Prenatal diagnosis of carbamoyl-phosphate synthetase deficiency by fetal liver biopsy

In a pregnancy at risk for carbamoyl-phosphate synthetase (CPS) deficiency, prenatal diagnosis was attempted by fetal liver biopsy, performed at 18 weeks of gestation. CPS activity was absent and the diagnosis was confirmed after termination of the pregnancy. The technique employed for fetal liver biopsy is described together with an evaluation of its possible role in prenatal diagnosis.     

Oxidative stress in duckweed (Lemna minor L.) caused by short-term cadmium exposure

The mechanisms of plant defence against cadmium toxicity have been studied by short-term exposure of Lemna minor L. (common duckweed) to concentrations of CdCl2 ranging from 0 to 500microM. High accumulation of cadmium was observed (12,320+/-2155microgg(-1) at 500microM CdCl2), which caused a gradual decrease of plant growth, increased lipid peroxidation, and weakened the entire antioxidative defence. Total glutathione concentration decreased significantly; however, the concentration of oxidized glutathione remained stable. The responses of four antioxidant enzymes showed that catalase was the most inhibited after CdCl2 exposure, ascorbate peroxidase and guaiacol peroxidase moderately, and glutathione reductase least. The total antioxidative potential revealed an induced antioxidative network at 0.1microM CdCl2 (137+/-13.2% of the control) and its reduction to only 47.4+/-4.0% of the control at higher cadmium concentrations. The possible application of the examined biomarkers in ecotoxicological research is discussed.     

Evaluation of various tests for the diagnosis of soil contamination by 2,4,5-trichlorophenol (2,4,5-TCP)

Soil response to contamination with 2,4,5-triclorophenol was studied to test the validity of the concept of Generic Reference Levels (GRL), the main criterion used to define soil contamination. Soil samples were artificially contaminated with doses between 0 and 5000 mg kg⁻¹ of 2,4,5-triclorophenol, and analysed by various tests. Where possible, the response of soils to the contaminant was modelled by a sigmoidal dose-response curve in order to estimate the ED₅₀ values. The tests provided different responses, but only microbial biomass-C and dehydrogenase and urease activities demonstrated soil deterioration in response to contamination. The results suggest that the diagnosis of soil contamination has been greatly simplified in the legislation by the provision of a single figure for each compound, and that the GRL concept could perhaps be substituted by measurement of ED₅₀ values, which better reflect the alteration of a soil due to the presence of a xenobiotic substance.     

Restoring biochemical activity and bacterial diversity in a trichloroethylene-contaminated soil: the reclamation effect of vermicomposted olive wastes

