Borčinová Martina, Pitkina Anastasiya, Marešová Helena, Štěpánek Václav, Palyzová Andrea, Kyslík Pavel
Department of Genetics and Microbiology, Faculty of Science, Charles University in Prague, CZ-12840, Prague 2, Czech Republic.
Institute of Microbiology, Czech Academy of Sciences, CZ-14220, Prague 4, Czech Republic.
Folia Microbiol (Praha). 2020 Dec;65(6):1061-1072. doi: 10.1007/s12223-020-00819-z. Epub 2020 Sep 8.
Ecosystems worldwide are exposed to pollutants connected to the industrial production of pharmaceuticals. The objective of this study was to study the composition and characteristics of the soil microbial communities that had been exposed to long-term selection pressure caused by the industrial production of penicillin G. Soil samples from four sites among the penicillin G production plant were analysed using 16S rRNA profiling via Illumina MiSeq platform and were compared with the control samples from four sites outside the plant. Total metagenomic DNA from the impacted soil was also used for the preparation of E. coli T1R-based fosmid library which was consequently qualitatively tested for the presence of penicillin G acylase (PGA)-encoding genes using the method of sequence homology. Analyses of alpha diversity revealed that the long-term antibiotic presence in the soil significantly increased the microbial diversity and richness in terms of Shannon diversity index (p = 0.002) and Chao estimates (p = 0.004). Principal component analysis showed that the two types of communities (on-site and control) could be separated at the phylum, class and genus level. The on-site soil was enriched in Betaproteobacteria, Deltaproteobacteria, Gemmatimonadetes, Acidobacteria and Planctomycetia, while a significant decrease in Actinobacteria was observed. Metagenomic fosmid library revealed high hit rates in identifying PGAs (14 different genes identified) and confirmed the biotechnological potential of soils impacted by anthropogenic activity. This study offers new insights into the changes in microbial communities of soils exposed to anthropogenic activity as well as indicates that those soils may represent a hotspot for biotechnologically interesting targets.
全球生态系统都受到与药品工业生产相关污染物的影响。本研究的目的是研究长期受到青霉素G工业生产所造成的选择压力影响的土壤微生物群落的组成和特征。通过Illumina MiSeq平台，利用16S rRNA分析对青霉素G生产厂内四个地点的土壤样本进行了分析，并与厂外四个地点的对照样本进行了比较。受影响土壤的总宏基因组DNA也被用于构建基于大肠杆菌T1R的fosmid文库，随后使用序列同源性方法对该文库进行了青霉素G酰化酶（PGA）编码基因存在情况的定性检测。α多样性分析表明，从香农多样性指数（p = 0.002）和Chao估计值（p = 0.004）来看，土壤中抗生素的长期存在显著增加了微生物的多样性和丰富度。主成分分析表明，这两类群落（现场和对照）在门、纲和属水平上可以区分开来。现场土壤中β-变形菌纲、δ-变形菌纲、芽单胞菌门、酸杆菌门和浮霉菌门的含量较高，而放线菌门的含量则显著下降。宏基因组fosmid文库在鉴定PGA方面显示出较高的命中率（鉴定出14个不同基因），并证实了受人为活动影响的土壤具有生物技术潜力。本研究为受人为活动影响的土壤微生物群落变化提供了新的见解，同时表明这些土壤可能是生物技术感兴趣目标的热点地区。
