三种海水蟹类血淋巴细胞的比较

采用Wright’s-Geimsa混合染料对三疣梭子蟹、锈斑蟳和锯缘青蟹的血淋巴细胞进行染色,光镜下对这三种海水蟹的血淋巴细胞进行了观察和比较。根据血淋巴细胞的核质比、细胞整体着色特点、细胞质中颗粒的存在与否、颗粒的数量和分布等,三种海水蟹的血淋巴细胞均可分为颗粒细胞、半颗粒细胞和透明细胞三种主要类型,三种蟹的同类血淋巴细胞在染色形态上稍有差别。三种蟹的颗粒细胞大小差异较大,而半颗粒细胞和透明细胞大小相近。三种蟹的血淋巴细胞均以半颗粒细胞为主,透明细胞在血淋巴细胞中所占比例最小。三种蟹的血淋巴细胞密度均存在明显的雌雄差异,锈斑蟳和锯缘青蟹血淋巴细胞分类计数也存在明显的雌雄差异。     

贝类精子质量检测与评价方法的研究Ⅴ:栉江珧精子线粒体活性的荧光检测

采用罗丹明123(Rh123)和碘化丙啶(PI)两种荧光染料对栉江珧(Atrina pectinata)精子线粒体活性进行检测,比较不同染液浓度和染色时间的检测效果,并用该方法检测和比较超低温冷冻保存对线粒体活性的影响。结果显示:Rh123和PI染色的最适染液浓度均为10μg/mL,染色时间控制在10 min之内;染色后具有线粒体活性的精子发出绿色荧光,死精子发出红色荧光,质膜受损但线粒体还未受损的精子发出红绿色荧光,表明Rh123和PI双荧光染色检测栉江珧精子线粒体活性是可行的;精液经冷冻后,线粒体活性为61.9%、质膜完整率56.8%,显著低于新鲜精液组(95.3%、93.5%),表明冷冻对精子结构会造成不同程度的损伤,Rh123和PI双荧光染色法可用来检测栉江珧精子质量。     

温度和盐度对4种壳色马氏珠母贝谷胱甘肽过氧化物酶基因表达的影响

谷胱甘肽过氧化物酶(GPX)基因表达可反映生物对环境压力的抗逆能力。采用中心复合设计和响应曲面分析法研究不同温度(18~34℃)、盐度(20~40)对4种壳色马氏珠母贝GPX基因表达量的影响。结果表明,马氏珠母贝4种壳色群体GPX基因表达量随着温度和盐度的升高,呈现先降低后升高的趋势。温度的一次、二次效应对4种壳色马氏珠母贝GPX基因表达的影响有统计学意义(P0.01)。盐度的一次效应仅对黑壳色马氏珠母贝GPX基因表达影响有统计学意义(P0.05);盐度的二次效应对白壳色马氏珠母贝GPX基因表达的影响有统计学意义(P0.05),对黑、红、黄壳色马氏珠母贝GPX基因表达的影响有统计学意义(P0.01)。温盐协同效应对黑、红、黄壳色马氏珠母贝GPX基因表达的影响有统计学意义(P0.05),且温度的效应大于盐度效应。经响应曲面分析法和优化,得到马氏珠母贝的GPX基因表达量在适宜的温度、盐度范围内呈峰值变化,白、黑、红、黄等4种壳色马氏珠母贝分别在(21.4℃,30.1)、(24.8℃,28.5)、(25.2℃,30.5)、(25.1℃,29.7)等温盐度组合条件下达到最优(低)值(0.072、0.065、0.057、0.078),满意度分别为100%、100%、97%、100%。所建立回归模型方程的决定系数R2范围为0.971 3~0.995 8,校正决定系数R2范围为0.942 6~0.991 5,预测决定系数R2范围为0.713 9~0.968 0,模型拟合度高,模型恰当,可用于预测温度、盐度对4种壳色马氏珠母贝GPX基因表达量的影响。     

