Development of an enzyme linked immunosorbent assay for the determination of phenothiazine drugs in meat and animal feeds

In this study, 2-chlorophenothiazine was used to synthesize a hapten for production of monoclonal antibody. The obtained monoclonal antibody showed high crossreactivities to chlorpromazine, promethazine and perphenazine, and showed low crossreactivities to acepromazine and fluphenazine. After evaluation of three coating antigens, a heterologous competitive indirect enzyme linked immunosorbent assay was developed to determine the five phenothiazines in animal feeds and the residues of chlorpromazine, promethazine and perphenazine in meat. The crossreactivities to the five analytes were in a range of 2.4%–98%. The limits of detection for the five drugs in feeds were in a range of 0.1–3.0 μg g^?1, and that for chlorpromazine, promethazine and perphenazine in meat were in a range of 0.5–0.8 ng g^?1. Their recoveries from standards fortified blank samples (chicken, pork and feeds) were in a range of 74.1%–96.5% with coefficients of variation of 6.4%–15.1%. Therefore, this method could be used as a rapid screen tool to determine phenothiazine drugs in animal feeds and animal derived foods.     

Assays of dioxins and dioxin-like compounds in actually contaminated soils using transgenic tobacco plants carrying a recombinant mouse aryl hydrocarbon receptor-mediated β-glucuronidase reporter gene expression system

The transgenic tobacco plant XD4V-26 carrying the recombinant mouse aryl hydrocarbon receptor XD4V-mediated β-glucuronidase (GUS) reporter gene expression system was used for assay of dioxins and dioxin-like compounds consisting of polychlorinated dibenzeno-p-dioxins, polychlorinated dibenzofurans, and coplanar polychlorinated biphenyls (Co-PCBs) in actually contaminated soils. The transgenic tobacco plant XD4V-26 showed a significant dose-dependent induced GUS activity when cultured on MS medium containing PCB126 [toxic equivalency factor (TEF) = 0.1]. In contrast, PCB169 and PCB180, which have 0.03 of TEF and unassigned TEF values, respectively, did not significantly induce GUS activity under the same conditions as with PCB126. When the tobacco plants were cultivated for up to 5 weeks on actually contaminated soils with dioxins and dioxin-like compounds collected from the periphery of an incinerator used for disposal of residential and industrial wastes, GUS activity in the leaves was dose-dependently increased. The plants clearly detected 360 pg-TEQ g^?1 of dioxins and dioxin-like compounds in this assay. There was a positive correlation between GUS activity and TEQ value of dioxins and dioxin-like compounds in the plants. This assay does not require any extraction and purification processes for the actually contaminated soil samples.     

The detection of dioxin- and estrogen-like pollutants in marine and freshwater fishes cultivated in Pearl River Delta, China

In this study we aimed to assess the dioxin- and estrogen-like activities of contaminants extracted from twenty species of freshwater and seawater fishes, using luciferase reporter assays. Transfected MCF7 cells were treated with sample extracts and luciferase activities were then measured at 24-h of post-treatment. The mean values of the detected dioxin- and estrogen-like activities in the freshwater fishes were 25.3 pg TEQ/g ww and 102.3 pM EEQ/g ww whereas in the seawater fishes, the values were 46.2 pg TEQ/g ww and 118.8 pM EEQ/g ww. Using sample-relevant dosage of estrogen, inductions of cell proliferation markers (i.e. retinoblastoma, cyclin D) and stimulations of cell growth were revealed by Western blotting, colony formation and BrdU uptake assays. A cotreatment with TCDD significantly reduced these effects. Using the sample extracts with different dioxin- and estrogen-like activities, similar observation was revealed. The data highlighted the mixture effect of food contaminants on human health.     

