圆背角无齿蚌血细胞培养

用新设计的培养基培养了圆背角无齿蚌的血细胞,在倒置相差镜下进行了活体观察及扫描电镜摄影,发现培养的血细胞无颗粒细胞、颗粒细胞、透明细胞和类淋巴细胞,前三者均能伸出长的伪足和突起,与瓶壁紧密贴附,呈体外培养的成纤维细胞型;后者呈圆形,不与瓶壁贴附;四者的比例约为4：2：3：1。在活体内注射或在培养基上加入PHA和ConA,培养2－7d中,每天取部分供加入秋水仙素,用空气干燥法制片,作染色体观察,但未观察到转化细胞和有丝分裂相。研究结果表明,圆背角无齿蚌的颗粒细胞、无细胞和透明细胞在体外贴附玻璃表面的特征与高等动物的巨噬细胞类似,而不贴瓶的圆形细胞与高等动物的淋巴细胞类似,但在体外培养均不能繁殖,它们可能是高度分化的细胞。     

背角无齿蚌心肌纤维的超微结构观察

本文研究了背角无齿蚌心肌纤维的超微结构,在电子显微镜下观察了心脏各部结构固定的超薄切片,分析和讨论了河蚌心肌纤维的粗丝与细丝的排列状况和徽细结构,测最了粗丝和细丝的直径,并对线粒体,脂褐素等细胞器的形态结构,分布等进行了描述,对心肌纤维的连接方式进行了观察,并与脊椎动物心肌纤维的超微结构进行了相应的比较。     

圆背角无齿蚌血细胞吞噬作用的研究

初步研究了蚌血细胞离体条件下的吞噬作用以及血清因子对其的影响。发现蚌血细胞在无血清的条件下能独立完成对大肠杆菌的识别、吞噬。蚌血清对大肠杆菌有调理作用,能促进蚌血细胞对其的吞噬。介蚌血细胞不能识别、吸附和吞噬鸡红细胞,显示出蚌免疫防御能力的局限性。还发现ＥＤＴＡ能有效地抑制蚌血细胞凝集,其凝集作用可能与Ｃａ＾２＋等阳离子有关。     

背角无齿蚌生殖细胞及钩介幼虫的扫描电镜观察

本文报告背角无齿蚌的精子、卵子、钩介幼虫的扫描电镜观察。精子头部呈椭圆形，由顶体帽、顶体后区、核区和颈部组成，尾细长呈线状，卵子圆形，有许多卵闰，在生殖腺中有各种不同发育程度的卵细胞，钩介幼虫在育儿囊（外鳃）中发育，有一系列变态过程，表明卵子是分期分批排出，钩介幼虫也是分批发育和成熟的。     

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。     

椭圆背角无齿蚌贝壳棱柱层的结构

用光学显微镜和扫描电子显微镜对椭圆背角无齿蚌Anadonta woodiana elliptica贝壳的横截面、纵截面和水平截面进行了观测。结果表明,边长为10～50μm、高度为35～47μm的多边棱柱体是贝壳棱柱层结构的基本单元。它们在贝壳的水平轴和横轴的二维空间方向逐步进行堆砌,以形成棱柱层。在堆砌时所形成的夹角,导致了贝壳在横轴和纵轴方向形成一定的弧度。在外套膜分泌形成贝壳的过程中,依次形成角质层、棱柱层、珍珠层。     

背角无齿蚌幼蚌对水体铜的吸收特征研究

为了探究在不同浓度铜(Cu)暴露下背角无齿蚌(Anodonta woodiana)对水体Cu的吸收特征,选定对Cu吸收能力更强的幼蚌作为实验对象,依据Cu对幼蚌96 h-EC50和我国渔业水质标准(GB11607—89)中Cu限量设定5个浓度梯度2.0、1.0、0.1、0.01和0.005 mg·L^-1,进行24 h Cu暴露实验及水体Cu含量测定。结果显示:随暴露浓度的升高,幼蚌Cu吸收效率迅速升高,最高值出现在2.0 mg·L^-1暴露组为(0.69±0.11)μg/(g·h);幼蚌Cu去除率总体呈现出降低趋势,其中0.005 mg·L^-1暴露组去除率最高为84.8%, 1.0 mg·L^-1暴露组去除率最低为28.9%。综上所述,背角无齿蚌幼蚌具有较强的Cu吸收能力,表明其在淡水渔业水域环境Cu污染防控方面以及开发作为监测评价淡水渔业水域环境Cu污染的模式生物方面具有非常高的应用潜力。     

背角无齿蚌珍珠囊形成过程的组织学和酶组织化学研究

本文采用Ｈ．Ｅ染色法和酶组织化学方法,对背角无齿蚌珍珠囊形成过程的组织学和酶组织化学的变化情况进行了研究,证实了“初生珍珠囊”的形成和溶解及“次生珍珠囊”的形成,并推没“次生珍珠囊”表皮细胞是由育珠蚌结缔组织转化而来的;观察了育珠手术后九种酶在珍珠囊形成的各个阶段和在珍珠囊和育珠蚌不同部位的变化情况,说明了珍珠囊珙成过程是与复杂的能量代谢,物质代谢及物质转运等有关的生理生化过程。     

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。     

Sixteen polymorphic microsatellites in bighead carp (Aristichthys nobilis) and cross-amplification in silver carp (Hypophthalmichthys molitrix)

A (GT)(n) enriched partial genomic library of bighead carp (Aristichthys nobilis) was constructed by employing the (fast isolation by AFLP of sequences containing repeats) FIASCO protocol. Sixteen loci exhibited polymorphism with two to seven alleles/locus (mean 3.263) in a test population and the observed heterozygosity ranging from 0.100 to 0.690 (mean 0.392). Eleven of the 16 bighead carp microsatellites were found to be also polymorphic in silver carp. These polymorphic loci should provide sufficient level of genetic diversity to evaluate population structure of bighead carp.     

