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1.
Acidification is considered the most important one of the primary chemical stress factors that impact on freshwater ecosystems. In unpolluted freshwater systems, the primary controls on the degree of acidification are factors such as the geological substrate of the catchment area, the presence of organic acids secreted by vegetation in the river system, and equilibrium exchange of carbon dioxide with the atmosphere. Anthropogenic factors that can impact on the degree of acidification of freshwater systems include agricultural, mining and industrial activities, either through direct runoff into river systems or through deposition of atmospheric pollutants from these sources. The capacity factors alkalinity and acidity, which represent the acid- and base-neutralizing capacity (ANC and BCN) of an aqueous system, have been used as more reliable measures of the acidic character of freshwater systems than pH. Unlike pH, ANC and BNC are not affected by parameters such as temperature and pressure. Therefore, ANC has been employed as a predictor of biological status in critical load assessments. Freshwater systems with ANC's eq/L isμeq/L are considered sensitive to acidification, ANC=0 μbelow 150 commonly used as the predictor for fish species such as trout in lakes, and an eq/L as more realistic for streams. Acid-neutralizing capacity μANC value of 40 (ANC) can be determined by titration with a strong acid to a preselected equivalence point. Alternatively, it can be calculated as the difference between base cations ([BC]) and strong acid anions ([SAA]): ANC=[BC]- [SAA]=[Ca^2+]+[Mg^2+]+[Na^+]+[K^+]-[SO4^2-]-[NO3^-]-[Cl^-] To date, there has been no attempt to establish the ANC of South Africa's freshwater ecosystems or variability therein, despite the fact that long-term water quality monitoring data exist for all the parameters needed to calculate it according to the above equations. As a result, the relationship between the acid neutralizing capacity of freshwater ecosystems in South Africa and biodiversity factors, such as fish status, is unknown. Results of the first comprehensive (country-wide scale) evaluation of the acid neutralizing capacity of river systems in South Africa will be presented. Long-term monitoring data obtained from the Department of Water Affairs and Forestry (DWAF) from most of South Africa's river systems were used to establish geographic and temporal variabilities in ANC. The results show that the Berg and Breede River systems are most susceptible to acidification, and that geological substrate appears to explain most of the geographic variabilities observed.  相似文献   
2.
通过制备针对东海原甲藻细胞破碎物的多克隆和单克隆抗体,建立了基于双抗体酶联免疫分析定量检测东海原甲藻(Prorocentrum donghaiense)的检测方法。利用该方法对单一藻种、混合藻种和现场样品中的东海原甲藻进行检测的结果与镜检结果相一致,最低检测限度为1×103 cells/m L。该方法的建立对中国近海赤潮暴发的实时监控具有重要意义。  相似文献   
3.
海豚链球菌(Streptococcus iniae)是引起鱼类链球菌病的主要致病菌,对牙鲆(Paralichthys olivaceus)等养殖鱼类造成巨大危害。本文制备了海豚链球菌强毒株NUF849的福尔马林灭活全菌(FKC)、胞外产物(ECPs)及全菌与胞外产物的混合物(FKC+ECPs),以其免疫牙鲆,在免疫后第42天分别以海豚链球菌NUF849和NUF812攻毒,并在免疫前以及免疫后第7、14、21、28、35、42天与攻毒后第7天取样,分析3种免疫原以及免疫后再攻毒所诱发的血清抗体应答。结果显示,免疫后牙鲆体内产生了分别针对FKC和ECPs的特异性抗体,且各免疫组中FKC诱导的抗体水平较ECPs高,2种抗原间存在一定程度的交叉反应;FKC+ECPs组的两种抗体水平显著高于其他组(P0.05);攻毒后第7天2种抗体水平都显著升高(P0.05),并且免疫原来源菌株NUF849攻毒组的2种抗体水平高于非免疫原来源菌株NUF812攻毒组。攻毒后存活牙鲆的血清对NUF849、NUF812全菌蛋白及其胞外产物进行Western-blot分析,结果显示抗血清与全菌蛋白的阳性反应条带位于25~100kDa,与胞外产物的阳性反应条带位于18~100kDa。本文结果表明灭活海豚链球菌与胞外产物都能够诱发牙鲆产生特异性抗体,二者混合物的免疫效果更好,且NUF849来源的免疫原可以刺激牙鲆产生针对NUF812菌株的交叉保护抗体,为牙鲆海豚链球菌疫苗成分的筛选提供了基础资料。  相似文献   
4.
在鲨鱼等软骨鱼类体内存在天然的缺失轻链、仅包含重链的抗体,源于这种重链抗体可变区的片段称为单域抗体(single domain antibody)。目前单域抗体研究中的抗原主要来源于水溶性蛋白或病原体,筛选膜蛋白鲨鱼源单域抗体的领域接近空白。本研究以重组表达的小球藻病毒玻尿酸合成酶(一种膜蛋白)为抗原,经过免疫、建库、淘选、验证等步骤获得了抗原特异性条纹斑竹鲨(Chiloscyllium plagiosum)单域抗体序列。随后利用大肠杆菌表达了该单域抗体,通过等温滴定量热技术(ITC)测定了其与抗原的亲和力,证实了以膜蛋白作为抗原制备鲨鱼源单域抗体的可行性。  相似文献   
5.
