Details
| Original language | English |
|---|---|
| Article number | 432 |
| Number of pages | 14 |
| Journal | Microorganisms |
| Volume | 8 |
| Issue number | 3 |
| Publication status | Published - 19 Mar 2020 |
Abstract
The stress response of ammonia-oxidizing bacteria (AOB) to oxygen deprivation limits AOB growth and leads to different nitrification pathways that cause the release of greenhouse gases. Measuring the stress response of AOB has proven to be a challenge due to the low growth rates of stressed AOB, making the sample volumes required to monitor the internal stress response of AOB prohibitive to repeated analysis. In a proof-of-concept study, confocal Raman microscopy with excitation resonant to the heme c moiety of cytochrome c was used to compare the cytochrome c content and activity of stressed and unstressed Nitrosomonas europaea (Nm 50), Nitrosomonas eutropha (Nm 57), Nitrosospira briensis (Nsp 10), and Nitrosospira sp. (Nsp 02) in vivo. Each analysis required no more than 1000 individual cells per sampling; thus, the monitoring of cultures with low cell concentrations was possible. The identified spectral marker delivered reproducible results within the signal-to-noise ratio of the underlying Raman spectra. Cytochrome c content was found to be elevated in oxygen-deprived and previously oxygen-deprived samples. In addition, cells with predominantly ferrous cytochrome c content were found in deprived Nitrosomonas eutropha and Nitrosospira samples, which may be indicative of ongoing electron storage at the time of measurement.
Keywords
- Confocal Raman microscopy, Cytochrome c, Nitrosomonas, Nitrosospira, Oxygen deprivation, Resonance Raman spectroscopy
ASJC Scopus subject areas
- Immunology and Microbiology(all)
- Microbiology
- Immunology and Microbiology(all)
- Virology
- Medicine(all)
- Microbiology (medical)
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In: Microorganisms, Vol. 8, No. 3, 432, 19.03.2020.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Small Sample Stress
T2 - Probing Oxygen-Deprived Ammonia-Oxidizing Bacteria with Raman Spectroscopy In Vivo
AU - Kniggendorf, Ann Kathrin
AU - Nogueira, Regina
AU - Bahmanabad, Somayeh Nasiri
AU - Pommerening-Röser, Andreas
AU - Roth, Bernhard Wilhelm
N1 - Funding Information: Funding: This research was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation), grant numbers 397827619 (Ann-Kathrin Kniggendorf) and 397827619 (Regina Nogueira). Bernhard Roth was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany’s Excellence Strategy within the Cluster of Excellence PhoenixD (EXC 2122, Project ID 390833453).
PY - 2020/3/19
Y1 - 2020/3/19
N2 - The stress response of ammonia-oxidizing bacteria (AOB) to oxygen deprivation limits AOB growth and leads to different nitrification pathways that cause the release of greenhouse gases. Measuring the stress response of AOB has proven to be a challenge due to the low growth rates of stressed AOB, making the sample volumes required to monitor the internal stress response of AOB prohibitive to repeated analysis. In a proof-of-concept study, confocal Raman microscopy with excitation resonant to the heme c moiety of cytochrome c was used to compare the cytochrome c content and activity of stressed and unstressed Nitrosomonas europaea (Nm 50), Nitrosomonas eutropha (Nm 57), Nitrosospira briensis (Nsp 10), and Nitrosospira sp. (Nsp 02) in vivo. Each analysis required no more than 1000 individual cells per sampling; thus, the monitoring of cultures with low cell concentrations was possible. The identified spectral marker delivered reproducible results within the signal-to-noise ratio of the underlying Raman spectra. Cytochrome c content was found to be elevated in oxygen-deprived and previously oxygen-deprived samples. In addition, cells with predominantly ferrous cytochrome c content were found in deprived Nitrosomonas eutropha and Nitrosospira samples, which may be indicative of ongoing electron storage at the time of measurement.
AB - The stress response of ammonia-oxidizing bacteria (AOB) to oxygen deprivation limits AOB growth and leads to different nitrification pathways that cause the release of greenhouse gases. Measuring the stress response of AOB has proven to be a challenge due to the low growth rates of stressed AOB, making the sample volumes required to monitor the internal stress response of AOB prohibitive to repeated analysis. In a proof-of-concept study, confocal Raman microscopy with excitation resonant to the heme c moiety of cytochrome c was used to compare the cytochrome c content and activity of stressed and unstressed Nitrosomonas europaea (Nm 50), Nitrosomonas eutropha (Nm 57), Nitrosospira briensis (Nsp 10), and Nitrosospira sp. (Nsp 02) in vivo. Each analysis required no more than 1000 individual cells per sampling; thus, the monitoring of cultures with low cell concentrations was possible. The identified spectral marker delivered reproducible results within the signal-to-noise ratio of the underlying Raman spectra. Cytochrome c content was found to be elevated in oxygen-deprived and previously oxygen-deprived samples. In addition, cells with predominantly ferrous cytochrome c content were found in deprived Nitrosomonas eutropha and Nitrosospira samples, which may be indicative of ongoing electron storage at the time of measurement.
KW - Confocal Raman microscopy
KW - Cytochrome c
KW - Nitrosomonas
KW - Nitrosospira
KW - Oxygen deprivation
KW - Resonance Raman spectroscopy
UR - http://www.scopus.com/inward/record.url?scp=85082670545&partnerID=8YFLogxK
U2 - 10.3390/microorganisms8030432
DO - 10.3390/microorganisms8030432
M3 - Article
AN - SCOPUS:85082670545
VL - 8
JO - Microorganisms
JF - Microorganisms
SN - 2076-2607
IS - 3
M1 - 432
ER -