Abstract: Bovine mastitis is one of the diseases which can cause heavy economic losses.In this paper, the antibacterial experiment, which included various kinds of factors, such as bacteriocin, H₂O₂, showed the antibacterial capacity of 6 strains of Lactobacillus acidophilus and its antibacterial material.It was showed that 6 strains of lactic acid bacteria had great antibacterial ability on Staphylococcus aureus, Lactobacillus acidophilus NCFM (H7) acellular supernatant fluid had the strongest antibacterial capacity to 4 strains of pathogenic bacteria which could cause bovine mastitis;bacteriocin produced from KLDS AD3 (H6) had the best inhibitory effect to Streptococcus agalactiae, bacteriocin produced from KLDS L6 (H3) had the best inhibitory effect to Streptococcus uberis and Escherichia coli, and its H₂O₂ had great antibacterial impact on Streptococcus uberis, Escherichia coli and Staphylococcus aureus, bacteriocin produced from KLDS AD2 (H5) also had the best inhibitory effect to Staphylococcus aureus.L6 (H3) bacteriostatic active macterial had great heat-resisting capacity, however, AD2 (H5) bacteriostatic active macterial had great thermal sensitivity.These substances had better antibacterial activity in acidic conditions.