Oligosaccharides are natural constituents of all bacteria, fungi, plants and placental mammals’ milk. The milk is a rich source of oligosaccharides with different novel oligosaccharides depends on the nature of their origin in which mammals the milk belongs. Buffalo milk oligosaccharides have resistivity against parasitic infections. Donkey milk oligosaccharides have ability to stimulate non-specific and specific immunological resistance. Goat milk oligosaccharides is useful for intestinal protection and repair after a damage caused by DSS (Dextron sodium sulphate)-induced colitis and their implication in human intestinal inflammation. Goat milk oligosaccharides have anti-inflammatory effects. The cows’ milk oligosaccharides reduce the adhesion of enterotoxic Escherichia coli strains of the calf. Remember all bioactive properties of different origin, Gaddi sheep's milk was collected and processed by Modified method of Kobata and Ginsburg and then it was purified by Sephadex G-25 Gel column. The homogeneity is confirmed by reverse phase high performance chromatography and supported by thin layer chromatography.The acetylation of oligosaccharides mixture followed by the silica gel chromatography led to isolation of a novel oligosaccharides. The antimicrobial activity of Gaddi sheep’s milk oligosaccharides was investigated against three bacterial namely Staphylococcus aureus, Escherichia coli and Salmonella Typhimurium and two fungal strains namely Candida albicans and Aspergillus niger are selected on the basis of their relevance as human pathogens. All samples of milk oligosaccharide exhibited antimicrobial activity against dermatomycotic fungi and foodborne pathogen bacteria.