The potential of Bacillus and Enterococcus probiotic strains to combat helicobacter pylori attachment to the biotic and abiotic surfaces / El potencial de las cepas probióticas de Bacillus y Enterococcus para combatir la adhesión de Helicobacter pylori a las superficies bióticas y abióticas

The prevention of biofilm formation plays a pivotal role in managing Helicobacter pylori inside the body and the environment. This study showed in vitro potentials of two recently isolated probiotic strains, Bacillus sp. 1630F and Enterococcus sp. 7C37, to form biofilm and combat H. pylori attachment to the abiotic and biotic surfaces. Lactobacillus casei and Bifidobacterium bifidum were used as the reference probiotics. The biofilm rates were the highest in the solid–liquid interface for Lactobacillus and Bifidobacterium and the air–liquid interface for Bacillus and Enterococcus. The highest tolerances to the environmental conditions were observed during the biofilm formations of Enterococcus and Bifidobacterium (pH), Enterococcus and Bacillus (bile), and Bifidobacterium and Lactobacillus (NaCl) on the polystyrene and glass substratum, respectively. Biofilms occurred more quickly by Bacillus and Enterococcus strains than reference strains on the polystyrene and glass substratum, respectively. Enterococcus (competition) and Bacillus (exclusion) achieved the most inhibition of H. pylori biofilm formations on the polystyrene and AGS cells, respectively. Expression of luxS was promoted by Bacillus (exclusion, 3.2 fold) and Enterococcus (competition, 2.0 fold). Expression of ropD was decreased when H. pylori biofilm was excluded by Bacillus (0.4 fold) and Enterococcus (0.2 fold) cells. This study demonstrated the ability of Bacillus and Enterococcus probiotic bacteria to form biofilm and combat H. pylori biofilm formation.(AU)

The potential of Bacillus and Enterococcus probiotic strains to combat helicobacter pylori attachment to the biotic and abiotic surfaces.

The prevention of biofilm formation plays a pivotal role in managing Helicobacter pylori inside the body and the environment. This study showed in vitro potentials of two recently isolated probiotic strains, Bacillus sp. 1630F and Enterococcus sp. 7C37, to form biofilm and combat H. pylori attachment to the abiotic and biotic surfaces. Lactobacillus casei and Bifidobacterium bifidum were used as the reference probiotics. The biofilm rates were the highest in the solid-liquid interface for Lactobacillus and Bifidobacterium and the air-liquid interface for Bacillus and Enterococcus. The highest tolerances to the environmental conditions were observed during the biofilm formations of Enterococcus and Bifidobacterium (pH), Enterococcus and Bacillus (bile), and Bifidobacterium and Lactobacillus (NaCl) on the polystyrene and glass substratum, respectively. Biofilms occurred more quickly by Bacillus and Enterococcus strains than reference strains on the polystyrene and glass substratum, respectively. Enterococcus (competition) and Bacillus (exclusion) achieved the most inhibition of H. pylori biofilm formations on the polystyrene and AGS cells, respectively. Expression of luxS was promoted by Bacillus (exclusion, 3.2 fold) and Enterococcus (competition, 2.0 fold). Expression of ropD was decreased when H. pylori biofilm was excluded by Bacillus (0.4 fold) and Enterococcus (0.2 fold) cells. This study demonstrated the ability of Bacillus and Enterococcus probiotic bacteria to form biofilm and combat H. pylori biofilm formation.

In vitro virulence activity of Pseudomonas aeruginosa, enhanced by either Acinetobacter baumannii or Enterococcus faecium through the polymicrobial interactions.

Microbes within an infection impact neighbors' pathogenicity. This study aimed to address in vitro virulence activity of Pseudomonas aeruginosa under the binary interaction with Acinetobacter baumannii or Enterococcus faecium, co-isolated from two chronic wound infections. The biofilm formation of Pseudomonas was enhanced 1.5- and 1.4-fold when it was simultaneously cultured with Acinetobacter and Enterococcus, respectively. Pseudomonas motility was increased by 1.9- and 1.5-fold (swimming), 3.6- and 1.9-fold (swarming), and 1.5- and 1.5-fold (twitching) in the dual cultures with Acinetobacter and Enterococcus, respectively. The synergistic hemolysis activity of Pseudomonas was observed with the heat-killed Acinetobacter and Enterococcus cells. The minimum inhibitory concentration of ciprofloxacin against Pseudomonas was increased from (µg mL-1) 25 to 400 in the individual and mixed cultures, respectively. The pyocyanin production by Pseudomonas in the single and mixed cultures with Acinetobacter and Enterococcus was (µg/mL) 1.8, 2.3, and 2.9, respectively. The expression of lasI, rhlI, and pqsR genes was up-regulated by 1.0-, 1.9-, and 16.3-fold, and 4.9-, 1.0-, and 9.3-fold when Pseudomonas was incubated with Acinetobacter and Enterococcus, respectively. Considering the entire community instead of a single pathogen may lead to a more effective therapeutic design for persistent infections caused by Pseudomonas.

