25–30 μg/ml and EDTA alone was also used at the same concentrations. 10 μl of the Candidal suspension with an approximate concentration of 1 × 103 was centrally inoculated in triplicate in each media and incubated at 35 °C. The colonies were observed daily and the growth was visually compared with

ceftriaxone treated control. Further to estimate the growth inhibition, the experiment was carried out by macro broth tube dilution method,10 in a set of tubes containing RPMI medium with PF-01367338 mouse different concentrations of ceftriaxone, Elores containing 6.25–30 μg/ml of EDTA. The test was conducted in two parts – one part of the experiment was carried with single treatment and CFU were enumerated and the second part was continued

with the replenishment of same concentration of the drug dissolved in RPMI medium to replenish the BIBW2992 research buy degraded drug and exhausted nutrients every 24 h. After overnight incubation, the organisms were enumerated by colony count. The sample was diluted and pour plated with Sabouraud’s Dextrose Agar to count the colony forming units per ml. Results were expressed as mean and standard deviation. The bacterial counts in the control and treatment groups were compared statistically by Dunnett’s test using GraphPad Instat 3 software and a P value of <0.05 was considered significant. The average inhibition zone diameters of test substances by disk diffusion tests obtained with Candida are shown in Table 1 and Fig. 1. Inhibition zone diameters were mostly dependent on concentration of EDTA and ceftriaxone present in the Elores regardless of sulbactam and also suggest that there might be some synergistic action between ceftriaxone and EDTA. The agar dilution method used to evaluate the antifungal activity on Candidal growth has shown that the growth was effectively suppressed TCL even at lower concentrations of 6.25 and 12.5 μg/ml of EDTA

in Elores (Fig. 2 and Fig. 3) compared to ceftriaxone alone. The tube method used for the estimation of growth suppression showed the similar pattern and was in agreement with the agar dilution method which was assessed by visual growth. The results presented in Table 2 show the log difference at different concentrations of EDTA and Elores containing equivalent amounts of EDTA after 24 h, 48 h (Fig. 2), 72 (Fig. 3) and 96 h with single treatment. The log difference was noted to be 2.56 for EDTA (P < 0.05) while 3.70 for Elores (P < 0.05) at the lowest concentrations and the log difference decreased with the time. Table 3 shows the log difference at different concentrations of EDTA and Elores containing equivalent amounts of EDTA for four consecutive days with replenishment of the drug every 24 h. The log difference in multiple treatment was 2.51(P < 0.05) and 3.69 (P < 0.05) after 24 h for EDTA and Elores respectively.