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。     

5-羟色胺对栉孔扇贝血淋巴中髓过氧化物酶活性和一氧化氮含量的影响

用质量浓度为0.05、0.5、5μg/ml的5-羟色胺与栉孔扇贝血淋巴孵育,采用生化方法,测定栉孔扇贝血清和血细胞中髓过氧化物酶活性和一氧化氮含量。结果表明,3个5-羟色胺质量浓度下各试验组的血清中髓过氧化物酶活性均低于对照组,差异显著;各试验组的血细胞中髓过氧化物酶活性均低于对照组,差异显著。5-羟色胺质量浓度为0.05μg/ml时,各试验组的血清一氧化氮含量低于对照组,差异不显著;5-羟色胺质量浓度为0.5、5μg/ml时,各试验组的血清一氧化氮含量高于对照组,差异显著。3个5-羟色胺质量浓度下,各试验组的血细胞一氧化氮含量均低于对照组,差异显著。说明5-羟色胺对栉孔扇贝血淋巴中的髓过氧化物酶活性和一氧化氮含量起到了调节作用。5-羟色胺可能在栉孔扇贝的免疫调节中发挥重要作用。     

栉孔扇贝血淋巴中酚氧化酶和髓过氧物酶活性↑（*）

用电镜细胞化学和分光光度技术对栉孔扇贝血淋巴中的氧化酶（ＰＯ）和髓过氧物酶（ＭＰＯ）进行了研究。结果表明：细胞质中直径分别为３００～４３０ｎｍ和１３０～２００ｎｍ的大、小２种颗粒呈ＰＯ强阳性,次级溶酶体和小颗粒呈ＭＰＯ强阳性。生化血清中存在ＰＯ和ＭＰＯ活性,血细胞仅有ＭＰＯ活性。以ＴＭＢ为底物测得血清中不依赖卤素的ＭＰＯ活力为１０７．２Ｕ,以二乙醇胺为底物测得血清中依赖卤素的ＴＭＢ为底物测得血清中     

浒苔多糖对华贵栉孔扇贝血淋巴中SOD酶和溶菌酶活性的影响

于华贵栉孔扇贝体内注射了0.1%,0.25%,0.5%3个不同浓度的浒苔多糖后,在24,48,72 h分别测定了华贵栉孔扇贝血淋巴中SOD酶和溶菌酶活力。结果表明:0.1%和0.25%实验组扇贝血淋巴中SOD酶活仅在注射24h后显著地高于对照组(P<0.05),而0.5%实验组在注射24 h后极显著高于对照组(P<0.01),而且在注射48h和72 h后仍显著高于对照组(P<0.05);0.1%实验组对扇贝血淋巴中溶菌酶的作用无显著影响,而0.25%和0.5%实验组对扇贝血淋巴中溶菌酶活力在不同测定时间都表现出明显的促进作用,其中0.5%实验组对血细胞中溶菌酶作用效果极显著(P<0.01);说明浒苔多糖具有增强华贵栉孔扇贝免疫活性的作用。     

浒苔多糖对扇贝SOD酶和溶菌酶活力的影响

华贵栉孔扇贝体内注射了体积分数0．1％、0．25％的浒苔多糖后，在第24、48、72小时分别测定了扇贝血淋巴中SOD酶和溶菌酶活力。实验组扇贝血清中SOD酶活力在不同测定时间都显著地高于对照组，而血细胞中SOD酶活力在注射24h后和48h后显著地高于对照组；实验组扇贝血淋巴中溶菌酶在注射24h后和48h后显著高于对照组。说明浒苔多糖具有增强华贵栉孔扇贝免疫活性的作用。     

栉孔扇贝血清中的免疫因子的研究↑（*）

本文对栉孔扇贝血清的细菌凝集活性、抑菌活性和溶菌活性进行研究。选用２４种细菌和真菌进行凝集试验,发现栉孔扇贝血清能够对金黄色葡萄球菌和代远洋弧菌产生凝集作用,血清的细菌凝集作用经数种细菌刺激没有发生改变,说明这种凝集作用是不可诱导的。栉孔扇贝的血清对某些细菌的生长有一定的抑制作用,但在体外彻底杀灭细菌的效果极差,经大肠杆菌和副溶血弧菌刺激后,抑菌效果基本不变。栉孔扇贝血清还具有一定的溶菌作用,而且大肠杆菌的刺激可以明显提高这种溶菌作用。     

