Lakhasly

Methods: - Qualitative evaluations were carried out in nutrient broth according to (Hafez et al., 2023).To determine the antimicrobial activity of this silver nano sample throughout the calculation the (%) reduction of colony forming unit (CFU) that appeared after the inoculating 25.0 mL small glass vials containing 5.0 mL sterile nutrient broth medium inoculated with 100.0 uL of pathogenic fungal yeast Candida albicans suspension contains 1.5 x 108 CFU inoculum (0.5 McFarland standard, 1.5 x 108 CFU /ml)(McFarland, 1907) which labeled control vial, then a 100.0 uL of the silver nano solution (after sonication for 10 minutes) was added to the second inoculated vial that labeled treated nano sample vial.The sample [(Silver nano)] was applied After the media cooled and solidified on a 0.6 cm well of the inoculated agar plates which was prepared previously by using the 0.6 cm cork borer applied Well Diffusion Method, in this method, each well was filled with 75.0 ul of the silver nano sample (after sonication for 10 minutes), then the inoculated plates were placed in the refrigerator for one hour to allow the nano sample to more diffuse, followed by incubation at 37 oC for 24 hours, and zones of inhibition (ZI) were measured in mm (Tohamy & El-Masry, 2024).Separately 20.0 mL nutrient agar medium was added to the previously inoculated plates, then incubated at 37 oC for 24 hours, after the proper incubation period, calculating the reduction growth rate R (%) for treated strain compared with the control fungal yeast Candida albicans strain (untreated) according to the following equation: (Hamoda et al., 2022).Relative CFU Reduction (%)] = (A- B /A) x 100

Methods: -
Qualitative evaluations were carried out in nutrient broth according to (Hafez et al., 2023). The inoculation of pathogenic microorganism used in this study was pathogenic fungal yeast [Candida albicans] prepared from fresh overnight broth cultures using nutrient broth medium that were incubated at 37°C (Hamoda et al., 2022 & Abdel Raoof et al., 2024). The inoculum size of this pathogenic strain was prepared and adjusted to approximately 0.5 McFarland standard (1.5 × 108 CFU /ml) (McFarland, 1907), 25.0 µL inoculum size of each microorganism strain was separately inoculated into each plate containing 25.0 mL of the sterile nutrient agar medium (NA). The sample [(Silver nano)] was applied After the media cooled and solidified on a 0.6 cm well of the inoculated agar plates which was prepared previously by using the 0.6 cm cork borer applied Well Diffusion Method, in this method, each well was filled with 75.0 µl of the silver nano sample (after sonication for 10 minutes), then the inoculated plates were placed in the refrigerator for one hour to allow the nano sample to more diffuse, followed by incubation at 37 ºC for 24 hours, and zones of inhibition (ZI) were measured in mm (Tohamy & El-Masry, 2024).

In the second experiment, the antimicrobial method was carried out using both nutrient agar and nutrient broth media, according to Hamoda et al., (2022). The % reduction of (% CFU-reduction) colony-forming unit was measured for this tested strain by counting the total viable colonies of that pathogenic fungal yeast Candida albicans that appeared in both the control and the treated vials separately after the proper incubation period. To determine the antimicrobial activity of this silver nano sample throughout the calculation the (%) reduction of colony forming unit (CFU) that appeared after the inoculating 25.0 mL small glass vials containing 5.0 mL sterile nutrient broth medium inoculated with 100.0 µL of pathogenic fungal yeast Candida albicans suspension contains 1.5 × 108 CFU inoculum (0.5 McFarland standard, 1.5 × 108 CFU /ml)(McFarland, 1907) which labeled control vial, then a 100.0 µL of the silver nano solution (after sonication for 10 minutes) was added to the second inoculated vial that labeled treated nano sample vial.

Then, after the previous proper incubation period from both the vials labeled stoke control and stoke treated nano sample respectively, aseptically 1.0 mL was transferred separately and passed to a new 9.0 mL sterile nutrient broth in 15.0 mL falcon tube, vortex well, the broth culture of this mixture was mixed well to give dilution factor 10-1 of control and treated nano sample respectively. The antimicrobial evaluation was determined by counting the colony forming unit (CFU) that appeared after inoculating the sterile petri-dishes with 1.0 mL from the the stoke control and stoke treated nano sample respectively in addition to 1.0 mL from the dilutions 10-1 of both control and treated nano sample. Separately 20.0 mL nutrient agar medium was added to the previously inoculated plates, then incubated at 37 ºC for 24 hours, after the proper incubation period, calculating the reduction growth rate R (%) for treated strain compared with the control fungal yeast Candida albicans strain (untreated) according to the following equation: (Hamoda et al., 2022).
Relative CFU Reduction (%)] = (A- B /A) × 100