4种壳色马氏珠母贝遗传规律研究

以黑、白、红、黄4种壳色马氏珠母贝选育系F6代为亲本,选取4种壳色最纯的雌雄个体各1个,采用解剖授精法和双列杂交的方法繁育子代,建立了6个杂交组合,包括4个自交组及12个正反交组。结果表明,马氏珠母贝壳色由背景色基因和前景色基因共同表达而成,位于不同的基因位点上;经过连续7代选育的4种壳色纯化度较高,壳色可以稳定遗传给下一代,其遗传规律比较复杂。黄色和白色为马氏珠母贝的背景色,白壳色贝存在母系遗传,若为隐性纯合含致死基因,黄色相对白色为显性;黑色和红色为马氏珠母贝前景色,相对无前景色为显性,黑色与红色共显。     

噬菌弧菌N1对淡水和海水养殖水体细菌群落影响PCR-DGGE分析

【目的】研究噬菌弧菌Bacteriovorax sp. N1在一个投放周期内对淡水和海水养殖环境中细菌、弧菌总数及细菌群落结构的影响。【方法】自市售微生态制剂分离蛭弧菌N1并进行分子鉴定,测定其裂解效果,制备高浓度N1菌液分别投放至淡水红鲤鱼(red carp)和海水仿刺参(Apostichopus japonicus)养殖水体,采用细菌平板计数法及PCR-变性梯度凝胶电泳(PCR-DGGE)技术分析48 h内水体环境中细菌、弧菌总数及细菌群落结构变化。【结果】经鉴定蛭弧菌N1为噬菌弧菌,其对大肠杆菌(Escherichia coli)、溶藻弧菌(Vibrio alginolyticus)、黄海希瓦氏菌(Shewanella smarisflavi)、灿烂弧菌(Vibrio splendidus)、哈氏弧菌(Vibrio harveyi)和金黄色葡萄球菌(Staphylococcus aureus)均有裂解效果。将噬菌弧菌N1投放进淡水和海水养殖环境的12～24 h,其能显著降低两种环境中弧菌含量(P 0.05)。DGGE分析显示添加噬菌弧菌N1后淡水组优势菌群弧菌属(Vibrio,a1)和不可培养杆菌属(Uncultured bacterium, a5)在12 h以后含量有明显的减少,假单胞菌属(Pseudomonas, a3)和红杆菌科Shimia属(Shimia, a6)菌含量增加。海水组优势菌群不可培养杆菌属(c2)菌在12 h时变成非优势菌群,而白杆菌属(Albirhodobacter,c1)增加成为优势菌群。噬菌弧菌N1在水中含量在24 h时降至最低。【结论】噬菌弧菌N1对海水和淡水环境中的弧菌属和不可培养杆菌属菌群有明显的裂解作用导致其含量下降,但也使假单胞菌属(Pseudomonas),红杆菌科Shimia属和白杆菌属(Albirhodobacter)菌群含量增加,但生物效应不明。为维持噬菌弧菌N1对弧菌的控制,需要24 h左右重新补充投放。     

石油污染土壤生物修复的强化技术

为探索石油污染土壤的高效修复方法,从实验室保存的优势菌中筛选得到4株降油效果最佳菌,采用摇床和恒温培养箱培养,对含油量为5%的石油污染土壤进行微生物菌剂强化处理和环境强化实验.微生物菌剂强化结果表明:4种菌和除油效果最好的A、C、D混合菌3 d可将石油烃依次降解24%、19.81%、22.55%、26.46%、39.67%;并对该菌群的最佳投加配比进行确定,A、C、D菌群数量的最佳配比为NA∶NC∶ND=1∶2∶0.5,3 d内菌群A、C、D在最佳接种配比情况下可将石油烃降解44.2%.环境强化实验结果表明:A、C、D菌群在最佳修复条件营养物质C∶N∶P为75∶8∶3、表面活性剂为0.5%、通气条件为6层纱布、电子受体H2O2的加入量为1.5%下,3 d内石油烃降解61.46%,比自然条件下修复的除油率4.7%提高了56.76%,较只进行菌种强化时最高除油率44.2%提高了约17%.     

