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This study investigates the impact of bromelain, a plant enzyme, on the physicochemical and sensory properties of horse meat, as well as the effects of different bacterial cultures (Lactococcus lactis, Lactococcus lactis subsp. lactis biovar diacetylactis, Lactobacillus acidophilus, and Bifidobacterium longum) on the inhibition of lipid oxidation and control of pH during chilled storage. Horse meat (longissimus dorsi) samples (n = 14) were treated with bromelain in two forms (powder and aqueous solution) and with three methods: immersion in enzyme solution, spreading enzyme powder on meat, and syringing enzyme solution into the meat. After fermentation, a part of the meat samples (n = 6) was treated with different bacteria compositions at a 5% weight ratio and stored at 0–2 °C for 6 days. Injecting 3–5% bromelain solutions was most effective at tenderizing the meat, reducing shear force by up to 56% after 8 h. This injection also maximized the water-holding capacity (78–81%) and minimized cooking losses (21–26%), compared to 38% for the control meat sample without treatment. Syringing with 3% bromelain yielded the highest sensory scores across the tenderness, flavor, and overall palatability parameters. The combination of L. acidophilus, Lc. lactis, and B. longum at a ratio of 1.5:1.5:2 was highly effective in reducing oxidative spoilage and optimizing pH levels, thereby ensuring extended meat storability. This study demonstrates that bromelain treatment is an effective method for improving the tenderness, WHC, and sensory properties of horse meat. The LAB combination showed efficient acid formation, crucial for enhancing meat preservation. [ABSTRACT FROM AUTHOR]