of the microbial community.Communicated by W.D. Grant     

The ecology of the extremely halophilic archaea

Abstract: The extremely halophilic archaea (family Halobacteriaceae) are the dominant heterotrophic organisms in hypersaline environments in which salt concentrations exceed 250–300 g l⁻¹. During the last decades our knowledge on the taxonomy, physiology and biochemistry of the Halobacterium group has greatly increased. However, our understanding of the ecology of the halophilic archaea lags far behind the progess made in the study of other aspects of their biology. A few hypersaline environments, such as the Dead Sea and solar salterns, have been studied more in depth, using techniques such as lipid analysis to obtain information on the types of organisms present and measurement of uptake of labeled substrates to quantify the dynamics of bacterial processes. The results of these studies, in combination with the information obtained from laboratory studies of representative isolates of the Halobacteriaceae, enable the beginning of an understanding of the functioning of the halophilic archaea in nature.     

Tolerance of extremely halophilic archaebacteria towards bromide

The tolerance of halophilic archaebacteria towards bromide was tested in view of the fact that bromide occurs in natural brines in concentrations of up to 66 mM. It was found that, while concentrations of up to 0.8–1M are tolerated well by all halobacterial types examined, great differences exist between species with respect to bromide tolerance. WhileHalobacterium (H. salinarium, H. halobium, andH. sodomense) andNatronobacterium species are only moderately tolerant,Haloarcula (H. vallismortis, H. marismortui), andHaloferax species (H. mediterranei, H. gibbonsii) tolerate higher concentrations.Haloferax volcanii proved extremely tolerant and showed growth in bromide media at very low chloride concentrations (below 50 mM). No correlation was found between bromide tolerance and the bromide concentration in the habitat from which the strains were isolated. Iodide proved much more toxic than bromide. Bromide-tolerant strains also proved relatively resistant to growth inhibition by iodide.     

DNA base composition of extremely halophilic cocci

Summary The percentage guanine+cytosine (GC) content of the DNA of 9 extremely halophilic cocci were determined from melting temperature (T _m) and the E ₂₆₀/E ₂₈₀ ratio at pH 3. The values found ranged from 60.5–65.8, with an average 62.6% GC.     

Cell wall of an extremely halophilic coccus

Summary Amino acid and amino sugar analyses have been carried out on hydrolyzed wall preparations from an extremely halophilic coccus. A limited number of ninhydrin-positive compounds were found. The amino acids glycine and glutamic acid or glutamine were present, as well as the amino sugars glucosamine and galactosamine. In addition, three unidentified compounds were detected. Other peptidoglycan components, as found in non-halophilic Gram-positive cocci, were absent.     

Salt tolerance of archaeal extremely halophilic lipid membranes

The membranes of extremely halophilic Archaea are characterized by the abundance of a diacidic phospholipid, archaetidylglycerol methylphosphate (PGP-Me), which accounts for 50-80 mol% of the polar lipids, and by the absence of phospholipids with choline, ethanolamine, inositol, and serine head groups. These membranes are stable in concentrated 3-5 m NaCl solutions, whereas membranes of non-halophilic Archaea, which do not contain PGP-Me, are unstable and leaky under such conditions. By x-ray diffraction and vesicle permeability measurements, we demonstrate that PGP-Me contributes in an essential way to membrane stability in hypersaline environments. Large unilamellar vesicles (LUV) prepared from the polar lipids of extreme halophiles, Halobacterium halobium and Halobacterium salinarum, retain entrapped carboxyfluorescein and resist aggregation in the whole range 0-4 m NaCl, similarly to LUV prepared from purified PGP-Me. By contrast, LUV made of polar lipid extracts from moderately halophilic and non-halophilic Archaea (Methanococcus jannaschii, Methanosarcina mazei, Methanobrevibacter smithii) are leaky and aggregate at high salt concentrations. However, adding PGP-Me to M. mazei lipids results in gradual enhancement of LUV stability, correlating with the PGP-Me content. The LUV data are substantiated by the x-ray results, which show that H. halobium and M. mazei lipids have dissimilar phase behavior and form different structures at high NaCl concentrations. H. halobium lipids maintain an expanded lamellar structure with spacing of 8.5-9 nm, which is stable up to at least 100 degrees C in 2 m NaCl and up to approximately 60 degrees C in 4 m NaCl. However, M. mazei lipids form non-lamellar structures, represented by the Pn3m cubic phase and the inverted hexagonal H(II) phase. From these data, the forces preventing membrane aggregation in halophilic Archaea appear to be steric repulsion, because of the large head group of PGP-Me, or possibly out-of-plane bilayer undulations, rather than electrostatic repulsion attributed to the doubly charged PGP-Me head group.     

Biodegradation of hydrocarbons by an extremely halophilic archaebacterium

An archaebacterium (strain EH4) able to biodegrade saturated and aromatic hydrocarbons has been isolated from a sail-marsh. Maximum growth on eicosane (62% of biodegradation, 10 h generation time) was reached in a medium prepared with a natural hypersaline water collected from a salt-marsh (3.5 mol/1 NaCl concentration). No growth on hydrocarbons was observed for NaCl concentration lower than 1.8 mol/1.