栉孔扇贝血淋巴中ACP和AKP活性及其电镜细胞化学研究↑（*）

用电镜细胞化学和分光光度技术对栉孔扇贝血淋巴中的酸性磷酸酶（ＡＣＰ） 和碱性磷酸酶（ＡＫＰ） 的研究结果表明,溶酶体呈强的ＡＣＰ 阳性;ＡＫＰ 阳性颗粒常沿细胞膜分布;部分细胞ＡＣＰ 阳性颗粒较多,且靠近细胞膜,胞浆中也有分布。生化测定结果表明,血细胞和血清中均存在ＡＣＰ和ＡＫＰ 活性,且血清中的２ 种酶的酶活力均高于血细胞中,血清中ＡＣＰ和ＡＫＰ 的最适ｐＨ 分别为４ ．４ 和１０ ．３ ,最适温度分别为３５ ℃和５５ ℃。     

对虾皮下及造血组织坏死杆状病毒的精细结构、核酸、多肽及血清学研究

对纯化的对虾皮下及造血组织坏死杆状病毒（ＨＨＮＢＶ）进行了超微结构、紫外吸收、核酸类型及多肽ＳＤＳ－ＰＡＧＥ的研究。同时用单克隆抗体对ＨＨＮＢＶ的两分离株和ＭＢＶ进行了血清学比较研究。用ＴＭＶ作为内标定物测得病毒粒子大小为１５０ｎｍ×４５０ｎｍ，其衣壳为两个帽状物端夹１３圈螺旋对称的园柱体结构。病毒在２６０ｎｍ处吸光系数为０．１１７ｍｇ／ｍＬ／ＯＤ２６０。病毒核酸为双链ＤＮＡ，分子量大于３５×１０６ｄ。病毒囊膜有１２种主要多肽。病毒衣壳有两种主要多肽。从寿光（ＨＨＮＢＶ—９３７）和青岛（ＨＨＮＢＶ—９３８）分离到的病毒种在ＳＤＳ—ＰＡＧＥ和抗ＨＨＮＢＶ—９３７单抗ＥＬＩＳＡ反应上没有显著差异。ＨＨＮＢＶ与斑节对虾杆状病毒（ＭＢＶ）在抗原决定簇上存在差别。     

INGESTIONANDEXCRETIONOFSEVERALANTIBACTERIALDRUGSBYTHEBRINESHRIMP（ARTEMIAPARTHENOGENETICA）1．TOXICITYT

ＩＮＧＥＳＴＩＯＮＡＮＤＥＸＣＲＥＴＩＯＮＯＦＳＥＶＥＲＡＬＡＮＴＩＢＡＣＴＥＲＩＡＬＤＲＵＧＳＢＹＴＨＥＢＲＩＮＥＳＨＲＩＭＰ（ＡＲＴＥＭＩＡＰＡＲＴＨＥＮＯＧＥＮＥＴＩＣＡ）１．ＴＯＸＩＣＩＴＹＴＯＡＲＴＥＭＩＡ，ＩＮＧＥＳＴＩＯＮＡＮＤＥＸＣＲ...     

淡水无核珍珠荧光光谱的研究↑（*）

对三角帆蚌珍珠、贝壳珍珠层及商品珍珠粉等进行了固体表面荧光测定。荧光图谱显示：在２８０ｎｍ激发波长下,３种样品都具有３４５ｎｍ和４６５ｎｍ的特征荧光峰,且３４５ｎｍ处荧光强度最大,并伴有荧光淬灭现象。这说明珍珠及其贝壳中含有色氨酸（Ｔｒｐ）。用５ｍｍｏｌ／ＬＢａ（ＯＨ）２碱法水解处理从珍珠及其贝壳中提取的壳角蛋白,测得水解液的内源荧光光谱和其中Ｔｒｐ的含量为００４％～００７％。     

在微机上建立渔捞产量数据库

在微机上建立渔捞产量数据库孔兰香，仝龄，陈绵通，戴芳群（中国水产科学研究院黄海水产研究所，青岛２６６０７１）关键词数据库，渔捞产量，马面，监测，预报ＤＡＴＡＢＡＳＥＯＦＦＩＳＨＣＡＴＣＨＯＮＩＢＭ－ＰＣＣＯＭＰＵＴＥＲＫｏｎｇＬａｎｘｉａｎｇＴｏｎｇ...     