DNA damage in Populus tremuloides clones exposed to elevated O3

The effects of elevated concentrations of atmospheric tropospheric ozone (O₃) on DNA damage in five trembling aspen (Populus tremuloides Michx.) clones growing in a free-air enrichment experiment in the presence and absence of elevated concentrations of carbon dioxide (CO₂) were examined. Growing season mean hourly O₃ concentrations were 36.3 and 47.3 ppb for ambient and elevated O₃ plots, respectively. The 4th highest daily maximum 8-h ambient and elevated O₃ concentrations were 79 and 89 ppb, respectively. Elevated CO₂ averaged 524 ppm (+150 ppm) over the growing season. Exposure to O₃ and CO₂ in combination with O₃ increased DNA damage levels above background as measured by the comet assay. Ozone-tolerant clones 271 and 8L showed the highest levels of DNA damage under elevated O₃ compared with ambient air; whereas less tolerant clone 216 and sensitive clones 42E and 259 had comparably lower levels of DNA damage with no significant differences between elevated O₃ and ambient air. Clone 8L was demonstrated to have the highest level of excision DNA repair. In addition, clone 271 had the highest level of oxidative damage as measured by lipid peroxidation. The results suggest that variation in cellular responses to DNA damage between aspen clones may contribute to O₃ tolerance or sensitivity.     

Studies on the toxic effects of pentachlorophenol on the biological activity of anaerobic granular sludge

In order to explore the pathway of the anaerobic biotreatment of the wastewater containing pentachlorophenol (PCP) and ensure the normal operation of Upflow Anaerobic Sludge Blanket (UASB) reactor, the anaerobic sludge under different acclimation conditions were selected to seed and start up UASB reactors. Anaerobic toxicity assays were employed to study the biological activity, the tolerance and the capacity to degrade PCP of different anaerobic granular sludge from UASB reactors. Results showed that the anaerobic granular sludge acclimated to chlorophenols (CPs) could degrade PCP more quickly (up to 9.50mg-PCPg(-1)TVSd(-1)). And the anaerobic granular sludge without acclimation to CPs had only a little activity of degrading PCP (less than 0.07mg-PCPg(-1)TVSd(-1)). Different PCP concentrations (2, 4, 6, 8mgL(-1)) had different inhibition effects on glucose utilization, volatile fatted acidity (VFA)-degrading and methanogens activity of PCP degradation anaerobic granular sludge, and the biological activity declined with the increase in PCP concentration. The methanogens activity suffered inhibition from PCP more easily. The different acclimation patterns of seeded sludge had distinctly different effects on biological activity of the degradation of PCP of anaerobic granular sludge from UASB reactors. The biological activity of the anaerobic granular sludge acclimated to PCP only was also inhibited. This inhibition was weak compared to that of anaerobic granular sludge acclimated to CPs, further, the activity could recover more quickly in this case. In the same reactor, the anaerobic granular sludge from the mid and base layers showed higher tolerance to PCP than that from super layer or if the sludge is unacclimated to CPs, and the corresponding recovery time of the biological activity in the mid and base layers were short. Acetate-utilizing methanogens and syntrophic propinate degraders were sensitive to PCP, compared to syntrophic butyrate degraders.     

失去记忆性贝毒ASP酶联免疫检测方法的研究

建立了赤潮毒素失去记忆性贝毒中主要成分软骨藻酸(domoic acid,DA)的竞争酶联免疫检测方法.通过将DA偶联到蛋白载体上,免疫BALB/c小鼠,免疫数次后,得到抗DA的多克隆抗血清,以DA-OVA为包被抗原,利用抗原抗体反应,建立间接竞争酶联免疫吸附技术(Enzyme-linked immunosorbant assay,ELISA)分析检测海水样品和海洋贝类中的赤潮毒素失去记忆性贝毒DA的方法.结果表明,该方法最低检出限10μg/L,海水样品平均回收率102.%,批内变异系数12.%;贝类样品平均回收率为111.%,批内变异系数4.8%.     