杀菌方式对即食调味克氏原螯虾尾贮藏品质的影响

【目的】分析电子束辐照(6 kGy)和高温杀菌(116℃、15 min)两种方式杀菌对即食调味克氏原螯虾尾贮藏过程中的品质影响。【方法】通过测定感官评价、pH、色度、总挥发性盐基氮(TVB-N)、硫代巴比妥酸反应物(TBARs)、羰基含量、挥发性成分等指标,分析两种杀菌方式对调味虾尾的品质变化的影响。【结果】辐照组虾肉亮度L^*值、红度a^*值显著高于其他两组(P <0.05),高温组的挥发性成分气味分布变化较小,至贮藏30 d时仍无明显异味。但辐照组在贮藏过程中硫化成分较高,高温组存在黑变现象,总色差值?E值显著较高高于其两组(P <0.05);辐照组和高温组TVB-N值和羰基含量增长速率均低于对照组,但TBARs值较高,其脂质氧化程度高。【结论】两种杀菌方式均可在一定程度上延缓克氏原螯虾尾贮藏中的感官劣变,辐照组蛋白质和脂质的氧化程度均显著低于高温组。综合考虑,辐照杀菌处理对即食克氏原螯虾尾贮藏过程中品质保持效果较佳。     

MASS BALANCES OF MAJOR CHEMICAL CONSTITUENTS IN BOHAI SEA WATER Ⅱ.REMOVAL PROCESSES

The removal processes of major seawater chemical constituents in the Bohai Sea include seawater ex-change between the Bohai Sea and the Northern Yellow Sea,cation exchange,interstitial water burialand spray,formation of illite,sepiolite,sulfate and sulfur,and carbonates.The mass balance calculationbased on these processes is very applicable to the Bohai Sea.     

Detection of the end point temperature of thermal denatured protein in fish and chicken meat through SDS-PAGE electrophoresis

Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was applied in the detection of the end point temperature (EPT) of thermal denatured protein in fish and meat in this study. It was also used in studying the thermal denatured temperature range of proteins in salmon and chicken meat. The results show that the temperature ranges of denatured proteins were from 65 ℃ to 75 ℃ , and these temperature ranges were influenced by the processing methods. Through SDS-PAGE, the features of repeated he...     

Diversity and bioactivity of actinomycetes from marine sediments of the Yellow Sea

Among the 116 actinomycetes collected from marine sediments of the Yellow Sea,56 grew slowly and appeared after 2?3 weeks of incubation.Among the 56 strains,only 3 required seawater(SW) for growth,and 21 grew well in the medium prepared with SW rather than distilled water(DW),while the remaining 32 grew well either with SW or with DW.Six representatives with different morphological characteristics,including 1 SW-requiring strain and 5 well-growing with SW strains,were selected for phy-logenetic analysis based on 16S rRNA gene.Two strains belong to Micrococcaceae and Nocardiopsaceae respectively.The other 4 strains belong to the family of Streptomycetaceae.In the analyzed 6 strains,one was related to Nocardiopsis spp.and the other three were related to Streptomyces spp.,representing new taxa.Bioactivity testing of fermentation products from 3 SW-requiring strains and 21 well-growing with SW strains revealed that 17 strains possessed remarkable activities against gram-positive pathogen or/and tumor cells,suggesting that they were prolific resources for natural drug discovery.     

