鱼类补体系统（Complement system of fish）的特点

本刊讯：鱼类补体系统由35种以上蛋白质裂解酶、酶抑制因子和受体构成。浙江大学动物科学学院张静等人研究报道，与哺乳动物相比，硬骨鱼类补体系统具有以下特点：     

热应激对鱼类生命活动影响的研究进展

本文综述了热应激对鱼类生命活动的影响,包括代谢与生长、抗氧化能力、热休克蛋白以及免疫相关研究成果。鱼受到热应激时,机体会产生对自身有害的活性氧,在一定的温度范围内,鱼类通过酶联通路和非酶联通路来消除体内多余的活性氧。大多水鱼类在适宜温度内,生长率与水温成正相关,超过适宜温度,鱼类代谢紊乱,生长缓慢甚至停止,长时间的热应激会导致鱼类死亡。研究鱼类的耐高温性,对创造更高的水产养殖经济效益和选育耐高温品种具有重要的理论价值和现实意义。     

异育银鲫血清蛋白质图谱的构建及生物信息学分析

异育银鲫是国内淡水养殖的优良品种之一，近年来，其病害问题日趋严重，极大影响了养殖效益。鱼类在感染病害后，可引起血清中某些蛋白质的表达水平发生变化，因此通过筛选某些特异蛋白质，将其作为生物标记物，有助于鱼类病害的识别及防治。利用蓝绿温和胶电泳结合色谱-质谱联用技术对异育银鲫血清蛋白质进行分离和鉴定，共鉴定成功721个血清蛋白质。生物信息学分析表明，这些蛋白质主要参与补体和凝血级联、代谢途径以及细菌感染途径等。本试验鉴定出多种非特异性免疫相关蛋白质，其中包含C3蛋白和α2-巨球蛋白，基于强度的绝对蛋白质定量分析表明，不同种类的补体C3蛋白和α2-巨球蛋白具有不同的表达量。部分潜在的血清标记物如血清转铁蛋白、肌酸激酶、α2-烯醇化酶以及补体因子Hf等在血清中均被成功鉴定。本试验结果可为异育银鲫血清蛋白质功能研究、蛋白质相互作用以及血清标记物筛选提供新的研究手段和方法，对于鱼类的疾病预防工作具有重要意义。     

中华倒刺鲃血清凝集素对细菌凝集作用的观察报告

随着水产养殖业的繁荣,水污染加剧,养殖鱼类频频发生各种疾病,因此研究鱼类抗菌相关的特别是非特异性免疫因子,已成为世界研究热点。目前研究的鱼类抗菌相关因子主要包括:凝集素、抗菌肽、干扰素、补体、白细胞介素、溶菌酶、转铁蛋白、鱼精蛋白、金属硫蛋白、免疫球蛋白等等。     

鱼类的非特异性免疫概述

对于鱼类免疫的研究很早就出现,鱼类是特异性免疫和非特异性免疫并存的脊椎动物。但与哺乳类相比,鱼类的特异免疫系统尚不够发达,特异性免疫机制还不完善。因此,在鱼类对外界刺激及病原生物侵袭的防御反应中,非特异性免疫起着重要作用。现已知参与鱼类非特异性免疫反应的因子主要包括巨噬细胞、粒细胞和细胞毒性细胞等一些具有吞噬作用的细胞,以及介导一系列免疫或应激反应的蛋白分子,如补体、细胞因子、趋化因子及溶菌酶和抗菌肽等。     

中草药对水产动物免疫作用研究进展

中草药具有免疫调节作用的主要成分包括生物碱、多糖、皂甙、蒽类、挥发油、有机酸等，中药免疫词节剂对机体的作用，主要通过激活网状内皮系统和补体、激活巨噬细胞和T、B淋巴细胞以及诱生多种细胞因子等途径。综述了中草药对鱼类、虾蟹的免疫作用。     

