Factors affecting the determination of respiratory activity on the basis of cyanoditolyl tetrazolium chloride reduction with membrane filtration.

Applied and Environmental Microbiology
B H PyleG A McFeters

Abstract

Deficiencies in traditional bacterial enumeration techniques which rely on colony formation have led to the use of total direct counting methods, such as the acridine orange direct count technique for the enumeration of planktonic bacteria. As total direct counts provide no information on the viability or activity of the organisms, demonstration of respiratory activity with the fluorochrome cyanoditolyl tetrazolium chloride (CTC) has been employed. We have modified this technique by performing filtration prior to CTC incubation. Cells captured on a polycarbonate membrane were incubated on absorbent pads saturated with medium containing CTC. Following counterstaining with DAPI (4(prm1),6-diamidino-2-phenylindole) total and respiring cells were enumerated by epifluorescence microscopy. Factors affecting CTC reduction by Klebsiella pneumoniae, Salmonella typhimurium, and Escherichia coli K-12 were investigated. With K. pneumoniae, nutrient additions to the CTC medium did not increase the number of respiring cells detected. CTC reduction by all three organisms decreased in response to an increase of the pH of the CTC medium above pH 6.5. Increasing phosphate concentrations contributed to this inhibitory effect. CTC-membrane filter ...Continue Reading

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Citations

Apr 5, 2003·Applied and Environmental Microbiology·Laura J RoseMatthew J Arduino
Dec 1, 1998·Journal of Applied Microbiology·G A McFetersS C Broadaway
Mar 28, 2002·Applied and Environmental Microbiology·Leonid A KulakovJohn F O'Hanlon
Jan 5, 2002·Journal of Food Protection·K Takeuchi, J F Frank
Oct 6, 2000·Annual Review of Microbiology·U SzewzykK H Schleifer
Dec 1, 1998·Journal of Applied Microbiology·D P Sartory, J Watkins
May 29, 1999·Journal of Applied Microbiology·D T Reynolds, C R Fricker
Feb 14, 2009·Langmuir : the ACS Journal of Surfaces and Colloids·Ye-Jin Eun, Douglas B Weibel
Dec 1, 1996·Applied and Environmental Microbiology·P MorinJ C Goldman
Dec 1, 1996·Applied and Environmental Microbiology·S UllrichM Mehrens
Jul 1, 1997·Applied and Environmental Microbiology·F JouxM Troussellier

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