Optimization of probios biomass production on low-cost food industrial waste-based media as anti- Salmonella solution

Abstract

This study uses industrial waste to enhance growth media in order to combat the growing challenge of multi-drug resistant salmonella in chicken. The purpose of this project is to provide the poultry industry with a practical solution to reduce MDR salmonella through the commercialization of probiotics (PASS: patent number 534/2022). Since cost-effectiveness is crucial, we are concentrating on creating and improving low-cost production media using specific industrial waste ingredients (whey, corn-steep liquor, and molasses) to help these probiotics become commercially viable. Utilization of these wastes for the purpose of developing growth media also contributes to environmental conservation by reducing pollution specifically of water bodies. An experimental trial was conducted to optimize the production medium for the enhanced production of Lactobacillus reuteri 104, Enterococcus faecium 113, and Enterococcus faecium 114 biomass. Further optimization was performed by using statistical methods such as box-Behnken design of response surface methodology to develop cost-effective media. According to our findings, 30% of molasses performed better than the control MRS medium. On whey, the growth pattern is dynamic and non-linear. Notably, for strain Enterococcus faecium 113, our results were superior to MRS at 10% w/v whey concentration, and for strains Lactobacillus reuteri 104 and Enterococcus faecium 114, they were comparable. In the case of CSL, we detected an increase in probiotics yield upto 10% v/v of CSL, there was a subsequent drop. Although growth is significantly reduced compared to MRS, it is still significant for high-volume production. After the evaluation of growth of probiotics on crude media, Box-Behnken response surface methodology was used to find the optimized media proportion, which generated different combinations of input values, i.e. 17 combinations of independent variables (molasses, whey and CSL). Growth parameters of probiotic strains (YOD 104, Ybiomass 104, YOD 113, Ybiomass 113, YOD 114, Ybiomass 114) were analyzed as a function of independent factors. Regression models were generated for the prediction of all the responses and explained statistically by analysis of variance (ANOVA). Multiple correlation coefficient values for all the models were close to one showing a good relation between predicted and experimental values. An optimized run was generated in the numerical optimization portion of RSM i.e. Run (22.5 5.25 10) with 22.5% v/v molasses, 5.25% w/v whey and 10% v/v CSL. A run was performed on optimized conditions to confirm the prediction value and a good agreement was found between predicted and experimental values. While tenfold cost reduction was found in case of waste based designed media as compared to MRS aligning the goal of developing economic media for commercialization of probiotics.