<abstract xmlns="http://www.w3.org/1999/xhtml">

<p>Bacteria play a fundamental role in the cycling of nutrients in aquatic environments. A precise distinction between active and inactive bacteria is crucial for the description of this process. We have evaluated the usefulness of Coomassie Blue G250 for fluorescent staining of protein containing potentially highly active bacteria. We found that the G250 solution has excitation and emission properties appropriate for direct epifluorescence microscopy observations. It enables fast and effective fluorescent visualization of living, protein-rich bacteria, both in freshwater environment and culture. Our results revealed that the number of G250-stained bacteria from eutrophic lake was positively correlated with other standard bacterial activity markers, like number of bacteria containing 16S rRNA, bacterial secondary production or maximal potential leucine-aminopeptidase activity. In case of the <italic>E. coli</italic> culture, the percentage of bacteria visualized with G250 was similar to that of bacteria which accumulated tetracycline. Compared to other common methods utilizing fluorogenic substances for bacteria staining, the approach we evaluated is inexpensive and less hazardous (for example mutagenic) to the environment and researchers. It can be regarded as an additional or alternative method for protein rich, active bacteria staining.</p>
</abstract>

Bacteria play a fundamental role in the cycling of nutrients in aquatic environments. A precise distinction between active and inactive bacteria is crucial for the description of this process. We have evaluated the usefulness of Coomassie Blue G250 for fluorescent staining of protein containing potentially highly active bacteria. We found that the G250 solution has excitation and emission properties appropriate for direct epifluorescence microscopy observations. It enables fast and effective fluorescent visualization of living, protein-rich bacteria, both in freshwater environment and culture. Our results revealed that the number of G250-stained bacteria from eutrophic lake was positively correlated with other standard bacterial activity markers, like number of bacteria containing 16S rRNA, bacterial secondary production or maximal potential leucine-aminopeptidase activity. In case of the E. coli culture, the percentage of bacteria visualized with G250 was similar to that of bacteria which accumulated tetracycline. Compared to other common methods utilizing fluorogenic substances for bacteria staining, the approach we evaluated is inexpensive and less hazardous (for example mutagenic) to the environment and researchers. It can be regarded as an additional or alternative method for protein rich, active bacteria staining.

Coomassie Blue G250 for Visualization of Active Bacteria from Lake Environment and Culture

Faculty of Biology, Biological and Chemical Research Centre

Polish Journal of Microbiology

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Bacteria play a fundamental role in the cycling of nutrients in aquatic environments. A precise distinction between active and inactive bacteria is crucial for...

Coomassie Blue G250 for Visualization of Active Bacteria from Lake Environment and Culture

Published Online: Sep 27, 2017

Page range: 365 - 373

Received: Jan 24, 2017

Accepted: Apr 12, 2017

DOI: https://doi.org/10.5604/01.3001.0010.4867

Keywords
active bacteria, aquatic bacteria, aquatic environment, bacterial culture, Coomassie Blue G250, fluorescence

© 2017 BARTOSZ KIERSZTYN et al., published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Coomassie Blue G250 for Visualization of Active Bacteria from Lake Environment and Culture

Published Online: Sep 27, 2017

Page range: 365 - 373

Received: Jan 24, 2017

Accepted: Apr 12, 2017

DOI: https://doi.org/10.5604/01.3001.0010.4867

Keywordsactive bacteria, aquatic bacteria, aquatic environment, bacterial culture, Coomassie Blue G250, fluorescence

© 2017 BARTOSZ KIERSZTYN et al., published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Keywords
active bacteria, aquatic bacteria, aquatic environment, bacterial culture, Coomassie Blue G250, fluorescence