鱼类利用脂肪节约蛋白的研究进展

通常,鱼类由饲料中摄入的部分蛋白是作为能量来源而被消耗掉的。如果饲料中脂肪含量适当增加,则可以减少蛋白的比例,从而降低饲料成本,这一研究已经成为国际水产养殖领域的热点。国外学者对不同的鱼类进行了大量的尝试性研究,结果表明多种鱼具有利用脂肪节约蛋白的能力。脂肪节约蛋白对鱼类的生长摄食状态、机体的营养组成、免疫功能等具有明显的影响。介绍了脂肪和蛋白之间的转化及能量关系,综述了脂肪节约蛋白对鱼体的各种影响,并对该研究方向存在的问题和发展前景提出了一些建议。     

鱼类免疫增强剂的种类及研究进展(一)

<正>现有的研究结果已经证明,能激活鱼、虾免疫系统的免疫增强剂有很多种。根据其来源,大致可以分为来自于细菌的肽聚糖和细菌脂多糖(LPS);放线菌的短肽;酵母菌和海藻的β-1,β-葡聚糖、β-1,6-葡聚糖、来自甲壳动物外壳的甲壳质、壳多糖等。将上述各种免疫增强剂投喂于鱼类时,可以提高溶菌酶和补体的活性,增加机体中补体的C3成分;不仅可以增强巨噬细胞和嗜中性细胞的吞噬活性,还可以提高这些细胞的杀菌     

爱德华氏菌灭活疫苗对黄颡鱼免疫相关基因表达及过氧化氢酶、超氧化歧化酶和补体C3活性的影响

为评价爱德华氏菌灭活疫苗对黄颡鱼的免疫效果,分析了灭活爱德华氏菌疫苗对黄颡鱼非特异性免疫应答相关的免疫基因表达和酶活性的影响。结果显示,黄颡鱼鳃组织中抗菌肽基因和肝组织中长型肽聚糖识别蛋白基因在免疫7、14、21、28d后的表达均显著上调,肠道中抗菌肽基因和脾脏组织中长型肽聚糖识别蛋白基因的表达在免疫7、14、21d亦显著上升。血清中过氧化氢酶、超氧化歧化酶和补体C3活性出现不同程度的提高,其中,黄颡鱼过氧化氢酶活力在免疫14、21d后显著高于对照组,超氧化歧化酶活力在免疫14、21、28d后显著高于对照组,在21d达到峰值,补体C3在免疫7、14、21、28d后均显著上升(P0.05),在免疫7d后血清中补体C3的含量达到高峰。试验结果表明,爱德华氏菌灭活疫苗可显著提高黄颡鱼免疫基因抗菌肽和长型肽聚糖识别蛋白的表达以及酶的活性,进一步揭示爱德华氏菌灭活疫苗可提高黄颡鱼的非特异性免疫能力。     

鲤鱼单卵微量蛋白电泳的研究

本文利用微量蛋白电泳方法，通过对3个鲤鱼不同养殖品种单个卵粒全蛋白和酯酶同工酶的电泳研究，分析了鲤鱼单卵酯酶同工酶特点。实验结果表明，不同鲤鱼品种酯酶同工酯有明显差异，这种差异不仅表现在酯酶酶带的有无，也表现在酯酶酶带相对活性的高低。另外，本文还探讨了微量蛋白电泳方法和显色技术对于研究鱼类胚胎发育和鱼类育种学的意义及应用价值。     

仿刺参补体B因子溶血活性初步研究

本文应用高等哺乳动物血清补体B因子溶血检测技术检测仿刺参个体体内补体B因子。通过溶血法检测了健康仿刺参以及人工诱导化皮仿刺参体腔液补体B因子活性。结果显示仿刺参体腔液有补体B因子存在，且含补体B因子仿刺参体腔液溶血活性明显高于去补体B因子体腔液的溶血活性。实验对仿刺参体腔液进行加热处理结果显示其补体B因子是对热不稳定的分子，此特性与高等哺乳动物B因子特征相同。仿刺参体内存在补体B因子，它与脊椎动物补体B因子性质相似，在免疫旁路途径中起重要作用。     

