Isolation and Characterization of Moderately Halophilic and Halotolerant Bacteria from the Freshwater Al-Asfar Lake, Al Ahsa, Saudi Arabia

I. Alshubaith¹ and D. J. Gilmour^2,

¹P.O. Box 60066 Hofuf, Al Ahsa, Saudi Arabia

²Department of Molecular Biology and Biotechnology, Firth Court, Western Bank, University of Sheffield, Sheffield S10 2TN, UK

Cite this paper:
I. Alshubaith and D. J. Gilmour. Isolation and Characterization of Moderately Halophilic and Halotolerant Bacteria from the Freshwater Al-Asfar Lake, Al Ahsa, Saudi Arabia. Journal of Applied & Environmental Microbiology. 2017; 5(2):79-85. doi: 10.12691/jaem-5-2-4

Abstract

A range of water samples from the Al-Asfar lake, Al Ahsa, Saudi Arabia were enriched with LB medium (plates and liquid medium) containing a range of salt concentrations up to 2 M NaCl. Three strains of halotolerant or moderately halophilic bacteria were isolated and identified by 16S rDNA sequencing as belonging to the genera Staphylococcus, Halobacillus and Halomonas. The first two organisms (S. warneri and Halobacillus sp.) were further charcaterized to understand their ability to grow at high salinities up to 3 M NaCl. Halobacillus sp. was shown to be moderately halophilic (optimum growth between 0.17 and 1 M NaCl), whereas S. warneri was shown to be halotolerant, optimum growth at 0.17 M NaCl. Nuclear magnetic resonance (NMR) was used to determine the compatible solutes accumulated by the two strains. Betaine was accumulated by both organisms and Halobacillus sp. also utilized glutamate at low salt concentrations. This work demonstrates the presence of moderately halophilic and halotolerant bacteria in a freshwater lake.

Keywords:
moderate halophiles freshwater Halobacillus Staphylococcus

This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

References:

