Isolation and characterization of halophilic bacteria from Urmia Lake in Iran

Urmia Lake is one of the most permanent hypersaline lakes in the world which is threatened by hypersalinity and serious dryness. In spite of its importance no paper has been published regarding bacterial community of this lake. Accordingly, the present study aimed to investigate the halophilic bacteria in the aforementioned lake. In so doing, thirty seven strains were isolated on six different culture media. The isolated strains were characterized using phenotypic and genotypic methods. Growth of the strains occurred at 2535 degrees C, pH 6-9 and 7 to 20% (w/v) NaCl indicating that most of the isolates were moderately halophiles. Catalase, oxidase and urease activities were found to be positive for the majority of the isolates. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the isolated bacteria belonged to two major taxa: Gammaproteobacteria (92%, including Salicola [46%], Pseudomonas [13.5%], Marinobacter [ 11%], Idiomarina [11%], and Halomonas [8%]) and Firmicutes (8%, including Bacillus [5%] and Halobacillus [3%]). In addition, a novel bacterium whose 16S rRNA gene sequence showed almost 98% sequence identity with the taxonomically troubled DSM 3050T, Halovibrio denitrificans HGD 3T and Halospina denitrificans HGD 1-3T, each, was isolated. 16S rRNA gene similarity levels along with phenotypic characteristics suggest that some of the isolated strains could be regarded as potential type strain for novel species, on which further studies are recommended.     

Screening and isolation of halophilic bacteria producing extracellular hydrolyses from Howz Soltan Lake,Iran

Screening of bacteria from different areas of Howz Soltan playa, a hypersaline lake in the central desert zone of Iran, led to the isolation of 231 moderately halophilic bacteria, which were able to grow optimally in media with 5–15% of salt, and 49 extremely halophilic microorganisms that required 20–25% of salt for optimal growth. These isolates produced a great variety of extracellular hydrolytic enzymes. A total of 195, 177, 100, 95, 92, 68, 65, 33, and 28 strains produced lipases, amylases, proteases, inulinases, xylanases, cellulases, pullulanases, DNases, and pectinases, respectively. In comparison with gram-negative bacteria, the gram-positive halophilic rods, showed more hydrolytic activities. Several combined activities were showed by some of these isolates. One strain presented 9 hydrolytic activities, 4 strains presented 8 hydrolytic activities, 10 strains presented 7 hydrolytic activities and 29 strains presented 6 hydrolytic activities. No halophilic isolate without hydrolytic activity has been found in this study. According to their phenotypic characteristics and comparative partial 16S rRNA sequence analysis, the halophilic strains were identified as members of the genera: Salicola, Halovibrio, Halomonas, Oceanobacillus, Thalassobacillus, Halobacillus, Virgibacillus, Gracilibacillus, Salinicoccus, and Piscibacillus. Most lipase and DNase producers were members of the genera Gracilibacillus and Halomonas, respectively, whereas most of the isolates able to produce hydrolytic enzymes such as amylase, protease, cellulose (CMCase) and inulinase, belonged to gram-positive genera, like Gracilibacillus, Thalassobacillus, Virgibacillus, and Halobacillus.     

Isolation and identification of moderately halophilic bacteria producing hydrolytic enzymes from the largest hypersaline playa in Iran

Iran has many hypersaline environments, both the permanent and seasonal ones. One of the seasonal hypersaline lakes in the central desert zone is Aran-Bidgol Lake in which microbial diversity has not been characterized, thus the potential usage of this microbial community in biotechnology remained unknown. In this study, screening the halophilic hydrolytic enzyme-producing bacteria from different areas of this lake led to isolation of 61 gram-positive and 22 gram-negative moderately halophilic bacteria. These bacterial isolates were shown to produce a wide variety of hydrolytic enzymes including DNase, inulinase, amylase, lipase, pectinase, protease, chitinase, pullulanase, cellulase, and xylanase. The most common enzymes were DNase and inulinase in gram-positive bacteria, lipase in gram-negative bacteria, and pullulanase and cellulase in gram-positive cocci. Interestingly, combined hydrolytic activates were observed in some isolates. According to their phenotypic characteristics and comparative partial 16S rRNA sequence analysis, the moderately halophilic strains belonged to the genera Halobacillus, Thalassobacillus, Bacillus, Salinicoccus, Idiomarina, Salicola, and Halomonas.     