Investigation of humoral immune factors from selected groups of southern bluefin tuna, Thunnus maccoyii (Castelnau): implications for aquaculture

Serum immunoglobulin, lysozyme and classical and alternative complement activity were analysed in different groups of wild and captive southern bluefin tuna (SBT), Thunnus maccoyii (Castelnau), from ambient water temperatures of 12 ± 1 and 20 ± 1 °C. Groups held captive for the longest time were found to have the highest levels of these humoral immune mediators, despite a drop in ambient temperature from 20 ± 1 to 12 ± 1 °C during the captivity period. Therefore, it may be that the immune response in these endothermic fish is not inhibited by low temperature to the extent seen in poikilothermic fish. Also, length of time in captivity appears to be associated with increased antigen exposure to maintain high levels of humoral immune mediators in these groups. Lysozyme activity was optimal at pH 5.8 and 6.2, suggesting that two isoforms, with different pH optima, are present. The SBT serum was found to lyse sheep erythrocytes by both classical and alternative complement pathways. Classical pathway activity occurred in the absence of prior sensitization with antiserum to sheep red blood cells, suggesting that natural antibodies may be present (or lectin or C-reactive protein mediated activation). Complement activity was relatively resistant to freezing at −20 °C but heating at between 45 and 50 °C for 20 min destroyed all complement activity.     

Opsonization by fish antibody and complement in the immune phagocytosis by peritoneal exudate cells isolated from salmonid fishes

Abstract. Opsonic activities of fish antibody and complement in the phagocytic responses of fish leucocytes have been investigated using salmonid peritoneal exudate cells (PEC) and viable cells of Aeromonas salmonicida as antigenic particles. PEC were isolated from liquid paraffin-injected coho salmon, masu salmon and rainbow trout. After incubation at 20°C for 2 h, phagocytosis by PEC was evaluated. Phagocytic activity was attributed to neutrophil-like and macrophage-like cells, though in the absence of immune serum activity was low. The addition of fish antibody (heal-inactivated antiserum) slightly increased the activity. On the addition of normal serum as a complement source phagocytic activity was dramatically accelerated. This acceleration was interpreted as opsonization as a result of the classical pathway activation of fish complement by an antigen-antibody reaction. The complement of mammals (goat, guinea pig and rabbit) did not possess opsonic activity in the presence of fish antibody.     

Effects of dietary blend of fish oil with corn oil on growth and non-specific immune responses of grouper, Epinephelus malabaricus

A growth trial was conducted to investigate the effects of fish oil and corn oil on the growth and non‐specific immune responses of juvenile grouper, Epinephelus malabaricus. Five semi‐purified diets were supplemented with 40 g kg⁻¹ of either fish oil (F4), corn oil (C4) or blend of fish oil with corn oil at ratio of 3 : 1 (F3C1); 1 : 1 (F2C2) and 1 : 3 (F1C3). Each diet was fed to triplicate groups of grouper (mean initial weight: 10.26 ± 0.14 g) in a recirculating rearing system for 8 weeks. Weight gain and feed efficiency of fish fed the F4, F3C1 and F2C2 diets were the highest (P < 0.05), followed by fish fed the F1C3 diet, and the lowest in fish fed the C4 diet. Fish fed the C4 diet had a lower survival rate than fish on other dietary treatments. Fatty acid composition of liver and muscle in fish generally reflected the composition of the diet. Leukocyte superoxide anion (O⁻₂) production ratio was the highest in fish fed the F3C1 and F2C2 diets, followed by fish fed the F4 and F1C3 diets, and the lowest in fish fed the C4 diet. Fish fed the F3C1 and F2C2 diets had higher plasma lysozyme activities than fish fed the F4 and C4 diets. Plasma alternative complement activity was higher in fish fed the F3C1, F2C2 and F1C3 diets than fish fed the F4 and C4 diets. These results suggest that grouper fed diets with 3 : 1 or 1 : 1 of fish oil to corn oil ratio had similar growth to the fish fed diet with fish oil. Blend of fish oil with corn oil in diet significantly enhanced non‐specific immune responses of grouper when the fed diet contained fish oil as the only lipid source.     

