Document Detail

Method for enumeration of 5-cyano-2,3-ditoyl tetrazolium chloride (CTC)-active cells and cell-specific CTC activity of benthic bacteria in riverine, estuarine and coastal sediments.
MedLine Citation:
PMID:  11118655     Owner:  NLM     Status:  MEDLINE    
Bacteria are the most abundant and active organisms in marine sediments and are critical for nutrient cycling and as a food source to many benthic and pelagic organisms. Bacteria are found both as free-living cells and as particle-associated cells, which can make investigations of these communities difficult. We found that common procedures for extracting bacteria from sediments leave the bacteria clay particle-associated and the clay particles clump, which reduce the reproducibility of direct counts. We optimized a sonication/surfactant method that produces a homogeneous suspension of bacterial cells against a uniform background of clay particles, which results in reproducible samples for epifluorescence microscopy. We developed a method to estimate CTC-positive cells and cell-specific CTC content in intact cores of surficial sediment communities from riverine, estuarine and coastal sites. Benthic bacterial abundances averaged 4.9x10(8) cells/g dry wt sediments in Apalachicola River, Florida sediments, 4.9-13.8x10(9) cells/g dry wt sediments in a variety of Apalachicola Bay sediments and 3.6x10(8) cells/g dry weight in shallow, anoxic Gulf of Mexico sediments. Percent CTC-positive cells ranged from low values of 9-10% CTC-positive cells in Apalachicola River and Apalachicola Bay sediments to high values of 25% CTC-positive cells in anoxic Gulf of Mexico sediments. After correction for abiotic CTC reduction and chlorophyll interference, estimates of cell-specific CTC reduction ranged from 0.15 to 0.55 fmol CTC(red)/active cell in the Apalachicola Bay sediments to 1.6 to 3.8 fmol CTC(red)/active cell in anoxic Gulf of Mexico sediments.
L M Proctor; A C Souza
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Journal of microbiological methods     Volume:  43     ISSN:  0167-7012     ISO Abbreviation:  J. Microbiol. Methods     Publication Date:  2001 Jan 
Date Detail:
Created Date:  2001-01-26     Completed Date:  2001-05-10     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8306883     Medline TA:  J Microbiol Methods     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  213-22     Citation Subset:  IM    
Department of Oceanography, Florida State University, Tallahassee, FL 32306-4320, USA.
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MeSH Terms
Bacteria / growth & development,  isolation & purification*,  metabolism*
Bacteriological Techniques
Colony Count, Microbial*
Geologic Sediments / microbiology*
Microscopy, Fluorescence
Oxygen Consumption
Tetrazolium Salts / metabolism*
Water Microbiology*
Reg. No./Substance:
0/Tetrazolium Salts; 102568-47-8/5-cyano-2,3-ditolyltetrazolium

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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