Cycoń Mariusz, Borymski Sławomir, Orlewska Kamila, Wąsik Tomasz J, Piotrowska-Seget Zofia
Department of Microbiology and Virology, School of Pharmacy with the Division of Laboratory Medicine, Medical University of Silesia Sosnowiec, Poland.
Department of Microbiology, University of Silesia Katowice, Poland.
Front Microbiol. 2016 Jun 28;7:1015. doi: 10.3389/fmicb.2016.01015. eCollection 2016.
The occurrence of antibiotics and antibiotic resistance genes in the environment has become a subject of growing concern. The extensive use of vancomycin and other pharmaceuticals may alter the biodiversity of soil microbial communities and select antibiotic-resistant bacteria. Therefore, the purpose of the study was to evaluate the impact of vancomycin and/or vancomycin-resistant Citrobacter freundii on soil microbial communities using the denaturing gradient gel electrophoresis (DGGE) and the phospholipid fatty acid (PLFA) approaches. The experiment had a completely randomized block design with the following treatments: control soil (C), soil with vancomycin (1 mg/kg soil-VA1), soil with vancomycin (10 mg/kg soil-VA10), soil with C. freundii (Cit), soil with vancomycin (1 mg/kg soil) and C. freundii (VA1+Cit), and soil with vancomycin (10 mg/kg soil) and C. freundii (VA10+Cit). A bacterial strain resistant to vancomycin was isolated from raw sewage collected from the municipal sewage treatment plant. The obtained results indicated that the antibiotic and/or the bacterial strain exerted a selective pressure that resulted in qualitative and quantitative changes in the population of soil microorganisms. However, a multivariate analysis showed that the genetic and structural diversity of the soil microbial community was primarily affected by the incubation time and to a lesser extent by the antibiotic and introduced bacteria. DGGE analysis clearly showed that certain species within the bacterial community were sensitive to vancomycin as was evidenced by a decrease in the values of S (richness) and H (Shannon-Wiener) indices. Moreover, a PLFA method-based analysis revealed alterations in the structure of the soil microbial community as indicated by changes in the biomass of the PLFA biomarkers specific for Gram-positive and Gram-negative bacteria as well as fungi. The changes observed in the community of soil microorganisms may decrease the rate of microbial-mediated processes, which can lead to a disturbance in the ecological balance of the soil ecosystem.
环境中抗生素及抗生素抗性基因的出现已成为一个日益受到关注的问题。万古霉素和其他药物的广泛使用可能会改变土壤微生物群落的生物多样性，并筛选出抗抗生素细菌。因此，本研究的目的是使用变性梯度凝胶电泳（DGGE）和磷脂脂肪酸（PLFA）方法，评估万古霉素和/或耐万古霉素弗氏柠檬酸杆菌对土壤微生物群落的影响。该实验采用完全随机区组设计，设置以下处理：对照土壤（C）、添加万古霉素的土壤（1毫克/千克土壤 - VA1）、添加万古霉素的土壤（10毫克/千克土壤 - VA10）、添加弗氏柠檬酸杆菌的土壤（Cit）、添加万古霉素（1毫克/千克土壤）和弗氏柠檬酸杆菌的土壤（VA1 + Cit），以及添加万古霉素（10毫克/千克土壤）和弗氏柠檬酸杆菌的土壤（VA10 + Cit）。从城市污水处理厂收集的原污水中分离出一株耐万古霉素的细菌菌株。所得结果表明，抗生素和/或该细菌菌株施加了选择压力，导致土壤微生物种群在定性和定量上发生变化。然而，多变量分析表明，土壤微生物群落的遗传和结构多样性主要受培养时间影响，其次才受抗生素和引入细菌的影响。DGGE分析清楚地表明，细菌群落中的某些物种对万古霉素敏感，S（丰富度）和H（香农 - 维纳）指数值下降证明了这一点。此外，基于PLFA方法的分析揭示了土壤微生物群落结构的改变，这表现为革兰氏阳性菌、革兰氏阴性菌以及真菌特有的PLFA生物标志物生物量的变化。在土壤微生物群落中观察到的这些变化可能会降低微生物介导过程的速率，进而导致土壤生态系统的生态平衡受到干扰。
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