, Volume 99, Issue 1, pp 43–53 | Cite as

Presence and potential role of thermophilic bacteria in temperate terrestrial environments

  • M. C. Portillo
  • M. Santana
  • J. M. Gonzalez
Original Paper


Organic sulfur and nitrogen are major reservoirs of these elements in terrestrial systems, although their cycling remains to be fully understood. Both sulfur and nitrogen mineralization are directly related to microbial metabolism. Mesophiles and thermophiles were isolated from temperate environments. Thermophilic isolates were classified within the Firmicutes, belonging to the Geobacillus, Brevibacillus, and Ureibacillus genera, and showed optimum growth temperatures between 50°C and 60°C. Sulfate and ammonium produced were higher during growth of thermophiles both for isolated strains and natural bacterial assemblages. They were positively related to organic nutrient load. Temperature also affected the release of sulfate and ammonium by thermophiles. Quantitative, real-time reverse-transcription polymerase chain reaction on environmental samples indicated that the examined thermophilic Firmicutes represented up to 3.4% of the total bacterial community RNA. Temperature measurements during summer days showed values above 40°C for more than 10 h a day in soils from southern Spain. These results support a potential role of thermophilic bacteria in temperate terrestrial environments by mineralizing organic sulfur and nitrogen ruled by the existence and length of warm periods.


Thermophiles Firmicutes Sulfate Ammonium Mineralization Temperate soils 



This work was supported through projects from the Spanish Ministry of Science and Innovation (CGL2009-12328/BOS; CSD2009-00006) and the regional governments of Aragón (PM055/2006) and Andalusia (BIO-288). The participation of Feder funds in these projects is acknowledged.


