Design and use of group-specific primers and probes for real-time quantitative PCR

Real-time quantitative polymerase chain reaction (qPCR) has gained popularity as a technique to detect and quantify a specific group of target microorganisms from various environmental samples including soil, water, sediments, and sludge. Although qPCR is a very useful technique for nucleic acid quantification, accurately quantifying the target microbial group strongly depends on the quality of the primer and probe used. Many aspects of conducting qPCR assays have become increasingly routine and automated; however, one of the most important aspects, designing and selecting primer and probe sets, is often a somewhat arcane process. In many cases, failed or non-specific amplification can be attributed to improperly designed primer-probe sets. This paper is intended to provide guidelines and general principles for designing group-specific primers and probes for qPCR assays. We demonstrate the effectiveness of these guidelines by reviewing the use of qPCR to study anaerobic processes and biologic nutrient removal processes. qPCR assays using group-specific primers and probes designed with this method, have been used to successfully quantify 16S ribosomal Ribonucleic Acid (16S rRNA) gene copy numbers from target methanogenic and ammonia-oxidizing bacteria in various laboratory- and full-scale biologic processes. Researchers with a good command of primer and probe design can use qPCR as a valuable tool to study biodiversity and to develop more efficient control strategies for biologic processes.     

均匀设计法优化造纸黑液的湿式氧化法处理工艺

通过均匀设计实验,对造纸黑液进行湿式氧化处理,数据回归得到CODCr、吸光度、浊度的回归方程,各回归方程显著,氧分压对造纸黑液各指标影响最大;研究反应过程中反应温度、氧分压、进水pH值、进水浓度、搅拌强度、反应时间对造纸黑液湿式氧化(WAO)处理的影响,6因素优化的操作条件依次为:180℃、3.0MPa、7.25、5500mg/L、500r/min、60min;在优化的操作条件下,造纸黑液的CODCr、CODCr去除率为1661mg/L、69.8%,吸光度、脱色率为0.63、96.3%,浊度、浊度去除率为661NTU、74.8%.均匀设计法在造纸黑液WAO处理中得到了较好的应用.     

Determination of cobalt by air-assisted liquid–liquid microextraction

Abstract

A rapid and selective technique for extraction, preconcentration and determination of trace amounts of cobalt in water and pharmaceutical samples by air-assisted liquid–liquid microextraction combined with flame atomic absorption spectrometry is proposed. 1-Nitroso-2-naphthol is used as a complexing agent and 1-octanol as an extraction solvent. Parameters relevant for analytical effectivity, i.e. pH of sample solution, concentration of complexing agent, volume of extraction solvent, and number of extraction cycles are optimized using a Box–Behnken design. At optimum conditions, a dynamic linear range of 5–600?µg L^?1 is obtained, with a limit of detection of 1.2?µg L^?1. The method is used for determination of Co(II) in environmental water and pharmaceutical samples.     

Removal of Remazol Brilliant Violet-5R dye using periwinkle shells

The purpose of this research is to obtain optimal processing conditions for the adsorption of Remazol Brilliant Violet-5R (RBV-5R) dye onto activated carbon prepared from periwinkle shells (PSAC) by chemical activation with KOH using response surface methodology. Central composite design (CCD) was used to determine the effects of three preparation variables; CO₂ activation temperature, CO₂ activation time and KOH:char impregnation ratio (IR) on two responses; percentage RBV-5R dye removal and PSAC yield. Based on the CCD, two quadratic models were developed for percentage RBV-5R dye removal and PSAC yield, respectively. The most influential factor on each experimental design response was identified from the analysis of variance (ANOVA). The optimum conditions for the adsorption of RBV-5R dye onto PSAC were CO₂ activation temperature of 811 °C, CO₂ activation time of 1.70 h and IR of 3.0, resulting in 81.28% RBV-5R dye removal and 28.18% PSAC yield. PSAC prepared under optimum conditions was mesoporous with a Brunauer–Emmett–Teller surface area of 1894 m²·g^?1, total pore volume of 1.107 cm³·g^?1 and average pore diameter of 2.32 nm. The surface morphology and functional groups of the activated carbon were respectively determined from the scanning electron microscopy and Fourier transform infrared analysis.     