[1]	Ventosa, A., Marquez, M. C., Garabito, M. J., and Arahal, D. R. (1998). Moderately halophilic gram-positive bacterial diversity in hypersaline environments, Extremophiles 2, 297-304.
[2]	Jose Leon, M., Martinez-Checa, F., Ventosa, A., and Sanchez-Porro, C. (2015). Idiomarina aquatica sp nov., a moderately halophilic bacterium isolated from salterns, International Journal of Systematic and Evolutionary Microbiology 65, 4595-4600.
[3]	Xue, Y., Ventosa, A., Wang, X., Ren, P., Zhou, P., and Ma, Y. (2008). Bacillus aidingensis sp nov., a moderately halophilic bacterium isolated from Ai-Ding salt lake in China, International Journal of Systematic and Evolutionary Microbiology 58, 2828-2832.
[4]	Gilmour, D. J. (1990). Halotolerent and halophilic microorganisms In Microbiology of Extreme Environments. Ed. Edwards, C., Open University Press, Milton Keynes, U.K., 147-177.
[5]	Oren, A. (2008). Nomenclature and taxonomy of halophilic archaea - comments on the proposal by DasSarma and DasSarma for nomenclatural changes within the order Halobacteriales, Int J Syst Evol Micr 58, 2245-2246.
[6]	Echigo, A., Hino, M., Fukushima, T., Mizuki, T., Kamekura, M., and Usami, R. (2005). Endospores of halophilic bacteria of the family Bacillaceae isolated from non-saline Japanese soil may be transported by Kosa event (Asian dust storm), Saline Systems 1, 8.
[7]	Echigo, A., Fukushima, T., Mizuki, T., Kamekura, M., and Usami, R. (2007). Halalkalibacillus halophilus gen. nov., sp nov., a novel moderately halophilic and alkallphilic bacterium isolated from a non-saline soil sample in Japan, International Journal of Systematic and Evolutionary Microbiology 57, 1081-1085.
[8]	Usami, R., Echigo, A., Fukushima, T., Mizuki, T., Yoshida, Y., and Kamekura, M. (2007). Alkalibacillus silvisoli sp nov, an alkaliphilic moderate halophile isolated from non-saline forest soil in Japan, International Journal of Systematic and Evolutionary Microbiology 57, 770-774.
[9]	Echigo, A., Minegishi, H., Mizuki, T., Kamekura, M., and Usami, R. (2010). Geomicrobium halophilum gen. nov., sp nov., a moderately halophilic and alkaliphilic bacterium isolated from soil, International Journal of Systematic and Evolutionary Microbiology 60, 990-995.
[10]	Echigo, A., Minegishi, H., Shimane, Y., Kamekura, M., and Usami, R. (2012). Natribacillus halophilus gen. nov., sp nov., a moderately halophilic and alkalitolerant bacterium isolated from soil, International Journal of Systematic and Evolutionary Microbiology 62, 289-294.
[11]	Chen, Y.-G., Hao, D.-F., Chen, Q.-H., Zhang, Y.-Q., Liu, J.-B., He, J.-W., Tang, S.-K., and Li, W.-J. (2011). Bacillus hunanensis sp nov., a slightly halophilic bacterium isolated from non-saline forest soil, Antonie Van Leeuwenhoek International Journal of General and Molecular Microbiology 99, 481-488.
[12]	Chen, Y.-G., Zhang, Y.-Q., Chen, Q.-H., Klenk, H.-P., He, J.-W., Tang, S.-K., Cui, X.-L., and Li, W.-J. (2011). Bacillus xiaoxiensis sp nov., a slightly halophilic bacterium isolated from non-saline forest soil, International Journal of Systematic and Evolutionary Microbiology 61, 2095-2100.
[13]	Weisburg, W. G., Barns, S. M., Pelletier, D. A., and Lane, D. J. (1991). 16S ribosomal DNA amplification for phylogenetic study, Journal of Bacteriology 173, 697-703.
[14]	Frings, E., Kunte, H. J., and Galinski, E. A. (1993). Compatible solutes in representatives of the genera, Brevibacterium and Corynebacterium: occurrence of tetrahydropyrimidines and glutamine, FEMS Microbiol. Lett. 109, 25-32.
[15]	Arahal, D. R., Vreeland, R. H., Litchfield, C. D., Mormile, M. R., Tindall, B. J., Oren, A., Bejar, V., Quesada, E., and Ventosa, A. (2007). Recommended minimal standards for describing new taxa of the family Halomonadaceae, International Journal of Systematic and Evolutionary Microbiology 57, 2436-2446.
[16]	da Costa, M. S., Santos, H., Galinski, E. A., and Antranikian, G. (1998). An overview of the role and diversity of compatible solutes in Bacteria and Archaea, Advances in Biochemical Engineering Biotechnology; Biotechnology of extremophiles, 117-153.
[17]	Empadinhas, N., and da Costa, M. S. (2008). Osmoadaptation mechanisms in prokaryotes: distribution of compatible solutes, International Microbiology 11, 151-161.
[18]	Spring, S., Ludwig, W., Marquez, M. C., Ventosa, A., and Schleifer, K. H. (1996). Halobacillus gen nov, with descriptions of Halobacillus litoralis sp nov and Halobacillus trueperi sp nov, and transfer of Sporosarcina halophila to Halobacillus halophilus comb nov, International Journal of Systematic Bacteriology 46, 492-496.
[19]	Barbosa, D. C., Bae, J. W., von der Weid, I., Vaisman, N., Nam, Y. D., Chang, H. W., Park, Y. H., and Seldin, L. (2006). Halobacillus blutaparonensis sp nov., a moderately halophilic bacterium isolated from Blutaparon portulacoides roots in Brazil, Journal of Microbiology and Biotechnology 16, 1862-1867.
[20]	Liu, W. Y., Zeng, J., Wang, L., Dou, Y. T., and Yang, S. S. (2005). Halobacillus dabanensis sp nov and Halobacillus aidingensis sp nov., isolated from salt lakes in Xinjiang, China, International Journal of Systematic and Evolutionary Microbiology 55, 1991-1996.
[21]	Cummings, S. P., and Gilmour, D. J. (1995). The effect of NaCl on the growth of a Halomonas species: accumulation and utilization of compatible solutes, Microbiology 141, 1413-1418.
[22]	Kunte, H. J. (2006). Osmoregulation in bacteria: compatible solute accumulation and osmosensing, Environmental Chemistry 3, 94-99.
[23]	Saum, S. H., and Müller, V. (2008). Regulation of osmoadaptation in the moderate halophile Halobacillus halophilus: chloride, glutamate and switching osmolyte strategies, Saline Systems 4.
[24]	Kamath, U., Singer, C., and Isenberg, H. (1992). Clinical significance of Staphylococcus warneri bacteremia, J Clin Microbiol 30, 261-264.
[25]	Kassem, I. I. (2009). Detection and characterization of staphylococcal pathogens in the environment: a community approach, PhD thesis, The University of Toledo.
[26]	Ivić, I., Karanović, J., and Pavičić-Ivelja, M. (2013). Sepsis with multiple abscesses caused by Staphylococcus warneri: a case report, Central European Journal of Medicine, 1-3.
[27]	Kropp, K. A., Lucid, A., Carroll, J., Belgrudov, V., Walsh, P., Kelly, B., Smith, C., Dickinson, P., O'Driscoll, A., Templeton, K., Ghazal, P., and Sleator, R. D. (2014). Draft Genome Sequence of a Staphylococcus warneri Strain Isolated from a Preterm Neonate Blood Sepsis Patient at the Royal Infirmary, Edinburgh, Scotland, Genome announcements 2.
[28]	Phukon, M., Sahu, P., Srinath, R., Nithya, A., and Babu, S. (2013). Unusual Occurrence of Staphylococcus warneri as Endophyte in Fresh Fruits along with Usual Bacillus spp, Journal of Food Safety 33, 102-106.
[29]	Miller, K. J., Zelt, S. C., and Bae, J.-H. (1991). Glycine betaine and proline are the principal compatible solutes ofStaphylococcus aureus, Current Microbiology 23, 131-137.