Strain-Specific Quantification of Native Probiotic Bacillus spp. and Their Effect on Liver Function and Gut Microflora of Experimental Rats

Abstract

Safety and efficacy was investigated for two candidate probiotic B. flexus MCC2427 and B. licheniformis MCC2512 via in vivo studies on albino Wistar rats. In acute toxicity assay, rats were fed with single dose of 10¹⁰ cfu mL^-1 of probiotics. The follow-up studies for next 14 days did not reveal any toxicity-related criteria indicating the non-toxicity nature of probiotics. In 90-day repeated dosage studies, the cultures were administered in three doses (10⁶, 10⁷, 10⁸ cfu mL^-1). Results showed no overt toxic effect and no drastic treatment-related changes pertaining to histopathology of vital organs. DNA fingerprinting indicated the lack of bacterial translocation. Superoxide dismutase and catalase activity indicated their antioxidant potential. Reduced serum cholesterol with improved HDL-cholesterol specified the cholesterol-reducing ability of the cultures, which was also apparent with increased excretion of cholic acid in feces. Both probiotic cultures positively altered the gut microbial environment, retained lactic acid bacterial effect, and simultaneously reduced pathogenic strains. A sensitive and rapid tool was developed using strain-specific qPCR primers, which facilitated appropriate estimation of test culture in feces. The data strongly advocate the safety of tested probiotics at levels used in the study.

Keywords: Bacillus; Fecal microflora; Liver fatty acid profile; Probiotic; qPCR.