Isolation of Thermotoga spp. and Fervidobacterium spp., and Characterization of 16S rRNA Genes of Order Thermotogales: Unique Lineage of Hyperthermophiles Thriving in 3 Hot Springs in Thailand

Main Article Content

Porranee Keawram
Veeranun Pongsapukdee
Wirojne Kanoksilapatham

Abstract

Order Thermotogales comprise mostly strictly anaerobic thermophilic and hyperthermophilic bacteria which thrive in high temperature habitats throughout the world. In order to study diversity of hyperthermophiles in Thailand’s hot springs, culture dependent technique was performed at 80 oC. Pure cultures of 19 isolates obtained from 3 hot springs were characterized based on 16S rRNA genes of order Thermotogales. Twelve and 7 isolates were identified as deepest blanches within two clades formed by divergent encapsulated rod shaped bacteria belonging to Fervidobacterium and Thermotoga, respectively. Phylogenetic evidences suggest that the Thailand strains from both branches are possible candidates for two novel species. Closely related strains within both genera were identified regardless of their habitats and years of sample collection, implying that these native inhabitants found in Thailand’s hot springs might have evolved from a unique common ancestor. Because all isolates require extremely high temperatures for growth, these isolates are likely sources of genes encoding numerous thermostable enzymes.

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How to Cite
Keawram, P., Pongsapukdee, V., & Kanoksilapatham, W. (2016). Isolation of Thermotoga spp. and Fervidobacterium spp., and Characterization of 16S rRNA Genes of Order Thermotogales: Unique Lineage of Hyperthermophiles Thriving in 3 Hot Springs in Thailand. Science, Engineering and Health Studies, 10(1), 9–20. https://doi.org/10.14456/sustj.2016.1
Section
Research Articles
Author Biography

Wirojne Kanoksilapatham, Department of Statistics, Faculty of Science, Silpakorn University, Nakhon Pathom, Thailand. Corresponding author's e-mail: [email protected]

Associate Professor

Department of Microbiology, Faculty of Science, Silpakorn University

References

Alain, K., Marteinsson, V. T., Miroshnichenko, M. L., Bonch-Osmolovskaya, E. A., Prieur, D., and Birrien, J-L. (2002). Marinitoga piezophila sp. nov., a rod-shaped, thermo-piezophilic bacterium isolated under high hydrostatic pressure from a deep-sea hydrothermal vent. International Journal of Systematic and Evolutionary Microbiology, 52(Pt 4): 1331-1339.

Andrews, K. T., and Patel, B. K. C. (1996). Fervidobacterium gondwanense sp. nov., a new thermophilic anaerobic bacterium isolated from nonvolcanically heated geothermal waters of the great artesian basin of Australia. International Journal of Systematic Bacteriology, 46(1): 265-269.

Antoine, E., Cilia, V., Meunier, J. R., Guezennec, J., Lesongeur, F., and Barbier, G. (1997). Thermosiphomelanesiensis sp. nov., a new thermophilic anaerobic bacterium belonging to the order Thermotogales, isolated from deep-sea hydrothermal vents in the southwestern Pacific Ocean. International Journal of Systematic Bacteriology, 47(4): 1118-1123.

Baker, G. C., and Cowan, D. A. (2004). 16S rDNA primers and the unbiased assessment of thermophile diversity. Biochemical Society Transactions, 32(2): 218-221.

Balk, M., Weijma, J., and Stams, A. J. (2002). Thermotoga lettingae sp. nov., a novel thermophilic, methanol-degrading bacterium isolated from a thermophilic anaerobic reactor. International Journal of Systematic and Evolutionary Microbiology, 52(Pt 4): 1361-1368.

Ben Hania, W., Postec, A., Aüllo, T., Ranchou-Peyruse, A., Erauso, G., Brochier-Armanet, C., Hamdi, M., Ollivier, B., Saint-Laurent, S., Magot, M., and Fardeau, M. L. (2013). Mesotoga infera sp. nov., a mesophilic member of the order Thermotogales, isolated from an underground gas storage aquifer. International Journal of Systematic and Evolutionary Microbiology, 63(Pt 8): 3003-3008.

