This study evaluated the effects of probiotic administration on growth performance, health status, and fecal microbiota in Holstein calves. Thirty animals were assigned to two groups: the treated group of animals, from 2 to 30 days of age, received a daily oral capsule containing Lactobacillus johnsonii, Ligilactobacillus murinus, Limosilactobacillus mucosae and Ligilactobacillus salivarius (1 × 109 CFU per capsule), whereas the control group received placebo. Health status was monitored using the Wisconsin scoring system, and body weight was recorded at 2 and 60 days of age. Fecal microbiota was characterized at 30 ± 3 days through 16S rRNA gene sequencing. No significant differences were observed between groups regarding diarrhea incidence, weight gain, or microbiota structure. A higher relative abundance of Lactobacillus and Collinsella was observed in the treated group.