免疫多糖对中国对虾血清溶菌酶、磷酸酶和过氧化物酶的作用

于１９９７年９月在青岛红鸟养殖场采集中国对虾,给对虾腹腔注射不同剂量的海藻多糖和北虫草我糖后,用生化法测定其血清中的溶菌酶,碱性磷酸酶,酸性磷酸酶和过氧化物酶活性的动力学变化。结果表明,注射１．０％北虫草多糖及１．０％海藻多糖４８ｈ后,对虾血清中的ＬＳＺ活性由对照组的０．２５Ｕ／ｍｌ分别增至０．８８Ｕ／ｍｌ和０．７５Ｕ／ｍｌＰＯＤ活性由对照组的０．１１Ｕ／ｍｇ分别增至２．２Ｕ／ｍｇ和１．９６Ｕ／ｍ     

Enzymatic methods for the determination of pollution in seawater using salt resistant alkaline phosphatase from eggs of the sea urchin Strongylocentrotus intermedius

A new salt resistant alkaline phosphatase from eggs of the sea urchin Strongylocentrotus intermedius (StAP) has been shown to have a unique property to hydrolyze substrate in seawater without loss of enzymatic activity. The enzyme has pH optimum at 8.0–8.5. Model experiments showed various concentrations of copper, zinc, cadmium and lead added to seawater or a standard buffer mixture to inhibit completely the enzyme activity at the concentrations of 15–150 μg/l. StAP sensitivity to the presence in seawater of metals, pesticides, detergents and oil products appears to be considerably less. Samples of seawater taken from aquatic areas of the Troitsy Bay of the Peter the Great Bay, Japan Sea have been shown to inhibit the enzyme activity; the same was shown for the samples of fresh waters. The phosphatase inhibition assay developed proved to be highly sensitive, technically easy-to use allowing to test a great number of samples.     

温度和盐度对华贵栉孔扇贝免疫相关酶的联合效应

为研究温度和盐度对华贵栉孔扇贝免疫相关酶活力的联合效应,采用中心复合设计(CCD)和响应曲面分析法(RSM)进行试验,在实验室条件下研究了温度(19~31℃),盐度(22~38)对华贵栉孔扇贝酸性磷酸酶(ACP)、碱性磷酸酶(AKP)、谷胱甘肽还原酶(GR)三种免疫相关酶活性的联合效应,旨在考察温度与盐度对ACP,AKP和GR活性的影响及华贵栉孔扇贝免疫水平较高时的最佳温度和盐度组合,为华贵栉孔扇贝人工养殖所需条件提供理论基础。结果表明:温度的一次效应和二次效应对ACP,AKP和GR活力影响显著(P0.05);盐度的一次效应和二次效应对ACP,AKP和GR的活力影响显著(P0.05);温度和盐度的交互作用对ACP,AKP和GR活力的影响显著(P0.05)。采用响应曲面法,建立了温度和盐度对华贵栉孔扇贝ACP,AKP和GR活性影响的模型方程,该模型方程决定系数分别为0.986 6,0.981 3和0.957 6(P0.01),预测系数分别为0.934 5,0.888 0和0.738 4,表明该模型可以用于预测华贵栉孔扇贝ACP,AKP和GR活力变化。通过模型优化和验证实验,得到在温度24.30℃、盐度30.55时,ACP和AKP活力达到最小值,分别为170.43,181.74U/mg,而GR活力最大为0.34U/mg,满意度达0.95。     

Combined effects of cadmium and butachlor on soil enzyme activities and microbial community structure

The combined effects of cadmium (Cd, 10 mg/kg of soil) and butachlor (5, 10 and 50 mg/kg of soil) on enzyme activities and microbial community structure were assessed in phaeozem soil. The result showed that phosphatase activities were decreased in soils with Cd (10 mg/kg of soil) alone whereas urease acitivities were unaffected by Cd. Urease and phosphatase activities were significantly reduced by high butachlor concentration (50 mg/kg of soil). When Cd and butachlor concentrations in soils were added at milligram ratio of 2:1 or 1:2, urease and phosphatase activities were decreased, while enzyme activities were greatly improved at the ratio of 1:5. This study indicates that the combined effects of Cd and butachlor on soil urease and phosphatase activities depend largely on the addition concentration ratios to soils. The random amplified polymorphic DNA (RAPD) analysis showed that the changes occurring in RAPD profiles of different treated samples included variation in loss of normal bands and appearance of new bands compared with the control soil. The RAPD fingerprints showed substantial differences between the control and treated soil samples, with apparent changes in the number and size of amplified DNA fragments. The results showed that the addition of high concentration butachlor and the combined applied Cd and butachlor significantly affected the diversity of microbial community. The present results suggest that RAPD analysis in conjunction with other biomarkers such as soil enzyme parameter etc. would prove a powerful ecotoxicological tool.     

