De Felice Bruna, Argenziano Carolina, Guida Marco, Trifuoggi Marco, Russo Francesca, Condorelli Valerio, Inglese Mafalda
Department of Life Sciences, University of Naples II, Via Vivaldi 43, Caserta, 81100, Italy.
Mol Biotechnol. 2009 Feb;41(2):123-32. doi: 10.1007/s12033-008-9112-1. Epub 2008 Oct 15.
Little is known about pharmaceutical and personal care products pollutants (PPCPs), but there is a growing interest in how they might impact the environment and microbial communities. The widespread use of Viagra (sildenafil citrate) has attracted great attention because of the high usage rate, the unpredictable disposal and the unknown potential effects on wildlife and the environment. Until now information regarding the impact of Viagra on microbial community in water environment has not been reported. In this research, for the first time, the genetic profile of the microbial community, developing in a Viagra polluted water environment, was evaluated by means of the 16S and 18S rRNA genes, for bacteria and fungi, respectively, amplified by polymerase chain reaction (PCR) and separated using the denaturing gradient gel electrophoresis (DGGE) technique. The DGGE results revealed a complex microbial community structure with most of the population persisting throughout the experimental period. DNA sequences from bands observed in the different denaturing gradient gel electrophoresis profiles exhibited the highest degree of identity to uncultured bacteria and fungi found previously mainly in polluted environmental and treating bioreactors. Biotransformation ability of sildenafil citrate by the microbial pool was studied and the capability of these microorganisms to detoxify a polluted water ecosystem was assessed. The bacterial and fungal population was able to degrade sildenafil citrate entirely. Additionally, assays conducted on Daphnia magna, algal growth inhibition assay and cell viability determination on HepG2 human cells showed that biotransformation products obtained from the bacterial growth was not toxic. The higher removal efficiency for sildenafil citrate and the lack of toxicity by the biotransformation products obtained showed that the microbial community identified here represented a composite population that might have biotechnological relevance to retrieve sildenafil citrate contaminated sites.
人们对药品和个人护理产品污染物(PPCPs)了解甚少,但对于它们如何影响环境和微生物群落的兴趣与日俱增。伟哥(枸橼酸西地那非)的广泛使用因其高使用率、不可预测的处置方式以及对野生动物和环境的潜在未知影响而备受关注。到目前为止,尚未有关于伟哥对水环境中微生物群落影响的相关报道。在本研究中,首次分别通过聚合酶链反应(PCR)扩增细菌的16S rRNA基因和真菌的18S rRNA基因,并使用变性梯度凝胶电泳(DGGE)技术进行分离,对在受伟哥污染的水环境中发育的微生物群落的遗传特征进行了评估。DGGE结果显示出复杂的微生物群落结构,大多数种群在整个实验期间持续存在。在不同变性梯度凝胶电泳图谱中观察到的条带的DNA序列与先前主要在受污染环境和处理生物反应器中发现的未培养细菌和真菌具有最高程度的同一性。研究了微生物群体对枸橼酸西地那非的生物转化能力,并评估了这些微生物对受污染水生态系统解毒的能力。细菌和真菌群体能够完全降解枸橼酸西地那非。此外,对大型溞进行的试验、藻类生长抑制试验以及对HepG2人细胞的细胞活力测定表明,从细菌生长中获得的生物转化产物无毒。枸橼酸西地那非的较高去除效率以及所获得的生物转化产物缺乏毒性表明,这里鉴定出的微生物群落代表了一个复合群体,可能在修复受枸橼酸西地那非污染的场地方面具有生物技术相关性。