扇贝血细胞胞内酶活性的细胞学观察

分别对海湾扇贝、虾夷扇贝、栉孔扇贝的3种血细胞胞内酶：碱性磷酸酶(ALP)、过氧化物酶(POD)、酚氧化酶(PO)进行了免疫细胞化学研究，并对血细胞酶反应阳性率作了统计。结果表明，3种扇贝血细胞ALP阳性率相差不大，海湾扇贝为46．6％，虾夷扇贝为42．4％，栉孔扇贝为36．8％；3种扇贝血细胞POD阳性率相差较大，栉孔扇贝阳性率高达50．2％，海湾扇贝为44．2％，虾夷扇贝仅为17．2％。3种扇贝血细胞PO孵育反应后，由于海湾扇贝、虾夷扇贝PO反应活性很低，瑞氏染液复染后，无法区分阴性、阳性细胞。实验中仅对栉孔扇贝PO阳性率进行了统计，栉孔扇贝血细胞PO阳性率为39．3％。     

病毒感染对栉孔扇贝血淋巴中过氧化氢酶和过氧化物酶活力的影响

用从病贝体内分离的非包涵体球形病毒感染栉孔扇贝,采用分光光度技术,分别于6,24,72h,测定栉孔扇贝血清和血细胞中过氧化氢酶(CAT)活力和过氧化物酶(POD)活力。结果表明,感染病毒后,血清中CAT活力在6,24,72h时,实验组均高于对照组,6h时差异极显著,24h时差异显著,72h时差异不显著;血细胞中CAT活力在6,24,72h时,实验组均高于对照组,6h和24h时差异极显著,72h时差异不显著。血清和血细胞中POD活力在6h和24h时,实验组均高于对照组,且均差异极显著。说明病毒感染后栉孔扇贝血淋巴中CAT和POD的活力增强。     

栉孔扇贝血淋巴中酯酶活性及其性质

采用紫外分光光度法测定栉孔扇贝（Chlamysfarreri）血淋巴中酯酶的活性，并研究底物、抑制剂、激活剂、温度、pH对酯酶活力的影响及酶的热稳定性。结果表明，栉孔扇贝血清与血细胞中均存在酯酶，且血细胞中的酶活力高于血清。血清与血细胞中酯酶的最适作用底物有所不同，血清中为α-醋酸萘酯，而血细胞中为α-丁酸萘酯。当以α-醋酸萘酯为底物时，血清中酯酶的最适温度为45℃，最适pH值为7．5；以α-丁酸萘酯为底物时，血清中酯酶的最适温度为40℃，最适pH值为8．0。以α-醋酸萘酯及α-丁酸萘酯为底物分别测定酶的热稳定时，酶活力变化相似，在20℃时活力最高，而后缓慢下降。栉孔扇贝血淋巴中酯酶属中温型中性酶类，低温下较稳定，底物专一性不强，血清中的酯酶可能来自血细胞，该酶在贝类的免疫防御中町发挥重要的作用。     

华贵栉孔扇贝肉及其酶解产物对半去势雄性大鼠生殖能力的影响

为探究华贵栉孔扇贝(Chlamys nobilis)肉及其酶解产物对改善男性生殖能力的功效,该研究以半去势SD雄性大鼠为动物实验模型,考察华贵栉孔扇贝肉及其酶解产物对半去势雄性大鼠的性行为表现、脏器系数、血清性激素水平、阴茎组织一氧化氮(NO)和总一氧化氮合酶(NOS)活力等的影响。结果表明,华贵栉孔扇贝肉及其酶解产物可缩短半去势雄性大鼠的勃起潜伏期(P0.01),增加扑捉次数(P0.05)、骑乘次数(P0.05)等性行为表现;显著提升半去势雄性大鼠的睾丸、前列腺-精囊腺系数(P0.05),能显著提高半去势大鼠血清睾酮(T)含量,稳定黄体生成素(LH)和促卵泡激素(FSH)水平(P0.05);同时能显著介导NO和NOS的生成。综上,华贵栉孔扇贝肉及其酶解产物对半去势雄性大鼠的生殖能力有一定改善作用,其改善机制可能与调节半去势雄性大鼠血清性激素、促进阴茎信号因子的生成有关。     

Growth,survival and immune activity of scallops,Chlamys farreri Jones et Preston,compared between suspended and bottom culture in Haizhou Bay,China

We examined the growth, survival and immune response of the scallop, Chlamys farreri, during a 1‐year period in deep water of Haizhou Bay. Scallops were cultured using two methods: (1) in lantern nets at a 5 m depth and (2) in a bottom culture system (sleeves) on the seabed at about a 25 m depth. Shell heights, meat dry weight and immune activities in the haemolymph (superoxide dismutase and myeloperoxidase) were measured bimonthly or quarterly from July 2007 to June 2008. Survival was measured at the end of the study and environmental parameters in the experimental layers were monitored during the experiment. The growth and immune activities of scallops were lower when the water temperature was high, which was consistent with the main mortality occurring in summer. The growth and immunity of scallops were higher in the suspended culture than in the bottom culture during the experiment, with the exception of shell growth during the last study period. Survival of scallops in the suspended culture (54.6±12.3%) was significantly lower than that in the bottom culture (86.8±3.5%) at the end of this study. We conclude from our results that the high mortality of C. farreri can be prevented by culturing them in a bottom culture system before November of the first year, and then transferring them to a suspended culture to improve scallop production.     

