Evaluation of the efficacy of different sanitizers on the surface of specific pathogens free eggs

Introduction: Specific pathogens free (SPF) eggs is a crucial raw material, with application in several Active Pharmaceutical Ingredients (API) of vaccines included in Brazilian National Immunization Program.Microbiological quantification is a key factor which provides the evaluation and quantification of microorganisms in raw materials, such as SPF eggs.Its sanitization demands a robust validation, which evaluates the time of contact, sanitizing concentration and its efficacy against different microorganisms.Objectives: Ensure the sanitizing efficacy for SPF eggs used in API production.Methodology: Four sanitizers were analyzed in this study: ethanol 70%; ethanol 70% with iodine 0.01%; hydrogen peroxide 6%; a solution containing peracetic acid 0.08%, hydrogen peroxide 1% and acetic acid 10%.Seven ATCC reference strains and two in-house strains were used in this assay.The SPF eggs were contaminated with a solution of each of these microorganisms diluted in PBS, in a concentration of 3 log 10 CFU; after drying, the eggs were exposed to the sanitizers.Three eggs were used as positive controls, and three eggs were used for each type of sanitizing.After the cleaning process, the eggs were placed in a bag with PBS for the microorganisms to be released in the solution.After, membrane filtration methodology was applied, and the membranes were transferred to TSA for bacteria and SDA for fungi.Residual activity of the sanitizing solution was also evaluated.For this purpose, the surface of the eggs was inoculated with the microorganisms after the cleaning process, and filtration method was applied.After incubation, the Colony Forming Units (CFU) were counted and the efficacy of each sanitizer was calculated using decimal reduction, in logarithmic scale, by the difference of positive control and postcleaning values. Results:The initial inoculums were not greater than 3.81 log 10 CFU, and a reduction of not less than 1.02 log 10 CFU was observed for each one of the microorganisms, for each sanitizing, assuring its efficacy, based on Parenteral Drug Association Technical Report N. 70, that proposes a reduction greater than 1.00 log 10 CFU/mL. Conclusion:The sanitizers included in this study showed efficacy for reducing bioburden of the ATCC strains: Aspergillus brasiliensis, Bacillus subtilis, Candida albicans, Escherichia coli, Pseudomonas aeruginosa, Salmonella Typhimurium and Staphylococcus aureus, and the in-house strains: Enterococcus faecalis and Acinetobacter seifertii.This study will allow a wider range of disinfectant solutions to be used for surface disinfection of SPF eggs, contributing to the CCS in the pharmaceutical industry.