  1. Allison SD, Martiny JBM (2008) Resistance, resilience, and redundancy in microbial communities. Proc Natl Acad Sci USA 105:11512–11519PubMedCrossRefGoogle Scholar
  2. Anandham R, Indiragandhi P, Madhaiyan M, Ryu KY, Jee HJ, Sa TM (2008) Chemolithoautotrophic oxidation of thiosulfate and phylogenetic distribution of sulfur oxidation gene (soxB) in rhizobacteria isolated from crop plants. Res Microbiol 159:579–589PubMedCrossRefGoogle Scholar
  3. Battisti DS, Naylor RL (2009) Historical warnings of future food insecurity with unprecedented seasonal heat. Science 323:240–244PubMedCrossRefGoogle Scholar
  4. Boyd WL, Boyd JW (1962) Viability of thermophiles and coliform bacteria in arctic soils and water. Can J Microbiol 8:189–192PubMedCrossRefGoogle Scholar
  5. Calderon F, Reeves JB, Collins HP, Paul EA (2011) Chemical differences in soil organic matter fractions determined by diffuse-reflectance mid-infrared spectroscopy. Soil Sci Soc Am J 75:568–579Google Scholar
  6. Cataldo DA, Haroon M, Schrader LE, Youngs VL (1975) Rapid colorimetric determination of nitrate in plant-tissue by nitration of salicylic-acid. Commun Soil Sci Plant Anal 6:71–80CrossRefGoogle Scholar
  7. Crawford JW, Harris JA, Ritz K, Young IM (2005) Towards an evolutionary ecology of life in soil. Trends Ecol Evol 20:81–87PubMedCrossRefGoogle Scholar
  8. Curtis TP, Sloan WT, Scannell JW (2002) Estimating prokaryotic diversity and its limits. Proc Natl Acad Sci USA 99:10494–10499PubMedCrossRefGoogle Scholar
  9. Diez B, Pedros-Alio C, Marsh TL, Massana R (2001) Application of denaturing gradient gel electrophoresis (DGGE) to study the diversity of marine picoeukaryotic assemblages and comparison of DGGE with other molecular techniques. Appl Environ Microbiol 67:2942–2951PubMedCrossRefGoogle Scholar
  10. Eriksen J (1996) Incorporation of S into soil organic matter in the field as determined by the natural abundance of stable S isotopes. Biol Fertil Soils 22:149–155CrossRefGoogle Scholar
  11. Eriksen J (2008) Soil sulfur cycling in temperate agricultural systems. In: Jez J (ed) Sulfur: a missing link between soils, crops and nutrition. American Society of Agronomy Inc., Madison, WI, pp 25–44Google Scholar
  12. Eschenlauer SCP, McKain N, Walker NA, McEwan NR, Newbold CJ, Wallace RJ (2002) Ammonia production by ruminal microorganisms and enumeration, isolation, and characterization of bacteria capable of growth on peptides and amino acids from the sheep rumen. Appl Environ Microbiol 68:4925–4931PubMedCrossRefGoogle Scholar
  13. Ghani A, McLaren RG, Swift RS (1993) Mobilization of recently-formed soil organic sulphur. Soil Biol Biochem 25:1739–1744CrossRefGoogle Scholar
  14. Gonzalez JM, Ortiz-Martinez A, Gonzalez-del Valle MA, Laiz L, Saiz-Jimenez C (2003) An efficient strategy for screening large cloned libraries of amplified 16S rDNA sequences from complex environmental communities. J Microbiol Meth 55:459–463CrossRefGoogle Scholar
  15. Gonzalez-Prieto SJ, Carballas T (1991) Composition of organic N in temperate humid region soils (NW Spain). Soil Biol Biochem 23:887–895CrossRefGoogle Scholar
  16. Grayston SJ, Germida JJ (1990) Influence of crop rhizosphere on populations and activity of heterotrophic sulfur-oxidizing microorganisms. Soil Biol Biochem 22:457–463CrossRefGoogle Scholar
  17. Gruber N, Galloway JN (2008) An earth-system perspective of the global nitrogen cycle. Nature 451:293–296PubMedCrossRefGoogle Scholar
  18. Hervás A, Camarero L, Reche I, Casamayor EO (2009) Viability and potential for immigration of airborne bacteria from Africa that reach high mountain lakes in Europe. Environ Microbiol 11:1612–1623PubMedCrossRefGoogle Scholar
  19. Huber C, Loy A, Nickel M, Arnosti C, Baranyi C, Brüchert V, Ferdelman T, Finster K, Christensen FM, de Rezende JR, Vandieken V, Jørgensen BB (2009) A constant flux of diverse thermophilic bacteria into the cold arctic seabed. Science 325:1541–1544CrossRefGoogle Scholar
  20. Huber C, Arnosti C, Brüchert V, Loy A, Vandieken V, Jørgensen BB (2010) Thermophilic anaerobes in arctic marine sediments induced to mineralize complex organic matter at high temperature. Environ Microbiol 12:1089–1104CrossRefGoogle Scholar
