Diversity of actinomycete and their metabolites isolated from Howz Soltan Lake, Iran.

Saline environments are largely unexplored sources of actinomycetes with the potential to produce biologically active secondary metabolites. A total of 34 actinomycete isolates from water, sediments and mostly rhizosphere (82%) were collected from different sites at Howz Soltan Lake in Iran. Based on phylogenetic analysis, the isolates belonged to the genera Streptomyces, Nocardia and Saccharomonospora. Cytotoxic assay revealed extract from isolate act9 as the most potent (19.716±5.72 µg/ml) against the MDA-MB-231 human breast cancer cell. Also, 38% of the isolates showed antimicrobial activity against some of the test microorganisms. The ethyl-acetate extract of isolate act18 showed the strongest antibacterial effect against Staphylococcus aureus and MRSA, and was further analyzed by GC/MS. Ar-tumerone (26.41%) and butyl isodecyl phthalate (21.77 %) were the main constituents detected in the extract. This is the first time Ar-tumerone is being detected in a prokaryote. Isolate act18 showed a high 16S rRNA sequence similarity to that of Streptomyces youssoufiensis DSM 41920. In addition, a number of the isolates produced different enzymes including lipase, amylase, protease, gelatinase, urease and lecithinase. Some of the isolates belonging to the genera Streptomyces and Nocardia exhibited plant growth promoting activity such as increased seed germination, stem length and the number of Echium leaves during the 20 days. Findings from this study indicated the diversity and biosynthetic potential of actinomycetes from saline environment.

Molecular investigation of bacterial communities during the manufacturing and ripening of semi-hard Iranian Liqvan cheese.

Liqvan (or Lighvan) is a traditional Iranian cheese from the East Azerbaijan province of Iran, which is made of raw ewe's milk without the addition of a starter. The grazing pastures, environmental conditions and the ancient regional production methods allocate a distinctive microbial ecology to this type of cheese, and these factors are consequently associated with the quality of the product. In this study, the microbiota of the milk, curd and cheese has been investigated using culture independent approaches. Denaturing gradient gel electrophoresis (DGGE) of the bacteria, 16S rRNA based high-throughput sequencing and enumeration of the live bacterial community by means of quantitative PCR (qPCR) have been used for this purpose. The results showed that the main bacterial population in the milk belonged to both microbial contaminants and lactic acid bacteria (LAB). However, both of these populations were totally replaced by LAB during ripening. The present survey contributes by describing the microbiota of this ancient cheese in more detail during fermentation and ripening.

Isolation and identification of culturable halophilic bacteria with producing hydrolytic enzyme from Incheh Broun hypersaline wetland in Iran.

Incheh Broun hypersaline wetland is located near the border of Turkmenistan in thenorth of Iran. This wetland is notable because of salinity (280g/l) and alteration in pH range (2.8 to 6.8). Eastern part of wetland is affected by wastewater of iodine extraction factory.  Samples were taken from soil, water and salt. Totally, 400 bacterial strains were isolated of which 194 strains were Gram-positive bacilli, 184 strains were Gram-negative rod and 22 strains were Gram-positive cocci. According to phylogenetic analysis of 16S rRNA, selected strains were placed in three taxonomic phyla including Firmicutes, Actinobacteria and Gammaproteobacteria. Optimum growth was evaluated for salt and 22 strains were found to be moderate halophile and 33 strains were halotolerant. Production of lipase, amylase, gelatinase and protease was examined. Gram-positive bacilli were the main producers of hydrolytic enzymes. Gelatinase and protease were the most frequent enzymes. Gram-positive cocci were the main producers of lipase but they didn't produce amylase.

Fungal Planet description sheets: 281-319.

