The catholyte stream (Aversol™ by Trustwater) Selleckchem Rigosertib has a high pH and is classified as an amphoteric surfactant, having reduced surface tension and mild detergent-like properties. Trustwater’s automated process uses this solution to maintain the Ecasol stream at a neutral pH. The new Ecasol solution was titrated at 700 ppm free available chlorine (FAC), with a pH of 6.7. The solution was delivered to the lab on the day of the experiment and was used immediately upon delivery, with a time from solution generation to lab experimentation of approximately 2 h. The stability of Ecasol depends upon storage conditions because it can lose up to

10% of its activity within 3 weeks of generation if it is not stored properly. Two concentrations of Ecasol, 150 ppm and 500 ppm FAC, were prepared by diluting the solution with deionized water. These testing concentrations selleck screening library were selected because 150 ppm is the most commonly

used concentration for food contact surface sanitization, based on the recommendation of 40CFR 180.940, and the concentration of 500 ppm was selected because Ecasol is a known sporocidal at this concentration. The test was performed in 6-well tissue culture plates, and the experiments were conducted in triplicate. The six wells of the plate were labeled A through F, and the FCV suspension was uniformly applied to the bottom of the six wells at 100 μL/well. The inoculum was allowed to dry for 30 min at room temperature (approximately 23 °C) in a type II biosafety cabinet. After the inoculum dried, the Ecasol solution

was Chlormezanone added to wells A–C at 5 mL/well. Wells D–F served as controls for each treated well (well D for well A, and so on), with 5 mL of phosphate buffered saline (PBS) per well. The plate was incubated at room temperature on an orbital shaker (at 120 rpm) for different time periods (1, 2, and 5 min for wells A and D, B and E, and C and F, respectively). After the appropriate contact times, the well contents were immediately diluted 10-fold using a maintenance medium to stop Ecasol activity at the indicated times. Serial 10-fold dilutions of these eluates were prepared in Eagle’s MEM, followed by inoculation of CRFK cells grown in 96-well microtiter plates, using four wells for each test dilution. The inoculated plates were incubated at 37 °C and examined daily for 4 days by microscope for FCV-induced cytopathic effects (CPE). The virus titers were calculated by the Reed and Muench method [13], and the log reductions were calculated by comparing the titers of the Ecasol-treated wells with those of the PBS-treated control wells. To determine the cytotoxicity of the Ecasol solution to the CRFK cells, 10-fold serial dilutions of Ecasol prepared in Eagle’s MEM were added to monolayers of CRFK cells prepared in a 96-well plate (4 wells/dilution).