Background, aim, and scope  In this work, the potential for using olive-mill solid waste as an organic amendment for biochemical and biological restoration of a trichloroethylene-contaminated soil, which has previously been stabilized through vermicomposting processes, has been explored. Materials and methods  Trichloroethylene-contaminated water was pumped into soil columns with a layer of vermicompost at 10-cm depth (biobarrier system). The impacts of the trichloroethylene on the microbial community were evaluated by determining: (1) the overall microbial activity (estimated as dehydrogenase activity) and enzyme activities related to the main nutrient cycles (β-glucosidase, o-diphenoloxidase, phosphatase, urease, and arylsulphatase activities). In addition, isoelectric focusing of the soil extracellular humic-β-glucosidase complexes was performed to study the enzymatically active humic matter related to the soil carbon cycle. (2) The soil bacterial diversity and the molecular mechanisms for the bacterial resistance to organic solvents were also determined. For this, polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) was used to detect changes in bacterial community structure and PCR-single-strand conformational polymorphism (SSCP) was developed and optimised for detection and discrimination of the resistance-nodulation-division (RND) genes amplified from the contaminated soils. Results  Vermicompost reduced, with respect to the unamended soil, about 30% of the trichloroethylene leaching during the first month of the experiment. Trichloroethylene had a marked negative effect on soil dehydrogenase, β-glucosidase, urease, phosphatase, and arylsulphatase activities. Nevertheless, the vermicompost tended to avoid this toxic effect. Vermicompost also displays stable humic-β-glucosidase complexes that increased the extracellular activity related to C-cycle in the contaminated soils. The isoelectric focusing technique showed a more biochemically active humic matter in the soil sampled under the vermicompost. The behaviour of the three main phyla of bacteria isolated from the DGGE bands was quite different. Bands corresponding to Actinobacteria disappeared, whereas those affiliated with Proteobacteria remained after the trichloroethylene contamination. The disappeared Actinobacteria became visible in the soil amended with the vermicompost. Bands corresponding to Bacteriodetes appeared only in columns of contaminated soils. In this study, six types of RND proteins were detected by PCR-SSCP in the natural soil, three in the trichloroethylene-contaminated soil and 7/5 in trichloroethylene-contaminated soil above/below the vermicompost in the biobarrier columns. Trichloroethylene tended to reduce or eliminate all the clones detected in the uncontaminated soil, whereas new efflux pumps appeared in the biobarrier columns. Discussion  Although enzymes incorporated into the humic substances of vermicomposted olive wastes are quite stable, trichloroethylene also inhibited the background levels of the soil extracellular β-glucosidase activity in the amended soils. The decrease was less severe in the biobarrier system, but in any case, no relation was found between the levels of trichloroethylene in soil and extracellular β-glucosidase activity, or between the latter and the quantity of humic carbon in soils. The isoelectric focusing technique was carried out in the humic fraction to determine whether the loss of activity occurred in overall extracellular β-glucosidase or in that linked to stable humic substances (humic–enzyme complexes). The contaminated soils showed the lower enzyme activities, whereas contaminated and amended soils presented greater quantity of focalised (and therefore stable) humic carbon and spectra heterogeneity: very different bands with higher enzyme activities. No clear relationship between trichloroethylene concentration in soil and diversity of the bacterial population was noted. Similar patterns could be found when the community structures of bacteria and microbial activity were considered. Since the use of the dehydrogenase assay has been recognised as a useful indicator of the overall measure of the intensity of microbial metabolism, these results could be attributed to PCR-DGGE methodology, since the method reveals the presence of dominant populations regardless of their metabolic state. Trichloroethylene maintained or even increased the number of clones with the DNA encoding for RND proteins, except for the contaminated soil located above the vermicompost. However, the main effect of trichloroethylene was to modify the structure of the community in contaminated soils, considering the type of efflux pumps encoded by the DNA extracted from soil bacteria. Conclusions  Trichloroethylene inhibited specific functions in soil and had a clear influence on the structure of the autochthonous bacterial community. The organic matter released by the vermicomposted olive waste tended to avoid the toxic effect of the contaminant. Trichloroethylene also inhibited the background levels of the soil extracellular β-glucosidase activity, even when vermicompost was present. In this case, the effect of the vermicompost was to provide and/or to stimulate the humic-β-glucosidase complexes located in the soil humic fraction >10⁴, increasing the resistance of the enzyme to the inhibition. The bacterial community from the soil presented significantly different mechanisms to resistance to solvents (RND proteins) under trichloroethylene conditions. The effect of the vermicompost was to induce these mechanisms in the autochthonous bacterial community and/or incorporated new bacterial species, able to grow in a trichloroethylene-contaminated ambient. Coupled biochemical and molecular methodologies are therefore helpful approaches in assessing the effect of an organic amendment on the biochemical and biological restoration of a trichloroethylene-contaminated soil. Recommendations and perspectives  Since the main biochemical and biological effects of the organic amendment on the contaminated soil seem to be the incorporation of biochemically active humic matter, as well as new bacterial species able to grow in a trichloroethylene-contaminated ambient, isoelectric focusing and PCR-SSCP methodologies should be considered as parts of an integrated approach to determine the success of a restoration scheme.     

28-Homobrassinolide mitigates boron induced toxicity through enhanced antioxidant system in Vigna radiata plants

The objective of this study was to establish relationship between boron induced oxidative stress and antioxidant system in Vigna radiata plants and also to investigate whether brassinosteroids will enhance the level of antioxidant system that could confer tolerance to the plants from the boron induced oxidative stress. The mung bean (V. radiata cv. T-44) plants were administered with 0.50, 1.0 and 2.0 mM boron at 6 d stage for 7 d along with nutrient solution. At 13 d stage, the seedlings were sprayed with deionized water (control) or 10⁻⁸ M of 28-homobrassinolide and plants were harvested at 21 d stage to assess growth, leaf gas-exchange traits and biochemical parameters. The boron treatments diminished growth, water relations and photosynthetic attributes along with nitrate reductase and carbonic anhydrase activity in the concentration dependent manner whereas, it enhanced lipid peroxidation, electrolyte leakage, accumulation of H₂O₂ as well as proline, and various antioxidant enzymes in the leaves of mung bean which were more pronounced at higher concentrations of boron. However, the follow-up application of 28-homobrassinolide to the boron stressed plants improved growth, water relations and photosynthesis and further enhanced the various antioxidant enzymes viz. catalase, peroxidase and superoxide dismutase and content of proline. The elevated level of antioxidant enzymes as well as proline could have conferred tolerance to the B-stressed plants resulting in improved growth, water relations and photosynthetic attributes.     