Novel species of fungi described in the present study include the following from South Africa: Alanphillipsia aloeicola from Aloe sp., Arxiella dolichandrae from Dolichandra unguiscati, Ganoderma austroafricanum from Jacaranda mimosifolia, Phacidiella podocarpi and Phaeosphaeria podocarpi from Podocarpus latifolius, Phyllosticta mimusopisicola from Mimusops zeyheri and Sphaerulina pelargonii from Pelargonium sp. Furthermore, Barssia maroccana is described from Cedrus atlantica (Morocco), Codinaea pini from Pinus patula (Uganda), Crucellisporiopsis marquesiae from Marquesia acuminata (Zambia), Dinemasporium ipomoeae from Ipomoea pes-caprae (Vietnam), Diaporthe phragmitis from Phragmites australis (China), Marasmius vladimirii from leaf litter (India), Melanconium hedericola from Hedera helix (Spain), Pluteus albotomentosus and Pluteus extremiorientalis from a mixed forest (Russia), Rachicladosporium eucalypti from Eucalyptus globulus (Ethiopia), Sistotrema epiphyllum from dead leaves of Fagus sylvatica in a forest (The Netherlands), Stagonospora chrysopyla from Scirpus microcarpus (USA) and Trichomerium dioscoreae from Dioscorea sp. (Japan). Novel species from Australia include: Corynespora endiandrae from Endiandra introrsa, Gonatophragmium triuniae from Triunia youngiana, Penicillium coccotrypicola from Archontophoenix cunninghamiana and Phytophthora moyootj from soil. Novelties from Iran include Neocamarosporium chichastianum from soil and Seimatosporium pistaciae from Pistacia vera. Xenosonderhenia eucalypti and Zasmidium eucalyptigenum are newly described from Eucalyptus urophylla in Indonesia. Diaporthe acaciarum and Roussoella acacia are newly described from Acacia tortilis in Tanzania. New species from Italy include Comoclathris spartii from Spartium junceum and Phoma tamaricicola from Tamarix gallica. Novel genera include (Ascomycetes): Acremoniopsis from forest soil and Collarina from water sediments (Spain), Phellinocrescentia from a Phellinus sp. (French Guiana), Neobambusicola from Strelitzia nicolai (South Africa), Neocladophialophora from Quercus robur (Germany), Neophysalospora from Corymbia henryi (Mozambique) and Xenophaeosphaeria from Grewia sp. (Tanzania). Morphological and culture characteristics along with ITS DNA barcodes are provided for all taxa.

Halovenus aranensis gen. nov., sp. nov., an extremely halophilic archaeon from Aran-Bidgol salt lake.

A novel red-pigmented halophilic archaeon, strain EB27(T), was isolated from Aran-Bidgol salt lake, a hypersaline playa in Iran. Cells of strain EB27(T) were non-motile and pleomorphic (rods to triangular or disc-shaped). Strain EB27(T) required at least 2.5 M NaCl and 0.1 M MgCl(2) for growth. Optimal growth was achieved at 4 M NaCl and 0.5 M MgCl(2). The optimum pH and temperature for growth were pH 7.5 and 40 °C; it was able to grow at pH 6.0-8.0 and 25-50 °C. 16S rRNA gene sequence analysis showed that strain EB27(T) is a member of the family Halobacteriaceae; however, levels of 16S rRNA gene sequence similarity were as low as 90.0, 89.3 and 89.1 % to the most closely related haloarchaeal taxa, namely Halalkalicoccus tibetensis DS12(T), Halosimplex carlsbadense 2-9-1(T) and Halorhabdus utahensis AX-2(T), respectively. The DNA G+C content of strain EB27(T) was 61 mol%. Strain EB27(T) contained phosphatidylglycerol and phosphatidylglycerol phosphate methyl ester, common phospholipids found in haloarchaea, together with two minor phospholipids. The only quinone present was MK-8(II-H(2)). Physiological, biochemical and phylogenetic differences between strain EB27(T) and recognized genera of extremely halophilic archaea suggest that this strain represents a novel species in a new genus within the family Halobacteriaceae, for which the name Halovenus aranensis gen. nov., sp. nov. is proposed. The type strain of Halovenus aranensis, the type species of the new genus, is strain EB27(T) ( = IBRC-M 10015(T) = CGMCC 1.11001(T)).

Haloarchaeobius iranensis gen. nov., sp. nov., an extremely halophilic archaeon isolated from a saline lake.