  21. Intergovernmental Panel on Climate Change (IPCC) (2007) Fourth assessment report: synthesis.
  22. Jarvis SC, Stockdale EA, Shepherd MA, Powlson DS (1996) Nitrogen mineralization in temperate agricultural soils: processes and measurement. Adv Agron 57:187–235CrossRefGoogle Scholar
  23. Jones SE, Lennon JT (2010) Dormancy contributes to the maintenance of microbial diversity. Proc Natl Acad Sci USA 107:5881–5886PubMedCrossRefGoogle Scholar
  24. Kartal B, Koleva M, Arsov R, van der Star W, Jetten MSM, Strous M (2006) Adaptation of a freshwater anammox population to high salinity wastewater. J Biotechnol 126:546–553PubMedCrossRefGoogle Scholar
  25. Kellogg CA, Griffin DW (2006) Aerobiology and the global transport of desert dust. Trends Ecol Evol 21:638–644PubMedCrossRefGoogle Scholar
  26. Kelly DP, Shergill JK, Lu W-P, Wood AP (1997) Oxidative metabolism of inorganic sulfur compounds by bacteria. Ant van Leeuw Intl J Microbiol 71:95–107CrossRefGoogle Scholar
  27. Kolmert A, Wikström P, Hallberg KB (2000) A fast and simple turbidimetric method for the determination of sulfate in sulfate-reducing bacterial cultures. J Microbiol Meth 41:179–184CrossRefGoogle Scholar
  28. Kreutzer K, Butterbach-Bahl K, Rennenberg H, Papen H (2009) The complete nitrogen cycle of an N-saturated spruce forest ecosystem. Plant Biol 11:643–649PubMedCrossRefGoogle Scholar
  29. Kuisiene N, Raugalas J, Stuknyte M, Chitavichius D (2007) Identification of the genus Geobacillus using genus-specific primers, based on the 16S-23S rRNA gene internal transcribed spacer. FEMS Microbiol Lett 277:165–172PubMedCrossRefGoogle Scholar
  30. Lane DJ (1991) 16S/23S rRNA sequencing. In: Stackebrandt E, Goodfellow M (eds) Nucleic acid techniques in bacterial systematic. Wiley, Chichester, pp 205–248Google Scholar
  31. Lobell DB, Burke MB (2008) Why are agricultural impacts of climate change so uncertain? The importance of temperature relative to precipitation. Environ Res Lett 3:034007CrossRefGoogle Scholar
  32. Madigan M, Martinko JM, Parker J (2003) Brock biology of microorganisms. Prentice Hall Inc., New JerseyGoogle Scholar
  33. Marchant R, Banat IM, Rahman TJ, Berzano M (2002) The frequency and characteristics of highly thermophilic bacteria in cool soil environments. Environ Microbiol 4:595–602PubMedCrossRefGoogle Scholar
  34. Marchant R, Franzetti A, Pavlostathis SG, Tas DO, Erdbrügger I, Unyayar A, Mazmanci MA, Banat IM (2008) Thermophilic bacteria in cool temperate soils: are they metabolically active or continually added by global atmospheric transport? Appl Microbiol Biotechnol 78:841–852PubMedCrossRefGoogle Scholar
  35. Martínez-Lladó X, Valderrama C, Rovira J, Marti V, Giménez J, de Pablo J (2011) Sorption and motility of Sb(V) in calcareous soils of Catalonia (NE Spain): Batch and column experiments. Geoderma 160:468–476CrossRefGoogle Scholar
  36. McCalley CK, Sparks JP (2009) Abiotic gas formation drives nitrogen loss from a desert ecosystem. Science 326:837–840PubMedCrossRefGoogle Scholar
  37. Melillo JM, Steudler PA, Aber JD, Newkirk K, Lux H, Bowles FP, Catricala C, Magill A, Ahrens T, Morrisseau S (2002) Soil warming and carbon-cycle feedbacks to the climate system. Science 298:2173–2176PubMedCrossRefGoogle Scholar
  38. Molin S, Givskov M (1999) Application of molecular tools for in situ monitoring of bacterial growth activity. Environ Microbiol 1:383–391PubMedCrossRefGoogle Scholar
  39. Mutzel A, Reinscheid UM, Antranikian G, Müller R (1996) Isolation and characterization of a thermophilic Bacillus strain, that degrades phenol and cresols as sole carbon and energy source at 70°C. Appl Microbiol Biotechnol 46:593–596CrossRefGoogle Scholar
  40. Navas A, Machin J (2002) Spatial distribution of heavy metals and arsenic in soils of Aragón (northeast Spain): controlling factors and environmental implications. Appl Geochem 17:961–973CrossRefGoogle Scholar
