Skip to main content
PMC home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1997 Feb;63(2):474–481. doi: 10.1128/aem.63.2.474-481.1997

Stable-Isotope Analysis of a Combined Nitrification-Denitrification Sustained by Thermophilic Methanotrophs under Low-Oxygen Conditions

R Pel, R Oldenhuis, W Brand, A Vos, J C Gottschal, K B Zwart
PMCID: PMC1389516  PMID: 16535510

Abstract

To simulate growth conditions experienced by microbiota at O(inf2)-limited interfaces of organic matter in compost, an experimental system capable of maintaining dual limitations of oxygen and carbon for extended periods, i.e., a pO(inf2)-auxostat, has been used. (sup15)N tracer studies on thermophilic (53(deg)C) decomposition processes occurring in manure-straw aggregates showed the emission of dinitrogen gas from the reactor as a result of simultaneous nitrification and denitrification at low pO(inf2) values (0.1 to 2.0%, vol/vol). The N loss was confirmed by nitrogen budget studies of the system. Depending on the imposed pO(inf2), 0.6 to 1.4 mmol of N/day (i.e., 20 to 40% of input N) was emitted as N(inf2). When the pO(inf2) was raised, the rates of both nitrification and denitrification increased instantaneously, indicating that ammonia oxidation was limited by oxygen. In auxostats permanently running at pO(inf2) >= 2% (vol/vol), the free ammonium pool was almost completely oxidized and was converted to nitrite plus nitrate and N(inf2) gas. Labelling of the auxostat with [(sup13)C]carbonate was conducted to reveal whether nitrification was of autotrophic or heterotrophic origin. Incorporation of (sup13)CO(inf2) into population-specific cellular compounds was evaluated by profiling the saponifiable phospholipid fatty acids (FAs) by using capillary gas chromatography and subsequently analyzing the (sup13)C/(sup12)C ratios of the individual FAs, after their combustion to CO(inf2), by isotope ratio mass spectrometry. Apart from the observed label incorporation into FAs originating from a microflora belonging to the genus Methylococcus (type X group), supporting nitrification of a methylotrophic nature, this analysis also corroborated the absence of truly autotrophic nitrifying populations. Nevertheless, the extent to which ammonia oxidation continued to exist in this thermophilic community suggested that a major energy gain could be associated with it.

