Author/Authors :
Faghri، Jamshid نويسنده , , Poursina، Farkhondeh نويسنده Department of Microbiology, Isfahan University of Medical Sciences, Isfahan, IR Iran , , MOGHIM، SHARAREH نويسنده , , Zarkesh-Esfahani، Sayyed Hamid نويسنده PhD, Department of Immunology, Medical School, Isfahan University of Medical Sciences, Isfahan , , Nasr Esfahani، Bahram نويسنده Department of Microbiology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, IR Iran , , Fazeli، Hossein نويسنده Department of Microbiology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, IR Iran , , Mirzaei، Nasrin نويسنده Department of Biology, Tonekabon branch, Islamic Azad University, Tonekabon, IR Iran , , Jamshidian، Azam نويسنده Department of Microbiology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, IR Iran , , Ghasemian Safaei، Hajieh نويسنده ,
Abstract :
Background:: Helicobacter pylori (H. pylori) is a spiral Gram negative bacteria that can transform to the coccoid form in adverse conditions.
Objectives:: The aim of this study was to determine the in vitro morphological and bactericidal effects of metronidazole, amoxicillin and clarithromycin on H. pylori.
Materials and Methods:: The standard strain 26695 of H. pylori was cultured on Brucella agar (BA) and the minimum inhibitory concentrations (MICs) of three antibiotics were determined by E-test method. The bacteria were exposed to antibiotics at 1/2 MIC, MIC and 2X MIC concentrations in Brucella broth (BB). Induced coccoid forms were confirmed by Gram staining and light microscopy. The viability of cells as well as the susceptibility of viable coccoids to antibiotics were examined using the flow cytometry method.
Results:: All of the three antibiotics at sub-MIC induced coccoid forms. The highest rates of coccoids ( > 90%) were induced at 0.008 ?g/mL concentration (1/2 MIC) of amoxicillin, 72 hours postexposure. Metronidazole and clarithromycin with 1/2 MIC (0.5 and 0.125 µg/mL respectively) induced lower rates of coccoid forms (60% and 40% respectively). Potent bactericidal effects on coccoids were observed with Metronidazole at 2X MIC and clarithromycin at MIC (0.25 µg/mL) (80 - 90%). Amoxicillin with MIC and 2X MIC had no bactericidal effect on coccoid forms.
Conclusions:: Despite the good in vitro bactericidal effect of amoxicillin on spiral forms of H. pylori, this antibiotic has little effect on induced coccoids that may develop after the inappropriate in vivo antibacterial treatment. Hence, for successful therapy, it is essential not only to eradicate the spiral forms, but to eliminate the viable coccoids.
