| Summary: | The continued exploration of the diversity of lactic acid bacteria in little-studied ecological niches represents a fundamental activity to understand the diffusion and biotechnological significance of this heterogeneous class of prokaryotes. In this study, <i>Lactiplantibacillus plantarum</i> (<i>Lpb. plantarum</i>) strains were isolated from Tunisian vegetable sources, including fermented olive and fermented pepper, and from dead locust intestines, which were subsequently evaluated for their antimicrobial activity against foodborne pathogenic bacteria, including <i>Escherichia coli</i> O157:H7 CECT 4267 and <i>Listeria monocytogenes</i> CECT 4031, as well as against some fungi, including <i>Penicillium expansum</i>, <i>Aspergilus niger</i>, and <i>Botrytis cinerea</i>. In addition, their resistance to oro-gastro-intestinal transit, aggregation capabilities, biofilm production capacity, adhesion to human enterocyte-like cells, and cytotoxicity to colorectal adenocarcinoma cell line were determined. Further, adhesion to tomatoes and the biocontrol potential of this model food matrix were analyzed. It was found that all the strains were able to inhibit the indicator growth, mostly through organic acid production. Furthermore, these strains showed promising probiotic traits, including in vitro tolerance to oro-gastrointestinal conditions, and adhesion to abiotic surfaces and Caco-2 cells. Moreover, all tested <i>Lpb. plantarum</i> strains were able to adhere to tomatoes with similar rates (4.0–6.0 LogCFU/g tomato). The co-culture of LAB strains with pathogens on tomatoes showed that <i>Lpb. plantarum</i> could be a good candidate to control pathogen growth. Nonetheless, further studies are needed to guarantee their use as probiotic strains for biocontrol on food matrices.
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