Isolation and characterization of microsatellite loci in western North American Anodonta species

We have developed and characterized 13 microsatellite loci from a group of Anodonta species in western North America, and demonstrated their utility in populations representing two major clades in this genus. Allelic diversity and polymorphic information content were high for all loci, although these characteristics varied across populations. Deviations from Hardy-Weinberg genotypic ratios were not detected, although the estimated frequency of null alleles was high in one population for one locus. This is the first set of microsatellite loci to be developed for freshwater mussels in western North America, and will be useful for describing gene flow patterns among populations.     

H+-ATPase of crude homogenate of the outer mantle epithelium of Anodonta cygnea

The outer mantle epithelium of the freshwater bivalve, Anodonta cygnea, is responsible for the mineralisation of the shell. Under short circuit conditions, it generates a current that is due to the operation of an electrogenic proton pump located in the apical barrier of that epithelium. Bafilomycin A1 and Concanamycin A inhibited the short circuit current. The IC50 and maximum inhibition dose were 0.17 and 0.5 microM for Bafilomycin A1, and 0.7 and 5 microM for Concanamycin A. The present work was undertaken to further characterise V-type ATPase of the outer mantle cells. The V-ATPase enzymatic activity of crude homogenate, measured as the amount of inorganic phosphorous released, due to ATP hydrolysis, in the presence of Na2SO3 (200 mM) was found to be 4.6+/-1.1 micromol Pi/mg protein/h, at 27 degrees C, pH 7.0-7.4 and ATP 4.5-6.0 microM. Bafilomycin A1 and Concanamycin A inhibit the V-ATPase activity with an IC50 of 14 and 8 nmol mg(-1), respectively. Dicyclohexylcarbodiimide (DCCD; 100 mM) and NaNO3 (100 microM) inhibited the V-type ATPase in what it seems a non-specific manner and NEM (100 mM) was unable to do it. Bafilomycin A1 (10 microM) and Concanamycin A (10 microM), inhibited 50-60% of the total activity.     

Population-specific responses to an invasive species

Predicting the impacts of non-native species remains a challenge. As populations of a species are genetically and phenotypically variable, the impact of non-native species on local taxa could crucially depend on population-specific traits and adaptations of both native and non-native species. Bitterling fishes are brood parasites of unionid mussels and unionid mussels produce larvae that parasitize fishes. We used common garden experiments to measure three key elements in the bitterling–mussel association among two populations of an invasive mussel (Anodonta woodiana) and four populations of European bitterling (Rhodeus amarus). The impact of the invasive mussel varied between geographically distinct R. amarus lineages and between local populations within lineages. The capacity of parasitic larvae of the invasive mussel to exploit R. amarus was higher in a Danubian than in a Baltic R. amarus lineage and in allopatric than in sympatric R. amarus populations. Maladaptive oviposition by R. amarus into A. woodiana varied among populations, with significant population-specific consequences for R. amarus recruitment. We suggest that variation in coevolutionary states may predispose different populations to divergent responses. Given that coevolutionary relationships are ubiquitous, population-specific attributes of invasive and native populations may play a critical role in the outcome of invasion. We argue for a shift from a species-centred to population-centred perspective of the impacts of invasions.     

河蚌C反应蛋白一级结构分析

经亲和层析纯化的河蚌 C反应蛋白 ( CRP)具有 SDS- PAGE纯度 ,用经改进的双偶联 Ed-man方法测得其 N端残基为谷氨酸 ,而不是高等动物 (人与家兔 ) C反应蛋白 N端的焦谷氨酸 .河蚌 C反应蛋白 N端的一级结构由固相 Edman方法测得 ,依次为 H2 N- E- T- A- Y- S- C- I- T- A- V- ;C端的一级结构由羧肽酶 A降解法测得 ,依次为 - L/V- S- S- T- Y- COOH,也不同于人和家兔的 C反应蛋白 .在河蚌 CRP的胰蛋白酶酶解肽段中 ,其 N端及 C端的结构也得到了证实 .河蚌 C反应蛋白经 V8蛋白内切酶酶解 ,溴化氰裂解 ,肽段经 HPLC反相柱分离 ,共得到 35个肽段 ,所有肽段的氨基酸序列均由气相氨基酸自动分析仪测得 .结合河蚌 C反应蛋白的胰蛋白酶酶解肽段的分析结果 ,其一级结构已初步拼接完成 .在其一级结构中发现有类似于其它 CRP的 Ca2 + 结合部位和磷酸胆碱结合部位 .河蚌 C反应蛋白的分子结构中存在微观不均一性 .从已知河蚌 C反应蛋白的分子特点 ,包括分子量 ,糖基化比例 ,一级结构不均一等特点 ,可以推测它与高等动物的免疫蛋白有许多相关之处 .对于河蚌 C反应蛋白分子结构的分析 ,将有助于免疫系统蛋白的发生 ,进化等方面的研究     