Strain EB21(T) was isolated from a brine sample from Aran-Bidgol salt lake, a saline playa in Iran. Strain EB21(T) was an orange-red-pigmented, motile rod and required at least 2 M NaCl but not MgCl(2) for growth. Optimal growth was achieved at 3.5 M NaCl and 0.2 M MgCl(2). The optimum pH and temperature for growth were pH 7.5 and 40 °C, while it was able to grow at pH 6.0-8.0 and 25-55 °C. Analysis of the 16S rRNA gene sequence revealed that strain EB21(T) is a member of the family Halobacteriaceae, showing low levels of similarity to other members of the family. The highest sequence similarities, 91.8, 91.7 and 91.5 %, were obtained with the 16S rRNA gene sequences of the type strains of Halobiforma lacisalsi, Haloterrigena thermotolerans and Halalkalicoccus tibetensis, respectively. Polar lipid analyses revealed that strain EB21(T) contains phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and phosphatidylglycerol sulfate. Three unidentified glycolipids and one minor phospholipid were also observed. The only quinone present was MK-8(II-H(2)). The G+C content of its DNA was 67.7 mol%. On the basis of the data obtained, the new isolate could not be classified in any recognized genus. Strain EB21(T) is thus considered to represent a novel species in a new genus within the family Halobacteriaceae, order Halobacteriales, for which the name Haloarchaeobius iranensis gen. nov., sp. nov. is proposed. The type strain of Haloarchaeobius iranensis is EB21(T) ( = IBRC-M 10013(T)  = KCTC 4048(T)).

Bacillus iranensis sp. nov., a moderate halophile from a hypersaline lake.

A Gram-positive, moderately halophilic rod, designated X5BT, was isolated from saline mud of the hypersaline lake Aran-Bidgol in Iran. Strain X5BT was a strictly aerobic, motile bacterium that produced ellipsoidal endospores at a central-subterminal position in non-swollen sporangia. The isolate grew at pH 7.0-10.0 (optimum pH 7.5), at 25-45 °C (optimum 35 °C) and with 2.5-15 % (w/v) NaCl (optimum 5-7.5 %). On the basis of 16S rRNA gene sequences, strain X5BT belonged to the genus Bacillus and showed highest similarity with Bacillus persepolensis HS136T (95.6 % 16S rRNA gene sequence similarity) and Bacillus salarius BH169T (95.5 %). The DNA G+C content was 42.4 mol%. The major cellular fatty acids were anteiso-C15:0 and iso-C15:0 and the polar lipid profile consisted of phosphatidylglycerol, diphosphatidylglycerol, three phospholipids and two glycolipids. The diamino acid found in the cell-wall peptidoglycan was meso-diaminopimelic acid and the isoprenoid quinones were MK-7 (92 %), MK-6 (6 %) and MK-5 (2 %). On the basis of phylogenetic, chemotaxonomic and phenotypic data, a novel species of the genus Bacillus is proposed, with the name Bacillus iranensis sp. nov. The type strain is X5BT (=IBRC 10446T=DSM 23995T).

Isolation of moderately halophilic pseudoalteromonas producing extracellular hydrolytic enzymes from persian gulf.

Extracellular hydrolytic enzymes such as amylases, proteases, lipases and DNases have quite diverse potential usages in different areas such as food industry, biomedical sciences and chemical industries, also it would be of great importance to have available enzymes showing optimal activities at different values of salt concentrations and temperature. Halophiles are the most likely source of such enzymes, because not only their enzymes are salt-tolerant, but many are also thermotolerant. The purpose of this study was isolation of hydrolytic extracellular enzyme producing halophilic bacteria from water and sediment of the Persian Gulf. Isolated bacteria from water and sediment were inoculated in media with concentration of 0-20% NaCl to determine the optimum salt concentration for growth, isolates were also inoculated in 4 types of solid medium containing substrates of 3 extracellular hydrolytic enzymes including amylase, Protease and Lipase, to determine the quantitative detection of enzyme production, selected strains after more accurate physiological and biochemical studies were identified regarding phylogeny and molecular characteristics using 16S rRNA technique. Isolated enzyme producing bacteria belong to Pseudoalteromonas genera.

Lentibacillus persicus sp. nov., a moderately halophilic species isolated from a saline lake.