Isolation and characterization of microsatellites in Chinese sturgeon,Acipenser sinensis

From (GATA)_n and (AAAG)_n enriched genomic libraries for the Chinese sturgeon (Acipenser sinensis), 50 primer pairs were developed using the fast isolation by AFLP of sequences containing repeats (FIASCO) protocol. Forty‐six primer pairs exhibited highly polymorphic with two to 11 alleles per locus, while the rest four displayed monomorphic. These markers yielded 246 alleles in a survey of eight specimens of wild A. sinensis. Average observed heterozygosity ranged from 0.13 to 1.00. These loci should provide sufficient levels of genetic diversity to allow parentage analysis for artificial stocking management and delineation of fine‐scale population structure.     

Multiplex microsatellite PCR panels and their application in genetic analyses of bighead carp (Hypophthalmichthys nobilis) and silver carp (H. molitrix)

In this study, three multiplex PCR panels involving twelve microsatellite loci were developed for cross-species amplification in bighead carp (Hypophthalmichthys nobilis) and silver carp (H. molitrix). The panels were verified using population genetic analyses, and species and simulated hybrid discriminations based on the genotype data of all individuals from two populations for each species. The number of alleles (Na), the observed and expected heterozygosity (Ho and He), and the polymorphic information content (PIC) of twelve loci were ranged from 5 to 20, 0.189 to 0.956, 0.177 to 901, and 0.169 to 0.887, respectively. Overall, almost all the microsatellite loci were detected as highly polymorphic across the two species. The populations of same species showed close genetic distances and similar genetic structures. Significant genetic differentiations were only detected between populations from different species, with a higher variation between species (21.17%) compared with that between populations within species (0.18%). In the species and simulated hybrid discrimination analyses, the individuals from different species were separated distinctly, and the simulated hybrids could almost be separated with the true individuals. In brief, the results indicated the high efficiency of the microsatellite multiplex panels developed in this study, which provide a practicable tool for further genetic analyses of H. nobilis and H. molitrix.     

人工养殖中华鲟幼鱼摄食不同饵料的转化效率与生长特性

利用水蚯蚓与人工饲料分别投喂7月龄中华鲟(Acipenser sinensis)幼鱼,研究不同饵料对中华鲟幼鱼转化效率与生长特性的影响.结果表明:经过60 d生长,水蚯蚓组、人工饲料组中华鲟幼鱼的平均体长分别从177.0、191.6 mm增至216.6、332.5 mm,体质量分别从32.73、44.39 g增至58.27、264.71 g;水蚯蚓组、人工饲料组中华鲟幼鱼的饵料转化率分别为6.67～39.33、0.89～1.26,特定生长率分别为0.19%～2.33%、1.38%～3.94%.生长效率分别为2.54%～14.99%、79.56%～204.44%;人工饲料组中华鲟幼鱼的生长速度快于水蚯蚓组,肥满度大于水蚯蚓组;2个试验组中华鲟幼鱼的体长与体质量均呈幂函数关系,各阶段幂指数b值均小于3,表明中华鲟幼鱼为异速生长;水蚯蚓组中华鲟幼鱼肌肉的水分湿质量含量为82.23%±0.79%,蛋白干质量含量为62.79%±1.34%,均略比人工饲料组高;综合饵料转化效率与各项生长指标,表明人工饲料比水蚯蚓更适宜作为中华鲟幼鱼阶段的人工养殖饵料.     

野生及人工养殖中华鲟幼鱼肌肉营养成分的比较

对野生及人工养殖中华鲟幼鱼的肌肉营养成分和营养品质进行了分析比较.结果表明:野生中华鲟幼鱼肌肉中水分、粗蛋白和粗灰分含量均显著高于人工养殖中华鲟(P＜0.05),而粗脂肪含量显著低于人工养殖中华鲟(P＜0.05).野生和人工养殖中华鲟幼鱼的氨基酸组成基本一致,均含有18种氨基酸,必需氨基酸指数(EAAI)分别为72.02、66.21,其构成比例符合联合国粮农组织/世界卫生组织(FAO/WHO)的标准.脂肪酸中二十碳五烯酸(EPA)与二十二碳六烯酸(DHA)的含量野生中华鲟显著高于人工养殖中华鲟(P＜0.05),分别为22.99%、7.15%.矿物质含量丰富,微量元素含量野生中华鲟明显高于人工养殖中华鲟[动物学报 53(3):502-510,2007].     

Ingestion of bacteria in suspension Indian major carps (Catla catla,Labeo rohita) and Chinese carps (Hypophthalmichthys molitrix,Aristichthys nobilis)

Single-species groups of fry of the planktivorous, filter-feeding carps (silver carp Hypophthalmichthys molitrix, bighead carp Aristichthys nobilis, rohu Labeo rohita and catla Catla catla) were exposed to the bacterium Chromobacterium violaceum to assess if they could detect or ingest unattached bacteria. The results indicate that planktivorous carp fry are able to ingest unattached bacteria and that the numbers ingested increase with time and with bacterial concentration in the media, although probably by passive means. It is also suggested that unattached bacteria are of little nutritional significance to carps.