PKA/ATGL signaling pathway is involved in ER stress-mediated lipolysis in adipocytes of grass carp (Ctenopharyngodon idella)

Fish Physiology and Biochemistry - The relationship between endoplasmic reticulum stress (ER stress) and lipolysis in mammals has been widely studied, but it is relatively scarce in fish. The...     

Glomerulonephritis and immunosuppression associated with dietary essential fatty acid deficiency in gilthead sea bream, Sparus aurata L., juveniles

An experiment was conducted to determine the effect of an essential fatty acid (EFA) deficient diet on growth, immune status and renal morphology of juvenile gilthead sea bream, using two diets: a control diet containing 2% (DW) of n-3 high unsaturated fatty acid (n-3 HUFA) and a diet formulated to be deficient in EFA for this species and containing 0.5% DW of n-3 HUFA (diet NFA). After 9 weeks of feeding the EFA-deficient diet fish showed a reduction in growth compared with the control group (107.48 +/- 9.14 and 123.14 +/-11.87 g final weight respectively). Fish fed the NFA diet showed a reduction in the erythrocyte volume together with increased erythrocyte fragility, haemoglobin content and red blood cell count in comparison with fish fed the control diet. EFA deficiency also reduced cellular immunity in terms of neutrophil activity and the number of circulating lymphocytes. The serum alternative complement pathway was markedly reduced in fish fed the EFA deficient diet. Fish fed the control diet showed glomeruli with a well-defined Bowman's space and normal renal tubes. However, up to 88% of fish fed the EFA-deficient diet showed alterations in renal morphology affecting at least 50% of the glomeruli, which showed extreme dilation of capillaries and occlusion of Bowsman's capsule. Mesangial proliferation and diffuse thickening of the capillary walls, as well as renal tube degeneration, were also observed.     

Brook charr, Salvelinus fontinalis (Mitchill), and cryptobiosis: a potential salmonid reservoir host for Cryptobia salmositica Katz, 1951

Abstract. Laboratory-raised Cryptobia -susceptible brook charr, Salvelinus fontinalis (Mitchill), and rainbow trout, Oncorhynchus mykiss (Walbaum), were vaccinated intraperitoneally with a live Cryptobia salmositica vaccine (250000 parasites per fish), and 4 weeks later were challenged with the pathogen (250000 parasites per fish). Unvaccinated and infected brook charr had high parasitaemias but no clinical signs of disease, while unvaccinated and infected rainbow trout had anaemia and general oedema. Vaccinated and challenged fish had very low parasitaemias compared to unvaccinated and infected brook charr and rainbow trout. Complement fixing antibodies were detected in vaccinated and challenged fish 2 weeks after challenge. Unvaccinated and infected brook charr had consistently higher litres of complement fixing antibody than unvaccinated and infected rainbow trout. Parasitaemias were lower in all fish in which titres of complement fixing antibody were high. In a second experiment, brook charr inoculated intraperitoneally or intramuscularly with 100000 C. salmositica per fish had high parasitaemias but no anaemia or other clinical signs. The results show that susceptible brook charr do not suffer from cryptobiosis and may serve as reservoir hosts for C. salmositica in areas where the disease is prevalent. Vaccination to reduce the parasitaemia when fish become infected may be a control strategy in these areas.     

