A phenol amine molecule from Salinivenus iranica acts as the inhibitor of cancer stem cells in breast cancer cell lines.

In recent years, the anticancer properties of metabolites from halophilic microorganisms have received a lot of attention. Twenty-nine halophilic bacterial strains were selected from a culture collection to test the effects of their supernatant metabolites on stem cell-like properties of six human cancer cell lines. Human fibroblasts were used as normal control. Sphere and colony formation assay were done to assess the stem cell-like properties. invasion and migration assay, and tumor development in mice model were done to assess the anti-tumorigenesis effect in vitro and in vivo. The metabolites from Salinivenus iranica demonstrated the most potent cytotoxic effect on breast cancer cell lines (IC50 = 100 µg/mL) among all strains, with no effect on normal cells. In MDA-MB-231 cells, the supernatant metabolites enhanced both early and late apoptosis (approximately 9.5% and 48.8%, respectively) and decreased the sphere and colony formation ability of breast cancer cells. Furthermore, after intratumor injection of metabolites, tumors developed in the mice models reduced dramatically, associated with increased pro-apoptotic caspase-3 expression. The purified cytotoxic molecule, a phenol amine with a molecular weight of 1961.73 Dalton (IC50 = 1 µg/mL), downregulated pluripotency gene SRY-Box Transcription Factor 2 (SOX-2) expression in breast cancer cells which is associated with resistance to conventional anticancer treatment. In conclusion, we suggested that the phenol amine molecule from Salinivenus iranica could be a potential anti-breast cancer component.

Screening of antagonistic yeast strains for postharvest control of Penicillium expansum causing blue mold decay in table grape.

Blue mold decay caused by Penicillium expansum is one of the most important postharvest diseases of grapes, leading to considerable economic losses. Regarding the increasing demand for pesticide-free foods, this study aimed to find potential yeast strains for biological control of blue mold on table grapes. A total of 50 yeast strains were screened for antagonistic activity against P. expansum using the dual culture method and six strains significantly inhibited the fungal growth. All six yeast strains (Coniochaeta euphorbiae, Auerobasidium mangrovei, Tranzscheliella sp., Geotrichum candidum, Basidioascus persicus, and Cryptococcus podzolicus) reduced the fungal growth (29.6-85.0%) and the decay degree of wounded grape berries inoculated with P. expansum while G. candidum was found to be the most efficient biocontrol agent. On the basis of antagonistic activity, the strains were further characterized by in vitro assays involving inhibition of conidial germination, production of volatile compounds, iron competition, production of hydrolytic enzymes, biofilm-forming capacity, and exhibited three or more putative mechanisms. To our knowledge, the yeasts are reported for the first time as potential biocontrol agents against the blue mold of grapes but more study is required to evaluate their efficiency related to field application.

Establishment and Preservation of Lymphoblastoid Cell Lines from Fresh and Frozen Whole Blood and Mononuclear Cells.

Although blood cells are interesting sources for genome investigations, one of the main problems in obtaining genomic DNA from blood is the restricted amount of DNA. This obstacle can be avoided by generating Epstein-Barr virus (EBV)-induced B cell lines. This study investigates the efficiency of four different methods to generate lymphoblastoid cell lines (LCLs). Blood samples (n = 120) were obtained from donors and categorized into four groups: fresh whole blood, frozen whole blood, fresh peripheral blood mononuclear cells (PBMCs), and frozen PBMCs. The samples were followed by EBV transformation to generate LCLs. Quality control and authentication of the cells were performed using multiplex PCR and short tandem repeat (STR) analyses. Finally, we assessed the success rate and amount of time to establish the cell lines in each group. The results showed that the cells were not contaminated nor were they misidentified or cross-contaminated with other cells. The success rate of LCLs generated from the whole blood groups was lower than the PBMC groups. The freezing procedures did not have any considerable effect on the establishment of lymphoblastoid cells. These established cells have been preserved in the human and animal cell bank of the Iranian Biological Resource Center (IBRC) and are available for researchers. Due to the management and transformation of a substantial number of blood samples, we recommend that researchers freeze PBMCs for further use with high efficiency and time-saving. We suggest that whole fresh blood should be directly transformed when the volume of the blood sample is less than 0.5 ml.