Cai, J., Wang, Y., Liu, D., Zeng, Y., and Xue, Y. (2007). Fervidobacterium changbaicum sp. nov., a novel thermophilic anaerobic bacterium isolated from a hot spring of the Changbai Mountains, China. International Journal of Systematic and Evolutionary Microbiology, 57(Pt10): 2333-2336.

Charbonnier, F., and Forterre, P. (1995). Purification of plasmids from thermophilic and hyperthermophilic archaea. In Archaea, a Laboratory Manual, Thermophiles (Robb, F. T. and Place A. R., eds.), pp. 87-90. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.

Davey, M. E., Wood, W. A., Key, R., Nakamura, K., and Stahl, D. (1993). Isolation of three species of Geotoga and Petrotoga: two new genera, representing a new lineage in the bacterial line of descent distantly related to the “Thermotogales”. Systematic and Applied Microbiology, 16(2): 191-200.

Dipippo, J. L., Nesbø, C. L., Dahle, H., Doolittle, W. F., Birkland, N.K., and Noll, K. M. (2009). Kosmotoga olearia gen. nov., sp. nov., a thermophilic, anaerobic heterotroph isolated from an oil production fluid. International Journal of Systematic and Evolutionary Microbiology, 59(Pt 12): 2991-3000.

Dotzauer, C., Ehrmann, M. A., and Vogel, R. F. (2002). Occurrence and detection of Thermoanaerobacterium and Thermoanaerobacterin canned food. Food Technology and Biotechnology, 40(1): 21-26.

Duckworth, A. W., Grant, W. D., Jones, B. E., and Van Steenbergen, R. P. (1996). Phylogenetic diversity of soda lake alkaliphiles. FEMS Microbiology Ecology, 19(3): 181-191.

Erbeznik, M., Jones, C. R., Dawson, K. A., and Strobel, H. J. (1997). Clostridium thermocellum JW20 (ATCC 31549) is a coculture with Thermoanaerobacter ethanolicus. Applied Environmental Microbioogy, 63(7): 2949-2951.

Fardeau, M. L., Ollivier, B., Patel, B. K. C., Magot, M., Thomas, P., Rimbault, A., Rocchiccioli, F., and Garcia, J. L. (1997).Thermotoga hypogea sp. nov., a xylanolytic, thermophilic bacterium from an oil-producing well. International Journal of Systematic Bacteriology, 47(4): 1013-1019.

Friedrich, A. B., and Antranikian, G. (1996). Keratin degradation by Fervidobacterium pennavorans, a novel thermophilic anaerobic species of the order Thermotogales. Applied and Environmental Microbiology, 62(8):2875- 2882.

Huber, R., Langworthy, T. A., Kölnig, H., Thomm, M., Woese, C. R., Sleytr, U.B., and Stetter, K. O. (1986). Thermotoga maritimasp. nov. represents a new genus of uniquely extremely thermophilic eubacteria growing up to 90°C. Archives of Microbiology, 144: 324-333.

Huber, R., Woese, C. R., Langworthy, T. A., Fricke, H., and Stetter, K. O. (1989). Thermosipho africanus gen. nov. represents a new genus of thermophilic eubacteria within the ‘‘Thermotogales’’. Systematic and Applied Microbiology, 12(1): 32-37.

Huber, R., Woese, C. R., Langworthy, T. A., Kristjansson, J. K., and Stetter, K. O. (1990). Fervidobacterium islandicum sp. nov., a new extremely thermophilic eubacterium belonging to the “Thermotogales”. Archives of Microbiology, 154: 105-111.

Jannasch, H. W., Huber, R., Belkin, S., and Stetter, K. O. (1988). Thermotoga neapolitana sp. nov. of the extremely thermophilic, eubacterial genus Thermotoga. Archives of Microbiology, 150(1): 103-104.

