Antibacterial Activity of Lemon Juice on Multi-Drug Resistant Klebsiella species Harboring blaOXA-48 Gene

Babita Malla, S. Khanal, S. K C, B. Yadav, D. Joshi
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Abstract

Klebsiella species cause a wide range of infections, and infections caused by multi-drug resistant species have significantly higher rates of morbidity and mortality. Extended spectrum β-lactamase ESBL prodan emergingd carbapenem resistant  K. pneumoniae is emerging problem in hospital settings worldwise. However, the distribution of blaOXA-48  in Klebsiella pneumoniae has rarely been described from Nepal.  This study was carried out to evaluate in vitro antibacterial activity of lemon juice against clinical isolates of Klebsiella having blaOXA-48 gene. A total of 667 clinical specimens were processed for the identification of Klebsiella species by morphological, cultural and biochemical tests. Methods of antimicrobial susceptibility testing and confirmation of extended spectrum beta lactamase (ESBL) were adopted from the guidelines of the Clinical Laboratory Standard Institute (2018), and blaOXA-48 gene was detected by polymerase chain reaction. The antibacterial activity of lemon juice was assayed using agar well diffusion, and the bactericidal effect of lemon juice was determined by the broth dilution method. Out of 204 culture positive isolates, 29.5% were Klebsiella species of which 16.7% were ESBL producers. Among 19 isolates positive to Modified Hodge test, 68.4% were detected with the plasmid mediated blaOXA-48 gene. The MIC of lemon juice ranged from 12.5-50% (v/v), and the bacteriostatic activity was observed after 6-8 hours of exposure. The tested varieties of lemon juice showed potential inhibitory activity against Klebsiella species indicating that lemon juice has the potential to be used as an antibacterial agent.
柠檬汁对携带blaxa -48基因的多重耐药克雷伯菌的抑菌活性研究
克雷伯氏菌种类引起广泛的感染,由多重耐药物种引起的感染具有明显更高的发病率和死亡率。广谱β-内酰胺酶ESBL原和新出现的耐碳青霉烯类肺炎克雷伯菌是全球医院环境中出现的新问题。然而,blaOXA-48在肺炎克雷伯菌中的分布很少有来自尼泊尔的描述。本研究旨在评价柠檬汁对临床分离的blaOXA-48克雷伯菌的体外抗菌活性。对667份临床标本进行形态、培养和生化鉴定。采用临床检验标准协会(2018)指南的药敏试验和扩展谱β -内酰胺酶(ESBL)确证方法,采用聚合酶链反应检测blaOXA-48基因。采用琼脂孔扩散法测定柠檬汁的抑菌活性,用肉汤稀释法测定柠檬汁的杀菌效果。204株培养阳性菌株中,克雷伯氏菌占29.5%,ESBL产生菌占16.7%。改良霍奇试验阳性的19株分离株中,68.4%的分离株携带质粒介导的blaOXA-48基因。柠檬汁的MIC值为12.5 ~ 50% (v/v),暴露6 ~ 8小时后观察到抑菌活性。柠檬汁对克雷伯氏菌具有潜在的抑菌活性,表明柠檬汁具有作为一种抗菌剂的潜力。
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