Morphological identification and molecular validation of anchovies (Engraulidae) in the Persian Gulf and Oman Sea.

Validation of species using independent lines of evidence is sometimes desirable when their identification using only one approach is difficult or questionable. The identification of anchovies (Engraulidae) are often challenging based on morphology because closely related species exhibit only slight morphological differentiation. This study utilized morphological characteristics and DNA barcodes for identification and validation of anchovies in the Persian Gulf and Oman Sea. Based on morphology, we identified eight species: Thryssa hamiltonii, T. setirostris, T. vitrirostris, T. whiteheadi, T. dussumieri, Encrasicholina punctifer, E. pseudoheteroloba and Stolephorus indicus. A 658 bp region of mitochondrial cytochrome oxidase subunit I (COI) was generated for 53 specimens from these eight species. From these sequences, we built a Maximum Likelihood phylogenetic tree. In this tree, each species forms a monophyletic group confirming our initial morphological identification. In addition, we provided (and registered in GenBank) the first barcode sequences for T. whiteheadi, an endemic species of this region. Interspecies genetic distances were comprised between 0.168 to 0.275. The largest genetic distance was found between T. vitrirostris and S. indicus and the smallest between T. dussumieri and T. whiteheadi. This study successfully identified eight species of anchovies in the Persian Gulf and Oman Sea based on both morphological and molecular characters.

Planomicrobium iranicum sp. nov., a novel slightly halophilic bacterium isolated from a hypersaline wetland.

A novel strain, designated as MX6T was isolated from Meighan wetland, in the centre of Iran. The cells were Gram-stain-positive, motile, coccoid to rod-shaped, oxidase- and catalase-positive. The strain grew optimally at 35 °C, 3 % (w/v) NaCl and pH 7-7.5. A polyphasic taxonomic study was undertaken in order to characterize the strain in detail. The results of phylogenetic analysis based on 16S rRNA gene sequences indicated that MX6T represented a member of the phylum Firmicutes, family Planococcaceae, genus Planomicrobium, and showed the highest similarity with Planomicrobium flavidum ISL-41T (98.2 %) and Planomicrobium psychrophilum CMC 53orT (98.0 %). The main polar lipids of MX6T consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and seven unidentified phospholipids and its DNA G+C content was 45.5 mol%. Major cellular fatty acids were anteiso-C15 : 0, C16 : 1ω7c alcohol, iso-C14 : 0, iso-C15 : 0 and iso-C16 : 0 and the predominant respiratory quinone was Q-8 (62 %). Experimental DNA-DNA hybridization between MX6T and Planomicrobium flavidum IBRC-M 11047T was 20 %, supporting the differential taxonomic status of MX6T as representing a different taxon. All these data indicate that MX6T represents a novel species of the genus Planomicrobium, for which the name Planomicrobium iranicum sp. nov. is proposed. The type strain is MX6T (=IBRC M 10928T=LMG 28548T).

PCR-DGGE analysis of fungal community in manufacturing process of a traditional Iranian cheese.

BACKGROUND AND OBJECTIVES: The microbial communities of traditional milk-based food are of great importance in its manufacturing process, especially when using raw milk with natural cultures. Liqvan (Lighvan or Levan) is a traditional Iranian buried cheese, which is made from raw ewe's milk without a starter addition. The aim of this study was to explore the fungal active population during this cheese manufacturing process by comparing DNA and RNA based culture independent method Denaturing Gradient Gel Electrophoresis (DGGE). MATERIALS AND METHODS: Four samples of each milk, curd and ripened cheese were collected from Liqvan village located in East Azerbaijan province of Iran. Total DNA and RNA of each sample were extracted and PCR amplicons of D1 region of 26S rRNA gene was targeted for DGGE analysis. This method applied at both DNA and RNA levels in order to examine which taxonomic groups of fungi are active at each step of ripening. RESULTS: DGGE profiles of yeast amplicons showed different results between extracted DNA and RNA during ripening process. However, the main group that is present in all stages of ripening process belongs to the genus Candida although Kluyveromyces, Pichia, Galactomyces, Saccharomyces and Cryptococcus are most abundant fungi. CONCLUSION: As no starter culture added to Liqvan cheese it seems fungal diversity are mainly rely on the indigenous microbiota of milk. Furthermore, the percentage of the dominant fungal genera from the total sequences differed among DNA and cDNA libraries.