重金属离子对凡纳滨对虾的致病机理研究

研究了重金属离子Ｃｄ２＋、Ｐｂ２＋、Ｃｒ３＋对凡纳滨对虾血清ＳＯＤ、ＣＡＴ活性的影响。结果表明,低、中等浓度的Ｃｄ２＋、Ｐｂ２＋、Ｃｒ３＋均对凡纳滨对虾血清ＳＯＤ活性表现为较强的诱导作用。在一定的时间和较高浓度下,Ｃｄ２＋、Ｐｂ２＋、Ｃｒ３＋均对凡纳滨对虾血清ＳＯＤ、ＣＡＴ活性表现出显著抑制作用,具有一定的时间、剂量效应关系。在较低的Ｃｄ２＋、Ｐｂ２＋、Ｃｒ３＋浓度下,ＳＯＤ、ＣＡＴ活性明显高于对照,且随着各处理浓度的增加和时间的延长（７２ｈ）,Ｃｄ２＋、Ｐｂ２＋、Ｃｒ３＋对血清ＳＯＤ、ＣＡＴ活性表现出更加明显的抑制作用,即在低剂量、短时间内会导致酶活性提高;而在高剂量或长时间作用下,对虾ＳＯＤ、ＣＡＴ活性表现为抑制作用,且表现出２种金属离子对血清ＳＯＤ、ＣＡＴ活性的抑制作用比单一的金属离子抑制作用更加显著,显示出Ｃｄ２＋、Ｐｂ２＋、Ｃｒ３＋复合污染具有明显协同作用。金属离子的胁迫下使ＳＯＤ、ＣＡＴ的活性比例失调,使对虾的生理代谢紊乱,加速对虾的衰老和死亡。     

食盐对草鱼血清超氧化物歧化酶和溶菌酶活性的影响

草鱼养殖过程中经常使用食盐药浴,以便在高渗环境下杀死病原体。研究食盐药浴后草鱼代谢酶变化,有助于了解食盐药浴对草鱼机体生理免疫的影响。将草鱼（Ｃｔｅｎｐｈａｒｙｎｇｏｄｏｎｉｄｅｌｌａ）分别暴露于食盐浓度为０、２％、３％、４％水体中浸浴１０ｍｉｎ,然后置入水簇箱中饲养,分别在药浴后１２ｈ、２４ｈ、３６ｈ取样,测定其血清中超氧化物歧化酶（ＳＯＤ）和溶菌酶（ＬＳＺ）活性变化。结果显示,与对照组相比,血清ＳＯＤ活性在１２ｈ时,２％组中显著上升（Ｐ＜０．０５）,３％、４％组显著下降（Ｐ＜０．０５）;２４ｈ取样时,２％、３％组中无显著差异（Ｐ＞０．０５）,４％组显著下降（Ｐ＜０．０５）;３６ｈ取样时,各组间均无显著差异（Ｐ＞０．０５）;血清ＬＳＺ活性各组间均无显著差异（Ｐ＞０．０５）。食盐能引起草鱼ＳＯＤ活性变化,而对溶菌酶活性影响不大;表明食盐对草鱼不同代谢酶有不同的反应模式,抗氧化酶ＳＯＤ对食盐应激有敏感的生理应答。     

角叉菜凝集素的分离纯化及其性质

角叉菜（Ｃｈｏｎｄｒｕｓ ｏｃｅｌｌａｔｕｓ）经磷酸盐缓冲液抽提,２０％～７５％硫酸铵分级沉淀,牛甲状腺球蛋白－Ｓｅｐｈａｒｏｓｅ４Ｂ亲和层析,可以从红藻角叉菜中纯化出角叉菜凝集素（ＣＯＬ）,在ＰＡＧＥ上显示单一蛋白染色带,在等电聚焦电泳上显示单一蛋白染色带,其ＰＩ为８．３０,纯化后的ＣＯＬ的最大此外吸收峰在２８５ｎｍ,用ＳｅｐｈａｄｅｘＧ－２００分子筛层析测得其分子量为４２５２。该凝集素可以凝集     

