High-throughput screening assays for antibacterial and antifungal activities of Lactobacillus species

•Antibacterial and antifungal assays have been using individual screening approaches.•Screening assay using microtiter plates is suitable for every lab.•2000–5000 individual interactions can be screened per day.•Assay is suggested as a primary screening in combination with traditional methods.

We describe high-throughput screening techniques to rapidly detect either antimicrobial activity, using an agar-well diffusion assay in microtiter plates, or antifungal activity using an agar-spot assay in 24-well plates. 504 Lactobacillus isolates were screened with minimal laboratory equipment and screening rates of 2000–5000 individual antimicrobial interactions.