几种养殖鲍同工酶生化遗传的比较研究

运用聚丙烯酰胺凝胶电泳对我国主要养殖鲍(皱纹盘鲍、盘鲍及九孔鲍)12种同工酶(LDH、ME、MDH、GPI、PGM、IDH、IDDH、SOD、EST、ODH、ADH、AAT)26个基因座位进行了分析．结果表明：EST、PGM可以作为区分三种(亚种)鲍的生化遗传标记；在所分析的位点中，未发现多态现象，表明三种养殖鲍杂合子缺失严重，其主要原因可能在于哑基因、近交与自然选择。     

九孔鲍MSTN基因cSNP多态性及与生长性状关联分析

九孔鲍(Haliotis diversicolor supertexta)是中国南方具有较高经济价值的养殖贝类。肌肉生长抑制素(Myostatin, MSTN)是转化生长因子β超家族(TGF-β)中参与动物肌肉生长的重要调控因子, 因此MSTN基因是动物遗传改良的重要候选基因。为探究九孔鲍MSTN基因的多态性及其生长相关性, 实验采用PCR产物直接测序方法对九孔鲍MSTN基因进行单核苷酸多态性(SNP)筛选, 对MSTN基因SNP与体质量、壳长、壳宽进行关联性分析, 运用实时荧光定量PCR (qRT-PCR)技术检测九孔鲍MSTN基因不同基因型的表达水平。结果显示: 在九孔鲍MSTN基因中共筛选出9个SNP位点。SNP位点与生长性状关联分析发现, 九孔鲍MSTN基因编码区SNP位点g909C>T发生C/T同义突变与九孔鲍体质量、壳长、壳宽显著相关。TT基因型九孔鲍的体质量显著高于CC基因型和TC基因型个体; TT基因型的壳长、壳宽显著高于CC基因型(p<0.05)。通过qRT-PCR检测显示, 在九孔鲍MSTN基因SNP位点g909C>T的3种基因型中, TC和CC基因型的基因表达量显著高于TT基因型(p<0.05)。研究结果表明MSTN基因SNP位点g909C>T可作为九孔鲍标记辅助选择育种重要候选标记。     

九孔鲍褐藻酸酶、琼脂酶及纤维素酶的提取纯化

采用（NH4）2SO4分段盐析和葡聚糖凝胶SephadexG-100柱层析纯化技术，从九孔鲍Haliotis diversicolor supertexta内脏器官中提取纯化褐藻酸酶、琼脂酶及纤维素酶，结果表明，在（NH4）2SO4分段盐析纯化中，褐藻酸酶和纤维素酶的最适分离饱和度为60%，而琼脂酶为70%，分段盐析的提纯倍数为（以粗酶提取液为参照）褐藻酸酶13.3 ,琼脂酶8.7和纤维素酶10.9。葡聚糖凝胶SephadexG-100层析分离过程中，褐藻酸酶，琼脂酶和纤维素酶的比活力高峰分别出现在洗脱液的64，48和80ml处，提纯倍数分别为褐藻酸酶80.9，琼脂酶68.0及纤维素酶15.2，上述提纯方法的研究结果将为这3种酶性质的进一步研究以及作为工具酶制剂产品的开发提供了工艺技术基础。     

氟苯尼考在九孔鲍体内的药代动力学初步研究

初步研究了新型抗菌兽药氟苯尼考(florfenicol)在健康九孔鲍体内的药代动力学特征;建立了高效液相色谱法(HPLC)检测鲍血淋巴中氟苯尼考含量的可靠方法,平均回收率为85.69%,最低检测限0.05μg/cm3.以20×10-6(m/m)剂量对九孔鲍腹足单次肌注,血药浓度-时间数据用非房室模型的统计矩原理(SMT)处理后的主要药代动力学参数为:血药浓度-时间曲线下面积(AUC)为21.25mg/(h.dm3),平均滞留时间(MRT)为2.41h,清除率(C lB)为0.94dm3/(kg.h),稳态表观分布容积[Vd(ss)]为2.27dm3/kg,消除半衰期(t1/2β)为3.61h.结果显示氟苯尼考在九孔鲍体内吸收良好,分布广泛,代谢消除快.     