Antibacterial and antibiofilm efficacy of Ag NPs, Ni NPs and Al2O3 NPs singly and in combination against multidrug-resistant Klebsiella pneumoniae isolates.

BACKGROUND: Although traditional antibiotic therapy provided an effective approach to combat pathogenic bacteria, the long-term and widespread use of antibiotic results in the evolution of multidrug-resistant bacteria. Recent progress in nanotechnology offers an alternative opportunity to discover and develop novel antibacterial agents. METHODS: A total of 51 K. pneumoniae strains were collected from several specimens of hospitalized patients and identified by two parallel methods (biochemical tests and Vitek-2 system). The antibiotic sensitivity of isolates was evaluated by disk diffusion antibiogram and Vitek-2 system. The biofilms formation ability of antibiotic-resistant strains was examined by microtiter plate and tube methods based on crystal violet staining. The molecular technique was used to determine key genes responsible for biofilms formation of clinical isolates. The antibacterial and antibiofilm activities of Ag NPs, Ni NPs, Al2O3 NPs singly (NPs) and in combination (cNPs) were investigated against selected strains using standard methods. Moreover, the cytotoxicity of NPs was evaluated on mouse neural crest-derived (Neuro-2A) cell line. RESULTS: The results of bacterial studies revealed that more than 80 % of the isolates were resistant to commonly used antibiotics and about 95 % of them were able to form biofilms. Moreover, the presence of fimA and mrkA genes were determined in all biofilm-producing strains. The results of antibacterial and antibiofilm activities of NPs and cNPs demonstrated the lower MIC and MBEC values for Al2O3 NPs singly as well as for Ag/Ni cNPs and Ag/Al2O3 cNPs in combination, respectively. Overall, the inhibitory effects of cNPs were superior to NPs against all strains. Furthermore, the results of the checkerboard assays showed that Ag NPs act synergistically with two other NPs against multidrug-resistant Klebsiella pneumoniae (MDR-K. pneumoniae) isolates. The in vitro cytotoxicity assay revealed no significant toxicity of NPs against Neuro-2A cells. CONCLUSION: In the present study, the combination of Ag NPs, Ni NPs, and Al2O3 NPs were used against MDR-K. pneumoniae strains and antibacterial and antibiofilm activities were observed for Ag/Ni cNPs and Ag/Al2O3 cNPs.

Cell-free extract assisted synthesis of ZnO nanoparticles using aquatic bacterial strains: Biological activities and toxicological evaluation.

The introduction of novel bacterial strains and the development of microbial approaches for nanoparticles biosynthesis could minimize the negative environmental impact and eliminate the concern and challenges of the available approaches. In this study, a biological method based on microbial cell-free extract was used for biosynthesis of ZnO NPs using two new aquatic bacteria, Marinobacter sp. 2C8 and Vibrio sp. VLA. The synthesized ZnO NPs were characterized by UV-Visible spectroscopy, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), transmission electron microscopy (TEM), atomic force microscope (AFM), dynamic light scattering (DLS) and zeta potential. The UV-Visible absorption peak was found to be at 266 and 250 nm for ZnO-2C8 NPs and ZnO-VLA NPs, respectively. FTIR study suggested that the hydroxyl, amine, and carboxyl groups of bacterial proteins are mainly responsible for stabilizing the biosynthesized ZnO NPs. The formation of hexagonal wurtzite structure of ZnO NPs was confirmed by the XRD pattern. The morphology of the nanoparticles was found to be spherical with the average particle size of about 10.23 ± 2.48 nm and 20.26 ± 4.44 nm for ZnO-2C8 NPs and ZnO-VLA NPs, respectively. The values of zeta potential indicate the high stability of the biosynthesized ZnO NP. Zeta potential values indicated the high stability of the biosynthesized ZnO NP and were obtained -20.54 ± 7.15 and -23.87 ± 2.29 mV for ZnO-2C8 NPs and ZnO-VLA NPs, respectively. The biosynthesized ZnO NPs had antibacterial activity against Gram-negative and Gram-positive strains and possessed excellent antibiofilm activity with the maximum inhibition of about 96.55% at 250 µg/mL. The DPPH activity of ZnO-2C8 NPs and ZnO-VLA NPs were found 88.9% and 85.7% for 2500 µg/mL concentration, respectively. The toxicity test revealed the biocompatibility of the biosynthesized ZnO NPs. The results suggested that this approach is a very good route for synthesizing ZnO NPs with potential applications in biotechnology.