Isolation and characterization of some moderately halophilic bacteria with lipase activity

Lipases are an important class of enzymes which catalyze the hydrolysis of long chain triglycerides and constitute the most prominent group of biocatalysts for biotechnological applications. There are a number of lipases, produced by some halophilic microorganisms. In this study, some lipase producing bacteria from the Maharla salt lake located in south of Iran were isolated. All isolates were screened for true lipase activity on plates containing olive oil. The lipase activity was measured using titrimetric methods. Among thirty three isolates, thirteen strains demonstrating orange zone around colonies under UV light, were selected for identification using the molecular methods and some morphological characteristics. The bacterium Bacillus vallismortis BCCS 007 with 3.41 ± 0.14 U/mL lipase activity was selected as the highest lipase producing isolate. This is the first report of isolation and molecular identification of lipase producing bacteria from the Maharla lake.     

江苏盐城盐场中度嗜盐菌的分离与鉴定

采用高盐的牛肉膏蛋白胨培养基(盐浓度为8%NaCl),研究江苏省盐城市盐场土壤里中度嗜盐菌的分布情况及种群特征。从盐城市的射阳、新滩、灌东三处盐场土壤中共采集和分离得到13株中度嗜盐菌。通过形态观察、生理生化分析、16S rRNA序列分析和系统进化分析等方法进行初步鉴定,结果表明:分离到的中度嗜盐菌分属3个属,Virgibacillus属4株、Halomonas属7株和Marinobacter属2株。研究结果揭示盐城市的盐场存在较为丰富的中度嗜盐菌,具有较高的研究和利用价值。     

Isolation and characterization of moderately halophilic bacteria from fully cured salted cod (bachalao)

A total of 128 Strains of moderately halophilic bacteria were isolated from wet, dry and desalted bachalao (salted codfish) as well as from fresh cod and curing salt. The viable count of these bacteria in fully cured wet and dry bachalao ranged from 10³ to 10⁷ per g. All strains were characterized with 40 phenotypic tests and clustered using the S _SM coefficient and UPGMA linking analysis. The strains clustered into five phena at 75% similarity, with 77 strains in phenon A and 37 in phenon E. Two main colony types, smooth and rough, were observed and correspond to phena A, B and C on one hand and phenon E on the other. These two types seem to represent the dominating bacterial flora in fully cured, wet and dry bachalao, respectively. Representative strains of the smooth colony type were characterized further and found to grow well in 0.1–4.5 mol I⁻¹ NaCl and at 15–37°C. They have not, at present, been assigned to any known bacterial species.     

Hydrogeochemistry of seasonal variation of Urmia Salt Lake, Iran

Urmia Lake has been designated as an international park by the United Nations. The lake occupies a 5700 km2 depression in northwestern Iran. Thirteen permanent rivers flow into the lake. Water level in the lake has been decreased 3.5 m in the last decade due to a shortage of precipitation and progressively dry climate. Geologically the lake basin is considered to be a graben of tectonic origin. Na, K, Ca, Li and Mg are the main cations with Cl, SO4, and HCO3 as the main anions. F & Br are the other main elements in the lake. A causeway crossing the lake is under construction, which may affect the lake's annual geochemistry. The main object of this project is mainly to consider the potential of K-mineral production along with ongoing salt production. Seven hundred and four samples were taken and partially analyzed for the main cations and anions. Surface water (0.5 m. depth) was analyzed for Na, K, Mg, Ca, Br and Li, and averaged 87.118 g/lit, 1.48 g/lit, 4.82 g/lit, 4.54 g/lit, 1.19 ppm and 12.7 ppm respectively for the western half of the lake. Sodium ranged between 84 to 91.2 g/lit, and showed higher concentrations in the south than in the north. This unexpected result may be caused by shallower depth in the south and a higher net evaporation effect. Calcium ranged between 4.2 to 5 g/lit, apparently slightly higher in the north. K is higher in the south, possibly due to rivers entering from south that may carry slightly higher K in solution. In the middle-range samples (0.5–5 m.), K averaged 1.43 g/lit and ranged from 1.40 to 1.46 g/lit. At this intermediate depth the distribution of K is clearly higher to the south of the causeway that is currently under construction. It is not clear whether this increase is the effect of the causeway or the effect of the salty Aji-Chay River to the east, and the Khoy salt domes to the north of the lake. At depth (5 m–10 m), K averaged 1.48 g/lit and ranged from 1.4 to 1.49 g/lit, differing only in the second decimal from the average of the middle and surface samples. Ignoring the small difference between the averages of the three sample depths, the distribution of K is highly homogeneous in the lake water due to the mixing process. Therefore causeway construction has not yet strongly affected K distribution, or it may be at the starting point. Magnesium concentration ranged from 4.6 to 5-g/lit, and was elevated in the south. This differs somewhat compared to calcium. Lithium, with an average of 12–13 ppm, is slightly higher in the south, and has not shown any significant variation in all three seasons. Iodine was below the detection limit in the lake. Urmia Lake, geochemically, is highly uniform both to the south and north of the causeway, in both the surface and deep brines. K and Mg, which average 1.48 and 6.6 g/lit in order, could be elements worth production in addition to the NaCl currently being produced from the lake. Br, F, Li and B in the limit of <50 ppm don't look to be in the economical range.     