Jayasinghearachchi, H. S., and Lal, B. (2011). Oceanotoga teriensis gen. nov., sp. nov., a thermophilic bacterium isolated from offshore oil-producing wells. International Journal of Systematic and Evolutionary Microbiology, 61(Pt 3): 554-560.

Jeanthon, C., Reysenbach, A. L., L’Haridon, S., Gambacorta, A., Pace, N. R., Glénat, P., and Prieur, D. (1995). Thermotoga subterranea sp. nov., a new thermophilic bacterium isolated from a continental oil reservoir. Archives of Microbiology, 164: 91-97.

Keawram, P., and Kanoksilapatham, W. (2013). Diversity of hyperthermophilic bacteriabelonging to order Thermotogales thriving in three hot springs in Thailand: Resources of genes encoding thermostable enzymes. Silpakorn University Science and Technology Journal, 7(2): 17-28.

L’Haridon, S., Miroshnichenko, M. L., Hippe, H., Fardeau, M. L., Bonch-Osmolovskaya, E., Stackebrandt, E., and Jeanthon, C. (2001). Thermosipho geolei sp. nov., a thermophilic bacterium isolated from a continental petroleum reservoir in Western Siberia. International Journal of Systematic and Evolutionary Microbiology, 51(4): 1327-1334.

Lien, T., Madsen, M., Rainey, F. A., and Birkeland, N. K. (1998). Petrotoga mobilis sp. nov., from a North Sea oil-production well. International Journal of Systematic Bacteriology, 48(Pt 3): 1007-1013.

Martens, M., Dawyndt, P., Coopman, R., Gillis, M., de Vos, P., and Willems, A. (2008). Advantages of multilocus sequence analysis for taxonomic studies: a case study using 10 housekeeping genes in the genus Ensifer (including former Sinorhizobium). Journal of Systematic and Evolutionary Microbiology, 58(1): 200-214.

Miranda-Tello, E., Fardeau, M. L., Joulian, C., Magot, M., Thomas, P., Tholozan, J. L., and Ollivier, B. (2007). Petrotoga halophila sp. nov., a thermophilic, moderately halophilic, fermentative bacterium isolated from an offshore oil well in Congo. Journal of Systematic and Evolutionary Microbiology, 57(Pt 1): 40-44.

Miranda-Tello, E., Fardeau, M. L., Thomas, P., Ramirez, F., Casalot, L., Cayol, J. L., Garcia, J. L., and Ollivier, B. (2004). Petrotoga mexicana sp. nov., a novel thermophilic, anaerobic and xylanolytic bacterium isolated from an oil-producing well in the Gulf of Mexico. Journal of Systematic and Evolutionary Microbiology, 54(Pt 1): 169-174.

Mori, Y. (1990). Characterization of a symbiotic coculture of Clostridium thermohydrosulfuricum YM3 and Clostridium thermocellum YM4. Applied Environmental Microbiology, 56(1): 37-42.

Mori, K., Yamazoe, A., Hosoyama, A., Ohji, S., Fujita, N., Ishibashi, J., Kimura, H., and Suzuki, K. (2014). Thermotoga profunda sp. nov. and Thermotoga caldifontis sp. nov., anaerobic thermophilic bacteria isolated from terrestrial hot springs. Journal of Systematic and Evolutionary Microbiology, 64(Pt 6): 2128-2136.

Nam, G. W., Lee, D. W., Lee, H. S., Lee, N. J., Kim, B. C., Choe, E. A., Hwang, J. K., Suhartono, M. T., and Pyun, Y. R. (2002) Native-feather degradation by Fervidobacterium islandicum AW-1, a newly isolated keratinase-producing thermophilic anaerobe. Archives of Microbiology, 178: 538-547.

Nesbø, C. L., Bradnan, D. M., Adebusuyi, A., Dlutek, M., Petrus, A. K., Foght, J., Doolittle, W. F., and Noll, K. M. (2012). Mesotoga prima gen. nov., sp. nov., the first described mesophilic species of the Thermotogales. Extremophiles, 16: 387-393.

