Determination of the carbon content of biomass—A prerequisite to estimate the complete biodegradation of polymers

This article presents a method to determine the carbon content of biomass, which is formed when degrading biodegradable polymers in an aerobic aqueous test system. Existing methods for determining the carbon content of biomass (e.g., fumigazation, protein assays, dry solids) have several disadvantages when applied for polymer degradation tests. In this work a protein assay based on the Lowry method was used. It was shown that the ratio between protein and carbon content is not constant but depends on the composition of the microbial population, the growth phase, and the substrate supply. This effect was used for the method presented in this article. For determining the carbon content of biomass the absorbance obtained by the Lowry test is correlated directly with the carbon content of biomass in dependence on the duration of the degradation test. The calibration curves are obtained by a mixed population of microorganisms during the course of a degradation test.     

Prescreening teratogenic potential of chlorinated drinking water disinfection by products by using Hydra regeneration assay

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用SOS/umu测试评价光电催化降解五氯酚过程的遗传毒性特征

采用SOS/umu测试研究了五氯酚水溶液在光电催化反应过程中的遗传毒性变化及降解产物对鼠伤寒沙门氏菌TA1535/pSK1002的细胞毒性,并与五氯酚在直接光解、光催化反应过程中的降解产物进行了比较.五氯酚光电催化降解过程中降解产物对测试菌种的细胞毒性逐渐降低,产物经代谢活化的细胞毒性低于直接光解、光催化降解五氯酚的产物.五氯酚经光电催化降解15￣45分钟的产物经代谢活化测试结果呈阳性,在60￣120分钟的降解产物呈阴性,说明五氯酚的光电催化降解过程中生成了间接遗传毒性物质,但该类物质能够在光电催化过程中被去除.而五氯酚经直接光解、光催化处理120分钟的产物均具有遗传毒性风险.本研究结果表明光电催化技术的环境安全性优于直接光解、光催化技术.此外,SOS/umu测试作为一种简单、灵敏的遗传毒性物质检测方法,适合于评价光电催化技术的遗传毒性特征,可以作为评价该技术环境安全性的生态毒理学方法之一.     

用CBMN法评价饮用水处理流程中有机提取物的细胞毒性

利用ＣＨＯ细胞胞质分裂阻断微核（ＣＢＭＮ）法,对某自来水厂７个处理工艺流程水中的有机提取物的细胞毒性进行了评价。在实验剂量范围内,比较了各样品的含微核的双核细胞率（ＢＮＭＮ）、核分裂指数（ＮＤＩ）和胞质分裂阻断增殖指数（ＣＢＰＩ）,实验表明,源水加氯后比源水的细胞性增大,经机加池和煤砂滤池处理后的水样毒性较大,其中煤砂滤池水毒性最大,机加池水次之,经炭吸附后的水样比炭吸附前水样的毒性下降,管网水的     

Genotoxic effect of polycyclic aromatic hydrocarbons in the metropolitan area of Porto Alegre, Brazil, evaluated by Helix aspersa (Müller, 1774)

The purpose of this study was to biomonitor metropolitan areas of Porto Alegre (Brazil) for PAHs associated with atmospheric particles and check their effects on the DNA of the land mollusk Helix aspersa. The sampling sites are located in an urban area with heavy traffic: (i) Canoas, (ii) Sapucaia do Sul, and (iii) FIERGS/Porto Alegre. The samples were collected during a continuous period of 24 hours during 15 days using Stacked Filter Units (SFU) on polycarbonate filters (two separated size fractions: PM_10-2.5 and PM_<2.5). The concentrations of 16 major PAHs were determined according to EPA. Comet assay on H. aspersa hemolymph cells was chosen for genotoxicity evaluation. This evaluation shows that, in general, the smaller PM-size fractions (PM_<2.5) have the highest genotoxicity and contain higher concentrations of extractable organic matter. In addition, associations between chemical characteristics and PM carcinogenicity tend to be stronger for the smaller PM-size fractions.     

