Rat cardiomyocyte H9c2(2-1)-based sulforhodamine B assay as a promising in vitro method to assess the biological component of effluent toxicity

The treatment of wastewaters is crucial to maintain the ecological status of receiving waters, and thereby guarantee the protection of aquatic life and human health. Wastewater quality evaluation is conventionally based on physicochemical parameters, but increasing attention has been paid to integrate physicochemical and biological data. Nevertheless, the regulatory use of fish in biological testing methods has been subject to various ethical and cost concerns, and in vitro cell-based assays have thus become an important topic of interest. Hence, the present study intends: (a) to evaluate the efficiency of two different sample pre-concentration techniques (lyophilisation and solid phase extraction) to assess the toxicity of municipal effluents on rat cardiomyoblast H9c2(2-1) cells, and (b) maximizing the use of the effluent sample collected, to estimate the environmental condition of the receiving environment. The gathered results demonstrate that the H9c2(2-1) sulforhodamine B-based assay is an appropriate in vitro method to assess biological effluent toxicity, and the best results were attained by lyophilising the sample as pre-treatment. Due to its response, the H9c2(2-1) cell line might be a possible alternative in vitro model for fish lethal testing to assess the toxicity of municipal effluents. The physicochemical status of the sample suggests a high potential for eutrophication, and iron exceeded the permissible level for wastewater discharge, possibly due to the addition of ferric chloride for wastewater treatment. In general, the levels of carbamazepine and sulfamethoxazole are higher than those reported for other countries, and both surpassed the aquatic protective values for long-term exposure.     

大肠杆菌JM105中硝酸盐还原酶的制备及其性质

近年来,硝酸盐和亚硝酸盐的污染已引起普遍关注。目前,蔬菜和粮食的NO3-含量过高主要由于农药和化肥的使用、工业废水或生活污水灌溉等,食品类主要来自腌制食品和食品添加剂或防腐剂等。另外,向乳制品或食品中参碱、食盐、化肥、脏水、碱性水等也是硝酸盐污染的来源之一。因此,迫切需要快速、简便、可用于现场的硝酸盐检测方法。这就需仰赖生物酶方法,而生物酶方法的关键是酶制剂的制备和检测方法的建立。本研究通过筛选、厌氧和硝酸盐诱导培养、超声波细胞破碎和差速离心提取等方法,从大肠杆菌(Escherichia coli)JM105细胞膜中制备了硝酸盐还原酶并对其性质进行了研究。结果表明:从大肠杆菌JM105中制得的酶制剂活力很高,且在酶过量的情况下可将NO3-完全转化为NO2-,在用磷酸缓冲液清洗并冷冻保藏过夜后不含有亚硝酸盐还原酶。在加入黄素单核苷酸辅酶(FMN)后,该酶的活力可提高64%,比活力达0.42U·mg-1蛋白。该酶十分稳定,在40℃下24h活力无影响,在浓度为1mmol·L-1的Cu2 、Fe3 、Ca2 、Zn2 、Mg2 和Mo6 存在下,其活力亦不改变。因此,该酶可用于测定食品、蔬菜和环境中硝酸盐的含量。     

A single chorionic gonadotropin assay for maternal serum screening for Down's syndrome

A simple enzyme immunoassay measuring human chorionic gonadotropin in undiluted maternal serum has been developed in order to be used as a prenatal screening test for Down' s syndrome. A retrospective study of maternal serum sampled during pregnancies associated with trisomy 21 shows that with a 5% amniocentesis rate determined on a single test, the detection rate of trisomy 21 would be around two-thirds of the affected pregnancies. A prospective study of 9040 pregnant women under 38 years has confirmed the usefulness of the assay.     

Microcystin-LR detection based on indirect competitive enzyme-linked immunosorbent assay

Microcystins (MCs) are a group of closely related toxic cyclic heptapeptides produced by common cyanobacteria, which cause lots of accidents and threatens human health. In this paper, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was established and used to detect microcystin-LR (MC-LR) in drinking and surface waters. The concentration of coating antigen was 5 μ/mL, the dilution of monoclonal antibody MC10E7 was 1:3 000, the dilution of enzyme tracer (goat anti-mouse IgG-peroxidase) was 1:3 000, the standard concentration of MC-LR ranged from 0.001 μg/L to 30 μg/L, and o-phenylenediamine was used as substrate. The assay showed high relativity with high performance liquid chromatography (HPLC) with a correlation coefficient of more than 99%. The relative standard deviation was less than 10%, the detection limit was achieved down to 0.01 μg/L and up to 5.1 μg/L. The quantitative detection range was from 0.03 μg/L to 3 μg/L, and the antibody had high specificity for [4-arginine] microcystins. It performed well in spite of the influence of the real samples. Translated from Environmental Science, 2006, 27(6): 1166–1170 [译自: 环境科学]     

稀土金属离子对梨形四膜虫的24小时生长影响

应用４种方法（细胞直计计数法、中性红试验、重蛋白及核酸含量测定）研究不同浓度稀土金属离子Ｌａ＾２＋、Ｓｍ＾３＋、Ｙ＾３＋、Ｇｄ＾３＋处理２４ｈ后，对梨形四膜虫的生长影响，结果表明，在低浓度时Ｌａ＾３＋、Ｓｍ＾３＋、Ｙ＾３＋，对四膜虫具有刺激生长的作用。随着浓度的升高，４种稀土金属离子均表现出毒性效应；高浓度时，抑制生长艇特别明显。计算Ｌａ＾３＋、Ｓｍ＾３＋、Ｙ＾３＋、Ｇｄ＾３＋的半数抑制浓度（ＩＣ     

