Abstract
In this investigation, suitable microbiological techniques and procedures were developed to study associative growth relationships among various probiotic cultures in order to maintain their intended component balance, without any one particular probiotic culture dominating others in a mixed culture by screening them ahead of time using several differential and selective bacteriological media to arrive at a highly-functional therapeutic pharmaceutical mixed probiotic culture. A simulated procedure has been developed mimicking the gradual pH transition of the human Gastro Intestinal tract starting from duodenum to the distal end of the colon. This procedure was developed to study the strain dominance phenomena of when mixed probiotic cultures are grown in a growth medium and sub-cultured through several transfers. The selection criteria applied is if a mixed probiotic culture that can maintain all its individual components through several transfers in the growth medium should be the choice of using it as a functional therapeutic probiotic blend. The results of the investigation proved that undue dominance does exist among strains of probiotics, which belongs not only to different genera, but also among members of the same species when they are grown together. The techniques outlined in this investigation can and has been applied clinically to study the composition of fecal Microbiota, to assess the GI tract health of the human being, and also to develop custom blends of proprietary pharmaceutically-superior therapeutic multiple mixed probiotic cultures to treat Dysbiosis, with choice than chance. The results also proved dominance among the members of mesophilic probiotic species of Lactococcus is more prevalent than thermophilic probiotics belonging to the genus Streptococcus and Lactobacillus. It illustrates that multiple mixed strain probiotic cultures have to be manufactured - for therapeutic purpose - only after determining their associative growth relationships in order to eliminate undue dominance of one particular probiotic culture over another in a mixed culture, and also subsequently in the GI tract of humans after ingestion. Using the microbiological techniques and procedures developed in this investigation, the compositions of novel mixed-strain probiotic cultures outlined in US Patent 11,077,052 B1 have been evaluated and proven that probiotic components used were compatible with no undue domination, thus successfully curing viral and bacterial infections through optimized Microbiota.