Determination of Trace Copper (Ⅱ) in Water Samples by Kinetic-spectrophotometry

A new kinetic-spectrophotometric method is proposed for the determination of copper ( Ⅱ ). The method is based on the catalytic effect of copper ( Ⅱ ) on the oxidation of weak acid brilliant blue dye (RAWL) by hydrogen peroxide. The copper ( Ⅱ ) can be determined spectrophotometrically by measuring the decrease of absorbance of RAWL at λ = 626 nm using the fix-time method. The optimum reaction conditions are as follows: pH 7.20, buffer solution NaOH-KH2PO4, RAWL (200 mgL-1) 5.00 mL, H2O2 (30％) 0.50 mL, reaction temperature 80 ℃ and reaction time 20 min. The linear range of this method is between 0 μg L-1 and 12 μg L-1 and the limit of detection is 0.011 μg L-1, the relative standard deviation (RSD) in five replicate determinations for 2 and 8 μg L-1 copper ( Ⅱ ) are 3.2％ and 2.3％, respectively. Twenty ions do not interfere in the determination of copper ( Ⅱ ). The method has been applied satisfactorily to the determination of copper ( Ⅱ ) in freshwater samples (tap water and Yellow River water from Lijin, Shandong, China) and seawater samples (from the South China Sea), the recovery rates are 98.0％, 102.5％ and 96.0％, respectively.     

EFFECTS OF CO-CULTURE AND SALINITY ON THE GROWTH AND AGAR YIELD OF GRACILARIA TENUISTIPITATA VAR LIUI ZHANG ET XIA

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Corrosion behavior of anodic oxidized TiO<Subscript>2</Subscript> film in seawater

TiO₂ films were formed on metallic titanium substrates by the anodic oxidation method in H₂SO₄ solution under the 80V D.C.. Phase component and microstructure were characterized by X-ray diffraction (XRD) and scanning electron microscopy (SEM). Water contact angles on titanium oxide film surface were measured under both dark and sunlight illumination conditions. Corrosion tests were carried out in seawater under different illumination conditions by electrochemistry impedance spectrum (EIS) and polarization curves. The result showed that the TiO₂ film prepared by the anodic oxidation method was anatase with a uniform structure and without obvious pores or cracks on its surface. The average water contact angle of the film was 116.4° in dark, in contrast to an angle of 42.7° under the UV illumination for 2 hours, which demonstrates good hydrophobic property. The anti-corrosion behavior of the TiO₂ film was declining with the extended immersion time. Under dark conditions, however, the hydrophobic TiO₂ film retarded the water infiltrating into the substrate. The impedance changed slowly and the corrosion current density was 2 orders of magnitude lower than that with the film illuminated by sunlight. All of those mentioned above indicate that the TiO₂ film possesses much better performance under dark condition, and it can be applied as an engineering material under dark seawater environment.     

Determination of Trace Copper （Ⅱ） in Water Samples by Kinetic-spectrophotometry

A new kinetic-spectrophotometric method is proposed for the determination of copper （ Ⅱ ）. The method is based on the catalytic effect of copper （ Ⅱ ） on the oxidation of weak acid brilliant blue dye （RAWL） by hydrogen peroxide. The copper （ Ⅱ ） can be determined spectrophotometrically by measuring the decrease of absorbance of RAWL at λ = 626 nm using the fix-time method. The optimum reaction conditions are as follows： pH 7.20, buffer solution NaOH-KH2PO4, RAWL （200 mgL-1） 5.00 mL, H2O2 （30%） 0.50 mL, reaction temperature 80 ？？ and reaction time 20 min. The linear range of this method is between 0 μg L^-1 and 12 μg L^-1 and the limit of detection is 0.011 μg L-1, the relative standard deviation （RSD） in five replicate determinations for 2 and 8 μg L-1 copper （ Ⅱ ） are 3.2% and 2.3%, respectively. Twenty ions do not interfere in the determination of copper （ Ⅱ ）. The method has been applied satisfactorily to the determination of copper （ Ⅱ ） in freshwater samples （tap water and Yellow River water from Lijin, Shandong, China） and seawater samples （from the South China Sea）, the recovery rates are 98.0%, 102.5% and 96.0%, respectively.