Potential of Extracellular Vesicles in Neurodegenerative Diseases: Diagnostic and Therapeutic Indications.

Extracellular vesicles (EVs) are membrane-bound vesicles, including exosomes and microvesicles. EVs are nanometer sized, found in physiological fluids such as urine, blood, cerebro-spinal fluid (CSF), with a capacity of transferring various biological materials such as microRNAs, proteins, and lipids among cells without direct cell-to-cell contact. Many cells in the nervous system have been shown to release EVs. These vesicles are involved in intercellular communication and a variety of biological processes such as modulation of immune response, signal transduction, and transport of genetic materials with low immunogenicity; therefore, they have also been recently investigated for the delivery of therapeutic molecules such as siRNAs and drugs in the treatment of diseases. In addition, since EV components reflect the physiological status of the cells and tissues producing them, they can be utilized as biomarkers for early detection of various diseases. In this review, we summarize EV application, in diagnosis as biomarker sources and as a carrier tool for drug delivery in EV-based therapies in neurodegenerative diseases.

Establishment and Characterization of Primary Cultures from Iranian Oral Squamous Cell Carcinoma Patients by Enzymatic Method and Explant Culture.

OBJECTIVES: Oral Squamous Cell Carcinoma (OSCC) is the most frequent oral cancer worldwide. It is known as the eighth most common cancer in men and as the fifth most common cancer in women. Cytogenetic and biochemical studies in recent decades have emphasized the necessity of providing an appropriate tool for such researches. Cancer cell culture is a useful tool for investigations on biochemical, genetic, molecular and immunological characteristics of different cancers, including oral cancer. Here, we explain the establishment process of five primary oral cancer cells derived from an Iranian population. MATERIALS AND METHODS: The specimens were obtained from five oral cancer patients. Enzymatic, explant culture and magnetic-activated cell sorting (MACS) methods were used for cell isolation. After quality control tests, characterization and authentication of primary oral cancer cells were performed by short tandem repeats (STR) profiling, chromosome analysis, species identification, and monitoring the growth, morphology and the expression of CD326 and CD133 markers. RESULTS: Five primary oral cancer cells were established from an Iranian population. The flow cytometry results showed that the isolated cells were positive for CD326 and CD133 markers. Furthermore, the cells were free from mycoplasma, bacterial and fungal contamination. No misidentified or cross-contaminated cells were detected by STR analysis. CONCLUSIONS: Human primary oral cancer cells provide an extremely useful platform for studying carcinogenesis pathways of oral cancer in Iranian population. They may be helpful in explaining the ethnic differences in cancer biology and the individuality in anticancer drug response in future studies.

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).

Halovivax cerinus sp. nov., an extremely halophilic archaeon from a hypersaline lake.

An extremely halophilic archaeon, strain IC35(T), was isolated from a mud sample of the Aran-Bidgol salt lake in Iran. The novel strain was cream, non-motile, rod-shaped and required at least 2.5 M NaCl, but not MgCl2, for growth. Optimal growth was achieved with 3.4 M NaCl and 0.1 M MgCl2. The optimum pH and temperature for growth were pH 7.0 (grew over a pH range of 6.5-9.0) and 40 °C (grew over a temperature range of 30-50 °C), respectively. Analysis of 16S rRNA gene sequences revealed that strain IC35(T) clustered with species of the genus Halovivax, with sequence similarities of 97.3%, 96.6% and 96.3%, respectively, to Halovivax limisalsi IC38(T), Halovivax asiaticus EJ-46(T) and Halovivax ruber XH-70(T). The rpoB' gene similarities between the novel strain and Halovivax limisalsi IBRC-M 10022(T), Halovivax ruber JCM 13892(T) and Halovivax asiaticus JCM 14624(T) were 90.2 %, 90.2% and 89.9%, respectively. The polar lipid pattern of strain IC35(T) consisted of phosphatidylglycerol and phosphatidylglycerol phosphate methyl ester; six unknown glycolipids and two minor phospholipids were also observed. The only quinone present was MK-8 (II-H2). The G+C content of the genomic DNA was 63.2 mol%. DNA-DNA hybridization studies (29% hybridization with Halovivax limisalsi IBRC-M 10022(T)), as well as biochemical and physiological characterization, allowed strain IC35(T) to be differentiated from other species of the genus Halovivax. A novel species, Halovivax cerinus sp. nov., is therefore proposed to accommodate this strain. The type strain is IC35(T) ( = IBRC-M 10256(T) = KCTC 4050(T)).

