The enteric pathogens and oxidative stress are known to generate intestinal inflammation, chronic gut ailments and oncogenesis. Modulation of the gut microbial peak populations through herbal agents, offers a promising therapeutic modality. Tribulus terrestris Linn. (Tt), a widely documented medicinal herb in Ayurveda, was investigated for antioxidant, anti inflammatory and antimicrobial activities in vitro. Fruit extract of Tt and quercetin, evaluated for free radical scavenging by DPPH method, revealed IC50 values as 98.83 and 24.77 μg/mL, respectively. Anti-inflammatory attributes of Tt fruit extract and indomethacin, a known anti-inflammatory drug, rendered IC50 values as 10.8 and 12.9 μg/mL against protein denaturation. MTT assay on HCT-15 cells revealed a decrease in viability from 78 to 22% against 30 and 70 μg/mL of Tt fruit extract, respectively. Zone of inhibition against E. coli increased from 0.19 to 9.82 cm2 at 200 and 1000 μg/mL of Tt, respectively. The fruit extract of Tt enhanced the growth of probiotic Lactobacillus rhamnosus (L.rh) by 19, 44 and 50 % over the control at 100, 150 and 200 μg/mL, respectively. This study indicated the potential of Tribulus terrestris fruit extract against inflammatory, oxidative and microbe generated pathogenic ailments in the digestive system.