Full Text

The Full Text of this article is available as a PDF (435.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alldredge A. L., Cohen Y. Can microscale chemical patches persist in the sea? Microelectrode study of marine snow, fecal pellets. Science. 1987 Feb 6;235(4789):689–691. doi: 10.1126/science.235.4789.689. [DOI] [PubMed] [Google Scholar]
  2. Bloem J., Veninga M., Shepherd J. Fully automatic determination of soil bacterium numbers, cell volumes, and frequencies of dividing cells by confocal laser scanning microscopy and image analysis. Appl Environ Microbiol. 1995 Mar;61(3):926–936. doi: 10.1128/aem.61.3.926-936.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bédard C., Knowles R. Physiology, biochemistry, and specific inhibitors of CH4, NH4+, and CO oxidation by methanotrophs and nitrifiers. Microbiol Rev. 1989 Mar;53(1):68–84. doi: 10.1128/mr.53.1.68-84.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Coffin R. B., Velinsky D. J., Devereux R., Price W. A., Cifuentes L. A. Stable carbon isotope analysis of nucleic acids to trace sources of dissolved substrates used by estuarine bacteria. Appl Environ Microbiol. 1990 Jul;56(7):2012–2020. doi: 10.1128/aem.56.7.2012-2020.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dunfield P., Knowles R. Kinetics of inhibition of methane oxidation by nitrate, nitrite, and ammonium in a humisol. Appl Environ Microbiol. 1995 Aug;61(8):3129–3135. doi: 10.1128/aem.61.8.3129-3135.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fiebig K., Gottschalk G. Methanogenesis from Choline by a Coculture of Desulfovibrio sp. and Methanosarcina barkeri. Appl Environ Microbiol. 1983 Jan;45(1):161–168. doi: 10.1128/aem.45.1.161-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Green J., Dalton H. Steady-state kinetic analysis of soluble methane mono-oxygenase from Methylococcus capsulatus (Bath). Biochem J. 1986 May 15;236(1):155–162. doi: 10.1042/bj2360155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. HUTTON W. E., ZOBELL C. E. Production of nitrite from ammonia by methane oxidizing bacteria. J Bacteriol. 1953 Feb;65(2):216–219. doi: 10.1128/jb.65.2.216-219.1953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jahnke L. L., Nichols P. D. Methyl sterol and cyclopropane fatty acid composition of Methylococcus capsulatus grown at low oxygen tensions. J Bacteriol. 1986 Jul;167(1):238–242. doi: 10.1128/jb.167.1.238-242.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. King G. M., Schnell S. Ammonium and Nitrite Inhibition of Methane Oxidation by Methylobacter albus BG8 and Methylosinus trichosporium OB3b at Low Methane Concentrations. Appl Environ Microbiol. 1994 Oct;60(10):3508–3513. doi: 10.1128/aem.60.10.3508-3513.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Malashenko Iu R., Sokolov I. G., Romanovskaia V. A., Shkurko Iu B. Elementy litotrofnogo metabolizma u obligatnogo metilotrofa Methylococcus thermophilus. Mikrobiologiia. 1979 Jul-Aug;48(4):592–598. [PubMed] [Google Scholar]
  12. McKinley V. L., Vestal J. R. Biokinetic analyses of adaptation and succession: microbial activity in composting municipal sewage sludge. Appl Environ Microbiol. 1984 May;47(5):933–941. doi: 10.1128/aem.47.5.933-941.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Nakasaki K., Sasaki M., Shoda M., Kubota H. Characteristics of Mesophilic Bacteria Isolated during Thermophilic Composting of Sewage Sludge. Appl Environ Microbiol. 1985 Jan;49(1):42–45. doi: 10.1128/aem.49.1.42-45.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Nakasaki K., Shoda M., Kubota H. Effect of temperature on composting of sewage sludge. Appl Environ Microbiol. 1985 Dec;50(6):1526–1530. doi: 10.1128/aem.50.6.1526-1530.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Robertson L. A., Cornelisse R., De Vos P., Hadioetomo R., Kuenen J. G. Aerobic denitrification in various heterotrophic nitrifiers. Antonie Van Leeuwenhoek. 1989 Nov;56(4):289–299. doi: 10.1007/BF00443743. [DOI] [PubMed] [Google Scholar]
  16. Robertson L. A., van Niel E. W., Torremans R. A., Kuenen J. G. Simultaneous Nitrification and Denitrification in Aerobic Chemostat Cultures of Thiosphaera pantotropha. Appl Environ Microbiol. 1988 Nov;54(11):2812–2818. doi: 10.1128/aem.54.11.2812-2818.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Roy R., Knowles R. Effects of methane metabolism on nitrification and nitrous oxide production in polluted freshwater sediment. Appl Environ Microbiol. 1994 Sep;60(9):3307–3314. doi: 10.1128/aem.60.9.3307-3314.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sokolov I. G., Romanovskaia V. A., Shkurko Iu V., Malashenko Iu R. Sravnitel'naia kharakteristika fermentnykh sistem metanispol'zuiushchikh bakterii, okisliaiushchikh NH2OH i CH3OH. Mikrobiologiia. 1980 Mar-Apr;49(2):202–209. [PubMed] [Google Scholar]
  19. Strom P. F. Effect of temperature on bacterial species diversity in thermophilic solid-waste composting. Appl Environ Microbiol. 1985 Oct;50(4):899–905. doi: 10.1128/aem.50.4.899-905.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES