¹UCSI University, Faculty of Applied Sciences, Department of Food Science and Nutrition, 56000, Cheras, Kuala Lumpur, MALAYSIA DOI : 10.9775/kvfd.2024.30895 The integration of aqueous clove extract (ACE) and spray-dried probiotics into an edible film presents numerous health advantages, such as enhanced probiotic stability and antibacterial efficacy against harmful bacteria like Klebsiella pneumoniae and Pseudomonas aeruginosa. This study aimed to formulate a probiotic edible film with antibacterial properties by incorporating milk powder containing ACE and probiotic strains, namely Streptococcus thermophilus ATCC 19258, Lactobacillus bulgaricus ATCC 11842, and Lactococcus lactis MG1363. The milk powders, containing 7.5% (w/v) ACE with various probiotic strains, were combined with 2% (w/v) pectin biopolymer and 60% (w/w) glycerol plasticizer to produce the probiotic edible film with ACE. Antibacterial effects were investigated using disc diffusion, while probiotic film viability was assessed through the pour plate method. The study revealed higher viability in the probiotic strains L. lactis and S. thermophilus within the edible film, compared to L. bulgaricus. The specific strains exhibited increased antibacterial activity against K. pneumoniae and P. aeruginosa, underscoring the critical role of probiotic strain selection in determining functional properties. Notably, the edible film containing L. lactis and S. thermophilus demonstrated substantial antibacterial activity against both pathogens. However, further research is imperative to optimize industrial-scale production methods, assess efficacy in diverse food items, evaluate sensory properties, gauge barrier attributes, and investigate the impact of storage conditions on stability and performance. Keywords : Probiotic edible film, S. thermophilus, L. bulgaricus, L. lactis, Viability, Antibacterial