The prevalence of antibiotic resistance is a serious global health problem and a threat to human health. Staphylococcus aureus is a Gram-positive bacterium that can cause various infections in humans, such as skin infections, sepsis, endocarditis, osteomyelitis, and toxins that cause food poisoning. One way to overcome the problem of antibiotic resistance is to use Lactic Acid Bacteria (LAB). LAB has many benefits, including being able to function in food fermentation, produce antimicrobial compounds, and act as probiotics. This study aims to determine the inhibitory effect of LAB from baby feces as a natural antibacterial agent against S. aureus. The method used in this study was the agar well diffusion method. The number of LAB isolates used was 8 consisting of seven cocci and two bacilli forms and also used 2 controls, namely, negative control NaCl 0.9% and Positive Control Ciprofloxacin. This study consisted of six stages including media preparation, preparation of test bacteria, sample preparation, making suspensions of S. aureus and LAB, making Mcfarland solution, and antibacterial testing using the agar well diffusion method for 3 repetitions of the treatment. The results obtained showed that all LAB isolates were able to produce an inhibition zone against Staphylococcus aureus bacteria with the results of the isolates in diameter 01A 10-5 (7.67mm), 01A 10-6(2) (6mm), 03A10-7(1) (6.67mm), 01F 10-6(2) (2mm), 01F10-7(2) (1.67mm), 02AF10-7(1) (4.33mm), 03AF10-7(2) (4mm), 04AF10-7 (3.67mm). Based on the results of the Mann-Whitney test, it was found that all LAB isolates had significant differences from the positive control Ciprofloxacin. So it can be seen that LAB from baby feces is less effective in replacing antibiotics.