Escherichia coli enhances the virulence factors of Candida albicans, the cause of vulvovaginal candidiasis, in a dual bacterial/fungal biofilm.

Co-infection with other microorganisms can promote the Candida albicans to be invasive. In this study, Escherichia coli and C. albicans were co-isolated from the women with candidiasis symptoms. The in vitro effects of E. coli on C. albicans hypha development, biofilm formation, antibiotic susceptibility, dispersion from the biofilm, expression of Als3, Hwp1, and Tup1 genes, and pathogenesis in Galleria mellonella were investigated. Electron microscopic images revealed that hypha induction was markedly increased in the bacteria-fungi co-culture. Biofilm formation was increased 2.2 fold in the presence of E. coli. The minimum inhibitory concentration of nystatin against Candida was increased from (µg mL-1) 25 to 50 in the dual biofilm. Candida dissemination was increased up to 2.7 fold from the mixed fungi/bacteria biofilm. The expression of ALS3 and HWP1 genes was increased (5.9 and 2.0 fold, respectively) while the TUP1 gene expression was decreased (0.4 fold) when C. albicans was incubated with E. coli. The simultaneous injection of C. albicans and E. coli to the insect larvae increased Galleria mortality up to 40%. This study demonstrated the effects of E. coli to promote fungi virulence factors, which suggest polymicrobial interaction should be considered during treatment of fungal infections.

Co-immobilized spore laccase/TiO2 nanoparticles in the alginate beads enhance dye removal by two-step decolorization.

Combinatorial application of different dye removal methods with specific features can lead to a novel and robust decolorizing system. In this study the bacterial spore laccase and TiO2 nanoparticles were co-entrapped to enhance dye degradation. The optimum entrapment conditions were achieved in the presence of alginate 2% (w/v) and Ca2+ (0.2M), Cu2+ (0.05M) and Zn2+ (0.25M) as matric polymer and counterions, respectively. Immobilized laccase showed a wide range of pH and temperature stability in comparison to the free spores. The entrapped degradation systems include single laccase, single TiO2, laccase + TiO2 (one-step remediation), TiO2/laccase (two-step remediation), and laccase/TiO2 (two-step remediation) that result to the 22%, 26% 45.6%, 47.6%, and 69.3% indigo carmine decolorization in 60 min. In the kinetic studies, the half-life of indigo carmine (25 mg/l) in the remediation processes containing laccase, TiO2, laccase + TiO2, TiO2/laccase, and laccase/TiO2 was calculated as 173, 138, 161, 115, and 57 min, respectively. The degradation products by co-entrapped system were not toxic against Sorghum vulgare. The results showed two-step decolorization by co-entrapped spore laccase and TiO2 nanoparticles, including the pretreatment of dye by laccase, and then, treatment by TiO2 has potential for degradation of indigo carmine.

Microbial cell lysate supernatant (CLS) alteration impact on platinum nanoparticles fabrication, characterization, antioxidant and antibacterial activity.