超大跨度渡槽脉动风频率特性分析

贵州龙场渡槽是我国目前最大的拱跨渡槽,其脉动风频率特性分析对设计工作具有极其重要的指导作用。计算了龙场渡槽在有水、无水工况下的前20阶自振频率,分析了相应的振型;采用极值Ⅰ型概率分布函数,计算得到龙场渡槽所在地的50年一遇最大设计风速为17.6m/s;针对1986～2010年间的3次强风风速和50年一遇最大设计风速,计算了龙场渡槽所在地典型高程处的脉动风功率谱密度。研究表明:在同一高度,随着年平均最大风速的增加,功率谱密度最大值逐渐增大,其对应的频率也在增大;对于同一风速,随着高度的增加,功率谱密度的最大值逐渐增大,其对应的频率也在增大。通过比较龙场渡槽自振频率与脉动风功率谱密度显著值所对应的频率,认为龙场渡槽在上述风速下不会发生共振现象。     

简易生活垃圾填埋场综合治理工程设计

随着越来越多标准化生活垃圾填埋场的建成和投入运行,原有简易生活垃圾填埋场的环境综合治理被各级环境管理部门日益重视,成为改善环境急需解决的问题。以西北某市简易生活垃圾填埋场综合整治工程为例,通过分析简易垃圾填埋场存在的主要环境隐患,有针对性地提出了解决垃圾渗沥液、填埋气、边坡安全等环境和安全问题的具体工程措施,尽量将简易垃圾填埋场对周围环境的影响降到最低程度。     

用钢管酸洗废水生产聚合硫酸铁技术的研究和应用

为了解决钢管酸洗废水污染环境和浪费资源难题,可用其生产聚合硫酸铁,从而实现循环经济。采用蒸发浓缩和催化聚合反应实现聚合硫酸铁的生成。其产品质量达《净水剂聚合硫酸铁》标准,可作为絮凝剂使用。研究和实践证明,用钢管酸洗废水制备聚合硫酸铁是可行的,具有明显的经济效益和环境效益,值得推广。     

光催化膜反应器的设计及其在水处理中的应用

作为一种新型的水处理装置,光催化膜反应器具有反应迅速、降解效率高、降解彻底、无二次污染等特点,在节水减排、微污染水处理、中水回用等方面具有重要的应用价值。总结目前国内外各种不同类型的光催化膜反应器的设计、特点及在水处理中的应用情况,并对其今后的发展提出建议。     

基于Box Behnken设计的煤导热系数分析与预测

为研究分析水分、固定碳及密度3个因素对煤导热系数的影响,选取多个矿井不同煤质的煤作为实验煤样,使用NETZSCH LFA457型激光导热系数测试仪进行测定,并引入二次响应曲面试验设计思想,采用Design Expert软件进行煤质指标的Box Behnken试验设计,按设计表筛选实验数据,构建了导热系数多因素、多水平影响下二次回归响应曲面模型,并对不同采样地点的煤导热系数进行了分析与预测。结果表明:二次回归方程P值小于0.000 1,极显著,失拟项为0.072 4,不显著,回归方程具有统计学意义;当水分在0.9%~10.9%,固定碳在49.3%~83.6%,密度在1.2~1.92 g·cm³之间,对煤的导热系数一次项影响程度为密度>固定碳>水分;二次项影响程度为固定碳和密度>水分和密度>水分和固定碳,且固定碳和密度之间存在交互作用,水分和密度、水分和固定碳之间无交互作用;应用响应曲面模型预测的煤导热系数误差为4.3%,满足精度要求。     

医疗废物高温蒸汽集中处置工艺设计

以某市医疗废物集中处置中心为例,介绍8 t/d规模的高温蒸汽系统工艺设计,包括工艺流程以及产生的有害物质的处理,并对医疗废物高温蒸汽处理系统的设计技术进行探讨。