城市污泥与园林废弃物混合堆肥施用对林地土壤微生物量碳、氮及酶活性的影响

研究城市污泥和园林废弃物混合堆肥施用对土壤微生物量碳(SMBC)、微生物量氮(SMBN)及酶活性的影响,可为林地施用混合堆肥产品的安全及功效性评价提供科学依据.本研究采用大田试验,选取紫穗槐为研究对象,将污泥和园林废弃物以3种比例(GF(纯园林废弃物)、SGA(V(污泥)∶V(园林废弃物)=1∶3)、SGB(V(污泥)∶V(园林废弃物)=1∶1))进行混合堆肥,每种堆肥产品分别按0、5、10、50 kg·株-1(分别记作CK、T5、T10、T50)均匀施入样地.施肥1年后,于田间采样测定土壤生物活性指标及理化性质,采用主成分分析及单、双因素方差分析法研究混合堆肥产品施用后对土壤酶活性及微生物量碳、氮含量的影响.结果表明:施入GF、SGA和SGB时,SMBC和SMBN均在施用量50 kg·株-1时达到最大.随着施用量的增加,土壤酶指数(SEI)总体上呈现逐步递增的趋势,施入GF、SGA、SGB时,各个施肥梯度处理的土壤酶指数较CK分别增加93.33%~186.67%、14.58%~54.17%、119.05%~204.76%.通过主成分分析,依据各处理综合得分情况,不同堆肥产品总体上表现为:SGBGFSGA.施入GF、SGA和SGB,不同处理综合得分均为T50T10T5CK.说明施用堆肥产品能够提高土壤微生物量,增加土壤酶活性,改善土壤的生物环境.     

季节性冻融对扎龙湿地土壤微生物群落结构和胞外酶活性的影响

选取扎龙湿地芦苇沼泽为研究对象,采用磷脂脂肪酸法和微孔板法研究了季节性冻融期土壤微生物群落结构和胞外酶活性的动态变化规律.结果表明,微生物总磷脂脂肪酸（PLFA）含量和细菌PLFA含量均呈先降低后升高趋势,而真菌PLFA含量呈先升高后降低趋势.冻融初期至融冻初期,β-葡萄糖苷酶显著降低了71.84%,β-N-乙酰氨基葡萄糖苷酶表现为先升高后降低趋势,酸性磷酸酶在融冻初期最高、冻融初期最低,过氧化物酶和多酚氧化酶峰值均出现在融冻后期.主成分分析表明,季节性冻融显著改变了土壤微生物群落结构.冗余分析发现,土壤含水率与总PLFA、细菌、革兰氏阳性菌、革兰氏阴性菌、放线菌、过氧化物酶和多酚氧化酶呈显著正相关（p<0.05）;土壤全磷、速效磷、速效氮与真菌和β-N-乙酰氨基葡萄糖苷酶呈显著正相关;β-葡萄糖苷酶主要受土壤碳氮比和有机质影响,而土壤微生物与胞外酶活性在不同冻融时期具有显著相关性（p<0.05）.因此,探究全球变化背景下土壤微生物和胞外酶对季节性冻融过程的响应机制,可为深入解析扎龙湿地土壤质量演变特征及湿地生态系统功能的恢复与保护提供科学依据.     

一株新颖的Providencia sp.菌降解吲哚及合成靛蓝的特性研究

吲哚是一种典型的氮杂环芳烃污染物,在焦化废水和畜牧废水中大量存在.本研究从近海泥沙中分离纯化得到一株高效吲哚降解菌DCX,经16S rRNA基因序列比对分析,鉴定其为普罗维登斯菌属(Providencia sp.).该菌株能够以吲哚为唯一碳源,在28 h内将100 mg·L~(-1)吲哚完全降解.液相色谱/飞行时间-质谱联用(LC/TOF-MS)的结果表明,靛蓝、靛红、靛红酸及邻氨基苯甲酸是菌株DCX降解吲哚过程中的中间产物.此外,本研究中发现,外加营养物质可以促进菌株DCX降解吲哚,特别是加入酵母浸粉后,反应体系中会产生大量靛蓝.利用表面响应法确定菌株DCX转化吲哚合成靛蓝的最优条件为:吲哚207.49 mg·L~(-1),酵母浸粉2.9 g·L-1,接菌量4.23%(V/V),并且在最优条件下,靛蓝产率达到最高(9.90%),比初始条件提高了4.38倍.     

复合垂直流构建湿地基质微生物类群及酶活性的空间分布

对构建湿地基质微生物类群数目及酶活性的空间分布特点进行了研究，为进一步深入研究构建湿地净化污水的机制提供了可能，研究表明：（1）不同月份构建湿地基质的微生物类群目是不相同的。（2）基层不同层次之间其微生物类群数目不相同，上层数目较中下层多，最多几乎达三个数量级，（3）下行流池中的微生物类群数目较上行流池多。（4）下行流池中的脲酶活性较上行流池高，且上层高于中下层。（5）上行流池中的磷酸酶活性较下行流池为高。     

Microvillar enzyme assays in amniotic fluid and fetal tissues at different stages of development

A systematic study of microvillar enzyme activities in the amniotic fluid in correlation with their values in different fetal tissues during development has been undertaken. Microvillar enzymes appeared in the amniotic fluid at the time of disappearance of the anal membrane, 12–13 weeks, and declined from the 18th week until the 24th week. The study of fetal tissues and fluids has shown that gamma-glutamyltranspeptidase is mainly of liver origin. The significant decrease of the activities of these amniotic fluid enzymes has been the basis of prenatal diagnosis of cystic fibrosis. These assays may be useful for the diagnosis of certain digestive tract abnormalities at later stages of pregnancy.