Characterization of extremely halophilic Archaea isolated from the Ayvalik Saltern, Turkey (original) (raw)

Phenotypic Characterization of Halophilic Bacteria Isolated From Çamaltı Saltern in Turkey

Electronic Journal …, 2010

Çamaltı Saltern is the largest sea saltern on the Aegean cost of Turkey. Water samples collected from 6 different pools changing in the degree of salinity were subjected to bacterial isolation. A total of 12 isolates were randomly chosen and their properties were investigated. Phenotypic properties of the isolates as well as their characteristics for seven enzymes showed that these bacteria represents probably different genus and species of either Archaeal or Bacterial domains. Activities of hydrolytic enzymes (gelatinase, DNase, Tween-80 and cellulase), activities which are responsible for the detrimental effects of the crude salt in food and leather products were recorded for some of the isolates. Fatty acid methyl esther profiles of 6 isolates belonged to Bacteria domain were different representing different genera and species of Bacterial domain. To the author's knowledge this is the first study on the phenotypic and enzymatic properties of prokaryotic populations of water samples obtained from pools of Çamaltı saltern.

Isolation and characterization of halophilic bacteria from Çamaltı Saltern Turkey

New Biotechnology, 2009

Knowledge of the functions, interactions, and diversity of extremely halophilic microorganisms mostly comes from the results of studies performed in diff erent salterns throughout the world. As model habitats, salterns allow the comparison of diff erent techniques used for qualitative and quantitative analysis of halophilic communities in these hypersaline environments. Çamaltı Saltern is the biggest coastal solar saltern located on the Aegean coast of Turkey, and it produces most of the salt consumed in the country. In the present study, detection of prokaryotic communities of the Çamaltı Saltern was performed using 2 culture-independent methods. Real-time polymerase chain reaction (RT-PCR) and fl uorescein in situ hybridization (FISH) techniques were evaluated to analyze the microbial populations of Çamaltı Saltern. Of the Çamaltı samples, 48% to 67% were hybridized with the EUB338 probe and 33% to 57% were hybridized with the ARC915 probe. Repeatability of the RT-PCR experiments with environmental DNA was considered insuffi cient. However, FISH analysis may be combined with RT-PCR and these 2 techniques may be used in tandem to rapidly reveal quantitative aspects of the microbial population of hypersaline environments.

Extremely halophilic Archaea from Tuz Lake, Turkey, and the adjacent Kaldirim and Kayacik salterns

World Journal of …, 2007

Tuz Lake is a hypersaline lake located in Central Anatolia, Turkey. The lake and its salterns, Kaldirim and Kayacik, are the major sources of solar salt for industrial applications in Turkey, especially in the food and leather industries. Use of the crude solar salt often results in microbial deterioration of the products. We therefore initiated a thorough characterization of the microbial communities in Tuz Lake and its adjacent salterns, and we present here the results of investigations on diversity of extremely halophilic Archaea. Twenty-seven colonies of aerobic red or pink Archaea (family Halobacteriaceae) were selected according to colony shape, size, consistency and pigmentation, and characterized according to their phenotypic characteristics, polar lipid contents, and antibiotic sensitivities. Furthermore, 16S rRNA genes of the isolates were screened by DGGE analysis and partially sequenced. Phylogenetic analysis showed that most isolates belonged to the genera Haloarcula, Halorubrum and Halobacterium. Haloarcula was found to be dominant both in Tuz Lake and in the saltern samples. Halorubrum species were isolated from Tuz Lake and from the Kaldirim saltern, and Halobacterium species were recovered from Tuz Lake and from the Kayacik saltern. All strains showed various activities of hydrolytic enzymes (proteases, amylases, cellulases, and others), activities which are responsible for the detrimental effects of the crude salt in food and leather products.

Enzyme Characteristics of Extremely Halophilic Archaeal Community in Tuzkoy Salt Mine, Turkey

World Journal of Microbiology and Biotechnology, 2004

Tuzkoy is a large and unpolluted salt mine in Central Anatolia, Turkey. High quality salt in this mine may be supplied to the table salt market after a simple process instead of complicated refining. However, the extremely halophilic microbial content of the salt has not yet been investigated which may result in detrimental effects in industrial processes such the food and leather industries, if unprocessed salt is directly used. Therefore, to identify and characterize the microbial contaminants in Tuzkoy salt, chemical and microbial analyses were conducted on salt crystal samples collected from three different locations of the mine. Generally 10 5 -10 6 colony forming units of extremely halophilic microorganisms were detected per gram of salt samples. Twelve colonies were selected randomly for further characterization. Phenotypic characterization, lipid analysis, antibiotic susceptibility tests and positive PCR amplification results with Archaea-specific primers confirmed that all strains were halophilic Archaea belong to family Halobacteriaceae. According to denaturing gradient gel electrophoresis (DGGE) results, 10 comparatively different strains were selected for DNA sequencing. DNA sequences and phylogenetic analyses revealed that the isolated strains were mainly species of the genera Halobacterium, Haloarcula, Natrinema and Halorubrum. In addition enzymatic activity tests were also conducted to evaluate the salt quality for industrial applications. Results of gelatinase, caseinase, amylase, cellulase and lipase activity tests revealed that the isolated strains produced hydrolytic enzymes, which could cause deterioration in salt-treated food and hide. It may be pointed out that cellulase activity in halophilic Archaea has not been reported previously. Moreover, b-galactosidase activity has been reported in some Haloferax and Halorubrum species but not in the genus Halobacterium.