  41. Neefs JM, van de Peer Y, Hendriks L, de Wachter R (1990) Compilation of small ribosomal subunit RNA sequences. Nucl Acids Res 18:2237–2317PubMedCrossRefGoogle Scholar
  42. Niknahad-Gharmakher H, Delfosse O, Chauveau-Duriot B, Andueza D, Cornu A (2009) Estimation of sulfur mineralization and relationships with nitrogen and carbon in soils. Biol Fert Soils 45:297–304CrossRefGoogle Scholar
  43. Ordóñez-Fernández R, González-Fernández P, Giráldez-Cervera JV, Perea-Torres F (2007) Soil properties and crop yields after 21 years of direct drilling trials in southern Spain. Soil Tillage Res 94:47–54CrossRefGoogle Scholar
  44. Pedrós-Alió C (2006) Marine microbial diversity: can it be determined? Trends Microbiol 14:257–263PubMedCrossRefGoogle Scholar
  45. Pedrós-Alió C (2007) Dipping into the rare biosphere. Science 315:192–193PubMedCrossRefGoogle Scholar
  46. Perfumo A, Marchant R (2010) Global transport of thermophilic bacteria in atmospheric dust. Environ Microbiol 2:333–339CrossRefGoogle Scholar
  47. Portillo MC, Gonzalez JM (2008) Microbial communities and immigration in volcanic environments of Canary Islands (Spain). Naturwissenschaften 95:307–315PubMedCrossRefGoogle Scholar
  48. Portillo MC, Sririn V, Kanoksilapatham W, Gonzalez JM (2009) Differential microbial communities in hot spring mats from western Thailand. Extremophiles 13:321–331PubMedCrossRefGoogle Scholar
  49. Rahman TJ, Marchant R, Banat IM (2004) Distribution and molecular investigation of highly thermophilic bacteria associated with cool soil environments. Biochem Soc Transactions 32:209–213CrossRefGoogle Scholar
  50. Rowan AK, Snape JR, Fearnside D, Barer MR, Curtis TP, Head IM (2003) Composition and diversity of ammonia-oxidising bacterial communities in wastewater treatment reactors of different design treating identical wastewater. FEMS Microbiol Ecol 43:195–206PubMedCrossRefGoogle Scholar
  51. Rutledge RG (2004) Sigmoidal curve-fitting redefines quantitative real-time PCR with the prospective of developing automated high-throughput applications. Nucl Acids Res 32:e178PubMedCrossRefGoogle Scholar
  52. Schlesinger WH (1997) Biogeochemistry, 2nd edn. Academic Press, San Diego, CAGoogle Scholar
  53. Schoenau JJ, Malhi SS (2008) Sulfur forms and cycling processes in soil and their relationship to sulfur fertility. In: Jez J (ed) Sulfur: a missing link between soils, crops and nutrition. American Society of Agronomy Inc., Madison, WI, pp 1–10Google Scholar
  54. Sleutel S, Leinweber P, van Ranst E, Kader MA, Jegajeevagan K (2011) Organic matter in clay density fractions from sandy cropland soils with differing land-use history. Soil Sci Soc Am J 75:521–532Google Scholar
  55. Sokal RR, Rohlf JF (1981) Biometry. Freeman WH & Co., New YorkGoogle Scholar
  56. Sparling GP, Searle PL (1993) Dimethyl sulphoxide reduction as a sensitive indicator of microbial activity in soil: the relationship with microbial biomass and mineralization of nitrogen and sulphur. Soil Biol Biochem 25:251–256CrossRefGoogle Scholar
  57. Tabatabai MA (1984) Importance of sulfur in crop production. Biogeochem 1:45–62CrossRefGoogle Scholar
  58. Torsvik V, Øvreås L, Thingstad TF (2002) Prokaryotic diversity—magnitude, dynamics, and controlling factors. Science 296:1064–1066PubMedCrossRefGoogle Scholar
  59. Wen G, Schoenau JJ, Mooleki SP, Inanaga S, Yamamoto T, Hamamura K, Inoue M, An P (2003) Effectiveness of an elemental sulfur fertilizer in an oilseed–cereal–legume rotation on the Canadian Prairies. J Plant Nutr Soil Sci 166:54–60CrossRefGoogle Scholar
  60. Whitehead TR, Cotta MA (2004) Isolation and identification of hyper-ammonia producing bacteria from swine manure storage pits. Curr Microbiol 48:20–26PubMedCrossRefGoogle Scholar
  61. Wiegel J, Ljungdahl LG, Rawson JR (1979) Isolation from soil and properties of the extreme thermophile Clostridium thermohydrosulfuricum. J Bacteriol 139:800–810PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  1. 1.IRNAS-CSICSevillaSpain
  2. 2.Department of Fitotecnia, Instituto de Ciências Agrárias e Ambientais MediterrânicasUniversidade de ÉvoraÉvoraPortugal

Personalised recommendations