Microbial mediated biological synthesis of nanoparticles is of enormous interest to modern nanotechnology due to its simplicity and eco-friendliness. In the present study, a novel green method for the synthesis of platinum nanoparticles (PtNPs) has been developed using bio-derived product-cell lysate supernatant (CLS) from various microorganisms including Gram-negative bacteria: Pseudomonas kunmingensis ADR19, Psychrobacter faecalis FZC6, Vibrio fischeri NRRL B-11177, Gram-positive bacteria: Jeotgalicoccus coquinae ZC15, Sporosarcina psychrophila KC19, Kocuria rosea MN23, genetically engineered bacterium: Pseudomonas putida KT2440 and yeast: Rhodotorula mucilaginosa CCV1. The biogenic PtNPs were characterized by UV-visible spectroscopy, X-ray diffraction (XRD), Fourier transform infrared (FTIR), transmission electron microscopy (TEM), high resolution transmission electron microscopy (HRTEM), and atomic force microscopy (AFM). The UV-visible spectra showed a red shift in the absorbance of H2PtCl6.6H2O from 260 nm to 330 nm for all prepared PtNPs. The XRD patterns of the samples indicated the formation of high purity of the cubic phase. The FTIR spectra and EDS profiles of the samples demonstrated the existence of proteins on fabricated and stabilized PtNPs. The TEM and AFM images analysis showed the synthesis of smallest PtNPs by a bacterium strain (FZC6) and yeast while genetically engineered bacteria produced the largest NPs. Also, the HRTEM analysis showed the high crystallinity of PtNPs and the interplanar spacing of 0.2 nm, corresponds to the (1 1 1) of plane of PtNPs. The results of zeta potential indicated the high stability of PtNPs in neutral pH. Moreover, the suitability of PtNPs antioxidant and antibacterial activity was correlated to the size and zeta potential of microbe used for NPs biosynthesis. In conclusion, it was found that the type of microorganisms can have influences on PtNPs characteristics and properties as Gram-negatives produced smaller PtNPs while more negatively charged NPs were obtained by Gram-positives. These findings could facilitate the selection of appropriate green approaches for more effective biotechnological production of PtNPs.

Exploring the potentials of halophilic prokaryotes from a solar saltern for synthesizing nanoparticles: The case of silver and selenium.

Halophiles are the organisms that thrive in extreme high salt environments. Despite the extensive studies on their biotechnological potentials, the ability of halophilic prokaryotes for the synthesis of nanoparticles has remained understudied. In this study, the archaeal and bacterial halophiles from a solar saltern were investigated for the intracellular/extracellular synthesis of silver and selenium nanoparticles. Silver nanoparticles were produced by the archaeal Haloferax sp. (AgNP-A, intracellular) and the bacterial Halomonas sp. (AgNP-B, extracellular), while the intracellular selenium nanoparticles were produced by the archaeal Halogeometricum sp. (SeNP-A) and the bacterial Bacillus sp. (SeNP-B). The nanoparticles were characterized by various techniques including UV-Vis spectroscopy, XRD, DLS, ICP-OES, Zeta potentials, FTIR, EDX, SEM, and TEM. The average particle size of AgNP-A and AgNP-B was 26.34 nm and 22 nm based on TEM analysis. Also, the characteristic Bragg peaks of face-centered cubic with crystallite domain sizes of 13.01 nm and 6.13 nm were observed in XRD analysis, respectively. Crystallographic characterization of SeNP-A and SeNP-B strains showed a hexagonal crystallite structure with domain sizes of 30.63 nm and 29.48 nm and average sizes of 111.6 nm and 141.6 nm according to TEM analysis, respectively. The polydispersity index of AgNP-A, AgNP-B, SeNP-A, and SeNP-B was determined as 0.26, 0.28, 0.27, and 0.36 and revealed high uniformity of the nanoparticles. All of the synthesized nanoparticles were stable and their zeta potentials were calculated as (mV): -33.12, -35.9, -31.2, and -29.34 for AgNP-A, AgNP-B, SeNP-A, and SeNP-B, respectively. The nanoparticles showed the antibacterial activity against various bacterial pathogens. The results of this study suggested that the (extremely) halophilic prokaryotes have great potentials for the green synthesis of nanoparticles.

Laccase mediator system obtained from a marine spore exhibits decolorization potential in harsh environmental conditions.

Laccases play a significant role in remedying dye pollutants. Most of these enzymes are originated from terrestrial fungi and bacteria, thus they are not proper to be used in the environments with neutral/alkaline pH, or they may require laborious extraction/purification steps. These limitations can be solved using marine spore laccases through high stability and easy to use application. In the current study, laccase activity of the marine spore -forming Bacillus sp. KC2 was measured according to the guaiacol and syringaldazine oxidation. Abiotic stresses like pH of 6, temperature of 37 °C and 0.3 mM CuSO4 (in comparison with optimal sporulation conditions: pH of 8, temperature of 20 °C and 0.0 mM CuSO4) enhanced laccase formation in sporal coat. Maximum activity of enzyme was observed at 50 °C and pH 7, which did not change in the alkaline pH and temperature range of 20-70 °C. Results indicated ions, inhibitors and solvent stability of the enzyme and its activity were stimulated by Co2+, Mn2+, PMSF, acetone, acetonitrile, ethanol, and methanol. The spore laccase could decolorize synthetic dyes from various chemical groups including azo (acid orange, amaranth, trypan blue, congo red, and amido black), indigo (indigo carmine), thiazine (methylene blue, and toluidine blue), and triarylmethane (malachite green) with ABTS/syringaldazine mediators after 5 h. Degradation products were not toxic against Sorghum vulgare and Artemia salina model organisms. The enzyme mediator system showed high potentials for dye bioremediation over a wide range of harsh conditions.

