Background and Objectives: Developing effective treatment outcomes for multidrug-resistantAcinetobacter baumannii(MRAB) infections, with colistin and tigecycline the current frontline therapies, is challenging, because of the risk of renal toxicity and low blood concentrations of active ingredients when administered intravenously. This study aimed to identify the effect of combination therapy using conventional antimicrobial agents that are used for treating drug-resistant bacteria and the additional synergistic effect of four probiotic culture extracts isolated from the human body andLactobacilluspreparations.Materials and Methods: The antimicrobial combination and synergistic effect of addingLactobacillusextract against 33 strains ofA. baumanniiisolated from pus, urine, and other specimens submitted to the Department of Laboratory Medicine of a university hospital, located in Gyeonggi-do, Korea, was investigated over a 3-year period between January 2017 and December 2019.Results: Antimicrobial susceptibility tests on bacteria isolated in clinical practice demonstrated that 26 strains (79%) were MRAB, while multi-locus sequence typing indicated that ST191 was the predominant type (45%;n= 15). Checkerboard test results demonstrated that combination therapy using meropenem and colistin had the highest synergistic effect (fractional inhibitory concentration index = 0.5), while the time–kill assay test usingLactobacillusspp. culture extract exhibited an inhibitory effect within 1 h and complete inhibition of MRAB within 3 h.Lactobacillus paracaseiexhibited the fastest antimicrobial reactivity and longest sustained antimicrobial activity.Conclusion: These findings provide useful foundational data for an appropriate combination of colistin with other antimicrobial agents for treating MRAB infection in clinical settings, and the use of various probiotic culture extracts to reduce the required dosage, and therefore toxicity of colistin.