神经肽Y(Neuropeptide Y,NPY)被普遍认为是一种重要的促食因子,在调节鱼类的摄食行为方面起着至关重要的作用。为进一步研究牙鲆NPY蛋白的生物学功能,作者克隆了牙鲆NPY成熟肽序列(m-NPY)及含有信号肽的全长序列(f-NPY),利用原核表达系统分别进行体外重组表达,筛选出诱导剂IPTG最佳诱导浓度为0.8 mmol/L及最佳诱导时间3 h;此外,根据牙鲆NPY第49-64位氨基酸序列制备了多克隆抗体并通过Western blot验证该多抗能够有效检验重组NPY及牙鲆体内NPY的表达。研究结果为研究重组牙鲆NPY蛋白在牙鲆水产养殖产业中的应用及检测提供了依据。  相似文献   
6.
大黄鱼病原副溶血弧菌单克隆抗体制备及其应用   总被引:5,自引:0,他引:5  
用甲醛灭活副溶血弧菌(Vibrio parahaemolyticus)和溶藻弧菌(V.alginolyticus)制成免疫原免疫BALB/c小鼠,利用淋巴细胞杂交瘤技术,获得1株特异的针对副溶血弧菌的单克隆抗体,命名为D6F3H5。腹水及培养上清液抗体的ELISA效价分别为:1∶5 120和1∶1 280,该单克隆抗体与其它细菌没有明显的交叉反应。利用该单克隆抗体和兔抗副溶血弧菌多克隆抗体,建立了检测副溶血弧菌的双抗体夹心ELISA方法。该方法对副溶血弧菌的最小检出量为5×104个/mL。用双抗体夹心ELISA检测大黄鱼(Pseudosciaena crocea)样品,14尾患病大黄鱼中有11尾检出副溶血弧菌,而10尾健康大黄鱼都没有检出副溶血弧菌。由此可见,本试验制备的高特异性的抗副溶血弧菌单克隆抗体,可以用于患病大黄鱼副溶血弧菌的快速诊断。  相似文献   
7.
Observations from the US Environmental Protection Agency's Episodic Response Project (ERP) in the North‐eastern United States are used to develop an empirical/mechanistic scheme for prediction of the minimum values of acid neutralizing capacity (ANC) during episodes. An acidification episode is defined as a hydrological event during which ANC decreases. The pre‐episode ANC is used to index the antecedent condition, and the stream flow increase reflects how much the relative contributions of sources of waters change during the episode. As much as 92% of the total variation in the minimum ANC in individual catchments can be explained (with levels of explanation >70% for nine of the 13 streams) by a multiple linear regression model that includes pre‐episode ANC and change in discharge as independent variables. The predictive scheme is demonstrated to be regionally robust, with the regional variance explained ranging from 77 to 83%. The scheme is not successful for each ERP stream, and reasons are suggested for the individual failures. The potential for applying the predictive scheme to other watersheds is demonstrated by testing the model with data from the Panola Mountain Research Watershed in the South‐eastern United States, where the variance explained by the model was 74%. The model can also be utilized to assess ‘chemically new’ and ‘chemically old’ water sources during acidification episodes. Copyright © 1999 John Wiley & Sons, Ltd.  相似文献   
8.
《海洋科学》2012,36(3)
以杂色鲍(Haliotis diversicolor Reeve)为研究对象,通过密度梯度离心方法纯化得到高纯度血蓝蛋白,以其作为抗原皮下注射免疫新西兰大白兔,从而获得高效价的兔源多克隆抗血清。进一步通过ProteinA抗原亲和纯化的方法对该抗血清纯化,最终获得效价更高、检测特异性更好的血蓝蛋白多克隆抗体。应用该抗体进行Western检测发现,鲍血淋巴中存在着多样的血蓝蛋白衍生产物;进一步结合质谱技术对其中35kDa条带进行鉴定发现,其来源于血蓝蛋白I型亚基的H结构域。  相似文献   
9.
将新加坡石斑鱼虹彩病毒(Singapore grouper iridovirus,SGIV)的ORF162的开放式阅读框插入pET-32a表达载体T7启动子控制下的6-His·Tag编码基因上游,构建SGIVORF162原核表达质粒pET-ORF162。表达质粒转化入大肠杆菌BL21(DE3)菌株,经IPTG诱导,成功表达SGIV ORF162融合蛋白。对IPTG浓度、诱导温度、诱导时间等诱导表达条件进行优化后,确定在0.7mmol/LIPTG、16℃条件下诱导14h时可溶性SGIV ORF162重组蛋白占重组蛋白总量的95%。经镍琼脂糖凝胶纯化,获得纯度为90%以上的SGIV ORF162蛋白。用纯化的SGIV ORF162蛋白免疫小鼠,获得高效特异的SGIV ORF162多克隆抗体。  相似文献   
10.
用鲶爱德华氏菌兔抗血清作为一抗,碱性磷酸酶(AP)标记的羊抗兔IgG作为酶标二抗,建立黄颡鱼"红头病"病原菌—鲶爱德华氏菌的间接酶联免疫(ELISA)快速检测法,并优化检测条件。抗原最佳包被浓度为107/mL,一抗工作的最佳稀释度为1∶211,病原菌的检测灵敏度为105/mL,交叉反应实验证明该方法特异性强,与迟钝爱德华氏菌、弧菌等13种标准菌株无交叉。应用上述技术对人工感染发病鱼中分离的优势菌进行检测,结果表明阳性检出率为80%;对自然发病黄颡鱼体内分离获得的20株优势菌检测结果表明,12株菌为鲶爱德华氏菌。  相似文献   
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