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Abstract: The root of Terminalia macroptera Guill. & Perr. (Combretaceae) is widely used in African traditional medicine to treat various infectious diseases, including stomach-associated diseases. This study investigates the in vitro activity of T. macroptera root extract against reference strains and clinical isolates of H. pylori and attempts to localize the extract bioactivity. T. macroptera hydroethanol (80% V/V) root extract (Tmr) activity was tested against three standard strains and sixty two clinical strains of H. pylori. Tmr liquid–liquid partition fractions were screened against twenty H. pylori strains. Qualitative analysis of Tmr and its fractions was performed by HPLC-UV/DAD. The antibiotic characterization of the H. pylori strains revealed that 20% of the tested clinical isolates were resistant to at least two of the three antibiotics belonging to the main groups of antibiotics used in multi-therapy to eradicate H. pylori infections. In contrast, Tmr showed anti-H. pylori activity against the majority (92%) of the tested strains (MIC50 and MIC90 =200μg/ml). The Tmr water liquid–liquid fraction (Tmr-3) and the precipitate obtained from this fraction (Tmr-5) were the most active tested samples, showing a MIC50 of 100μg/ml. The present work proves the in vitro activity of T. macroptera against H. pylori, thus confirming the utility of this traditional medicinal plant to treat stomach complaints due to H. pylori infection. The main compounds of Tmr and of Tmr-3 were the ellagitannins terchebulin and punicalagin. These compounds can be considered as markers of T. macroptera root active extracts against H. pylori. [Copyright &y& Elsevier]