Urmia Lake (Northwest Iran): a brief review

Lake Urmia (or Ormiyeh) is one of the largest hypersaline lakes in the world and the habitat of a unique bisexual Artemia species (A. urmiana). Despite this, and several other values of the lake, little literature on it has been published. The present paper is an attempt to provide a brief review on various aspects of the lake. Urmia Lake, located in northwestern Iran, is an oligotrophic lake of thalassohaline origin with a total surface area between 4750 and 6100 km² and a maximum depth of 16 m at an altitude of 1250 m. The lake is divided into north and south parts separated by a causeway in which a 1500-m gap provides little exchange of water between the two parts. Due to drought and increased demands for agricultural water in the lake's basin, the salinity of the lake has risen to more than 300 g/L during recent years, and large areas of the lake bed have been desiccated. Therefore, management and conservation of this incomparable ecosystem should be considered to improve the current condition by fisheries research institutes.     

Cold-active halophilic bacteria from the ice-sealed Lake Vida,Antarctica

Lake Vida is a large, permanently ice-covered lake in the Victoria Valley of the McMurdo Dry Valleys, Antarctica and is unique among Dry Valley lakes because it is ice-sealed, with an ice-cover of nearly 19 m. Enrichment cultures of melt-water from Lake Vida 15.9 m ice yielded five pure cultures of aerobic, heterotrophic bacteria. Of these, one strain grew at −8°C and the four others at −4°C. All isolates were either halotolerant or halophilic, with two strains capable of growth at 15% NaCl. Phylogenetic analysis revealed the Lake Vida isolates to be Gammaproteobacteria, related to species of Psychrobacter and Marinobacter. This is the first report of pure cultures of bacteria from Lake Vida, and the isolates displayed a phenotype consistent with life in a cold hypersaline environment.     

Isolation and characterization of cellulolytic bacteria from the Stain house Lake,Antarctica

The main aim was to evaluate the occurrence of cellulolytic bacteria from the Stain house Lake, located at Admiralty Bay, Antarctica. Thick cotton string served as a cellulose bait for the isolation of bacteria. A total of 52 bacterial isolates were recovered and tested for their cellulase activity, and two of them, isolates CMAA 1184 and CMAA 1185, showed significant cellulolytic activity on carboxymethylcellulose agar plates. Phylogenetic analysis placed the isolates into the Bacillus 16S ribosomal RNA gene subclade. Both isolates produced a cold-active cellulase which may play a crucial role in this extreme environment.     

