Faghri Jamshid, Poursina Farkhondeh, Moghim Sharareh, Zarkesh Esfahani Hamid, Nasr Esfahani Bahram, Fazeli Hossein, Mirzaei Nasrin, Jamshidian Azam, Ghasemian Safaei Hajieh
Department of Microbiology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, IR Iran.
Department of Immunology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, IR Iran.
Jundishapur J Microbiol. 2014 Jan;7(1):e8704. doi: 10.5812/jjm.8704. Epub 2014 Jan 1.
Helicobacter pylori (H. pylori) is a spiral Gram negative bacteria that can transform to the coccoid form in adverse conditions.
The aim of this study was to determine the in vitro morphological and bactericidal effects of metronidazole, amoxicillin and clarithromycin on H. pylori.
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.
All of the three antibiotics at sub-MIC induced coccoid forms. The highest rates of coccoids (> 90%) were induced at 0.008 μg/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.
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.
幽门螺杆菌(H. pylori)是一种螺旋形革兰氏阴性菌,在不利条件下可转变为球形。
本研究旨在确定甲硝唑、阿莫西林和克拉霉素对幽门螺杆菌的体外形态学和杀菌作用。
幽门螺杆菌标准菌株26695在布鲁氏菌琼脂(BA)上培养,采用E-test法测定三种抗生素的最低抑菌浓度(MIC)。将细菌在布鲁氏菌肉汤(BB)中分别暴露于1/2 MIC、MIC和2倍MIC浓度的抗生素中。通过革兰氏染色和光学显微镜确认诱导形成的球形菌。使用流式细胞术检测细胞活力以及存活球形菌对抗生素的敏感性。
所有三种抗生素在亚MIC浓度下均诱导形成球形菌。暴露72小时后,在0.008μg/mL浓度(1/2 MIC)的阿莫西林作用下,诱导形成球形菌的比例最高(>90%)。1/2 MIC(分别为0.5和0.125μg/mL)的甲硝唑和克拉霉素诱导形成球形菌的比例较低(分别为60%和40%)。观察到2倍MIC的甲硝唑和MIC(0.25μg/mL)的克拉霉素对球形菌有较强的杀菌作用(80 - 90%)。MIC和2倍MIC的阿莫西林对球形菌无杀菌作用。
尽管阿莫西林对幽门螺杆菌螺旋形菌体外杀菌效果良好,但这种抗生素对体内不适当抗菌治疗后可能产生的诱导球形菌几乎没有作用。因此,为了成功治疗,不仅要根除螺旋形菌,还要消除存活的球形菌。