Z-Scan and Absorption Study of Crocin and Rhodamine B Decolorization by Bacterial Laccase.

Dye decolorization is currently measured by the reduction of dye absorption, based on its linear optical properties in solution. However, this method is not suitable for high dye concentrations with linear attributes in the absorption spectra. In this work, nonlinear optical properties are used to study dye decolorization of Crocin and Rhodamine B treated with a bacterial laccase. For this purpose, the Z-scan was used to obtain the nonlinear refractive indices and nonlinear absorption coefficient of Rhodamine B and Crocin in the presence and absence of the bacterial laccase enzyme. The results showed that nonlinear optical properties could be used to study dye decolorization in solutions.

Characterization of a bi-functional cellulase produced by a gut bacterial resident of Rosaceae branch borer beetle, Osphranteria coerulescens (Coleoptera: Cerambycidae).

A cellulolytic bacterium was obtained from the digestive tract of Osphranteria coerulescens. The breakdown of woody and cellulosic substances by this insect may be relative in part to its symbiont bacteria. Under optimal cultural conditions the novel isolate produced 5.35U/ml cellulase after 72h. The enzyme was purified to 36 fold with a 0.59% yield and showed a specific activity of 9.0U/mg. It presented its maximum activity at 60°C and pH 5, while it was stable in a wide range of temperature from 20 to 60°C and pH from 5 to 10. The purified enzyme had a molecular weight of 42.50kDa based on SDS-PAGE and zymogram analyses. It demonstrated high ions and solvent stability and its activity was stimulated by Mn2+, Na+, DMSO and chloroform. The enzyme could hydrolyze CMC, avicel, cellulose and sawdust. TLC analysis represented the cellobiose as the hydrolytic product of CMC. With regard to endo/exo glucanase activity and wide pH, temperature and solvent stability, it has potential for industrial application.

Oceanobacillus longus sp. nov., a moderately halophilic bacterium isolated from a salt lake.

A Gram-stain-positive, endospore-forming, long rod-shaped, strictly aerobic, moderately halophilic bacterium, designated strain T9BT, was isolated from a brine sample of the hypersaline lake Aran-Bidgol in Iran. Cells of strain T9BT were motile and produced colonies with a brown pigment. Growth occurred between 1.0 and 20 % (w/v) NaCl and the isolate grew optimally at 5.0 % (v/w) NaCl. The optimum pH and temperature for growth of the strain were pH 7.0 and 35 °C, while it was able to grow over pH and temperature ranges of pH 6.0-9.0 and 25-45 °C. Phylogenetic analysis based on the comparison of 16S rRNA gene sequences revealed that strain T9BT is a member of the genus Oceanobacillus. The closest relative to this strain was Oceanobacillus rekensis PT-11T with a similarity of 97.4 %, followed by Oceanobacillus profundus CL-MP28T and Oceanobacillus polygoni SA9T with 97.3 and 97.1 % similarity, respectively. The major cellular fatty acids of the isolate were anteiso-C15 : 0, iso-C14 : 0 and iso-C16 : 0. The polar lipids of strain T9BT consisted of phosphatidylglycerol, diphosphatidylglycerol, three phospholipids and one aminoglycolipid. It contained MK-7 as the predominant menaquinone and meso-diaminopimelic acid in the cell-wall peptidoglycan. The G+C content of the genomic DNA of this strain was 42.9 mol%. Phylogenetic analysis, DNA-DNA hybridization data and phenotypic characteristics allowed strain T9BT to be differentiated from other members of the genus Oceanobacillus. A novel species, Oceanobacillus longus sp. nov., is therefore proposed to accommodate this strain. The type strain is T9BT (=IBRC-M 10703T=LMG 29250T).

Oceanobacillus halophilus sp. nov., a novel moderately halophilic bacterium from a hypersaline lake.

