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Related Experiment Video

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Small Sample Stress: Probing Oxygen-Deprived Ammonia-Oxidizing Bacteria with Raman Spectroscopy In Vivo.

Ann-Kathrin Kniggendorf1, Regina Nogueira2, Somayeh Nasiri Bahmanabad2

  • 1Hannover Centre for Optical Technologies, Gottfried Wilhelm Leibniz University of Hannover, Nienburger Str. 17, 30167 Hannover, Germany.

Microorganisms
|March 25, 2020
PubMed
Summary
This summary is machine-generated.

Confocal Raman microscopy enables in vivo monitoring of ammonia-oxidizing bacteria stress responses. This technique quantizes cytochrome c, revealing elevated levels in oxygen-deprived bacteria, aiding nitrification pathway studies.

Keywords:
NitrosomonasNitrosospiraconfocal Raman microscopycytochrome coxygen deprivationresonance Raman spectroscopy

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Area of Science:

  • Microbiology
  • Environmental Science
  • Biochemistry

Background:

  • Ammonia-oxidizing bacteria (AOB) are crucial for nitrification, but their stress response to oxygen deprivation impacts nitrogen cycling and greenhouse gas emissions.
  • Assessing AOB stress is difficult due to low cell densities and high sample volume requirements for traditional methods.
  • Understanding AOB stress responses is vital for managing nitrification processes and mitigating environmental impacts.

Purpose of the Study:

  • To develop and validate a non-invasive method for quantifying in vivo stress responses in ammonia-oxidizing bacteria.
  • To compare cytochrome c content and activity in stressed versus unstressed AOB species.
  • To establish a spectral marker for monitoring AOB physiological states.

Main Methods:

  • Confocal Raman microscopy utilizing excitation resonant to the heme c moiety of cytochrome c.
  • In vivo analysis of four AOB strains: *Nitrosomonas europaea* (Nm 50), *Nitrosomonas eutropha* (Nm 57), *Nitrosospira briensis* (Nsp 10), and *Nitrosospira* sp. (Nsp 02).
  • Analysis of minimal sample volumes (≤1000 cells per sampling) to enable monitoring of low-concentration cultures.

Main Results:

  • A reproducible spectral marker for cytochrome c was identified, allowing for reliable measurements within the signal-to-noise ratio.
  • Cytochrome c content was significantly elevated in oxygen-deprived and previously oxygen-deprived AOB samples.
  • Ferrous cytochrome c was predominantly observed in oxygen-deprived *Nitrosomonas eutropha* and *Nitrosospira* samples, suggesting potential electron storage.

Conclusions:

  • Confocal Raman microscopy offers a sensitive, low-volume method for assessing AOB stress responses in real-time.
  • Elevated cytochrome c levels indicate a stress response to oxygen deprivation in AOB.
  • The presence of ferrous cytochrome c may signify active electron storage mechanisms under oxygen-deprived conditions in specific AOB species.