Bacterial Diversity in Çamalti Saltern, Turkey

PubMed, 2015

A combination of culture-dependent and culture-independent approaches was employed to identify the bacterial diversity of Çamalti solar saltern in Turkey. The bacterial communities of Çamalti Saltern were analyzed by molecular techniques that included denaturing gradient gel electrophoresis of 16S rRNA gene fragments PCR amplified from DNA extracted from the water samples of the saltern and 16S rRNA gene library analysis. A total of 42 isolates were identified at the genus/species level and 17 of them were found to belong to the Bacteria domain. All bacterial isolates were phylogenetically related to Halobacillus, Virgibacillus and Halomonas genus. A total of 50 clones from 16S rRNA gene library were analyzed by ARDRA. 16S rRNA sequence analysisof these clones revealed that most (85%) of the bacterial clones were related to Salinibacter genus members of the Bacteroidetes. The sequences of DGGE bands were related to the uncultured Salinibacter, uncultured halophilic bacterium and Halomonas sp. This work highlights the halophilic bacterial diversity of Çamalti marine solar saltern.

Characterization of halophilic Archaea isolated from different hypersaline ecosystems

Annals of Microbiology, 2006

Halobacterial representatives were isolated from salted fish, naturally occurring salt pans as well as artificial saline pools in the Mediterranean area. The isolation techniques experimented proved successful, allowing halophilic archaea to be retrieved in almost 72% of analysed samples. About 65% of strains could be presumptively ascribed to the species Haloarcula marismortui by RFLP (Restriction Fragment Length Polymorphism) grouping and 16S rRNA gene sequence comparison. Nevertheless, cluster analysis of RAPD-PCR patterns revealed a wide heterogeneity among isolated strains. Biochemical features of technological interest, such as proteolytic, lipolytic and decarboxylase activity, were investigated in order to elucidate the role of archaea during ripening of salted anchovies. Proteolytic activity was only evidenced on the sarcoplasmic fraction extracted from fresh anchovies and for only 4 strains out of 21. No strain revealed either lipolytic capability on cod liver oil or proteolytic activity on fresh anchovies myofibrillar extract. Finally, only one strain exhibited decarboxylase activity, minimizing the potential responsibility of cultivable archaea microflora in the spoilage of salted anchovies.

Relative abundance of halophilic archaea and bacteria in diverse salt-fermented fish products

LWT, 2019

Fermented fish that contain over 20% salt are expected to harbor extremely halophilic microorganisms. In the present study, salt-fermented fish samples (n = 105) which included fermented Indian shad, mackerel, anchovies, seer fish, pomfret and others collected from different locations in India were analyzed for the presence of extremely halophilic microorganisms using culture methods. The salt (NaCl) content in majority of the products was over 20% and the moisture content ranged from 28 to 57%. Three different halophilic media, each with 20 and 25% salt, were used for the enumeration and isolation of halophilic archaea and bacteria. Of 1635 isolates obtained from different media, 1198 and 437 were confirmed to be halophilic archaea and bacteria, respectively using 16S rRNA genes-specific PCR. Overall, archaea and bacteria represented 74.5% and 25.4% of the total halophilic microbial communities, respectively suggesting the predominance of archaea across a wide range of fermented fish products. The results of this study emphasize that the halophilic archaea have a dominant role in the fermentation process of fish and may contribute significantly to flavor, health benefits and bioactivities associated with fermented fish products.

Diversity of Extremely Halophilic Archaeal and Bacterial Communities from Commercial Salts