A moderately halophilic bacterium was isolated from a brine sample of a hypersaline lake, Aran-Bidgol, in Iran. The strain, designated J8BT, was Gram-stain-positive, endospore-forming, rod-shaped, strictly aerobic, motile and produced cream colonies. Strain J8BT grew in NaCl at between 3.0-15.0 % (w/v) (optimally at 7.5 % NaCl, w/v), between pH 6.5-9.0 (optimally at pH 8.0) and between 20-45 °C (optimally at 35 °C). Phylogenetic analysis, based on 16S rRNA gene sequences, revealed that strain J8BT is a member of the genus Oceanobacillus and most closely related to Oceanobacillus profundus CL-MP28T, Oceanobacillus polygoni SA9T and Oceanobacillus oncorhynchi R-2T (96.9 %, 96.3 % and 96.2 % similarities, respectively). The level of DNA-DNA relatedness between the novel isolate and O. profundus IBRC-M 10567T was 10 %. The major cellular fatty acids of the isolate were anteiso-C15 : 0, iso-C15 : 0 and anteiso-C17 : 0. The polar lipid pattern of strain J8BT consisted of phosphatidylglycerol, diphosphatidylglycerol, five phospholipids, two aminolipids and two glycoaminolipids. It contained MK-7 as the predominant menaquinone and meso-diaminopimelic acid in the cell-wall peptidoglycan. The G+C content of the genomic DNA of this strain was 39.2 mol%. Phenotypic characteristics, phylogenetic analysis and DNA-DNA relatedness data suggest that this strain represents a novel species of the genus Oceanobacillus, for which the name Oceanobacillus halophilus sp. nov. is proposed. The type strain is strain J8BT ( = IBRC-M 10444T = DSM 23996T).

Halosiccatus urmianus gen. nov., sp. nov., a haloarchaeon from a salt lake.

A novel, orange-pigmented, halophilic archaeon, strain DC8T, was isolated from Urmia salt lake in north-west Iran. The cells of strain DC8T were non-motile and pleomorphic, from small rods to triangular or disc shaped. The novel strain needed at least 2.5 M NaCl and 0.02 M MgCl2 for growth. Optimal growth was achieved at 4.0 M NaCl and 0.1 M MgCl2. The optimum pH and temperature for growth were pH 7.5 and 45 °C, respectively, and it was able to grow over a pH range of 7.0 to 8.5 and a temperature range of 25 to 55 °C. Analysis of the 16S rRNA gene sequence showed that strain DC8T was a member of the family Halobacteriaceae; however, its similarity was as low as 90.1 %, 89.3 % and 89.1 % to the most closely related haloarchaeal taxa, including type species of members of the genera Halosimplex, Halobaculum and Halomicrobium, respectively. The G+C content of its DNA was 68.1 mol%. Polar lipid analyses revealed that strain DC8T contained phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and phosphatidic acid. One unknown phospholipid, two major glycolipids and one minor glycolipid were also detected. The only quinone present was MK-8 (II-H2). The physiological, biochemical and phylogenetic differences between strain DC8T and other extremely halophilic archaeal genera with validly published names supported that this strain represents a novel species of a new genus within the family Halobacteriaceae, for which the name Halosiccatus urmianus gen. nov., sp. nov. is proposed. The type strain is strain DC8T ( = IBRC-M 10911T = CECT 8793T).

Halovarius luteus gen. nov., sp. nov., an extremely halophilic archaeon from a salt lake.

An extremely halophilic archaeon, strain DA50T, was isolated from a brine sample of Urmia lake, a hypersaline environment in north-west Iran. Strain DA50T was orange-pigmented, motile, pleomorphic and required at least 2.5 M NaCl but not MgCl2 for growth. Optimal growth was achieved at 4.0 M NaCl and 0.3 M MgCl2. The optimum pH and temperature for growth were pH 7.0 and 45 °C, while it was able to grow over a pH range of 6.5-8.0 and a temperature range of 25-50 °C. Analysis of 16S rRNA gene sequences revealed that strain DA50T is a member of the family Halobacteriaceae, showing a low level of similarity with other members of this family. Highest similarities, 94.4, 94.0 and 93.9 %, were obtained with the 16S rRNA gene sequences of the type strains of Natrialba aegyptia, Halobiforma lacisalsi and Halovivax asiaticus, respectively. Polar lipid analyses revealed that strain DA50T contains phosphatidylglycerol and phosphatidylglycerol phosphate methyl ester. Four unidentified glycolipids and two minor phospholipids were also observed. The only quinone present was MK-8(II-H2). The G+C content of its DNA was 62.3 mol%. On the basis of the data obtained, the new isolate could not be classified in any recognized genus. Strain DA50T is thus considered to represent a novel species of a new genus within the family Halobacteriaceae, order Halobacteriales, for which the name Halovarius luteus gen. nov., sp. nov. is proposed. The type strain of Halovarius luteus is DA50T ( = IBRC-M 10912T = CECT 8510T).