One independent variable rate equation describing utilization of biodegradable organic matter in activated sludge processes

This investigation aims to represent aerobic utilization of biodegradable organic matter present in wastewater by a rate equation. This rate equation can then be used to develop a substrate utilization (removal) kinetic model for unsteady state activated sludge process. To achieve this objective, theoretical utilization of biodegradable organic matter in batch process and growth pattern theory were studied. Also, experimental data representing removal of organic matter in different types of wastewaters were collected and analyzed for batch and continuous activated sludge assays. A rate equation was proposed to describe the utilization of biodegradable organic matter based on theoretical analysis of batch process. This rate equation was then verified through differential and integral analysis of the experimental data. Furthermore, a substrate kinetic model for batch and continuous processes was developed. The developed rate equation will facilitate the analysis and design of sequencing batch reactor (SBR) technology for biological treatment of wastewater.     

类酶联免疫分析法可行性的研究

多环芳烃作为持久性高毒性的有机污染物,一直受到广泛关注。酶联免疫分析技术因特异性强、快速、便捷等优点而广泛使用。建立了芳香烃受体介导的类酶联免疫竞争方法(类ELISA),用于检测环境中多环芳烃类物质的含量。在实验室条件下,分别检测了5种多环芳烃萘、苊、菲、荧蒽和芘,同时比较与高效液相色谱法(HPLC)和气相色谱-质谱联用法(GC/MS)在灵敏度、准确性和重现性等标的差异。HPLC检出限为2.70~10.71 ng/mL;GC/MS的检出限为2.60~24.40 ng/mL;类ELISA的检出限在1.24~3.10ng/mL。HPLC的加标回收率为58.03%~106.69%;GC/MS的加标回收率为88.16%~154.83%;类ELISA的加标回收率为74.67%~121.84%。HPLC的变异系数为4.26~9.03;GC/MS的变异系数为1.44~5.82;类ELISA的变异系数为2.56~8.16。结果表明,类ELISA方法灵敏度较好,准确性较高;而GC/MS方法重现性较好,检测更为稳定。     

Evaluation of drinking water treatment combined filter backwash water recycling technology based on comet and micronucleus assay

Based on the fact that recycling of combined filter backwash water（CFBW）directly to drinking water treatment plants（WTP）is considered to be a feasible method to enhance pollutant removal efficiency,we were motivated to evaluate the genotoxicity of water samples from two pilot-scale drinking water treatment systems,one with recycling of combined backwash water,the other one with a conventional process.An integrated approach of the comet and micronucleus（MN）assays was used with zebrafish（Danio rerio）to investigate the water genotoxicity in this study.The total organic carbon（TOC）,dissolved organic carbon（DOC）,and trihalomethane formation potential（THMFP）,of the recycling process were lower than that of the conventional process.All the results showed that there was no statistically significant difference（P>0.05）between the conventional and recycling processes,and indicated that the genotoxicity of water samples from the recycling process did not accumulate in 15 day continuous recycling trial.It was worth noting that there was correlation between the concentrations of TOC,DOC,UV₂₅₄,and THMFPs in water and the DNA damage score,with corresponding R² values of 0.68,0.63,0.28,and 0.64.Nevertheless,both DNA strand breaks and MN frequency of all water samples after disinfection were higher than that of water samples from the two treatment units,which meant that the disinfection by-products（DBPs）formed by disinfection could increase the DNA damage.Both the comet and MN tests suggest that the recycling process did not increase the genotoxicity risk,compared to the traditional process.     

二苯甲酮对斑点叉尾鮰鱼卵巢细胞生长的影响

试验通过直接、间接两种染毒手段来探讨二苯甲酮(benzophenone)对斑点叉尾鮰鱼卵巢细胞(CCO)生长的影响,以MTT法为手段测定染毒前后CCO细胞的存活数量和各阶段的生长状态,结果发现:当二苯甲酮以1～10μg/mL的浓度直接作用于CCO细胞时,CCO细胞生长在不同阶段均受到抑制作用:在指数增长期细胞生长速率减小,稳定期的存活量约为正常条件下的72%～91%,并且随着二苯甲酮浓度的增大,对CCO生长的抑制效应也相应的加剧;但是1μg/mL浓度的二苯甲酮在经过斑点叉尾鮰鱼原代肝细胞代谢后间接作用时,其代谢产物对CCO的生长在各阶段又呈现明显的促进作用,稳定期细胞数量约为正常条件下的109%。     

Development of competitive immunoassays to hydroxyl containing fungicide metabolites

This paper describes the isolation of monoclonal antibodies and the development of competitive immunoassays to pesticide metabolites of the fungicides imazalil, carbendazim and thiabendazole. The metabolite specific hydroxyl residues were used as the reactive group with which to link the metabolite to the carrier proteins Keyhole Limpet Haemocyanin (KLH) and Bovine Serum Albumin (BSA). In each case immune responses in mice were raised and monoclonal antibodies were produced. Antibodies were developed into competitive ELISAs to the appropriate metabolite. The antibody raised to a metabolite of imazalil was optimised into a competitive ELISA format which had an assay IC50 of 7.5 μg/L and a limit of detection (LOD) of 1.1 μg/L. A single antibody isolated against the metabolite of carbendazim had assay IC50s of 3.2 and 2.7 μg/L for the metabolites of carbendazim and thiabendazole respectively with an LOD of 0.38 μg/L for both. These sensitive immunoassays may have application in the monitoring of human exposure to these fungicide residues either by occupational or non-occupational routes.