Application of semipermeable membrane device (SPMD) to assess air genotoxicity in an occupational environment

Semipermeable membrane device (SPMD) is a passive sampler that sequesters lipophilic contaminants, mimicking the bioconcentration in the fatty tissue of organisms. This study was designed to assess the use of SPMD and biological tests (Comet assay and Ames test) for air monitoring. For this purpose an occupational environment with expected polycyclic aromatic hydrocarbons (PAHs) contamination (coke plant) was selected for a case study. The SPMDs were deployed in five occupational contaminated sites and in a control site. The SPMD dialysates were chemically analysed and examined for in vitro DNA-damaging activity in human cells (Jurkat) by Comet assay and for mutagenicity with the Ames test (TA98 strain, w/o S9). Total suspended particulates were also collected and analysed (GC–MS). No biological effect of SPMD extract was revealed in the control site. On the other hand, air samples collected with SPMDs within the coke plant showed variable degrees of genotoxic and mutagenic activity. The highest effects were associated with the highest PAH level recovered in the SPMDs extracts and in particulate samples.Results obtained support the sensitivity of biological tests associated to SPMD sampling for evaluating the health risk of potentially contaminated work environments highlighting the usefulness of SPMDs for environmental air quality monitoring.     

Genotoxicity evaluation of water soil leachates by Ames test, comet assay, and preliminary Tradescantia micronucleus assay

Combining genotoxicity/mutagenicity tests and physico-chemical methodologies can be useful for determining the potential genotoxic contaminants in soil samples. The aim of our study was to evaluate the genotoxicity of soil by applying an integrated physico-chemical-biological approach. Soil samples were collected at six sampling points in a Slovenian industrial and agricultural region where contamination by heavy metals and sulphur dioxide (SO₂) are primarily caused by a nearby power plant. The in vitro alkaline version of the comet assay on water soil leachates was performed with Caco-2 and HepG2 cells. A parallel genotoxicity evaluation of the samples was performed by Ames test using Salmonella typhimurium and the Tradescantia micronucleus test. Pedological analyses, heavy metal content determination, and different physico-chemical analyses, were also performed utilizing standard methodology. Water leachates of soil samples were prepared according to standard methods. Since only a battery of biotests with prokaryotic and eukaryotic organisms or cells can accurately estimate the effects of (geno)toxicants in soil samples and water soil leachates, a combination of three bioassays, with cells or organisms belonging to different trophic levels, was used. Genotoxicity of all six water soil leachates was proven by the comet assay on both human cell lines, however no positive results were detected by bacterial assay, Ames test. The Tradescantia micronucleus assay showed increase in micronuclei formation for three samples. According to these results we can assume that the comet assay was the most sensitive assay, followed by the micronucleus test. The Ames test does not appear to be sensitive enough for water soil leachates genotoxicity evaluations where heavy metal contamination is anticipated.     

