To improve the efficacy of preoperative antibiotics used in elective caesarean section (CS), we aimed to identify the bacteria contaminating the surgical site during this surgery. A study was conducted on 76 Belgian Blue cows. Bacteriology was performed on cotton swab sampled from the visceral and parietal peritoneum of each cow during the CS. Most of samples showed a negative culture (55/76; 72.37%), 19/76 (25%) were positive (p < 0.0001) and two samples were contaminated. In total, 32 isolates belonging to 18 species were identified. Most of them are aerobic (17/18; 94.44%) and half of them were gram-negative (G-). The most encountered bacteria were Acinetobacter sp. (6/32; 18.75%), Pseudomonas sp. (4/32; 12.5%), Aerococcus viridans (4/32; 12.5%), Psychrobacter sp. (3/32; 9.37%), and Escherichia coli (2/32; 6.25%). Among the identified isolates, 31/32 (96.87%) were aerobic and 1/32 (3.12%) was anaerobic (p = 0.0001). Furthermore, 20/32 (62.50%) strains were G− while 12/32 (37.5%) were gram-positive (G+) (p = 0.012). In fact, most of cultured strains were aerobic G− (20/32), 11/32 were aerobic G+ and 1/32 is anaerobic G+ (p < 0.0001). In conclusion, most of samples showed a negative bacteriology; however, aerobic G− strains were the most identified in positive swabs. Therefore, preoperative antibiotics should be aimed against these bacteria.