The lactoperoxidase system (LPS) is an antimicrobial system naturally present in milk that has been used as a preservative to inhibit microbial growth in raw milk when refrigeration is not viable. Traditionally, the Food and Agriculture Organization (FAO) has recommended the addition of sodium percarbonate and thiocyanate for LPS activation in such instances. Lactose oxidase oxidizes lactose to lactobionic acid, producing H₂O₂ that could act as an antimicrobial or potentially activate the LPS. The aim of this research was to determine if LO can be added as a preservative to inhibit microbial growth in raw milk when refrigeration is limited. A solution of LO was added to raw milk at a concentration of 0.12 g/L both individually (LOX) and combined with NaSCN at 14 mg/L (LOX-T), while the FAO method (FM) consisted of adding 14 mg/L of NaSCN and 30 mg/L of sodium percarbonate. A total of 4 treatments were evaluated including the control (C), all began within 2 h after milking. A total aerobic bacterial growth curve was determined for each treatment, with samples stored at 21°C and sampled 10 times over 24 h. The experiment was carried out in triplicate, and a Buchanan model was applied to each growth curve to calculate the lag phase and growth rate during exponential phase. A one-way ANOVA and Tukey’s test were performed to determine differences in counts among treatments. The lag phase for LOX-T (11.3 h) was significantly longer than C (4.0 h) and LO (5.4 h), but no different from FM (9.4 h). The average growth rate was 0.48 h⁻¹, and no significant difference was determined among treatments. The total bacterial count after 24 h for LOX-T (5.3 cfu/mL) was significantly lower than C (7.2 cfu/mL), LOX (7.1 cfu/mL), and FM (6.1 cfu/mL) (P < 0.0001). Overall, LOX-T shows the most potential for preservation of raw milk stored under abused temperature and its inhibition effect is comparable to the method of reference (FM). Results from this study suggest that LO is an alternative source of H₂O₂ that enhances the microbial inhibition achieved by the LPS. LO could be used to create enzyme-based preservation technologies for applications where cold chain access is limited.