Our aim was to determine the effects of probiotic supplementation (Bifidobacterium longum 35624; 1 billion CFU·d⁻¹) on exercise performance, immune modulation, and cognitive outlook in collegiate female athletes during six weeks of offseason training. Seventeen National Collegiate Athletic Association (NCAA) Division 1 collegiate female swimmers participated in this two-group matched, double-blind, placebo controlled design. Via stratified randomization, participants were assigned to probiotic (B. longum 35624; n = 8) or placebo (n = 9) groups. Pre, mid, and post-training, all participants completed exercise performance testing (aerobic/anaerobic swim time trials and force plate vertical jump) as well as provided serum (cytokine and gastrointestinal inflammatory markers) and salivary immunoglobulin A samples. Recovery-stress questionnaire for athletes (RESTQ-Sport) was administered at baseline and conclusion of each week. Data were analyzed by analysis of covariance (ANCOVA) by time point with the respective baseline values of each dependent variable being the covariate. No significant differences in exercise performance and biochemical markers were observed between groups following offseason training. Recovery-Stress Questionnaire for Athletes (RESTQ-sport) values in B. longum 35624 group had significantly higher (i.e., more desired; p < 0.05) values in sport recovery (weeks five and six) than placebo. Probiotic supplementation in collegiate female swimmers did not affect exercise performance or immune function throughout offseason training, but did indicate alterations in cognitive outlook.