Reed beds receiving industrial sludge containing nitroaromatic compounds

Goal, Scope and Background Sweden has prohibited the deposition of organic waste since January, 2005. Since 1 million tons of sludge is produced every year in Sweden and the capacity for incineration does not fill the demands, other methods of sludge management have to be introduced to a larger degree. One common method in the USA and parts of Europe is the use of wetlands to treat wastewater and sewage sludge. The capacity of reed beds to affect the toxicity of a complex mixture of nitroaromatics in sludge, however, is not fully elucidated. In this study, an industrial sludge containing explosives and pharmaceutical residues was therefore treated in artificial reed beds and the change in toxicity was studied. Nitroaromatic compounds, which are the main ingredients of many pharmaceuticals and explosives, are well known to cause cytotoxicity and genotoxicity. Recently performed studies have also showed that embryos of zebrafish (Danio rerio) are sensitive to nitroaromatic compounds. Therefore, we tested the sludge passing through constructed wetlands in order to detect any changes in levels of embryotoxicity, genotoxicity and dioxin-like activity (AhR-agonists). We also compared unplanted and planted systems in order to examine the impact of the root system on the fate of the toxicants. Methods An industrial sludge containing a complex mixture of nitroaromatics was added daily to small-scale constructed wetlands (vertical flow), both unplanted and planted with Phragmites australis. Sludge with an average dry weight of 1.25%, was added with an average hydraulic loading rate of 1.2 L/day. Outgoing water was collected daily and stored at −20°C. The artificial wetland sediment was Soxhlet extracted, followed by clean-up with multi-layer silica, or extracted by ultrasonic treatment, yielding one organic extract and one water extract of the same sample. Genotoxicity of the extracts was measured according to the ISO protocol for the umu-C genotoxicity assay (ISO/TC 147/SC 5/WG9 N8), using Salmonella typhimurium TA1535/pSK1002 as test organism. Embryotoxicity and teratogenicity were studied using the fish egg assay with zebrafish (Danio rerio) and the dioxin-like activity was measured using the DR-CALUX assay. Chemical analyses of nitroaromatic compounds were performed using Solid Phase Micro Extraction (SPME) and GC-MS. Results Organic extracts of the bed material showed toxic potential in all three toxicity tests after two years of sludge loading. There was a difference between the planted and the unplanted beds, where the toxicity of organic extracts overall was higher in the bed material from the planted beds. The higher toxicity of the planted beds could have been caused by the higher levels of total carbon in the planted beds, which binds organic toxicants, and by enrichment caused by lower volumes of outgoing water from the planted beds. Discussion Developmental disorders were observed in zebrafish exposed directly in contact to bed material from unplanted beds, but not in fish exposed to bed material from planted beds. Hatching rates were slightly lower in zebrafish exposed to outgoing water from unplanted beds than in embryos exposed to outgoing water from planted beds. Genotoxicity in the outgoing water was below detection limit for both planted and unplanted beds. Most of the added toxicants via the sludge were unaccounted for in the outgoing water, suggesting that the beds had toxicant removal potential, although the mechanisms behind this remain unknown. Conclusions During the experimental period, the beds received a sludge volume (dry weight) of around three times their own volume. In spite of this, the toxicity in the bed material was lower than in the sludge. Thus, the beds were probably able to actually decrease the toxicity of the added, sludge-associated toxicants. When testing the acetone extracts of the bed material, the planted bed showed a higher toxicity than the unplanted beds in all three toxicity tests. The toxicity of water extracts from the unplanted beds, detected by the fish egg assay, were higher than the water extracts from the planted beds. No genotoxicity was detected in outgoing water from either planted or unplanted beds. All this together indicates that the planted reed beds retained semi-lipophilic acetone-soluble toxic compounds from the sludge better than the unplanted beds, which tended to leak out more of the water soluble toxic compounds in the outgoing water. The compounds identified by SPME/GC in the outgoing water were not in sufficient concentrations to have caused induction in the genotoxicity test. Recommendations and Perspectives This study has pointed out the benefits of using constructed wetlands receiving an industrial sludge containing a complex mixture of nitroaromatics to reduce toxicity in the outgoing water. The water from planted, constructed wetlands could therefore be directed to a recipient without further cleaning. The bed material should be investigated over a longer period of time in order to evaluate potential accumulation and leakage prior to proper usage or storage. The plants should be investigated in order to examine uptake and possible release when the plant biomass is degraded. This article has been developed on the basis of a presentation given at the Annual meeting of SETAC Europe German Language Branch 2004 in Aachen. ESS-Submission Editor: Dr. Ludek Blaha (blaha@recetox.muni.cz)     

剑尾鱼卵黄蛋白原的ELISA检测

以卵黄脂磷蛋白(lipovitellin,Lv)抗血清为抗体,以纯化的卵黄蛋白原(vitellogenin,Vtg)为抗原,建立了间接酶联免疫吸附反应(ELISA)方法检测剑尾鱼(Xiphophorus helleri)雄鱼整体匀浆液中ρ(Vtg).结果表明,该方法的检测灵敏度为7.8 ng/mL,批内变异系数为5.094%,批间变异系数为5.540%,工作范围为32.5～2 000 ng/mL,在该范围内,标准曲线具有良好的线性和重复性.由于该方法可直接在1块或在不同的酶标板上准确地进行比较,因而可利用ELISA方法测定经诱导雄鱼整体匀浆液中ρ(Vtg)水平.结果显示,50 μg/L 17-β雌二醇(E₂)诱导21 d的雄性剑尾鱼有明显的Vtg产生;10 μg/L E₂诱导剑尾鱼亦有Vtg产生,但较50 μg/L E₂诱导组低;1 μg/L E₂诱导组及对照组剑尾鱼没有Vtg产生,检测孔OD450/对照OD₄₅₀(P/N)值小于2.1.      

油田硫酸盐还原菌快速定量检测方法

现有油田废水中硫酸盐还原菌检测周期长、检测费用较高,本研究应用聚合酶链式反应(PCR)技术与倍比稀释法(MPN)相结合的DSR-MPN-PCR法,对硫酸盐还原菌进行快速定量检测.从废水中制备了直接用于PCR扩增的菌液,保证了定量准确性;建立以硫酸盐还原菌亚硫酸盐还原酶基因(Dsr)为靶位点的通用探针DSR1F和DSR5R的反应体系和扩增条件.结果表明,该方法检测灵敏度明显比液体稀释培养法高2个数量级,真实地表征了废水中实际的SRB菌数量,整个操作过程需要3～4　h,检测结果非常稳定,降低了检测费用,可以在生产中应用.