Mastitis, which affects the milk production of dairy animals, is usually due to mammary gland invasion by bacterial pathogens. Emergence of antimicrobial resistance in bacteria and side effects associated with the use of anti-inflammatory cortisones in mastitis prompted for use of alternate/complementary therapeutics. As the plant Premna integrifolia was reported to exhibit antibacterial, anti-inflammatory/ immunomodulatory properties, its leaf and root aqueous extracts were tested for their antibacterial activity against Staphylococcus aureus and Escherichia coli, either individually or in combination with the antibiotics. The anti-inflammatory properties of the extracts were also tested against the bubaline mammary epithelial cells (MEC) infected with S. aureus and E. coli. In microbroth dilution assays for assessing minimum inhibitory concentration (MIC) in vitro, the leaf and root extracts of Premna integrifolia didn't exhibit any antimicrobial activity against S. aureus but showed significant antimicrobial activity on E. coli. In combination with the plant extract, the sensitivity of S. aureus to amoxicillin is not only increased but also the S. aureus isolates that were resistant to amoxicillin also became sensitive. The Premna integrifolia leaf and root extracts, however, showed antagonism on antimicrobial activity of enrofloxacin in combination. In addition the aqueous root extract of Premna integrifolia exhibited anti-inflammatory activity through down regulation of cytokines IL-6, IL-8 and TNF-α in S. aureus and IL-6 and IL-8 in E. coli infected MEC. These studies reveal antimicrobial activity of leaf and root extracts of Premna integrifolia on E. coli. In combination with amoxicillin these plant extracts increased the sensitivity of S. aureus to amoxicillin. The anti-inflammatory activity of root extract of Premna integrifolia on MEC infected with S. aureus and E. coli is also demonstrated in these studies.