A Gram-staining-positive, moderately halophilic bacterium, designated strain Amb31(T), was isolated from water of the hypersaline lake Aran-Bidgol in Iran and characterized taxonomically using a polyphasic approach. Cells were rods, motile and able to produce ellipsoidal endospores at a central position in swollen sporangia. Strain Amb31(T) was facultatively anaerobic and catalase- and oxidase-positive. The strain grew in a complex medium supplemented with 3-25 % (w/v) NaCl (optimum 7.5-10 %). Optimal growth was at 30-35 degrees C and pH 7.5. Phylogenetic analysis based on 16S rRNA gene sequence comparisons showed that strain Amb31(T) belonged to the genus Lentibacillus; it exhibited 16S rRNA gene sequence similarity values of 96.8 and 96.4 % to Lentibacillus salicampi SF-20(T) and Lentibacillus salinarum AHS-1(T), respectively, and values of 95.9-94.7 % to the type strains of other recognized species of Lentibacillus. The cell-wall peptidoglycan of strain Amb31(T) was based on meso-diaminopimelic acid and MK-7 was the respiratory isoprenoid quinone. The major fatty acids were anteiso-C(15 : 0) (44.7 %), iso-C(16 : 0) (21.4 %) and anteiso-C(17 : 0) (15.9 %) and the polar lipid pattern consisted of phosphatidylglycerol, diphosphatidylglycerol, five phospholipids and a glycolipid. The DNA G+C content was 44.1 mol%. All these features confirmed the placement of strain Amb31(T) within the genus Lentibacillus and the strain could be clearly differentiated from strains of the other species of Lentibacillus on the basis of several phenotypic, genotypic and chemotaxonomic features. DNA-DNA relatedness with the type strain of the most closely related strain, L. salicampi DSM 16425(T), was 28 %. Therefore, strain Amb31(T) represents a novel species of the genus Lentibacillus, for which the name Lentibacillus persicus sp. nov. is proposed. The type strain is Amb31(T) (=CCM 7683(T) =CECT 7524(T) =DSM 22530(T) =LMG 25304(T)).

Piscibacillus halophilus sp. nov., a moderately halophilic bacterium from a hypersaline Iranian lake.

A Gram-positive, moderately halophilic bacterium, designated strain HS224(T), was isolated from the hypersaline lake Howz-Soltan in Iran. Cells of strain HS224(T) were rod-shaped, motile and produced oval endospores. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain HS224(T) was affiliated to the genus Piscibacillus, exhibiting 98.5 % sequence similarity to the type strain of Piscibacillus salipiscarius. Strain HS224(T) was also related closely to the type strains of Aquisalibacillus elongatus (98.0 % 16S rRNA gene sequence similarity), Filobacillus milosensis (97.9 %) and Tenuibacillus multivorans (97.0 %). Strain HS224(T) was able to grow at NaCl concentrations of 1-20 % (w/v), with optimum growth occurring at 10 % (w/v) NaCl. The optimum temperature and pH for growth were 35 degrees C and pH 7.5. Major polar lipids were phosphatidylglycerol and diphosphatidylglycerol, the isoprenoid quinone was MK-7 and the peptidoglycan type was A1gamma, with meso-diaminopimelic acid as the diagnostic diamino acid; these characteristics were shared with P. salipiscarius. The major cellular fatty acids of strain HS224(T) were anteiso-C(15 : 0), iso-C(15 : 0), anteiso-C(17 : 0) and iso-C(16 : 0). The G+C content of the DNA was 37.5 mol%. The level of DNA-DNA relatedness between strain HS224(T) and P. salipiscarius JCM 13188(T) was 30.8 %. It is evident from the genotypic, chemotaxonomic and phenotypic data presented that strain HS224(T) represents a novel species of the genus Piscibacillus, for which the name Piscibacillus halophilus sp. nov. is proposed. The type strain is HS224(T) (=CCM 7596(T)=DSM 21633(T)=JCM 15721(T)=LMG 24786(T)).

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

A Gram-positive, moderately halophilic, endospore-forming bacterium, designated strain HS136T, was isolated from the hypersaline lake Howz-Soltan in Iran. Cells were motile rods, producing ellipsoidal endospores at a central-subterminal position in non-swollen sporangia. Strain HS136T, a strictly aerobic bacterium, grew between pH 7.0 and 10.0 (optimal growth at pH 8.0-8.5), between 25 and 45 degrees C (optimal growth at 40 degrees C) and at salinities of 5-20% (w/v) NaCl, growing optimally at 10% (w/v) NaCl. On the basis of 16S rRNA gene sequence analysis, strain HS136T was shown to belong to the genus Bacillus within the phylum Firmicutes and showed closest phylogenetic similarity to Bacillus salarius BH169T (95.2%) and Bacillus qingdaonensis CM1T (94.5%). The DNA G+C content of this new isolate was 37.1 mol%. The major cellular fatty acids of strain HS136T were iso-C15:0, anteiso-C15:0 and anteiso-C17:0, and its polar lipid pattern consisted of phosphatidylglycerol and diphosphatidylglycerol. The isoprenoid quinone was MK-7. The peptidoglycan type is A1gamma, with meso-diaminopimelic acid as the diagnostic diamino acid. On the basis of polyphasic evidence from this study, Bacillus persepolensis sp. nov. is proposed, with strain HS136T (=CCM 7595T=DSM 21632T=JCM 15720T=LMG 25222T) as the type strain.

Thalassobacillus cyri sp. nov., a moderately halophilic Gram-positive bacterium from a hypersaline lake.

A Gram-positive, moderately halophilic bacterium, designated strain HS286(T), was isolated from water of the hypersaline Lake Howz-Soltan in Iran. Cells were strictly aerobic, rod-shaped, motile and able to produce ellipsoidal endospores at a central-subterminal position in swollen sporangia. Isolate HS286(T) grew in a complex medium supplemented with 1-15 % (w/v) NaCl, with optimum growth at 8.0 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequence comparisons showed that strain HS286(T) was closely related to Thalassobacillus devorans G-19.1(T) (99.4 % gene sequence similarity). The other closest species were Halobacillus yeomjeoni MSS-402(T) (96.9 %) and other species of the genus Halobacillus (with 96.7-93.5 % similarity). Strain HS286(T) had cell-wall peptidoglycan based on meso-diaminopimelic acid and MK-7 as the respiratory isoprenoid quinone. The major fatty acids were anteiso-C(15 : 0) (43.8 %), iso-C(16 : 0) (21.4 %), iso-C(14 : 0) (9.4 %), anteiso-C(17 : 0) (8.7 %) and iso-C(15 : 0) (7.0 %) and the polar lipid pattern consisted of diphosphatidylglycerol, phosphatidylglycerol, two phospholipids and a glycolipid. The DNA G+C content was 43.0 mol%. All of these features confirmed the placement of isolate HS286(T) within the genus Thalassobacillus. However DNA-DNA hybridization between strain HS286(T) and the only recognized species of the genus Thalassobacillus, T. devorans G-19.1(T), was 27.3 %, showing unequivocally that the novel isolate constituted a new genospecies. Strain HS286(T) could be clearly differentiated from T. devorans and other phylogenetic neighbours on the basis of several phenotypic, genotypic and chemotaxonomic features. Therefore, strain HS286(T) constitutes a novel species, for which the name Thalassobacillus cyri sp. nov. is proposed. The type strain is HS286(T) (=CCM 7597(T)=JCM 15722(T)).

Bioemulsifier production by a halothermophilic Bacillus strain with potential applications in microbially enhanced oil recovery.

A halothermotolerant Gram-positive spore-forming bacterium was isolated from petroleum reservoirs in Iran and identified as Bacillus licheniformis sp. strain ACO1 by phenotypic characterization and 16S rRNA analysis. It showed a high capacity for bioemulsifier production and grew up to 60 degrees C with NaCl at 180 g l(-1). The optimum NaCl concentration, pH and temperature for bioemulsifier production were 4% (w/v), 8.0, and 45 degrees C, respectively. Although ACO1 did not utilize hydrocarbons, it had a high emulsifying activity (E (24) = 65 +/- 5%) on different hydrophobic substrates. Emulsification was optimal while growing on yeast extract as the sole carbon source and NaNO(3) as the nitrogen source. The efficiency of the residual oil recovery increased by 22% after in situ growth of B. licheniformis ACO1 in a sand-pack model saturated with liquid paraffin.

Decolorization of textile azo dyes by newly isolated halophilic and halotolerant bacteria.