Nunoura, T., Oida, H., Miyazaki, M., Suzuki, Y., Takai, K., and Horikoshi, K. (2007). Marinitoga okinawensis sp. nov., a novel thermophilic and anaerobic heterotroph isolated from a deep-sea hydrothermal field, Southern Okinawa Trough. Journal of Systematic and Evolutionary Microbiology, 57(3): 467-471.

Patel, B. K. C., Morgan, H. W., and Daniel, R. M. (1985). Fervidobacterium nodosum gen. nov. and spec. nov., a new chemoorganotrophic, caldoactive, anaerobic bacterium. Archives of Microbiology, 141: 63-69.

Podosokorskaya, O. A., Merkel, A. Y., Kolganova, T. V., Chernyh, N. A., Miroshnichenko, M. L., Bonch-Osmolovskaya, E. A., and Kublanov, I. V. (2011). Fervidobacterium riparium sp. nov. a thermophilic anaerobic cellulolytic bacterium isolated from a hot spring. International Journal of Systematic and Evolutionary Microbiology, 61(11): 2697-2701.

Postec, A., Le Breton, C., Fardeau, M. L., Lesongeur, F., Pignet, P., Querellou, J., Ollivier, B., and Godfroy, A. (2005). Marinitoga hydrogenitoleranssp. nov., a novel member of the order Thermotogales isolated from a black smoker chimney on the Mid-Atlantic Ridge. Journal of Systematic and Evolutionary Microbiology, 55(3): 1217-1221.

Ravot, G., Magot, M., Fardeau, M. L., Patel, B. K., Prensier, G., Egan, A., Garcia, J. L., and Ollivier, B. (1995). Thermotoga elfii sp. nov., a novel thermophilic bacterium from an African oil-producing well. Journal of Systematic and Evolutionary Microbiology, 45(2): 308-314.

Stackebrandt, E., and Geobel, B. M. (1994). Taxonomic note: a place for DNA-DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology. International Journal of Systematic Bacteriology, 44(4): 846-849.

Takahata, Y., Nishijima, M., Hoaki, T., and Maruyama, T. (2001). Thermotoga petrophila sp. nov. and Thermotoga naphthophila sp. nov., two hyperthermophilic bacteria from the Kubiki oil reservoir in Niigata, Japan. International Journal of Systematic and Evolutionary Microbiology, 51(5): 1901-1909.

Takai, K., and Horikoshi, K. (2000). Thermosipho japonicus sp. nov., an extremely thermophilic bacterium isolated from a deep-sea hydrothermal vent in Japan. Extremophiles, 4: 9-17.

Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M., and Kumar, S. (2011). MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution, 28(10): 2731-2739.

Thompson, J. D., Higgins, D. G., and Gibson, T. J. (1994). The CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position specific gap penalties and weight matrix choice. Nucleic Acids Research, 22(22): 4673-4680.

Tindall, B.J., Rosselló-Móra, R., Busse, H. J., Ludwig, W., and Kämpfer, P. (2010). Notes on the characterization of prokaryote strains for taxonomic purposes. International Journal of Systematic and Evolutionary Microbiology, 60(Pt 1): 249-266.

Wery, N., Lesongeur, F., Pignet, P., Derennes, V., Cambon-Bonavita, M.A., Godfroy, A., and Barbier, G. (2001). Marinitoga camini gen. nov., sp. nov., a rod-shaped bacterium belonging to the order Thermotogales, isolated from a deep-sea hydrothermal vent. International Journal of Systematic and Evolutionary Microbiology, 51(2): 495-504

Windberger, E. Huber, R. Trincone, A. Fricke, H., and Stetter, K.O. (1989). Thermotoga thermarumsp. nov. and Thermotoga neapolitana occurring in African continental solfataric springs. Archives of Microbiology, 151: 506-512.