Activation of intestinal epithelial cells in Atlantic cod,Gadus morhua,induced by algal derivatives

This study investigated the influences of two algal derivatives, alginic acid and fucoidan (FUC) on the activities of the intestinal epithelial cells (IEC) of Atlantic cod. The cells were isolated from three regions of the intestines, and primary cultures were incubated with the derivatives followed by temporal analysis of cellular activities. Epithelial cell survival in the fore and hind intestine was not significantly affected by the algal derivatives at both time points (3 and 24 h postincubation, hpi). Cellular survival in the mid‐intestine decreased significantly at 3 hpi. Stimulation of myeloperoxidase (MPO) activity of IEC induced by the algal derivatives was observed to be region dependent. Myeloperoxidase activity in the cells from fore‐ and mid‐intestines was not significantly affected by the two algal derivatives. Both algal derivatives significantly stimulated the activity at 3 hpi in the hind intestinal cells, and FUC‐exposed cells remained significantly elevated until 24 hpi. The results showed that algal derivatives could be applied as potential immunostimulants in cod with stimulatory action of MPO activity in the mucosal immune cells of the hind intestine.     

Dietary beta-1,3 glucan potentiates innate immunity and disease resistance of Asian catfish, Clarias batrachus (L.)

This study investigated the effects of short and prolonged administration of a yeast beta-glucan on non-specific immune parameters, growth rate and the disease resistance of Asian catfish, Clarias batrachus. Fish fed with a basal diet (control) and test diet (basal diet supplemented with 0.1% glucan) for 1, 2 and 3 weeks were assayed for superoxide production, serum myeloperoxidase (MPO) content, natural haemagglutinin level, complement and lysozyme activities. Fish were weighed at weekly intervals and specific growth rate (SGR, % increase in body weight per day) was determined. After each week, fish were challenged with Aeromonas hydrophila to measure the level of protection. Results showed that glucan administration at 0.1% in feed, significantly (P<0.05) enhanced MPO and lysozyme levels, superoxide production, haemagglutination titre and level of protection against A. hydrophila challenge, irrespective of length of exposure. The alternative complement activity and SGR were not affected by the dietary supplementation of yeast glucan. As glucan feeding at 0.1% for 1 week is able to enhance the non-specific immunity and disease resistance of catfish efficiently, short-term feeding might be used in farmed catfish diets to enhance disease resistance.     

The in vitro effects of divalent metal ions on the activities of immune‐related enzymes in coelomic fluid from the sea cucumber Apostichopus japonicus

The immune‐related enzymes in marine animals are very sensitive to divalent metal ions. To investigate the roles divalent metal ions play in the influence on the immunity of sea cucumber Apostichopus japonicus, one of the most important commercial species in Asian countries, the effects of eight divalent metal ions at concentrations of 2.5, 5, 10, 15, 20, 25 and 30 mmol L^?1 on the activities of superoxide dismutase (SOD), phenoloxidase (PO), acid phosphatase (ACP), alkaline phosphatase (AKP) and myeloperoxidase (MPO) in coelomic fluid were determined with the nitro blue tetrazolium chloride (NBT) method, dopachrome formation method, p‐nitrophenyl phosphate (pNPP) method and 3,3′,5,5′‐tetramethyl benzidine (TMB) method. The results indicated that Mg²⁺ enhanced the activities of SOD, PO, ACP and AKP significantly and showed no obvious effect on MPO activity; Zn²⁺ increased the activities of SOD, ACP and AKP, and showed no obvious effect on the activities of PO and MPO; Cu²⁺ enhanced the activities of ACP, AKP and MPO and activated SOD and PO at a certain concentration range; Ca²⁺ and Mn²⁺ inhibited the activities of ACP and AKP; Fe²⁺ had strong inhibitory effect on SOD activity; Pb²⁺ showed inhibitions on the activities of SOD, PO, ACP and AKP; and Cd²⁺ inhibited MPO activity greatly. The data obtained in this study collectively suggest that Mg²⁺, Zn²⁺ and Cu²⁺ have potential in promotion of A. japonicus immunity, while Ca²⁺, Fe²⁺, Pb²⁺, Cd²⁺ and Mn²⁺ might be limiting factors to the immune response of A. japonicus.     

