Bacteria are known for their potential to survive in harsh environmental conditions. The main mechanism involved in this adaptation is the ability to produce and maintain biofilm. This helps the bacteria in surface attachment upon which they survive in a complex matrix produced by the bacterial communities. It has been observed that a large number of the bacterial infections are associated with biofilm formation which is a protective matrix secreted by the bacterial colonies for better survival as compared to their planktonic counterparts. It helps the bacteria to survive in the stressed environment against antimicrobials and normal defence system of the body. It acts as a protective sheath that results in delayed antimicrobial penetration in the biofilm matrix, altered growth rate and physiological changes. These fatal bacterial biofilm-associated infections have been one of the leading causes of livestock death globally. These biofilm-producing bacteria have been detected in the respiratory, urinary, cardiovascular, reproductive, digestive and central nervous system of livestock like cows, buffaloes, sheep, horses and camels. In certain cases, biofilm helps in bacterial colonization and invasion and induces diseases like bovine myocarditis or respiratory diseases, moon blindness, wound infection, endometritis in mares and cows and mastitis in cows. They play an active role in enhancing bacterial virulence using various mechanisms. This chapter mainly focuses on the important bacterial infections of livestock in the Indian subcontinent, role of biofilm in inducing virulence resulting in antibiotic resistance along with other mechanism of invasion and spread of infection followed by probable methods to prevent biofilm-associated diseases in livestock.