| Use of a fluorescent redox probe for direct visualization of actively respiring bacteria. | |
| | |
MedLine Citation:
|
PMID: 1622256 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
The redox dye 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) was employed for direct epifluorescent microscopic enumeration of respiring bacteria in environmental samples. Oxidized CTC is nearly colorless and is nonfluorescent; however, the compound is readily reduced via electron transport activity to fluorescent, insoluble CTC-formazan, which accumulates intracellularly. Bacteria containing CTC-formazan were visualized by epifluorescence microscopy in wet-mount preparations, on polycarbonate membrane filter surfaces, or in biofilms associated with optically opaque surfaces. Counterstaining of CTC-treated samples with the DNA-specific fluorochrome 4',6-diamidino-2-phenylindole allowed enumeration of active and total bacterial subpopulations within the same preparation. Municipal wastewater, groundwater, and seawater samples supplied with exogenous nutrients yielded CTC counts that were generally lower than total 4',6-diamidino-2-phenylindole counts but typically equal to or greater than standard heterotrophic (aerobic) plate counts. In unsupplemented water samples, CTC counts were typically lower than those obtained with the heterotrophic plate count method. Reduction of CTC by planktonic or biofilm-associated bacteria was suppressed by formaldehyde, presumably because of inhibition of electron transport activity and other metabolic processes. Because of their bright red fluorescence (emission maximum, 602 nm), actively respiring bacteria were readily distinguishable from abiotic particles and other background substances, which typically fluoresced at shorter wavelengths. The use of CTC greatly facilitated microscopic detection and enumeration of metabolically active (i.e., respiring) bacteria in environmental samples. |
| | |
Authors:
|
G G Rodriguez; D Phipps; K Ishiguro; H F Ridgway |
Related Documents
:
|
18058176 - The identification of 2,4-diacetylphloroglucinol as an antifungal metabolite produced b... 18567526 - Sterilization of oil-field re-injection water using combination treatment of pulsed ele... 23363246 - Fungal diversity from various marine habitats deduced through culture-independent studies. 15049446 - Response to trisodium phosphate treatment of salmonella chester attached to fresh-cut g... 20347196 - Impact of soil organic matter on the distribution of polycyclic aromatic hydrocarbons (... 19618376 - Changes in 13c/12c of oil palm leaves to understand carbon use during their passage fro... |
Publication Detail:
|
Type: Journal Article |
Journal Detail:
|
Title: Applied and environmental microbiology Volume: 58 ISSN: 0099-2240 ISO Abbreviation: Appl. Environ. Microbiol. Publication Date: 1992 Jun |
Date Detail:
|
Created Date: 1992-08-06 Completed Date: 1992-08-06 Revised Date: 2010-09-09 |
Medline Journal Info:
|
Nlm Unique ID: 7605801 Medline TA: Appl Environ Microbiol Country: UNITED STATES |
Other Details:
|
Languages: eng Pagination: 1801-8 Citation Subset: IM |
Affiliation:
|
Biotechnology Research Department, Orange County Water District, Fountain Valley, California 92728-8300. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Bacteria
/
isolation & purification,
metabolism* Fluorescent Dyes Kinetics Microscopy, Fluorescence Oxidation-Reduction Oxygen Consumption Tetrazolium Salts Water Microbiology |
| Chemical | |
Reg. No./Substance:
|
0/Fluorescent Dyes; 0/Tetrazolium Salts; 102568-47-8/5-cyano-2,3-ditolyltetrazolium; 146-68-9/iodonitrotetrazolium |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: High-pressure, high-temperature bioreactor for comparing effects of hyperbaric and hydrostatic press...
Next Document: Cell-associated pectinolytic and cellulolytic enzymes in Rhizobium leguminosarum biovar trifolii.