Salinispirillum marinum gen. nov., sp. nov., a haloalkaliphilic bacterium in the family 'Saccharospirillaceae'.

A novel Gram-staining-negative, motile, non-pigmented, facultatively anaerobic, spirillum-shaped, halophilic and alkaliphilic bacterium, designated strain GCWy1(T), was isolated from water of the coastal-marine wetland Gomishan in Iran. The strain was able to grow at NaCl concentrations of 1-10% (w/v) and optimal growth was achieved at 3% (w/v). The optimum pH and temperature for growth were pH 8.5 and 30 °C, while the strain was able to grow at pH 7.5-10 and 4-40 °C. Phylogenetic analysis based on the comparison of the 16S rRNA gene sequence placed the isolate within the class Gammaproteobacteria as a separate deep branch, with 92.1% or lower sequence similarity to representatives of the genera Saccharospirillum and Reinekea and less than 91.0% sequence similarity with other remotely related genera. The major cellular fatty acids of the isolate were C(18 : 1)ω7c, C(16:0) and C(17 : 0), and the major components of its polar lipid profile were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. The cells of strain GCWy1(T) contained the isoprenoid quinones Q-9 and Q-8 (81% and 2%, respectively). The G+C content of the genomic DNA of this strain was 52.3 mol%. On the basis of 16S rRNA gene sequence analysis in combination with chemotaxonomic and phenotypic data, strain GCWy1(T) represents a novel species in a new genus in the family 'Saccharospirillaceae', order Oceanospirillales, for which the name Salinispirillum marinum gen. nov., sp. nov. is proposed. The type strain of the type species is GCWy1(T) ( = IBRC-M 10765(T) =CECT 8342(T)).

Halorientalis persicus sp. nov., an extremely halophilic archaeon isolated from a salt lake and emended description of the genus Halorientalis.

An extremely halophilic archaeon, strain D108(T), was isolated from a brine sample of Aran-Bidgol salt lake in Iran. The novel strain was cream-pigmented, motile, pleomorphic rod-shaped and required at least 2.5 M NaCl but not MgCl2 for growth. Optimal growth was achieved with 4.3 M NaCl and 0.1 M MgCl2. The optimum pH and temperature for growth were pH 7.5 and 40 °C, respectively, and the strain was able to grow over a pH range of 6.5 to 9.0, and a temperature range of 30 to 50 °C. Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain D108(T) clustered with the type strain of the sole species of the genus Halorientalis, Halorientalis regularis TNN28(T), with a sequence similarity of 98.8 %. The polar lipid pattern of strain D108(T) consisted of phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, one phosphoglycolipid and two glycolipids. The only quinone present was MK-8(II-H2). The DNA G+C content of strain D108(T) was 62.8 mol%. DNA-DNA hybridization studies (45 % with Halorientalis regularis IBRC-M 10760(T)), as well as biochemical and physiological characterization, allowed strain D108(T) to be differentiated from Halorientalis regularis. A novel species of the genus Halorientalis, Halorientalis persicus sp. nov., is therefore proposed to accommodate this strain. The type strain is D108(T) ( = IBRC-M 10043(T) = CECT 8375(T)). An emended description of the genus Halorientalis is also proposed.

Alloactinosynnema iranicum sp. nov., a rare actinomycete isolated from a hypersaline wetland, and emended description of the genus Alloactinosynnema.