九孔鲍养殖中的脓疱病病因分析和药物敏感性研究

在福建沿海养殖的九孔鲍不论是成鲍还是幼鲍均经常发生脓疱病,死亡率可达 50％-60％,其症状与皱纹盘鲍脓疱病相似,但对药物的敏感性却不同．研究发现: 导致福建省的九孔鲍发生脓疱病的主要致病菌为溶藻弧菌(Vibrio alginolyticus)而非 河流弧菌(V．fluvialis)．其对多数药物不敏感,在43种抗菌药物中只对呋喃妥因、氯霉 素、痢特灵等中度敏感,而对青霉素、复方新诺明和头孢类药物则完全不敏感．由于上述 药物已被列为禁用渔药而不宜使用,故生产中应以新研制的无公害防治药物替代．     

6—DMAP抑制九孔鲍受精卵第二极体的释放诱导三倍体

当50％九孔鲍受精卵出现第一极体后，用6－DMAP不同的浓度进行不同的持续时间诱导三倍体。结果表明，6－DMAP的诱导浓度为300μmol/dm^3、诱导持续时间为10min时，其诱导的三倍体率为90％以上，并且胚胎的孵化率高达85％，幼体的畸形率较低，为50％－55％。用该方法多次进行了九孔鲍三倍体生产诱导和苗种培育试验。本文还对6－DMAP的作用机理以及九孔鲍三倍体诱导中的诱导因子等问题进行了讨论。     

Toxic effects of several phthalate esters on the embryos and larvae of abalone Haliotis diversicolor supertexta

As the most widely used plasticizers in the world, phthalate esters (PAEs) are potential endocrine disruption compounds (EDCs). In the present study, the toxicity of dimethyl phthalate (DMP),diethyl phthalate (DEP), dibutyl phthalate (DBP), di (2-ethylhexyl) phthalate (DEHP) on embryogenesis and larvae development of the marine univalve Haliotis diversicolor supertexta was examined in laboratory. The results show that the malformation of embryos appeared during the experiment, such as embryos died or lysed, small transparent flocculent rings studded on the periphery of the embryo, and the larvae could failed to hatch. In embryo toxic test, embryos incubated at the highest concentration of DMP,DEP and DBP solutions showed significantly high abnormal rate compared with the control, while DEHP solutions displayed no significant difference. In larval toxic test, in all concentrations of DMP, DEP and DBP solutions, larval settlement rates were low significantly than that of the control. Similarly, DEHP solutions show nearly no effect on the larval settlement. The order of toxicity on embryos and larvae is DBP>DEP>DMP>DEHP. Being a simple and easy stimulation to indoor spawn, sensitive to environmental factors, and short culture time, the embryos of 1-1. diversicolor supertexta can be used to indicate toxicity of the PAEs.     

The hydrogen peroxide impact on larval settlement and metamorphosis of abalone Haliotis diversicolor supertexta

Abalone Haliotis diversicolor supertexta is an important economic mollusk.The settlement and metamorphosis are two critical stages during its development period,which has direct influence on abalone survival and production.The influence of reactive oxygen species(hydrogen peroxide) on abalone embryo and juvenile development were examined in this study.Larvae of Haliotis diversicolor supertexta were induced to settlement and metamorphose by exposure to seawater supplemented with hydrogen peroxide.They had the best performance at 800 μmol/L.The concentration of 1 000 μmol/L or higher was toxic to the larvae,as the larvae could settle down only at benthic diatom plates without complete metamorphosis.In addition,H2O2 adding time was critical to the larval performance.24h after two-day post-fertilization was proved to be the optimal adding time.In this paper,two action mechanisms of hydrogen peroxide are discussed:(1) hydrogen peroxide has direct toxicity to ciliated cells,thus cause apoptosis;(2) hydrogen peroxide,as a product from catecholamines' autoxidation process in vivo,can reverse this process to produce neuro-transmitters to induce abalone metamorphosis.     

金属离子和pH值对九孔鲍几种消化酶活力的影响

研究了金属离子对九孔鲍(Haliotis diversicolor supertepla)褐藻酸酶、琼脂酶及纤维素酶活力的影响。结果表明，3种酶的最适pH不同。分别为褐藻酸酶8．8、琼脂酶4．2、纤维素酶4．5-5．0。MgSO4为褐藻酸酶的激活剂、BaCl2为琼脂酶的激活剂、MnCl2是纤维素酶的激活剂。     

九孔鲍精子短期保存技术研究

应用单因素试验和三因素正交试验研究了温度、二甲基亚砜(DMSO)浓度及精子密度等因素对九孔鲍(Haliotis diversicolor supertexta)精子短期保存的影响。单因素试验结果表明:DMSO、维生素C、甘油、葡萄糖等添加剂中仅5%和7.5%DMSO有利于精子活力的保持;3个试验温度中28℃精子活力下降速度最快,其次为1℃,最慢是5℃;稀释度越高精子活力下降速度越快。正交试验中,九孔鲍精液分别在25个保存条件下保存96 h,保存期间每隔一段时间取样观察精子活力。25个处理的活力变化数据用生长函数拟合,根据拟合方程计算出精子活力半衰期,并对结果进行直观分析及方差分析。结果表明,较理想的精子保存条件是:温度为7℃,DMSO质量分数为3%,精子密度为7.5×108个/mL;各因素对精子活力半衰期的影响按精子密度、温度、DMSO密度顺序递减;精子密度对精子活力半衰期有显著影响(P<0.05),温度及DMSO浓度的影响不显著(P>0.05)。