Background: Gastrointestinal (GI) infections are a major cause of morbidity and hospitalization in children, particularly in the age group of 1 to 5 years. With increasing concerns over antibiotic resistance and recurrent diarrheal episodes, probiotics have emerged as a promising preventive strategy. This study aimed to evaluate the role of probiotic supplementation in reducing the incidence of gastrointestinal infections among children attending a tertiary care center in Karnataka. Methods: A prospective observational study was conducted over a period of two years, from February 2022 to February 2024, involving 1000 children aged 1 to 5 years. Participants were divided into two groups: those receiving regular probiotic supplementation and those not receiving probiotics. Clinical data including frequency, duration, and severity of GI infections were recorded. Data were analyzed to compare the incidence of GI infections between the two groups. Result: The study observed a significant reduction in the number of gastrointestinal infection episodes among children who received probiotics regularly as compared to those who did not. Additionally, the probiotic group showed reduced severity and shorter duration of symptoms. Hospital admission rates due to GI infections were also lower in the probiotic group. No significant adverse effects related to probiotic use were reported during the study period. Conclusion: Probiotic supplementation in children aged 1 to 5 years significantly reduces the incidence, severity, and duration of gastrointestinal infections. Probiotics may be considered as a safe and effective adjunctive preventive measure in pediatric populations at risk for GI infections, especially in settings with high burden and limited resources.