Cryptobia (Trypanoplasma) salmositica and salmonid cryptobiosis

Salmonid cryptobiosis is caused by Cryptobia (Trypanoplasma) salmositica. The haemoflagellate has been reported from all species of Pacific Oncorhynchus spp. on the west coast of North America. It is normally transmitted by the freshwater leech, Piscicola salmositica, in streams and rivers, and sculpins, Cottus spp., are considered important reservoir hosts. The pathogen can also survive on the body surface of fish because it has a contractile vacuole to osmoregulate when the fish is in fresh water. This allows for direct transmission between fish, especially in aquaculture facilities. The parasite divides rapidly by binary fission in the blood to cause disease, the severity of which is directly related to parasitaemia. Cryptobia salmositica has a mitochondrium and it normally undergoes aerobic respiration; however, if its mitochondrium is damaged it will switch to glycolysis. Its glycolytic enzymes and catalase are contained in glycosomes. Cysteine protease is a metabolic enzyme, and its neutralization inhibits oxygen consumption and multiplication of the parasite. An important virulent factor in cryptobiosis is a secretory metalloprotease. The protective mechanism involves production of complement fixing antibodies, phagocytosis by macrophages, and cell-mediated cytotoxicity. Recovered fish are protected, probably for life as the immunity is non-sterile. Clinical signs of the disease include anaemia, anorexia, splenomegaly, general oedema and abdominal distension with ascites. The metabolism and swimming performance of infected fish are significantly reduced and the bioenergetic cost of the disease is very considerable. Fish are susceptible to hypoxia and their immune system is depressed during acute cryptobiosis. Severity of the disease and mortality rates vary significantly between species and stocks of salmon. Protective strategies include selective breeding of Cryptobia-resistant fish. This is innate resistance to infection and it is controlled by a dominant Mendelian locus. In these fish the parasite is lysed via the alternative pathway of complement activation. In Cryptobia-tolerant fish (infected with the pathogen but which do not suffer from disease) the metalloprotease secreted by the parasite is neutralized by alpha2 macroglobulin. Hence, the production of a transgenic Cryptobia-tolerant salmon is an option. This strategy has the advantage in that human intervention (e.g. vaccination, chemotherapy) is not required once the transgenic fish is produced. Acquired immunity is another option; a single dose of the attenuated live vaccine protects fish for at least 2 years. The protective mechanism in vaccinated fish is similar to that in recovered fish. The trypanocidal drug, isometamidium chloride, is an effective therapeutic and prophylactic agent. It accumulates in the mitochondrium of the parasite and significantly disrupts aerobic respiration by causing lesions in the organelle. Efficacy of the drug is significantly increased after its conjugation to antibodies. This immuno-chemotherapeutic strategy has the advantage in that it will lower the drug dosage and hence side-effects of chemotherapy. It will probably reduce the accumulation of the drug in fish, an important consideration in food fish.     

Increasing Fish Oil Levels in Commercial Diets Influences Hematological and Immunological Responses of Channel Catfish,Ictalurus punctatus

Growth performance, immune responses and disease resistance were studied in juvenile channel catfish, Ictalurus punctatus, fed a commercial diet (35.3% crude protein and 5.6% lipid) supplemented with menhaden fish oil at levels of 0, 3, 6, and 9% for 15 wk. Dietary fish oil levels did not significantly influence growth performance of catfish. Fatty acid compositions of whole‐body and liver reflected dietary fatty acid composition. No differences were found in hematological values, except that fish fed the 9% fish oil diet had significantly lower hematocrit. The resistance of erythrocytes to hemolysis in hypotonic solutions increased with increasing fish oil levels and the highest resistance was seen with the 9% fish oil diet. Fish fed 6 and 9% added fish oil diets had significantly higher serum protein levels than that of control fish. Serum lysozyme activity of fish fed 3 and 6% added fish oil diet was significantly higher than that of the control. Complement activity and chemotaxis ratio significantly decreased in fish fed diets with 6 or 9% added fish oil. The 3% added fish oil diet, however, had significantly highest natural hemolytic complement activity. Mortality from Edwardsiella ictaluri 14 d postchallenge and antibody titers to E. ictaluri did not differ among treatments.