The present investigation was carried out to isolate the Bacillus species. From clinical and subclinical Bovine mastitis milk and to determine antimicrobial susceptibility. The samples were collected from mannargudi Taluk, Thiruvarur (Dt) Tamilnadu.from January 2016 to March 2016. A total of 41 Milk samples suffering from mastitis were screened and a total 9 Bacillus species. Were recovered. The isolated were subjected to the antibiotics resistance screening. The antibiotic resistance test showed that the isolated Bacillus species were resistant Methicillin (100%) followed by Penicillin G (91.40%), Oxacillin (80.54%), Cefixime (54.75%), Ampicillin (50.67%), Ceftriaxone (35.29) , Streptomycin (28.50%), Erythromycin (20.36%), Amikacin (17.64%), Gentamicin (12.21%), Cefpodoxime (8.59%), (12.21%), Tetracycline (7.69%), Chloramphenicol (6.33%), Azithromycin (5.42%), Ciprofloxacin (4.07%), Ofloxacin (2.26%) and all Bacillus specie were susceptible to vancomycin. The present study demonstrated the presence of alarming level of resistance of frequently and commonly used antimicrobial agents to the isolated bacteria. Therefore , an examination of the antibiotics resistance profiles of the isolated must be done earlier to the use of antibiotics in both to choose appropriate antibiotic for treatment and prevention of Bovine mastitis.