盐度对杂交鳢幼鱼生长及血液免疫指标的影响

在３５ｄ的试验周期内,研究了不同盐度（２、４、６、８）对杂交鳢（斑鳢♀×乌鳢♂）幼鱼生长及血液红细胞数量（ＲＢＣ）、白细胞数量（ＷＢＣ）、溶菌酶（ＬＺＭ）含量、超氧化物歧化酶活性（ＳＯＤ）和过氧化氢酶（ＣＡＴ）活性等免疫因子的影响。结果显示,随着盐度的升高,杂交鳢增重率分别为６７．８１％、６８．２６％、７１．１４％、３２．９６％和１４．２６％,存活率分别为１００％、１００％、１００％、１００％和９３．３％。盐度２、４组增重率和对照组差异不显著,盐度６、８组增重率和对照组差异显著（Ｐ＜０．０５）。随着盐度的升高,血液红细胞数量呈下降的趋势,对照组和其他组差异显著（Ｐ＜０．０５）,而白细胞数量先急剧上升后又恢复到对照组水平,盐度６组达到最高。血液中溶菌酶浓度随着盐度变化出现波动,盐度６组达到最高,且与对照组差异显著（Ｐ＜０．０５）,盐度８组溶菌酶浓度有所回降,但仍显著高于对照组（Ｐ＜０．０５）。超氧化物歧化酶活性变化趋势与溶菌酶浓度变化趋势相似,各组超氧化物歧化酶活性均高于对照组且差异显著（Ｐ＜０．０５）。当盐度升至６时,超氧化物歧化酶活力升至最高。过氧化氢酶活性随着盐度的增加逐步升高,盐度８组达到最高,且各组与对照组差异显著（Ｐ＜０．０５）。     

用偶氮磺（Ⅲ）测定甘露醇中微量硫酸根的分光光度法

用偶氮磺（Ⅲ）测定甘露醇中微量硫酸根的分光光度法辛福言（中国水产科学研究院黄海水产研究所，青岛２６６０７１）关键词偶氮磺（Ⅲ），甘露醇，分光光度法ＤＥＴＥＲＭＩＮＡＴＩＯＮＯＦＴＲＡＣＥＡＭＯＵＮＴＳＳＵＬＦＡＴＥＩＯＮＩＮＭＡＮＮＩＴＯＬＵＭＢＹＩ...     

乙酰胆碱对栉孔扇贝血淋巴中过氧化物酶活性和一氧化氮含量的影响

栉孔扇贝的血淋巴在不同质量浓度的乙酰胆碱作用下于体外孵育3h,测定栉孔扇贝的血清、血细胞中依赖卤素的髓性过氧化物酶、不依赖卤素的髓性过氧化物酶和一氧化氮活性的活力变化。结果表明,乙酰胆碱对于这两种酶和一氧化氮均有一定的影响,髓性过氧化物酶和一氧化氮活性大部分均为先升后降。据此,乙酰胆碱对栉孔扇贝血淋巴中的几种氧化酶有一定的作用。     

泥蚶血细胞耐饥饿及抗菌力特性的研究

显微镜下观察泥蚶血细胞抗菌能力以及在不同饥饿条件下细胞形态结构的变化。结果表明:经饥饿处理后,泥蚶血细胞随着时间的迁移,出现细胞形态不规则、胞质色泽变浅、胞质颗粒物减少、胞膜边缘光滑度减小等现象。血细胞对大肠杆菌、奥斯陆莫拉氏菌、脲放线杆菌均具较高的噬菌率,噬菌实验过程中血细胞形态结构以及细胞组分不断产生变化。细胞抗菌活性检测结果表明:血细胞对三者都具有一定的抗菌活性,活性依次为:大肠杆菌>奥斯陆莫拉氏菌>脲放线杆菌。     

