Measuring the Impact of Hydrocarbons on Rates of Nitrogen Fixation

  • Florin MusatEmail author
  • Niculina Musat
Part of the Springer Protocols Handbooks book series (SPH)


Crude oils consist of complex mixtures of hydrocarbons, chemicals which are composed exclusively of carbon and hydrogen atoms. With a very low nitrogen content, crude oil contamination through natural seepage or anthropogenic activities leads to an overload of the affected environment with organic carbon. Degradation of oil hydrocarbons by microorganisms is therefore limited by the availability of fixed nitrogen. Studies of bioremediation of crude oil spills have shown that addition of nitrogen fertilizers or nitrogen salts led to an enhancement of crude oil degradation. Fixed nitrogen in crude oil-contaminated environments could also be provided by N2 fixation, a process carried out by diverse groups of chemoautotrophic, heterotrophic, and phototrophic microorganisms. N2-fixing heterotrophic bacteria have been isolated from environments contaminated with crude oil. Some of these bacteria were able to fix N2 while growing with hydrocarbons as sole substrates. Microcosm studies showed that crude oil alters the N2-fixing microbial populations or offers a substratum for the growth of N2-fixing phototrophic microorganisms. Also, crude oil can indirectly stimulate the growth of N2-fixing microorganisms by reducing the grazing pressure. In this chapter we describe methods to measure N2 fixation rates using sediment or water microcosms. We present methods for preparation of microcosms with addition of crude oil. We describe two methods to measure the bulk N2 fixation by the entire microcosm: the acetylene reduction assay and the analysis of bulk N isotope ratios following incubations with 15N2. Also, we present a method to determine N2 fixation at cellular level, based on nanoSIMS analysis of individual cells from microcosms incubated with 15N2.


15N assimilation 15N isotope labeling Acetylene reduction assay Crude oil NanoSIMS Nitrogen fixation Single cell 


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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  1. 1.Department of Isotope BiogeochemistryHelmholtz Centre for Environmental Research – UFZLeipzigGermany

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