Studies were carried out on the decolorization of textile azo dyes by newly isolated halophilic and halotolerant bacteria. Among the 27 strains of halophilic and halotolerant bacteria isolated from effluents of textile industries, three showed remarkable ability in decolorizing the widely utilized azo dyes. Phenotypic characterization and phylogenetic analysis based on 16S rDNA sequence comparisons indicate that these strains belonged to the genus Halomonas. The three strains were able to decolorize azo dyes in a wide range of NaCl concentration (up to 20%w/v), temperature (25-40 degrees C), and pH (5-11) after 4 days of incubation in static culture. They could decolorize the mixture of dyes as well as pure dyes. These strains also readily grew in and decolorized the high concentrations of dye (5000 ppm) and could tolerate up to 10,000 ppm of the dye. UV-Vis analyses before and after decolorization and the colorless bacterial biomass after decolorization suggested that decolorization was due to biodegradation, rather than inactive surface adsorption. Analytical studies based on HPLC showed that the principal decolorization was reduction of the azo bond, followed by cleavage of the reduced bond.

Assaying the presence of histone-like protein HU in Halobacillus karajensis.

Histone-Like Proteins (HLPs) in bacteria are small basic proteins that contribute to the control of gene expression, recombination, DNA replication and compressing the bacterial DNA in the nucleoid. Among the HLPs, HU protein as a dimer plays an important role in binding to DNA and bending it. In this study, we showed that a 9.5-10 kDa protein with the same electrophoretic mobility as HU exists in Halobacillus karajensis which is a novel gram positive moderate halophile bacterium that was recently isolated from surface saline soil of the Karaj Region, Iran. The genes encoding HU protein were also assayed during this study by Polymerase Chain Reaction.

Halobacillus karajensis sp. nov., a novel moderate halophile.

A moderately halophilic, gram-positive, spore-forming bacterium was isolated from surface saline soil of the Karaj region, Iran. The strain, designated MA-2T, was strictly aerobic with rod-shaped cells that occurred singly, in pairs or short chains. It contained L-Om-D-Asp-type peptidoglycan and the major respiratory lipoquinone was MK-7. It was non-motile and had an ellipsoidal endospore located centrally or subterminally. Growth occurred at 10-49 degrees C and in the pH range 6.0-9.6. Strain MA-2T grew at salinities of 1-24% (w/v) NaCl, showing optimal growth at 10% (w/v). The DNA G + C content was 41.3 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain MA-2T was associated with Bacillus rRNA group 1. The micro-organisms showing the closest phylogenetic relationship to strain MA-2T were Halobacillus litoralis and Halobacillus trueperi. On the basis of phenotypic and chemotaxonomic characteristics, 16S rRNA gene sequence analysis and DNA-DNA similarity data, it is proposed that strain MA-2T (= DSM 14948T = LMG 21515T) should be placed in the genus Halobacillus as the type strain of a novel species, Halobacillus karajensis sp. nov.

Production of amylase by newly isolated moderate halophile, Halobacillus sp. strain MA-2.

Production of extracellular amylase was demonstrated under stress conditions of high temperature and high salinity in aerobically cultivated culture of a newly isolated moderately halophilic bacterium of spore-forming Halobacillus sp. strain MA-2 in medium containing starch, peptone, beef extract, and NaCl. The maximum amylase production was secreted in the presence of 15% (w/v) Na(2)SO(4) (3.2 U ml(-1)). The isolate was capable of producing amylase in the presence of NaCl, NaCH(3)COOH, or KCl, with the results NaCl>NaCH(3)COOH>KCl. Maximum amylase activity was exhibited in the medium containing 5% (w/v) NaCl (2.4 U ml(-1)). Various carbon sources induced enzyme production. The potential of different carbohydrates in the amylase production was in the order: dextrin>starch>maltose>lactose>glucose>sucrose. In the presence of sodium arsenate (100 mM), maximum production of the enzyme was observed at 3.0 U ml(-1). Copper sulfate (0.1 mM) decreased the amylase production considerately, while lead nitrate had no significant enhancement on amylase production (p<0.05). The pH, temperature, and aeration optima for enzyme production were 7.8, 30 degrees C, and 200 rpm, respectively, while the optimum pH and temperature for enzyme activity was 7.5-8.5 and 50 degrees C, respectively.