Characterization of basidiomycetous yeasts in hypersaline soils of the Urmia Lake National Park,Iran

Urmia Lake, located in northwest Iran, is an oligotrophic and extremely hypersaline habitat that supports diverse forms of life. Owing to its unique biodiversity and special environmental conditions, Urmia Lake National Park has been designated as one of the biosphere reserves by UNESCO. This study was aimed to characterize basidiomycetous yeasts in hypersaline soils surrounding the Urmia Lake National Park using a polyphasic combination of molecular and physiological data. Soil samples were collected from eight sites in Lake Basin and six islands insides the lake. Yeast strains were identified by sequencing the D1/D2 domains of the 26S rRNA gene. When D1/D2 domain sequencing did not resolve the identity of the species, strain identification was obtained by ITS 1 & 2 sequencing. Twenty-one species belonging to the genera Cystobasidium, Holtermanniella, Naganishia, Rhodotorula, Saitozyma, Solicoccozyma, Tausonia, Vanrija, and Vishniacozyma were identified. Solicoccozyma aeria represented the dominant species. The ability of isolates to grow at 10 and 15 % of NaCl was checked; about two-thirds of the strains grew at 10 %, while about 13 % of the isolates grew in medium with 15 % NaCl. this study is the first study on the culturable yeast diversity in hypersaline soils surrounding an Asian lake.     

Production,characterization and bioactivities of biosurfactants from newly isolated strictly halophilic bacteria

Eight distinct halophilic bacteria isolated from extreme saline soil samples of Khewra Salt Mines, Pakistan, were investigated for biosurfactant production. Isolates were identified by physiological, phenotypic and genetic characterization. Using 16S rDNA sequence analysis, the strains MB590, MB591, MB593, MB594, MB595 and MB596 were identified as Halomonas elongata, MB588 as Halobacillus karajiensis, and MB589 as Alkalibacillus almallahensis. Preliminary screening of biosurfactant production in halophilic bacteria was done by multiple screening assays. All biosurfactants showed significant emulsification properties and remarkably low surface tension values (up to 16.5 dynes/cm). Structural characterization of partially purified biosurfactants using Gas chromatography–mass spectrometry (GC–MS) and Fourier-transform infrared spectroscopy (FTIR) techniques indicated different fatty acids, glycolipid derivatives and a novel antimicrobial peptide furanomycin. These biosurfactants exhibited strong bioactivities against bacterial/fungal pathogens i.e. Klebsiella pneumoniae (86.5 %), Staphylococcus aureus (97.75 %), Bacillus subtilis (97 %), Enterococcus faecalis (97.6 %), E. coli (54.5 %), Aspergillus niger (87.75 %), Aspergillus fumigatus (93.1 %), Aspergillus flavus (80.4 %), and Fusarium solani (89.05 %). Additionally, biosurfactants also showed 85 % free radical scavenging activity indicating their antioxidant potential. The present study revealed the potential of halophilic bacterial biosurfactants as effective antimicrobial agents against various pathogens, and their possible applications in the biomedical field.     

新疆艾丁湖及邻近地区嗜盐菌和耐盐菌的数量分布

以盐（NaCI）为限制性生物因子,对艾丁湖地区原核生物的分布状态进行调查,从距艾丁湖中心1．5km处出发,定距离,分季节取土样及水样（共取样20个）。对样品菌群数量统计表明,在湖水中,春秋两嗜盐菌的数量高于耐盐菌。冬季湖水温度大大降低,盐结晶沉淀,湖水中的盐浓度降低,使得湖水中的耐盐菌的数量高于嗜盐菌的数量。土壤中的有要物比湖水中的丰富,温度也较稳定,因而耐盐菌的数量高于嗜盐菌的数量。距湖岩越远,土壤中对盐依赖性强的细菌越少,反之对盐依赖性弱的菌越多。由样品中分离得到细菌173株,放线菌12株,以不同浓度NaCI为唯一生长限制因子对所分离到的细菌进行生理生化实验,获得嗜盐菌61株,其中除3株可视为新种外（新种的鉴定将另外报告）,其它均为嗜盐菌的已知产生菌种,现仅报道艾丁湖及邻近地区以盐为限制因子的原核微生物分布。     