A Gram-staining-positive actinobacterial strain, Chem10(T), was isolated from soil around Inche-Broun hypersaline wetland in the north of Iran. Strain Chem10(T) was strictly aerobic, and catalase- and oxidase-positive. The isolate grew with 0-3 % NaCl, at 20-40 °C and at pH 6.0-8.0. The optimum temperature and pH for growth were 30 °C and pH 7.0, respectively. The cell wall of strain Chem10(T) contained meso-diaminopimelic acid as diamino acid and galactose, ribose and arabinose as whole-cell sugars. The polar lipid pattern contained diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. Strain Chem10(T) synthesized cellular fatty acids of the straight-chain saturated and mono-unsaturated, and iso- and anteiso-branched types C14 : 0, C16 : 0, iso-C16 : 1, anteiso-C17 : 0, iso-C16 : 0, iso-C14 : 0 and iso-C15 : 0, and the major respiratory quinone was MK-9(H4). The G+C content of the genomic DNA was 70.7 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain Chem10(T) belonged to the family Pseudonocardiaceae and showed the closest phylogenetic similarity to Alloactinosynnema album KCTC 19294(T) (98.3 %) and Actinokineospora cibodasensis DSM 45658(T) (97.9 %). DNA-DNA relatedness values between the novel strain and strains Alloactinosynnema album KCTC 19294(T) and Actinokineospora cibodasensis DSM 45658(T) were only 52 % and 23 %, respectively. On the basis of phylogenetic analysis, phenotypic characteristics and DNA-DNA hybridization data, a novel species of the genus Alloactinosynnema is proposed, Alloactinosynnema iranicum sp. nov. The type strain is Chem10(T) ( = IBRC-M 10403(T) = CECT 8209(T)). In addition, an emended description of the genus Alloactinosynnema is proposed.

Halopenitus malekzadehii sp. nov., an extremely halophilic archaeon isolated from a salt lake.

Strain CC65(T), a novel extremely halophilic archaeon, was isolated from a brine sample of a salt lake in Iran. The novel strain was light yellow-pigmented, non-motile, pleomorphic and required at least 1.7 M NaCl and 0.02 M MgCl2 for growth. Optimal growth was achieved at 3.5 M NaCl and 0.4 M MgCl2. The optimum pH and temperature for growth were pH 7.5 and 40 °C, respectively, while it was able to grow over a pH and a temperature range of pH 6.5-9.0 and 30-50 °C, respectively. Analysis of 16S rRNA gene sequence revealed that strain CC65(T) clustered with the sole member of the genus Halopenitus, Halopenitus persicus DC30(T) with a sequence similarity of 98.0%. The polar lipid profile of strain CC65(T) consisted of phosphatidylglycerol and phosphatidylglycerol phosphate methyl ester. An unidentified glycolipid and two minor phospholipids were also observed. The only quinone present was MK-8(II-H2). The DNA G+C content of strain CC65(T) was 63.8 mol%. On the basis of the biochemical and physiological characteristics, as well as DNA-DNA hybridization (44% with Halopenitus persicus IBRC 10041(T)), strain CC65(T) is classified as a novel species of the genus Halopenitus, for which the name Halopenitus malekzadehii sp. nov. is proposed. The type strain is CC65(T) ( = IBRC-M 10418(T) =KCTC 4045(T)).

Bacillus salsus sp. nov., a halophilic bacterium from a hypersaline lake.

A Gram-staining-positive, endospore-forming, rod-shaped, strictly aerobic, slightly halophilic bacterium, designated strain A24(T), was isolated from the hypersaline lake Aran-Bidgol in Iran. Cells of strain A24(T) were motile rods and produced oval endospores at a terminal position in swollen sporangia. Strain A24(T) was catalase and oxidase positive. Growth occurred with between 0.5 and 7.5% (w/v) NaCl and the isolate grew optimally at 3% (v/w) NaCl. The optimum temperature and pH for growth were 35 °C and pH 8.0, respectively. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain A24(T) belonged to the genus Bacillus within the phylum Firmicutes and showed the closest phylogenetic similarity with the species Bacillus alkalitelluris BA288(T) (97.2%), Bacillus herbersteinensis D-1,5a(T) (96.0%) and Bacillus litoralis SW-211(T) (95.6%). The G+C content of the genomic DNA of this strain was 35.9 mol%. The polar lipid pattern of strain A24(T) consisted of phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and two unknown phospholipids. The major cellular fatty acids of strain A24(T) were anteiso-C(15:0) and iso-C(15:0). The respiratory quinones were MK-7 (94%) and MK-6 (4%). The peptidoglycan contained meso-diaminopimelic acid as the diagnostic diamino acid. All these features confirm the placement of isolate A24(T) within the genus Bacillus. DNA-DNA hybridization experiments revealed a relatedness of 8% between strain A24(T) and Bacillus alkalitelluris IBRC-M 10596(T), supporting its placement as a novel species. Phenotypic characteristics, phylogenetic analysis and DNA-DNA relatedness data suggest that this strain represents a novel species of the genus Bacillus, for which the name Bacillus salsus sp. nov. is proposed. The type strain is strain A24(T) ( = IBRC-M 10078 (T) = KCTC 13816(T)).

