Aims: To evaluate the protective efficacy of Pediococcus pentosaceus GS4 in cadmium (Cd)-induced toxicity in Swiss albino mice. Methods and Results: The previously reported probiotic P. pentosaceus GS4 interaction with Cd was evaluated experimentally. Swiss albino mice were segregated randomly into three different groups containing nine animals each. The binding ability of P. pentosaceus GS4 and Cd was demonstrated by atomic absorption spectrometry, Fourier transform infrared spectroscopy and scanning electron microscopy analysis respectively. Cd challenge was attenuated by the P. pentosaceus GS4 treatment and was evaluated by estimating Cd level in liver and faeces, corresponding biomarkers for toxicity in liver and intestines and histopathological analysis with brush border membrane (BBM) integrity. Results showed that P. pentosaceus GS4 bound Cd effectively. Pediococcus pentosaceus GS4 intervention significantly enhanced faecal evacuation of Cd with an effect of reduced tissue deposition. Histologically, Cd-induced gut was found significantly replenished with lactobacilli in addition to reduced hyperplasia, lowered lymphocytes infiltration and enhancement of BBM-based disaccharidases, proving its role in protecting membrane integrity. Conclusions: Probiotic P. pentosaceus GS4 efficiently alleviated toxicity from vital organs such as liver and intestine caused by Cd exposure in a murine model. Significance and Impact of the Study: Intervention of P. pentosaceus GS4 alleviated Cd-induced toxicity, and thus it may be employed therapeutically to protect Cd toxicity. © 2019 The Society for Applied Microbiology