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Background  The growth-inhibiting and morphostructural effects of seven constituents identified in  Asarum heterotropoides  root on 14 intestinal bacteria were compared with those of the fluoroquinolone antibiotic ciprofloxacin.    Method  A microtiter plate-based bioassay in sterile 96-well plates was used to evaluate the minimal inhibitory concentrations (MICs) of the test materials against the organisms.    Results  δ-3-Carene ( 5 ) exhibited the most potent growth inhibition of Gram-positive bacteria ( Clostridium difficile  ATCC 9689,  Clostridium paraputrificum  ATCC 25780,  Clostridium perfringens  ATCC 13124, and  Staphylococcus aureus  ATCC 12600) and Gram-negative bacteria ( Escherichia coli  ATCC 11775 and  Bacteroides fragilis  ATCC 25285) (minimal inhibitory concentrations (MIC), 0.18–0.70 mg/mL) except for  Salmonella enterica  serovar Typhimurium ATCC 13311 (MIC, 2.94 mg/mL). The MIC of methyleugenol ( 2 ), 1,8-cineole ( 3 ), α-asarone ( 4 ), (−)-asarinin ( 6 ), and pellitorine ( 7 ) was between 1.47 and 2.94 mg/mL against all test bacteria (except for compound  2  against  C. difficile  (0.70 mg/mL); compounds  1  (23.50 mg/mL) and  4  (5.80 mg/mL) against  C. paraputricum ; compounds  2  (5.80 mg/mL),  4  (12.0 mg/mL), and  7  (0.70 mg/mL) against  C. perfringens ); compound  1  against  E. coli  (7.20 mg/mL) and  S. enterica  serovar Typhimurium (12.0 mg/mL). Overall, all of the constituents were less potent at inhibiting microbial growth than ciprofloxacin (MIC, 0.063–0.25 mg/ mL). The lactic acid-producing bacteria (four bifidobacteria and two lactobacilli) and one acidulating bacterium  Clostridium butyricum  ATCC 25779 were less sensitive and more susceptible than the five harmful bacteria and two nonpathogenic bacteria ( B. fragilis  and  E. coli ) to the constituents and to ciprofloxacin, respectively. Beneficial Gram-positive bacteria and harmful and nonpathogenic Gram-negative bacteria were observed to have different degrees of antimicrobial susceptibility to the constituents, although the antimicrobial susceptibility of the harmful Gram-positive bacteria and the harmful and nonpathogenic Gram-negative bacteria was not observed. Scanning electron microscopy observations showed different degrees of physical damage and morphological alteration to both Gram-positive and Gram-negative bacteria treated with α-asarone, δ-3-carene, pellitorine, or ciprofloxacin, indicating that they do not share a common mode of action.    Conclusion   A. heterotropoides  root-derived materials described merit further study as potential antibacterial products or lead molecules for the prevention or eradication from humans from diseases caused by harmful intestinal bacteria.

Growth-Inhibiting and morphostructural effects of constituents identified in Asarum heterotropoides root on human intestinal bacteria