背角无齿蚌碱性磷酸酶的功能基团研究

在一定条件下分别采用PMSF、DTT、PCMB、NBS、TNBS、SUAN、BrAc及IBr等化学修饰剂选择修饰背角无齿蚌碱性磷酸酶的多种氨基酸残基,并测定其酶活力变化。结果表明,PMSF、NBS、TNBS、SUAN、DTT的修饰能显著抑制酶的活力,活力的降低与修饰剂的浓度相关。BrAc、IAc、PCMB的修饰不表现对酶的抑制作用。作者初步认为,Ser、Lys和Trp残基是背角无齿蚌碱性磷酸酶的必需功能基团,部分二硫键时保护酶的催化功能也是必需的。     

常用鱼药漂白粉和氯杀宁对背角无齿蚌的毒性影响

试验以洱海流域的背角无齿蚌为研究对象,确定常用鱼药:漂白粉、氯杀宁对湖泊大型底柄动物背角无齿蚌的毒性影响.试验得出漂白粉作用下,背角无齿蚌24 h、48 h、72 h、96 h、120 h、144 h、168 h的半致死率;氯杀宁作用下,背角无齿蚌24 h、48 h、72 h、96 h的半致死率;漂白粉有效氯安全浓度95.25～108.86 mg/L;氯杀宁有效氯安全浓度3.32～3.55 mg/L.通过显微镜观察,漂白粉和氯杀宁的主要作用部位为背角无齿蚌的呼吸系统,在其鳃上可见明显病变.     

背角无齿蚌对养殖池塘底泥释放重金属的净化效果

通过建立微型生态系统, 分析养殖池塘底泥释放重金属的特征及背角无齿蚌(Anodonta woodiana)对底泥释放重金属的净化效果。底泥对Al、Cr、Mn、Fe、Co、Ni、Cu、Zn、As、Mo和Pb的最大释放量分别为636、1.5、70.9、34951、10.3、36.9、34.0、53.2、72.4、48.8和3.0 μg·kg-1 dw; 蚌能够对Al、Cr、Mn、Co、Cu、Zn、As和Mo产生净化作用(P<0.05), 最大去除率分别可达到84.7%、98.0%、33.3%、14.3%、23.5%、69.4%、50.0%和13.0%, 响应面优化分析显示养殖密度和处理时间分别为40 只·m-3和24.49 d、25 只·m-3和23.96 d, Al和As去除率可提升至93.8%和60.5%; Al、Cr、Fe、Co、Cu、Zn、As和Mo的净化效果与养殖数量相关, Al、Cr、Mn、Fe、Co、Ni、Cu、Zn、As、Mo和Pb的净化效果与处理时间相关, Cr、Co、Ni、Cu和Zn的净化效果与两者交互作用相关(P<0.05)。提示背角无齿蚌有潜力防控池塘底泥重金属污染。     

不同生境蚶形无齿蚌的形态观察

对不同生境下蚶形无齿蚌壳的形态、育儿囊的类型和结构以及钩介幼虫等进行了比较研究。结果表明,在不同生境下,蚶形无齿蚌个体大小有很大差异,壳具有高度的可塑性;育儿囊由两片外鳃构成,为外鳃类的同生型,钩介幼虫在育儿囊内呈散乱状态存在;在小同区域,钩介幼虫的大小不同,但其壳高与壳长的比例却是一致的,且不同生境钩介幼虫的超微结构相同,均为有钩型。     

背角无齿蚌四种组织细胞原代培养的比较

以背角无齿蚌（Anodonta woodiana）血细胞（含颗粒细胞和透明细胞）、消化腺细胞（含上皮样细胞和圆形小细胞）、外套膜细胞（含上皮样细胞、透明细胞、圆形大细胞和圆形小细胞）、斧足细胞（含上皮样细胞、透明细胞、圆形大细胞和圆形透亮细胞）为材料,比较了上述各类细胞的原代培养特征以及不同条件下的细胞存活率,优化了细胞制备及细胞活性鉴定的方法。结果显示,上述各类组织细胞存活率受不同培养温度、培养时间和培养基种类的影响,其中,血细胞96 h在20℃下、培养基2中的存活率最高,为94.67%±0.47%;消化腺细胞48 h在20℃下、培养基3中的存活率最高,为93.67%±1.70%;外套膜细胞48h在15℃下、培养基1中的存活率最高,为93.67%±1.7%;斧足细胞48h在15℃下、培养基1的存活率最高,为94.33%±0.94%。相较于25℃,在20℃和15℃下,4种组织细胞96h的存活率更为理想。本研究旨在为背角无齿蚌组织细胞系的建立及贝类细胞水平毒理学的研究提供基础数据。