Probiotics, specifically
Lacticaseibacillus rhamnosus
, have garnered attention for their potential health benefits. This study focuses on evaluating the probiotic properties of candidate probiotics
L. rhamnosus
IDCC 3201 (3201) using the
Caenorhabditis elegans
surrogate animal model, a well-established in vivo system for studying host–bacteria interactions. The adhesive ability to the host’s gastrointestinal tract is a crucial criterion for selecting potential probiotic bacteria. Our findings demonstrated that 3201 exhibits significantly higher adhesive capabilities compared with
Escherichia coli
OP50 (OP50), a standard laboratory food source for
C. elegans
and is comparable with the widely recognized probiotic
L. rhamnosus
GG (LGG). In lifespan assay, 3201 significantly increased the longevity of
C. elegans
compared with OP50. In addition, preconditioning with 3201 enhanced
C. elegans
immune response against four different foodborne pathogenic bacteria. To uncover the molecular basis of these effects, transcriptome analysis elucidated that 3201 modulates specific gene expression related to the innate immune response in
C. elegans
. C-type lectin-related genes and lysozyme-related genes, crucial components of the immune system, showed significant upregulation after feeding 3201 compared with OP50. These results suggested that preconditioning with 3201 may enhance the immune response against pathogens. Metabolome analysis revealed increased levels of fumaric acid and succinic acid, metabolites of the citric acid cycle, in
C. elegans
fed with 3201 compared with OP50. Furthermore, there was an increase in the levels of lactic acid, a well-known antimicrobial compound. This rise in lactic acid levels may have contributed to the robust defense mechanisms against pathogens. In conclusion, this study demonstrated the probiotic properties of the candidate probiotic
L. rhamnosus
IDCC 3201 by using multi-omics analysis.