en Anti-microbial agents, resistance and treatment protocols Anti-microbial agents, resistance and treatment protocols Original article Original article <div style="text-align: justify;"><b><span style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""><span style="color:black">Introduction:</span></span></span></span></b> <span style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""><span style="color:black">The people of Forumad, a village in Semnan Province, Iran, have long used a polyherbal formulation called <i>Sofoof</i> to prevent and treat gastrointestinal disorders. This formulation comprises 11 medicinal plants: <i>Carum carvi</i>, <i>Cuminum cyminum</i>, <i>Pimpinella anisum</i>, <i>Anethum graveolens</i>, <i>Achillea millefolium</i>, <i>Glycyrrhiza glabra</i>, <i>Artemisia absinthium</i>, <i>Terminalia chebula</i>, <i>Peganum harmala</i>, <i>Ferula communis</i>, and <i>Trachyspermum ammi</i>. This study aimed to investigate the antibacterial activity of the <i>Sofoof</i> polyherbal formulation and its individual plant constituents against six species of gastrointestinal pathogenic bacteria</span></span></span></span><span style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""><span style="color:black">. </span></span></span></span><b><span style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""><span style="color:black">Methods<span style="background:white">:</span></span></span></span></span></b><span style="font-size:10.0pt"><span style="background:white"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""><span style="color:black"> Extraction was performed using </span></span></span></span></span><span style="font-size:10.0pt"><span style="background:white"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""><span style="color:black">an infusion method with a solvent system of ethanol, 1-propanol, and water (1:1:1, <i>v/v/v</i>). Antibacterial activity was assessed using the agar well-diffusion assay, and the macrodilution method was used to determine the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC). <b>Results:</b> Extracts of <i>T. chebula</i>, <i>P. harmala</i>, and <i>C. cyminum</i> showed significant antibacterial activity. At a concentration of 31.25 mg/mL, the inhibition zone diameters of <i>T. chebula</i> extract against <i>Escherichia coli</i>,<i> Salmonella typhimurium</i>,<i> Enterococcus faecalis</i>,<i> Shigella dysenteriae</i>,<i> Staphylococcus aureus</i>,<i> </i>and<i> Bacillus cereus </i>were 13, 10, 13, 12, 29, and 24.5 mm, respectively. The MICs for <i>T. chebula</i> against these bacteria ranged from 0.98 mg/mL (<i>Bacillus cereus</i>) to 31.25 mg/mL (<i>Shigella dysenteriae</i>). Notably, the extract of the <i>Sofoof</i> polyherbal formulation (comprising 11 plants) showed lower antibacterial activity than the individual extracts of <i>T. chebula</i> and <i>P. harmala</i></span></span></span></span></span><span style="font-size:10.0pt"><span style="background:white"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""><span style="color:black">. </span></span></span></span></span><b><span style="font-size:10.0pt"><span style="background:white"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""><span style="color:black">Conclusion:</span></span></span></span></span></b> <span style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""><span style="color:black">This study validates the traditional use of the <i>Sofoof</i><b> </b>polyherbal formulation and identifies <i>T. chebula</i>, <i>P. harmala</i>, and <i>C. cyminum</i> as its most active antibacterial constituents. Gram-positive bacteria demonstrated<b> </b>greater susceptibility to the extracts than Gram-negative bacteria. The lower activity of the <i>Sofoof</i><b> </b>polyherbal formulation compared to certain individual plant extracts suggests possible antagonistic interactions, warranting further studies to elucidate constituent interactions<b> </b>within polyherbal formulations</span></span></span></span><span style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""><span style="color:black">.</span></span></span></span></div> &nbsp; Antibacterial activity, Gastrointestinal pathogenic bacteria, Medicinal plants, Sofoof polyherbal formulation 264 273 http://jommid.pasteur.ac.ir/browse.php?a_code=A-10-641-1&slc_lang=en&sid=1 Mohammad Javad Nosrati Mjavad.nosrati71@gmail.com 0009-0006-1939-7484 No Department of Microbiology, GKM.C., Islamic Azad University, Minudasht, Iran Hadi Koohsari hadikoohsari@yahoo.com 0000-0002-0676-2546 Yes Department of Microbiology, Az.C., Islamic Azad University, Azadshahr, Iran Morteza Noryan Noryan1972@gmail.com 0000-0001-5276-0640 No Department of Plant Breeding, Az.C., Islamic Azad University, Azadshahr, Iran