艾丁湖可培养嗜盐菌多样性及功能酶、抗菌活性筛选

【目的】探究艾丁湖可培养嗜盐菌的多样性、功能酶活性以及抗菌活性,进一步了解其次级代谢产物情况,为新型生物活性物质的发掘提供依据。【方法】选用以20种糖及糖的衍生物作为唯一碳源的寡营养培养基从艾丁湖5个样点中分离得到298株嗜盐菌,根据形态特征去重复后,选取62株菌运用16S r RNA基因系统发育分析的方法研究样品中嗜盐菌的多样性;从不同类群选取22株代表菌株,采用点接法进行3种功能酶的筛选,运用平板对峙法检测代表菌株对12种病原菌的抗菌活性。【结果】从5%、10%和15%3个盐浓度中分别分得221、54和23株嗜盐菌,获得的嗜盐菌分布在9个科18个属;其中放线菌分布于4个属,细菌分布于14个属;Nocardiopsis和Pontibacillus属为艾丁湖可培养嗜盐菌的优势类群,分别占17.7%和16.1%;有15株嗜盐菌相似性低于98.5%,可能为潜在新种。所选取的22株代表菌株中,分别有68.2%、22.7%和72.7%的实验菌株具有蛋白酶、淀粉酶和酯酶活性;45.5%的代表菌株对12种病原菌表现出了抗至少1种病原菌的活性,其中一株Nocardiopsis属放线菌能抗9种病原菌,表现出了广谱的抗菌活性。【结论】新疆艾丁湖土样中嗜盐菌的多样性较丰富,而且具有较好的生物活性,为后续进一步研究其次级代谢产物提供了依据。     

Exploration,antifungal and antiaflatoxigenic activity of halophilic bacteria communities from saline soils of Howze-Soltan playa in Iran

In the present study, halophilic bacteria communities were explored in saline soils of Howze-Soltan playa in Iran with special attention to their biological activity against an aflatoxigenic Aspergillus parasiticus NRRL 2999. Halophilic bacteria were isolated from a total of 20 saline soils using specific culture media and identified by 16S rRNA sequencing in neighbor-joining tree analysis. Antifungal and antiaflatoxigenic activities of the bacteria were screened by a nor-mutant A. parasiticus NRRL 2999 using visual agar plate assay and confirmed by high-performance liquid chromatography. Among a total of 177 halophilic bacteria belonging to 11 genera, 121 isolates (68.3%) inhibited A. parasiticus growth and/or aflatoxin production. The most potent inhibitory bacteria of the genera Bacillus, Paenibacillus and Staphylococcus were distributed in three main phylogenetic clusters as evidenced by 16S rRNA sequence analysis. A. parasiticus growth was inhibited by 0.7–92.7%, while AFB₁ and AFG₁ productions were suppressed by 15.1–98.9 and 57.0–99.6%, respectively. Taken together, halophilic bacteria identified in this study may be considered as potential sources of novel bioactive metabolites as well as promising candidates to develop new biocontrol agents for managing toxigenic fungi growth and subsequent aflatoxin contamination of food and feed in practice.

     

Isolation and characterization of glycolipids from some photosynthetic bacteria

Glycosyl glycerides have been found in substantial amounts in Chloropseudomonas ethylicum but could not be detected in two strains of Rhodopseudomonas palustris. Rhodospirillum molischianum possibly contains small amounts of monoglycosyl diglyceride. The glycolipids of C. ethylicum have been separated into two components. One of these, glycolipid I, is a monogalactosyl diglyceride. Glycolipid II, upon acid hydrolysis, yields galactose, rhamnose, and a third, unidentified sugar. The glycolipids or total lipids of the photosynthetic bacteria examined contained saturated and monounsaturated, but none of the more highly unsaturated, fatty acids.     

Isolation and characterization of quinoline-degrading bacteria from subsurface sediments

Two gram-negative, motile bacteria isolated from deep subsurface sediments mineralized the nitrogen-containing polyaromatic hydrocarbon quinoline under aerobic conditions and transformed quinoline to soluble intermediates under anaerobic conditions. Many aromatic compounds were also able to serve as the sole source of carbon and energy under aerobic conditions. Rapid aerobic mineralization of quinoline at concentrations as low as 0.002 microgram ml-1 indicates that these organisms possess a high-affinity uptake and utilization system, which may reflect the oligotrophic nature of deep subsurface environments. Both bacteria harbored four plasmids of identical size, ranging from 50 to 440 kilobases.