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Pearl millet dietary fibre fermentation using four co-culture of three probiotic strains Lactobacillus rhamnosus, Bifidobacterium longum, Bifidobacterium bifidus was studied for production of short chain fatty acids (SCFA). All co-cultures were grown on specific medium containing pearl millet fibre fractions; IDF, SDF, TDF as a main carbon source. SCFA production of the probiotics co-cultures was measured at 0, 6, 24, and 48 h using gas liquid chromatography. SCFA formation among the fibre fractions followed the pattern of TDF>SDF>IDF irrespective of co-cultures, indicated that TDF is the best possible dietary fibre for SCFA production. During all co-cultures fermentation with pearl millet fibre fractions, quantity of different SCFA’s produced was acetate>propionate >butyrate. The SCFA quantity was observed low with co-culture of probiotic comprised of same genus (p < 0.05). During 48 h fermentation, co-cultures (BL+BB, BB+LR and BL+LR) digested 81-88 % of the millet fibre fractions. Bifidobacterium bifidum + Bifidobacterium longum + Lactobacillus rhamnosus co-culture was found most efficient during pearl millet fermentation which digested the 97% fibre fractions and produced more SCFA than other combinations in the study. These findings conferred pearl millet dietary fibre fractions as new potential prebiotic for tested probiotic co-cultures.