人工养殖栉孔扇贝和海湾扇贝血淋巴中部分免疫因子的比较研究

采用分光光度技术对人工养殖的栉孔扇贝和海湾扇贝血淋巴中部分免疫因子进行了比较研究。结果表明,海湾扇贝血清和血细胞中溶菌酶、非特异性酯酶(NSE)、过氧化氢酶(CAT)、髓性过氧化物酶(MPO)、一氧化氮(NO)、一氧化氮合酶(NOS)、活性氧和H2O2的活力或含量均高于栉孔扇贝,两种扇贝除血清中MPO活力差异不显著外,其他各组均差异极显著或显著。表明人工养殖海湾扇贝较栉孔扇贝的免疫力强。     

凡纳滨对虾白斑综合征血液病理研究

对自然发病、投喂和注射感染的凡纳滨对虾白斑综合征（white spot syndrome，WSS）血液病理进行研究，结果发现：不同感染方式患病对虾的血液病理变化相似，表现为：1．患病对虾血细胞总数、透明细胞数量极显著减少，小颗粒细胞、大颗粒细胞极显著增加。2．显微病理变化主要表现为血涂片中血细胞明显减少且分布不均匀，破损或解体的细胞增多，呈典型的溶血状态。3．超微病理变化表现为，大部分血细胞坏死，少数血细胞呈不典型的凋亡。患病对虾的血细胞核中可见大量白斑综合征病毒（white spot syndrome virus，WSSV）粒子。病理变化表明血细胞是WSSV的主要靶细胞。     

三疣梭子蟹血淋巴免疫功能的初步研究

利用API ZYM试剂盒和酶活力测试盒检测了三疣梭子蟹血细胞及血清中的19种酶类及血清中溶菌酶、碱性磷酸酶、酸性磷酸酶、超氧化物歧化酶和过氧化物酶的活性。试验结果表明,在雄蟹血清和血细胞中均检测到19种酶,在雌蟹血清和血细胞中分别检测到18种和16种。碱性磷酸酶和酸性磷酸酶的活性很低。在此基础上,研究了三疣梭子蟹血清和血细胞溶解物(HLS)对溶藻弧菌的抗菌活力,结果显示血清样品对溶藻弧菌的抗菌活力较强,但其体外作用的持续时间较短;而血细胞溶菌酶对溶藻弧菌的抗菌活力较弱。     

Identification of a mannose‐binding lectin‐like protein recognized by the anti‐CD25 antibody in haemocytes from Cherax quadricarinatus

The crustacean haemolymph contains three main cell populations; however, it is not clear which mechanisms participate in the regulation of cells related to innate immunity. This work aimed to identify potential interleukin‐like receptors that could regulate cellular responses in Cherax quadricarinatus. By histochemical analysis with murine anti‐CD25 staining (targeting the α‐chain of the IL‐2 receptor), we identified that this antibody recognizes cytoplasmic granules in semigranular and granular haemocytes. In haemocytes stimulated with phorbol myristate acetate (PMA), increased fluorescence was observed in these cytoplasmic granules, whereas staining with a human IL‐2 antibody after stimulation with 1–10 ng/ml PMA revealed no overexpression of the receptor or oxidative burst in haemocytes. Two‐dimensional Western blot analysis of haemocyte lysates showed that anti‐CD25 identified a 27.4‐kDa protein with an isoelectric point (pI) of 7.7 and a 46‐kDa protein with a pI of 6.9. De novo sequencing of these proteins identified that they had 32% homology with a mannose‐binding lectin (MBL) from Pacifastacus leniusculus. Our results indicate that a mannose‐binding lectin‐like protein could exert a protective effect that prevents damage from other activated immune responses.     