提高扇贝制品安全水分含量的初步研究

取活体栉孔扇贝(壳高55—65mm)，剖取贝柱或汽蒸开壳后取贝肉，制成3类不同水分活度(aw)的试样，测定其制品水分含量和水分活度的关系以及贮藏试验中变质菌类型。结果表明，对于L—aw型制品(0．60＜aw＜0．70)，传统制品干贝的安全水分含量为17．3％以下，添加糖、盐等调味品后制品安全水分含量提高到20．3％，使用山梨酸钾作防霉剂进一步提高到24．1％。对于M—aw型制品(0．70＜aw＜0．90)，采取真空包装，制品安全水分含量可提高到38％。对于H—aw型制品(0．90＜aw＜0．95)，采取降低aw和pH、热加工、真空包装、二次低温加热杀菌的联合措施，制品安全水分含量可达50％。因此，采用适当的科学方法，扇贝制品的安全水分含量是可以提高的。     

温度波动对不同规格虾夷扇贝(Patinopecten yessoensis)生理和免疫指标的影响

为探讨黄海冷水团锋面温度波动对底播虾夷扇贝(Patinopecten yessoensis)的影向,采用室内模拟的方法,研究了温度波动对虾夷扇贝生理和免疫指标的影响.实验温度波动范围为15-10-15℃,升降温幅度为5℃/2 h,共进行了4次温度波动,分别测定了3个规格虾夷扇贝死亡率、耗氧率、排氨率以及血液中超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活力等生理、免疫指标的变化情况.结果显示,温度波动4次后,大、中、小3个规格虾夷扇贝的死亡率均较低,分别为4％、6％、6％,其中,大规格虾夷扇贝的死亡率低于中、小规格,并且大规格虾夷扇贝在前2次温度波动时出现死亡,第3次温度波动后,不再出现死亡.3个规格组在B1(波动1次)时的耗氧率与初始相比,均为降低;随着波动次数的增加,耗氧率逐渐增加而高于初始水平.除小规格组的B1外,排氨率均随温度波动次数的增加而降低;多重比较分析结果显示,大规格组的B3(波动3次)显著低于波动初始(P＜0.05);中规格组的B4(波动4次)显著低于波动初始(P＜0.05).虾夷扇贝的免疫指标对温度波动更为敏感:温度波动1次或者2次时,3个规格组的SOD和CAT活性显著降低(P＜0.05).上述结果表明,适宜温度范围内的温度波动,也会对虾夷扇贝的生理、免疫指标产生不同程度的影响.     

微胶囊剂型黄芪多糖对刺参生长性能、免疫力及抗病力的影响

在基础饲料中添加不同剂量(1%、3%、5%)的微胶囊剂型黄芪多糖(Astragalus polysac-charudes,APS)(载药量为17%),制成3种配合饲料,连续投喂初始体重为22.0±1.0g的刺参Apostichopus japonicus35d,探讨微胶囊剂型黄芪多糖(以下简称"微胶囊")对刺参生长性能、非特异性免疫及抗病力的影响。结果显示,35d后刺参最高特定生长率(Specific growth rate,SGR)为0.51%/d(5%添加组)(P<0.05);微胶囊添加量为3%时免疫效果最佳,刺参体腔液中酸性磷酸酶ACP、超氧化物歧化酶SOD、髓过氧化物酶MPO活性显著高于对照组(P<0.05)。试验期间每隔7d测定刺参体腔液样品,其中ACP活性在第28天最高,为11.2U/100ml(P<0.05);SOD活性在第21天最高,为63.3U/ml(P<0.05);MPO活性在第21天最高,为40MPO单位/ml(P>0.05);免疫试验结束后对刺参进行灿烂弧菌Vibrio splendidus攻毒实验,14d内观察得出,3%微胶囊添加组累积死亡率明显低于对照组(P<0.05)。由此可得,微胶囊剂型黄芪多糖对仿刺参生长性能、免疫力及抗病力方面均具有显著的增强效果,以3%添加量效果最佳,且添加量、作用时间、免疫效果之间存在相关性。     

铜污染对扇贝内脏团抗氧化酶活性的影响

在实验室条件下,将栉孔扇贝置于不同质量浓度的Cu2+溶液中10 d,测定其内脏团过氧化氢酶(CAT),谷胱甘肽过氧化物酶(GSH-PX)和超氧化物歧化酶(SOD)的活性.试验结果表明,随着Cu2+质量浓度的提高,CAT、SOD、GSH-PX的活性表现为"抑制-诱导-抑制"的规律,但整体仍是抑制的特性.扇贝内脏团组织CAT、GSH-PX、SOD对铜的早期污染均具指示作用,其中最为灵敏的指标是CAT,其次是SOD和GSH-PX .