间接免疫荧光抗体技术检测凡纳滨对虾红体病病原——副溶血弧菌

建立了凡纳滨对虾红体病病原--副溶血弧菌的间接免疫荧光抗体检测方法.确定免疫血清和荧光抗体的最适工作浓度分别为1∶ 1000和1∶ 200;交叉反应和阻断试验表明该方法具有较高的特异性.用该方法检测了10份人工感染后发病的凡纳滨对虾和10份感染后外观健康的凡纳滨对虾,阳性检出率分别为100%和60%,表明该技术不仅能够检测已发病的凡纳滨对虾,而且能够检测带菌的凡纳滨对虾,这对于水产养殖业中疾病的早期诊断有着重要意义,而且该方法具有快速、特异、简单的优点,适合于现场应用推广.     

Assessment of DNA damage and cholinesterase activity in soybean farmers in southern Brazil: High versus low pesticide exposure

Abstract

The aim of this study was to evaluate the DNA damage in soybean growers during two agricultural periods of a crop season (high and low exposure) and a control group, as well as butyrylcholinesterase (BChE) activity during these exposure periods in order to estimate the degree of BChE inhibition for the exposed group. DNA damage in peripheral whole blood was evaluated by the comet assay and plasma BChE activity was accessed as a measure of exposure to cholinesterase inhibitors. None of the soybean growers reported using full Personal Protective Equipment (PPE). BChE was lower in high exposure period than in low exposure period and DNA damage index was significantly increased in the high exposure period than in the low exposure period. In addition, DNA damage in both exposure periods was higher than control group. No correlation was found between exposure time and DNA damage and BChE activity. However, negative correlation was observed between DNA damage in high and low exposure periods. The results indicate that soybean growers are exposed to cholinesterase inhibitors and to pesticides mixtures with genotoxic potential.     

官厅水库水体中微囊藻毒素及其与微囊藻细胞密度相关性研究

2004-2005年对连年发生比较严重水华的官厅水库进行了2 a的浮游植物动态监测,并在2005年6-8月应用酶联免疫吸附法检测了主要水华藻类--铜绿微囊藻藻毒素质量浓度及其季节变化,初步探讨了微囊藻毒素质量浓度与铜绿微囊藻细胞密度之间的关系.实验结果表明,水库中的浮游植物细胞密度高,2004年平均值为1 308.35×104 cells/L,2005年达1 599.49×104 cells/L,同时呈现出明显的季节变化,在夏末秋初形成高峰.产毒的铜绿微囊藻为夏季最主要的优势水华种类,其细胞密度在7月达到最高,为4 748.58×104 cells/L;微囊藻毒素的质量浓度在7月亦达到最高值,为20 μg/L,都显著高于6月和8月(p＜0.05).水体中微囊藻毒素质量浓度与微囊藻细胞密度呈正相关关系(R=0.926).其结果为全面了解官厅水库水质状况以及对水污染进行治理,恢复其饮用水源的功能提供了科学依据.     

土壤微生物生物量及多样性测定方法评述

土壤微生物作为土壤中重要的分解者,在养分的循环与转化中起着重要的作用。另外,由于其与土壤理化性质相比,对外界环境的变化更为敏感,因此,土壤微生物性质常被用作表征土壤质量的灵敏性指标,而土壤微生物生物量与微生物多样性是土壤微生物学研究中应用最广泛的指标。土壤环境复杂,微生物种类丰富多样,需要合适的研究方法才能真正打开土壤系统这个黑箱,自19世纪60年代以后,土壤微生物学在研究方法上取得了重要的进展,本文综述了微生物生物量及多样性的概念及微生物生物量和多样性研究方法的发展状况,重点就已有应用比较多的研究方法如测定微生物生物量的熏蒸法,测定微生物多样性的平板计数法、Biolog微平板计数法、磷脂脂肪酸法及分子生物学等研究方法进行了详细介绍及优缺点分析。结合现有微生物生物量及多样性的测定方法,在未来的研究中,一方面应该引入更多的新技术,发展新方法,以期探索更多的未知微生物种群;其次应该根据实验要求,完善和改进现有研究方法,使其更为统一和准确;再者,在微生物数据的分析上应该探索和借鉴更多的分析方法和手段,从而更深入更全面的解读实验当中所得到的微生物的信息。