Frontiers in Microbiology, 2017

Salting is one of the oldest food preservation techniques. However, salt is also the source of living halophilic microorganisms that may affect human health. In order to determine the microbial communities of commercial salts, an investigation were done using amplicon sequencing approach in four commercial salts: Ethiopian Afdera salt (EAS), Ethiopian rock salt (ERS), Korean Jangpan salt (KJS), and Korean Topan salt (KTS). Using domain-specific primers, a region of the 16S rRNA gene was amplified and sequenced using a Roche 454 instrument. The results indicated that these microbial communities contained 48.22-61.4% Bacteria, 37.72-51.26% Archaea, 0.51-0.86% Eukarya, and 0.005-0.009% unclassified reads. Among bacteria, the communities in these salts were dominated by the phyla Proteobacteria, Bacteroidetes, Actinobacteria, and Firmicutes. Of the archaea, 91.58% belonged to the class Halobacteria, whereas the remaining 7.58, 0.83, and 0.01% were Nanoarchaea, Methanobacteria, and Thermococci, respectively. This comparison of microbial diversity in salts from two countries showed the presence of many archaeal and bacterial genera that occurred in salt samples from one country but not the other. The bacterial genera Enterobacter and Halovibrio were found only in Korean and Ethiopian salts, respectively. This study indicated the occurrence and diversity of halophilic bacteria and archaea in commercial salts that could be important in the gastrointestinal tract after ingestion.

Extremely Halophilic Bacterial Communities in fiereflikoçhisar Salt Lake in Turkey

fiereflikoçhisar Lake is the largest salt lake in central Turkey. This lake is a major source of solar salt for food, hide and other industries locally. Due to the economic importance of salt obtained from this lake, a microbial survey has been conducted. Six salt and three brine samples were obtained from the lake for physico-chemical and microbiological analyses. Physico-chemical analysis showed that the salts and brines contain sufficient ions and hardness to support extremely halophilic bacteria. The salt collected from the lake contained 10 4 -10 6 colony-forming units of extremely halophilic bacteria per gram and brine taken from the lake contained 10 3 -10 5 colony-forming units of extremely halophilic bacteria per ml. Colonial pigmentation from these samples ranged from bloodred to pale-pink. A total of 82 extremely halophilic aerobic strains were isolated from the salt and brine samples, 32 of which were randomly selected strains examined in greater detail. While brick-red colonies outnumbered all other colony pigmentations, the lake does appear to support a diverse bacterial community. Most colonies were 1 to 2 mm in diameter, circular, convex, glistening and entire. Optimum growth occurred at 25% (w/v) NaCl at 40 ºC and a pH of 7.5. Seventeen strains required at least 10% (w/v) NaCl for growth. Most cells of the strains were pleomorphic and stained Gram-negative. All strains were motile. Some of these strains produced industrially important enzymes such as lipases, gelatinases, cellulases and β-galactosidases. All of the strains hydrolysed Tween 80. Twenty-three strains produced gelatinase enzymes. Cellulase enzymes were produced by 14 of these strains. Only one strain showed positive β-galactosidase activity. Ten strains produced indol from tryptophan. Twenty-three strains showed positive methyl-red reactions. All strains showed negative Voges-Proskauer reactions. Thirty-two strains were tested by the disc diffusion method for their sensitivity to 13 different antibiotics. Most of these strains were resistant to ampicillin (10 µg), chloramphenicol (30 µg), cefadroxil (30 µg), erythromycin (15 µg), penicillin G (10U) and streptomycin (25 µg). All strains were resistant to amikacin (30 µg), ciprofloxacin (5 µg), neomycin (30 µg) and spiramycin (100 µg). Most strains were sensitive to bacitracin (10 U) and novobiocin (5 µg). Half of these strains were sensitive to sulfamethoxazole and trimethoprim (25 µg). These studies showed that fiereflikoçhisar Salt Lake is an optimal environment for extremely halophilic bacteria and the lake contains a viable, diverse, potentially and industrially important bacterial community. The extremely halophilic proteolytic bacterial contents of the salt and brine samples were too high to use directly in hide preservation. The presence of proteolytic strains in the lake's salt may cause an important quality problem in the leather business.

Taxonomic Analysis of Extremely Halophilic Archaea Isolated from 56-Years-Old Dead Sea Brine Samples

Systematic and Applied Microbiology, 2000

A taxonomic study comprising both phenotypic and genotypic characterization, has been carried out on a total of 158 extremely halophilic aerobic archaeal strains. These strains were isolated from enrichments prepared from Dead Sea water samples dating from 1936 that were collected by B. E. Volcani for the demonstration of microbial life in the Dead Sea. The isolates were examined for 126 morphological, physiological, biochemical and nutritional tests. Numerical analysis of the data, by using the SJ coefficient and UPGMA clustering method, showed that the isolates clustered into six phenons. Twenty-two out of the 158 strains used in this study were characterized previously (ARAHAL et ai., 1996) and were placed into five phenotypic groups. The genotypic study included both the determination of the guanineplus-cytosine content of the DNA and DNA-DNA hybridization studies. For this purpose, representative strains from the six phenons were chosen. These groups were found to represent some members of three different genera -Haloarcula (phenons A, B, and C), Haloferax (phenons D and E) and Halobacterium (phenon F) -of the family Halobacteriaceae, some of them never reported to occur in the Dead Sea, such as Haloarcula hispanica, while Haloferax volcanii (phenons D and E) was described in the Dead Sea by studies carried out several decades later than Volcani's work.