富营养水体磷酸酶动力学参数的季节变化——水体酶研究之一

本文报道了武汉东湖富营养湖区(东湖Ⅰ站)水体磷酸酶动力学参数(最大反应速度V_(max)和米氏常数K_m)的季节变化规律,在6月、8月和11月(或邻近时间),湖水溶解正磷酸盐的浓度较低而叶绿素的浓度达到高峰值,同时,V_(max)达到最大值,K_m值明显减小。因此,浮游植物可以通过提高酶的反应速度以及增强酶与底物的亲和能力这两种方式有效地利用有机磷。实验结果表明,湖水中对紫外线敏感的磷化合物可能是磷酸酶的竞争性抑制剂。     

Origin and characteristics of the zooplankton phosphatase activity in a coastal ecosystem of the Mediterranean sea (Toulon Bay)

In Toulon Bay (France), very high phosphatase activities have been found in the zooplankton fraction>90 microm. This work was intended to specify their origin. For that purpose, larvae, juvenile and adult Crustacea (Copepods: Calanoids, Cyclopoids, Branchiopods: Cladocera, and Cirripeds) were isolated. Their activities were measured using paranitrophenyl phosphate dissolved in sea water in order to calculate Km (the enzyme half saturation concentration) and Vmax (the reaction rate when the enzyme is saturated with substrate). Vmax were referred to protein contents of the isolated organisms to calculate specific activities. For all zooplankton groups high and low affinity phosphatase activities were found. The low affinity enzyme was responsible for at least 70% of the total phosphatase activity. Its specific activity was higher for larvae than for copepodites and adults. In Cirriped nauplii this activity was particularly high with values which were several hundred times higher than that in other Crustacea. These enzymes had optimum pH close to 8.4, magnesium requirement and were competitively inhibited by orthophosphate. Experiments with intact and lysed Cirriped nauplii confirmed that living organisms had only a weak external activity and showed that most of the activity of these larvae was primarily intracellular.     

Measurement of Low-level Extracellular Enzyme Activity in Natural Waters Using Fluorigenic Model Substrates

Activities of the extracellular enzymes ß-D-glucosidase, phosphatase, and leucine aminopeptidase were investigated in unpolluted waters to test the suitability of fluorigenic model substrates for measuring enzyme activities in water samples expected to sustain low levels of activity. The waters tested were surface running waters, shallow groundwaters, and deep groundwaters from water extraction wells. Enzyme activity was detected in all the waters tested, although some of the hydrolytic rates were at the lower limit of current analytical capability. Rates varied between 1.1 and 291 nmol/(L·h) for ß-D-glucosidase, between 9.2 and 320 nmol/(L·h) for phosphatase, and between 5.8 and 1560 nmol/(L·h) for leucine aminopeptidase. The method thus proved to be suitable for measuring low levels of extracellular enzyme activity in natural waters, but in any case, it is necessary to adapt the technique to the type of water investigated.     

P-immobilisation and phosphatase activities in lake sediment following treatment with nitrate and iron

The influence of a further developed inlake restoration method on the P-immobilisation and microbial activities, especially under anoxic conditions was investigated. The impact of nitrate and iron dosing with a newly developed nitrate storage compound (Depox^®Fe) was tested in enclosures in the eutrophic dimictic Lake Dagowsee, Germany. Additions of 50 g m⁻² of NO₃⁻–N and 66 g m⁻² of Fe³⁺ ensured availability of nitrate at the sediment surface during a 2-months period.As a result the phosphate release from the anoxic sediments was completely suppressed even 1 year after the application. The hypolimnetic deoxygenation was unaffected by the Depox^®Fe addition. However, sulfur reduction and methanogenesis were inhibited and the phosphatase activity increased.