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

• Published in: Polish journal of microbiology Vol. 66; no. 3; pp. 365 - 373
• Main Authors: Kiersztyn, Bartosz, Siuda, Waldemar, Chróst, Ryszard
• Format: Journal Article
• Language: English
• Published: Poland Exeley Inc 01-09-2017; De Gruyter Poland; Sciendo
• Subjects: active bacteria; Aminopeptidase; aquatic bacteria; Aquatic environment; Aquatic Organisms - metabolism; Bacteria; bacterial culture; Bacterial Proteins - metabolism; Biology; Biotechnology; Chemical properties; Chemical research; Coomassie Blue G250; E coli; Ecology; Escherichia coli - classification; Escherichia coli - metabolism; Eutrophic lakes; Eutrophication; Fluorescence; Fluorescent Dyes - chemistry; Freshwater environments; Lakes - microbiology; Leucine; Leucyl Aminopeptidase - metabolism; Microbial colonies; Microscopy, Fluorescence; Nutrient cycles; Nutrients; Phenyl compounds; Proteins; Research centers; RNA, Ribosomal, 16S - genetics; Rosaniline Dyes - chemistry; rRNA 16S; Staining; Staining and Labeling - methods; Visualization; Poland; active bacteria; Coomassie Blue G250; aquatic environment; bacterial culture; fluorescence; aquatic bacteria
• ISSN: 1733-1331; 2544-4646; 2544-4646
• DOI: 10.5604/01.3001.0010.4867

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.