Animal and in-vitro evidence on Heyndrickxia coagulans MTCC 25308.

Ethanol-induced gastric ulcer model in Wistar rats.

Wistar rats received Bacospore® (100 or 200 mg/kg) or ranitidine (20 mg/kg) orally for 7 days. After 24 h of fasting, gastric ulcers were induced with absolute ethanol (1 mL/200 g, p.o.). Gastric lesions, ulcer index, ulcer protection, and histopathology were assessed.

Marked gastroprotection was observed, with significant reduction in ulcer index and ~80.4% inhibition of ulcer formation, likely through enhancement of mucosal defence and preservation of epithelial integrity.

Castor oil-induced diarrhoea model in male Wistar rats.

Bacospore® was administered at 100 and 200 mg/kg, with loperamide (1 mg/kg) as the standard. Wet stool weight recorded at 2 and 4 hours.

26.6% inhibition of diarrhoea (loperamide: 42.2%) at 200 mg/kg, with wet stool weight at 4 h reduced to 1.16 ± 0.24 g vs. 1.58 ± 0.20 g in the control group.

In-vitro, enzyme-based biochemical assay.

Bacospore® tested at 12.5–200 µg/mL with EDTA as a positive control.

Showed dose-dependent collagenase inhibition, with 85.40% inhibition at 200 µg/mL.

In-vitro bile salt hydrolase (BSH) enzyme assay for cholesterol-lowering activity.

Bacospore® concentrations of 31.25–1,000 µg/mL were tested, with residual cholesterol measured enzymatically using bile salts as the reference.

Concentration-dependent cholesterol inhibition, increasing from 38.77% at 125 µg/mL to 97.72% at 1,000 µg/mL, compared with 99.41% for the bile-salt reference.

3T3-L1 preadipocyte cells and cell-free enzyme assays

MTT assay for cell viability, Oil Red O staining and PPAR-γ ELISA for adipogenesis, DPPH assay for antioxidant activity, and α-amylase and α-glucosidase inhibition assays.

Bacillus coagulans showed efficient antioxidant activity, with an IC₅₀ of 54.19 µg/mL. It maintained cell viability across the tested concentration range (7.8–1,000 µg/mL), with viability remaining around or above 100% at most concentrations. It also showed moderate anti-adipogenic and α-amylase inhibitory activity.