Antibacterial and Antibiofilm Activities of Eco-Friendly Hydrogel Moringa Oleifera–chitosan Formulation Against Escherichia Coli and Staphylococcus Aureus

Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) represent the dominant etiological agents of pyometra in cattle, and their strong biofilm-forming ability critically contributes to therapeutic failure and escalating antimicrobial resistance. This study aimed to develop and evaluate an eco-friendly hydrogel chitosan formulation containing Moringa oleifera (M. oleifera) leaf extract as an alternative antibacterial and antibiofilm strategy. The ethanolic extract of M. oleifera was characterized using GC–MS. Hydrogel formulations (10, 20, 30, and, 40%) were prepared and assessed for bacterial growth inhibition and biofilm degradation over 24, 48, and, 72 hours using microplate spectrophotometry. The formulations exhibited clear concentration- and time-dependent activities. The 10% formulation produced the strongest inhibition against E. coli, whereas S. aureus responded most effectively to the 30% formulation, which maintained stable inhibitory effects across all incubation periods. Biofilm assays revealed transient suppression in E. coli, with maximal degradation at 48 hours, while S. aureus showed more sustained inhibition, particularly under the 30% formulation. Correlation analysis demonstrated a stronger alignment between growth inhibition and biofilm reduction in E. coli, indicating more synchronized antibacterial and antibiofilm effects (p < 0.05). The M. oleifera–chitosan hydrogel formulation exhibited promising dual activity against planktonic and biofilm-associated bacteria, highlighting its potential as an eco-friendly therapeutic candidate for the management of bovine uterine infections and for reducing reliance on conventional antibiotics.