Cm0115

All surgical guides were investigated at different time intervals; after one day, one week, and two weeks from the production stage. Each first half of the three study groups was immersed in one of the three disinfectants; 100%VCO, 2% GA, and 70% EA for 20 min, left to dry for an additional 10 min then soaked in sterile glass containers containing 100 ml. of sterile distilled water for 10 min, and each second half of the three control groups was immersed in sterile glass containers containing 100 ml. of sterile distilled water for 20 min. Three samples were pipetted and cultured on three microbiological media, in which 50 µl (µl) were spread over the surface of Blood (Oxoid, CM0271) and MacConkey agar plates (Oxoid, CM0115) to be incubated at 37 °C for 24 h, and over the surface of Sabouraud dextrose agar plates (HiMEDIA, M063) to be incubated at 37 °C for 48–72 h. After the incubation period, all plates were examined, and the microbial count was done and expressed as colony-forming units per plate (CFU/plate). The percentage (%) of reduction was calculated by the following equation [18 (link)]: $$\%\text{of}\text{reduction}=\text{No}\text{of}\text{CFU}/\text{control}\text{plate}-\text{CFU}/\text{study}\text{plate}/\text{No}\text{of}\text{CFU}/\text{control}\text{plate}\times 100$$

The U-CC samples were subject to microbiological analyses (total viable bacteria (TBC), total viable lactic acid bacteria (LAB) and total viable enterobacteria (TEC)). Microbiological analyses of the U-CC were performed after 1, 3, 4, 7 and 10 days of storage in a refrigerator at +4 °C.
For analysis, 10 g of U-CC was homogenized with 90 mL of saline (9 g/L NaCl solution). Serial dilutions of 10⁴–10⁸ with saline were used for the sample preparation. Evaluation of TBC on plate count agar (PCA, CM0325, Oxoid Ltd., Basingstoke, UK), LAB on The Man Ragosa Sharpe agar (MRS, CM0361, Oxoid Ltd., Basingstoke, UK) and TEC on MacConkey agar (CM0115, Oxoid Ltd., Basingstoke, UK) was undertaken. The number of viable microorganisms was counted in the dilutions containing between 30 and 300 colonies and expressed as log₁₀ of colony-forming units per gram (CFUs/g) [32 (link)]. All results were expressed as the mean value of three determinations and standard error.

Extended-spectrum β-lactamases/AmpC-producing E. coli strains derived from the strain collection of the Institute for Animal Hygiene and Environmental Health, Freie Universitaet Berlin. The isolates were obtained from 7 different pig farms (n = 104) and 22 turkey farms (n = 51) in Germany. Each isolate originated from an individual sample including feces, boot swabs, manure, air or dust. ESBL/AmpC E. coli were selected using MacConkey agar (Oxoid, CM 0115, Wesel, Germany) supplemented with 1 mg/L cefotaxime, followed by species confirmation using MALDI-TOF identification (MALDI Microflex LT and Biotyper database, Bruker Daltonics, Bremen, Germany). The presence of the β-lactamase genes bla_CTX, bla_TEM, bla_SHV and the CIT-type AmpCs (e.g., CMY-2) was confirmed by real-time PCR as described by Roschanski et al. (2014) (link).