Assessment of viability of yeast by flow cytometry using Light Green as vital dye exclusion.

A comparative study of two different staining methods for estimation of yeast's viability is assessed by microbial flow cytometry. Propidium iodide (PI) a well known nucleic-acid binding vital dye exclusion is compared with light green (LG), a cytoplasmic function related vital dye widely used in histological, histochemical and immunocytochemical staining. The yeast model used was heat-killed Candida guilliermondii cells. The stained heat-killed bacterial population showed on the dot-plot flow cytometer analysis, a clear separation towards the green-orange (350-600 nm) for LG, and orange-red (550-725 nm) for PI on the fluorescence spectrum-zone channels. Both dyes showed by non-parametric Kolmcgorov-Smirnov, 99% of probability of difference between heat-killed and live microorganism's histogram-population, with a D = ranking from 0.50-0.64 for LG to 0.91-0.98 for PI. Higher cost and carcinogenic risk when handling PI, make LG a better choice for flow cytometry yeast viability tests.

[Bactericidal effect of hydrated lime in aqueous solution]. / Efecto bactericida de la cal hidratada en solución acuosa.

This study determined the bactericidal effect of the supernatants of saturated solutions of common lime and of micronized calcium hydroxide (Ca(OH)2) (1500 mg/L), which was used as a control, compared with disinfectants made of solutions of 0.33% colloidal silver (0.0016 mg/L), toluene sulfachloramine (41 mg/L) with sodium bicarbonate (9 mg/L), and sodium hypochlorite (5 mg/L). The test involved four strains of Vibrio cholerae 01, V. parahaemolyticus, Escherichia coli, Salmonella typhimurium, Shigella flexneri, Sh. sonnei, and Sa. enterititidis. These bacteria were inoculated into the bactericidal substances listed above and, after different incubation times, the number of surviving bacteria was determined in vitro by using a counting plate. The results were expressed in colony-forming units (CFU). An in situ estimate was made of the amount of V. cholerae on 35 strawberries and 35 radishes (having a weight of about 10 g per unit) after they were washed under a flow of potable water, submerged in the supernatant of the saturated lime solution (1.5 g/L), or both. The greatest bactericidal effect was obtained against V. cholerae 01 and was observed in 3 minutes. Other enterobacteria were resistant to the effect for up to 30 minutes.