克氏原螯虾血细胞及免疫功能的初步研究

甲壳动物血细胞在宿主免疫应答中发挥着重要的作用,其中包括识别、吞噬、包囊黑色素形成、胞毒作用和胞间的信息传递。不同类型的血细胞其防御机制不同,因此血细胞的形态和分类是甲壳动物免疫学的基础。通常情况,十足目甲壳动物的血细胞类型主要依据细胞质中颗粒物质的有无和颗粒物质的相对大小进行分类。研究采用Giemsa染色法、H.E染色法和电子显微技术法对克氏原螯虾血细胞进行分类,观察到3类血细胞：透明细胞、小颗粒细胞、大颗粒细胞,它们所占的比例分别为19.9%,46.1%,34.1%。在电镜下,这3类血细胞的超微结构极为容易辨识。透明细胞通常在3类细胞中最小,核质（N/C）比最高,但无明显的颗粒;相比之下,小颗粒细胞和大颗粒细胞内部的颗粒数量逐渐增多。利用体内和体外培养技术研究克氏原螯虾血细胞对外源体（以Sephadex A-25色谱硅珠作为抗原）的包囊作用,初步探讨克氏原螯虾血细胞在对外源体免疫应答过程中的识别机理。将Sephadex A-25色谱硅珠植入克氏原螯虾体内,抗原物周围被大量血细胞包囊,通过切片技术观察到抗原物周围的血细胞绝大部分为小颗粒细胞,证实在包囊过程中起主要作用的血细胞是小颗粒细胞,即小颗粒细胞在包囊反应中比较活跃。研究结果为以后进一步研究克氏原螯虾的细胞免疫机制和提高其抗病能力提供了科学依据。     

Characterization of haemocytes of the Indian edible oyster, Crassostrea madrasensis (Preston)

The haemocytes of the Indian edible oyster Crassostrea madrasensis were characterized using light and electron microscopy. The light microscopic study was conducted by staining a monolayer of the haemocytes with Geimsa. Cells without granules and with a large nucleus occupying much of the cytoplasmic area were grouped as hyalinocytes. Those with lesser amounts of basophilic cytoplasmic granules were characterized as semigranulocytes and those with large amounts of a mixture of acidophilic and basophilic granules were termed as granulocytes. Ultrastructural studies also revealed the presence of three types of haemocytes. Scanning electron microscopic studies were used to study the spreading behaviour of the haemocytes. Cytochemical studies revealed the presence of acidphosphatase, peroxidase and prophenol oxidase in the cells.     

Functional characterization of Farfantepenaeus californiensis, Litopenaeus vannamei and L. stylirostris haemocyte separated using density gradient centrifugation

Haemocytes from penaeid shrimp (Farfantepenaeus californiensis, Litopenaeus vannamei and L. stylirostris) were separated using a discontinuous Percoll gradient. Shrimp haemocytes were spontaneously adhered to glass, allowing slide preparations for staining and microscopic differential counting. Like other crustaceans, shrimp has three main populations differing in presence and size of cytoplasmic granules and each population seems to be biochemical or functionally compromised. Prophenoloxidase (proPO )activity was mainly located in large granules haemocytes (75%) while the small granules cells participate with 25%, but seem to be responsible for encapsulation. Haemocyte discrimination ability was tested using Sephadex? (Seph?), DEAE‐Seph? and CM‐Seph?. Only DEAE‐Seph? was encapsulated by shrimp haemocytes and provoked the release of proPO activating system, indicating the role of particle charge in the activation of shrimp immune response.     

Comparison of immune responses and antioxidant status of different generations of growth‐selected Portunus trituberculatus families

The purpose of this study was to investigate the effects of inbreeding on the immune responses and antioxidant status of Portunus trituberculatus juveniles. Results showed that inbreeding affected the total haemocyte count, and phagocytic, pro‐phenoloxidase (propo), phenoloxidase and antibacterial activities decreased after the seventh generation. Antioxidant status showed a similar pattern: total antioxidant capacity, superoxide dismutase (SOD) activity and GSH/GSSG in the cell‐free haemolymph and hepatopancreas decreased, while catalase activity in the cell‐free haemolymph increased (P < 0.05). There were no significant differences in α₂‐macroglobulin and bacteriolytic activities in the cell‐free haemolymph and glutathione peroxidase activity in the cell‐free haemolymph and hepatopancreas among nine inbreeding generations. Gene expression levels of proPO and crustin in haemocytes and SOD in haemocytes and hepatopancreas also decreased significantly as the inbreeding generations increased. The results suggest that a high level of inbreeding could severely reduce the physiological health of P. trituberculatus. Our obtained data would be particularly useful for P. trituberculatus breeding programmes.