Bacillus halosaccharovorans sp. nov., a moderately halophilic bacterium from a hypersaline lake.

A novel Gram-stain-positive, moderately halophilic bacterium, designated strain E33(T), was isolated from water of the hypersaline lake Aran-Bidgol in Iran and characterized taxonomically using a polyphasic approach. Cells of strain E33(T) were motile rods and produced ellipsoidal endospores at a central or subterminal position in swollen sporangia. Strain E33(T) was a strictly aerobic bacterium, catalase- and oxidase-positive. The strain was able to grow at NaCl concentrations of 0.5-25 % (w/v), with optimum growth occurring at 5-15 % (w/v) NaCl. The optimum temperature and pH for growth were 40 °C and pH 7.5-8.0, respectively. On the basis of 16S rRNA gene sequence analysis, strain E33(T) was shown to belong to the genus Bacillus within the phylum Firmicutes and showed the closest phylogenetic similarity with the species Bacillus niabensis 4T19(T) (99.2 %), Bacillus herbersteinensis D-1-5a(T) (97.3 %) and Bacillus litoralis SW-211(T) (97.2 %). The DNA G+C content of the type strain of the novel species was 42.6 mol%. The major cellular fatty acids of strain E33(T) were anteiso-C15 : 0 and iso-C15 : 0, and the polar lipid pattern consisted of diphosphatidylglycerol, phosphatidylglycerol, two unknown glycolipids, an unknown lipid and an unknown phospholipid. The isoprenoid quinones were MK-7 (97 %), MK-6 (2 %) and MK-8 (0.5 %). The peptidoglycan contained meso-diaminopimelic acid as the diagnostic diamino acid. All these features confirm the placement of isolate E33(T) within the genus Bacillus. DNA-DNA hybridization experiments revealed low levels of relatedness between strain E33(T) and Bacillus niabensis IBRC-M 10590(T) (22 %), Bacillus herbersteinensis CCM 7228(T) (38 %) and Bacillus litoralis DSM 16303(T) (19 %). On the basis of polyphasic evidence from this study, a novel species of the genus Bacillus, Bacillus halosaccharovorans sp. nov. is proposed, with strain E33(T) (= IBRC-M 10095(T) = DSM 25387(T)) as the type strain.

Limimonas halophila gen. nov., sp. nov., an extremely halophilic bacterium in the family Rhodospirillaceae.

A novel, Gram-staining-negative, non-pigmented, rod-shaped, strictly aerobic, extremely halophilic bacterium, designated strain IA16(T), was isolated from the mud of the hypersaline Lake Aran-Bidgol, in Iran. Cells of strain IA16(T) were not motile. Growth occurred with 2.5-5.2 M NaCl (optimum 3.4 M), at pH 6.0-8.0 (optimum pH 7.0) and at 30-50 °C (optimum 40 °C). Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain IA16(T) belonged in the family Rhodospirillaceae and that its closest relatives were Rhodovibrio sodomensis DSM 9895(T) (91.6 % sequence similarity), Rhodovibrio salinarum NCIMB 2243(T) (91.2 %), Pelagibius litoralis CL-UU02(T) (88.9 %) and Fodinicurvata sediminis YIM D82(T) (88.7 %). The novel strain's major cellular fatty acids were C19 : 0 cyclo ω7c and C18 : 0 and its polar lipid profile comprised phosphatidylglycerol, diphosphatidylglycerol, four unidentified phospholipids, three unidentified aminolipids and two other unidentified lipids. The cells of strain IA16(T) contained the ubiquinone Q-10. The G+C content of the novel strain's genomic DNA was 67.0 mol%. The physiological, biochemical and phylogenetic differences between strain IA16(T) and other previously described taxa indicate that the strain represents a novel species in a new genus within the family Rhodospirillaceae, for which the name Limimonas halophila gen. nov., sp. nov. is proposed. The type strain of Limimonas halophila is IA16(T) ( = IBRC-M 10018(T)  = DSM 25584(T)).