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.     

Discriminant analysis for assessing the value of amniotic fluid microvillar enzymes in the prenatal diagnosis of cystic fibrosis

We have analysed the sensitivity, specificity, and reliability of biochemical diagnosis based on microvillar membrane enzyme assay and using discriminant analysis in amniotic fluid samples obtained from 54 pregnancies at high risk for cystic fibrosis and 125 normal pregnancies. Our results show that amniotic fluid trehalase, alkaline phosphatase, alkaline phosphatase isoenzymes and gamma-glutamyltransferase enzyme activities measured during 16–20 gestational weeks, in spite of their non-specificity for cystic fibrosis, have a very good predictive value for fetal cystic fibrosis or exclude the possibility of the disease. Overall enzyme activity analysis provided over 90 per cent reliability of the method.     

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。     

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.     

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.     

Phytotoxicity and antioxidative enzymes of green microalga (Desmodesmus subspicatus) and duckweed (Lemna minor) exposed to herbicides MCPA,chloridazon and their mixtures

In this study, we evaluate the toxicity of MCPA (auxin-like growth inhibitor), chloridazon (CHD) (PSII-inhibitor) and their mixtures to floating plants and planktonic algae. Toxicity of MCPA (4-chloro-2-methylphenoxyacetic acid) and CHD (5-amino-4-chloro-2-phenyl-3(2H)-pyridazinone) was first assessed in two growth inhibition tests with Lemna minor (ISO/DIS 20079) and Desmodesmus subspicatus (ISO 8692). Next, herbicide mixtures at concentrations corresponding to the EC values were used to assess their interactive effects, and the biomarkers were: for duckweed fresh weight, frond area, chlorophyll content and number of fronds, and for algae cell count and cell volume. The 3d EC₁₀ and EC₅₀ values using cell counts of D. subspicatus were 142.7 and 529.1 mg/L for MCPA and 1.7 and 5.1 mg/L for CHD. The 7d EC₁₀ and EC₅₀ values using frond number of L. minor amounted to 0.8 and 5.4 mg/L for MCPA and 0.7 and 10.4 mg/L for CHD. Higher sensitivity of reproductive (number of cells/fronds) than growth processes (cell volume/frond area) to herbicides applied individually and in mixtures was especially pronounced in the responses of Desmodesmus. Herbicide interactions were assessed by the two-way ANOVA and Abbott's formula. Generally, an antagonistic interaction with Lemna was revealed by MCPA and chloridazon, whereas additive effect of both herbicides was observed for Desmodesmus. A significant stimulation of SOD and APX activity by binary mixtures was noted in algal cells mainly after 24 and 48 hours of exposure. The extremely high stimulation of the activity of both enzymes was induced by the combination EC₁₀CHD + EC₅₀MCPA (48 h). Presumably due to oxidative stress, the treatment with CHD at concentration EC₅₀ after 72h was lethal for algae grown in aerated cultures, in contrast to standardized test conditions. Taking into account the consequences of risk assessment for herbicide mixtures we can state that a relatively low toxicity, as well as the lack of significant synergy between MCPA and CHD to non-target plants appears to be the most important result.     

Repeated batch and continuous degradation of chlorpyrifos by Pseudomonas putida

The present study was undertaken with the objective of studying repeated batch and continuous degradation of chlorpyrifos (O,O-diethyl O-3,5,6-trichloropyridin-2-yl phosphorothioate) using Ca-alginate immobilized cells of Pseudomonas putida isolated from an agricultural soil, and to study the genes and enzymes involved in degradation. The study was carried out to reduce the toxicity of chlorpyrifos by degrading it to less toxic metabolites. Long-term stability of pesticide degradation was studied during repeated batch degradation of chlorpyrifos, which was carried out over a period of 50 days. Immobilized cells were able to show 65% degradation of chlorpyrifos at the end of the 50th cycle with a cell leakage of 112 × 10³ cfu mL^?1. During continuous treatment, 100% degradation was observed at 100 mL h^?1 flow rate with 2% chlorpyrifos, and with 10% concentration of chlorpyrifos 98% and 80% degradation was recorded at 20 mL h^?1 and 100 mL h^?1 flow rate respectively. The products of degradation detected by liquid chromatography–mass spectrometry analysis were 3,5,6-trichloro-2-pyridinol and chlorpyrifos oxon. Plasmid curing experiments with ethidium bromide indicated that genes responsible for the degradation of chlorpyrifos are present on the chromosome and not on the plasmid. The results of Polymerase chain reaction indicate that a ~890-bp product expected for mpd gene was present in Ps. putida. Enzymatic degradation studies indicated that the enzymes involved in the degradation of chlorpyrifos are membrane-bound. The study indicates that immobilized cells of Ps. putida have the potential to be used in bioremediation of water contaminated with chlorpyrifos.     

Ozone tolerance in Phaseolus vulgaris depends on more than one mechanism

Two bean cultivars with different sensitivity to ozone, i.e. the O₃-sensitive Cannellino and the O₃-tolerant Top Crop, were exposed to acute O₃-stress (165 nL L⁻¹) with the aim of evaluating physiological and biochemical traits that may confer O₃-tolerance. Stomatal conductance was smaller and the ability to dissipate excess energy, via regulated and unregulated nonphotochemical quenching mechanisms was greater in Top Crop than in Cannellino. These morphological and physiological-traits allowed the O₃-tolerant cultivar to compensate for the light-induced declines in Φ_PSII, to preserve photosystem II from excitation-energy, and likely to prevent the generation of ROS to a superior degree than the O₃-sensitive cultivar. Furthermore, the potential capacities to reducing the superoxide anion and H₂O₂ were significantly greater in Top Crop than in Cannellino. These findings are consistent with the early accumulation of H₂O₂, the almost complete disruption of cell structure, and irreversible damages to the photosynthetic apparatus observed in the O₃-sensitive cultivar.     

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.     

产漆酶食用菌的粗酶液对多环芳烃降解研究

漆酶是一种含铜的多酚氧化酶,可以降解多种有机污染物,其中包括多环芳烃。但是由于纯漆酶的价格昂贵,限制了在环境修复中的应用。采用液态和固态2种方式培养了食用菌Pleurotus ostreatus和Coprinus comatus,并考察了固态培养的粗酶液对多环芳烃的降解和去毒能力,结果发现2种真菌均可产漆酶,但不产木质素过氧化物酶和锰过氧化物酶,其中固态培养的漆酶产量高于液态培养,且C.comatus的漆酶产量大于P.ostreatus。2种真菌固态培养的粗酶液对多环芳烃均有一定的降解和去毒作用,且P.ostreatus粗酶液的漆酶活性虽然较低,但是对多环芳烃降解率和去毒效果却优于C.comatus,这可能是P.ostreatus的粗酶液中含有较多的调节物质造成的。P.ostreatus的固态发酵粗酶液对蒽、苯并[a]芘、苯并[a]蒽、苊等均有较高的降解率(50%以上),但是对生物有效性高的萘的降解率却较低,这种降解特征可能与底物的电离势大小有关。