Kapoor V, Elk M, Li X, Santo Domingo J W
Oak Ridge Institute for Science and Education, Oak Ridge, TN, USA.
U.S. Environmental Protection Agency, Office of Research and Development, Cincinnati, OH, USA.
Lett Appl Microbiol. 2016 Aug;63(2):155-61. doi: 10.1111/lam.12603.
The effect of cyanide (CN(-) ) on nitrification was examined with samples from nitrifying bacterial enrichments using two different approaches: by measuring substrate (ammonia) specific oxygen uptake rates (SOUR), and by using RT-qPCR to quantify the transcripts of functional genes involved in nitrification. The nitrifying bioreactor was operated as a continuous reactor with a 24 h hydraulic retention time. The samples were exposed in batch vessels to cyanide for a period of 12 h. The concentrations of CN(-) used in the batch assays were 0·03, 0·06, 0·1 and 1·0 mg l(-1) . There was considerable decrease in SOUR with increasing dosages of CN(-) . A decrease of more than 50% in nitrification activity was observed at 0·1 mg l(-1) CN(-) . Based on the RT-qPCR data, there was notable reduction in the transcript levels of amoA and hao for increasing CN(-) dosage, which corresponded well with the ammonia oxidation activity measured via SOUR. The inhibitory effect of cyanide may be attributed to the affinity of cyanide to bind ferric haeme proteins, which disrupt protein structure and function. The correspondence between the relative expression of functional genes and SOUR shown in this study demonstrates the efficacy of RNA-based function-specific assays for better understanding of the effect of toxic compounds on nitrification activity in wastewater.
The effect of cyanide on nitrifying bacteria was characterized by measuring physiological and transcriptional response. Cyanide was inhibitory to nitrification at concentrations that may be found in industrial waste. The RNA-based function-specific assays represent a mechanistic approach for better understanding the effect of toxic compounds on nitrification activity in wastewater. Moreover, the relative abundance of RNA transcripts can be used to closely track in situ nitrifying bacterial activity which can be used to predict inhibition events, thereby providing a metric to potentially improve performance of wastewater nitrifying systems.
使用两种不同方法,通过硝化细菌富集培养物的样本研究了氰化物(CN⁻)对硝化作用的影响:一是测量底物(氨)的特定氧摄取率(SOUR),二是使用RT-qPCR对参与硝化作用的功能基因转录本进行定量。硝化生物反应器作为连续反应器运行,水力停留时间为24小时。将样本在分批容器中暴露于氰化物12小时。分批试验中使用的CN⁻浓度分别为0.03、0.06、0.1和1.0 mg l⁻¹。随着CN⁻剂量增加,SOUR显著下降。在CN⁻浓度为0.1 mg l⁻¹时,观察到硝化活性下降超过50%。基于RT-qPCR数据,随着CN⁻剂量增加,amoA和hao的转录水平显著降低,这与通过SOUR测量的氨氧化活性高度吻合。氰化物的抑制作用可能归因于其与铁血红素蛋白结合 的亲和力,这会破坏蛋白质结构和功能。本研究中功能基因的相对表达与SOUR之间的对应关系表明,基于RNA的功能特异性检测方法对于更好地理解有毒化合物对废水中硝化活性的影响是有效的。
通过测量生理和转录反应来表征氰化物对硝化细菌的影响。氰化物在工业废水中可能存在的浓度下对硝化作用具有抑制性。基于RNA的功能特异性检测方法是一种机制性方法,有助于更好地理解有毒化合物对废水中硝化活性的影响。此外,RNA转录本的相对丰度可用于密切跟踪原位硝化细菌活性,这可用于预测抑制事件,从而提供一个指标来潜在地改善废水硝化系统的性能。