耿小雪,周怡,徐玮,张文兵,麦康森

• 1:中国海洋大学水产学院水产动物营养与饲料农业部重点实验室海水养殖教育部重点实验室

摘要(Abstract)：

为研究饲料中添加毕赤酵母表达的翻译控制肿瘤蛋白fortilin对凡纳滨对虾免疫反应和抗白斑综合征病毒(WSSV)感染能力的影响。以酵母表达上清冻干粉和酵母培养物冻干粉2种形式在基础饲料中分别添加100mg/kg的重组fortilin蛋白和空质粒重组蛋白,制成4种实验饲料,以基础饲料作为空白对照,设计5个实验组:基础饲料组(C)、酵母表达上清冻干粉组(P-1)、酵母培养物冻干粉组(P-2)、fortilin酵母上清冻干粉组(F-1)和fortilin酵母培养冻干粉组(F-2)。用5种饲料分别饲喂初始体重(4.87±0.26)g的凡纳滨对虾20d,随后进行WSSV攻毒实验。养殖实验表明,F-1组和F-2组中凡纳滨对虾血淋巴的血细胞数(THC)、呼吸爆发活性(RB)和血清酚氧化酶活性(PO)显著高于C组(P<0.05)。F-2组的THC、RB、PO和超氧化物歧化酶(SOD)活性均显著高于P-2组(P<0.05)。F-1组的PO和一氧化氮合酶(NOS)活性显著高于P-1组(P<0.05)。攻毒实验显示,饲料中添加fortilin蛋白显著降低了凡纳滨对虾的死亡率(P<0.05):F-1和F-2组的累计死亡率分别为19.2%和15.4%,P-1和P-2组的累计死亡率分别为56.0%和47.8%,C组累计死亡率为84.6%。研究结果表明,凡纳滨对虾摄食添加重组fortilin蛋白的饲料后,对虾的免疫力和抗WSSV感染的能力显著增强。

关键词(KeyWords)： 凡纳滨对虾;fortilin蛋白;白斑综合征病毒;免疫;饲料

Abstract:

Keywords:

基金项目(Foundation): 国家公益性行业(农业)专项经费项目(201103034)资助~~

作者(Author): 耿小雪,周怡,徐玮,张文兵,麦康森

DOI: 10.16441/j.cnki.hdxb.20150269

参考文献(References)：

• [1]农业部渔业局.中国渔业统计年鉴[M].北京:中国农业出版社,2014:28-30.Fisheries Service of Ministry of Agriculture.China Fishery Statistical Yearbook[M].Beijing:China Agriculture Press,2014:28-30.
• [2]Escobedo-Bonilla C M.Application of RNA interference(RNAi)against viral infections in shrimp:a review[J].Antivirals&Antiretrovirals,2011,9:2.
• [3]Yenofsky R,Cereghini S,Krowczynska A,et al.Regulation of mRNA utilization in mouse erythroleukemia cells induced to differentiate by exposure to dimethyl sulfoxide[J].Molecular and Cellular Biology,1983,3(7):1197-1203.
• [4]Bohm H,Benndorf R,Gaestel M,et al.The growth-related protein P23of the Ehrlich ascites tumor:translational control,cloning and primary structure[J].Biochemistry International,1989,19(2):277-286.
• [5]Xu A,Bellamy A R,Taylor J A.Expression of translationally controlled tumor protein is regulated by calcium at both the transcriptional and post transcriptional level[J].Biochemistry Journal,1999,342(3):683-689.
• [6]Gachet Y,Tournier S,Lee M,et al.The growth-related,translationally controlled protein P23has properties of a tubulin binding protein an associates transiently with microtubules during the cell cycle[J].Journal of Cell Science,1999,112:1257-1271.
• [7]Bommer U A,Thiele B J.The translationally controlled tumor protein(TCTP)[J].International Journal of Biochemistry and Cell Biology,2004,36(3):379-385.
• [8]Bangrak P,Graidist P,Chotigeat W,et al.Molecular cloning and expression of a mammalian homologue of a translationally controlled tumor protein(TCTP)gene fromPenaeus monodonshrimp[J].Journal of Biotechnology,2004,108(3):219-226.
• [9]Chen D,He N,Lei K,et al.Genomic organization of the translationally controlled tumor protein(TCTP)gene from shrimp Marsupenaeus japonicas[J].Molecular Biology Reports,2009,36(5):1135-1140.
• [10]Wang S,Zhao X F,Wang J X.Molecular cloning and characterization of the translationally controlled tumor protein from Fenneropenaeus chinensis[J].Molecular Biology Reports,2009,36(7):1683-1693.
• [11]Nupan B,Phongdara A,Saengsakda M,et al.Shrimp Pm-fortilin inhibits the expression of early and late genes of white spot syndrome virus(WSSV)in an insect cell model[J].Developmental&Comparative Immunology,2011,35(4):469-475.
• [12]Tonganunt M,Nupan B,Saengsakda M,et al.The role of Pmfortilin in protecting shrimp from white spot syndrome virus(WSSV)infection[J].Fish&Shellfish Immunology,2008,25(5):633-637.
• [13]Musthaq S K S,Kwang J.Evolution of specific immunity in shrimp-A vaccination perspective against white spot syndrome virus[J].Developmental&Comparative Immunology,2014,48(2):342-353.
• [14]Pathan M,Gireesh-Babu P,Pavan-Kumar A,et al.In vivo therapeutic efficacy of recombinant Penaeus monodon antiviral protein(rPmAV)administered in three different forms to WSSV infected Penaeus monodon[J].Aquaculture,2013,376-379:64-67.
• [15]周怡,张文兵,麦康森.凡纳滨对虾翻译控制肿瘤蛋白Fortilin的毕赤酵母表达及其对血细胞免疫反应的影响[J].中国海洋大学学报(自然科学版),2012,42(1-2):54-58.Zhou Y,Zhang W B,Mai K S.Secretion expression of fortilin in yeast Pichia pastoris and its effects on immune responses of hemocytes in white shrimp Litopenaeus vannamei[J].Journal of Ocean University of China,2012,42(1-2):54-58.
• [16]Bai N,Zhang W B,Mai K S,et al.Effects of discontinuous administration ofβ-glucan and glycyrrhizin on the growth and immunity of white shrimp Litopenaeus vannamei[J].Aquaculture,2010,306(1-4):218-224.
• [17]Xie X X,Li H Y,Xu L M,et al.A simple and efficient method for purification of intact white spot syndrome virus(WSSV)viral particles[J].Virus Research,2005,108(1):63-67.
• [18]Vargas-Albores F,Guzmán M A,Ochoa J L.An anticoagulant solution for haemolymph collection and prophenoloxidase studies of penaeid shrimp(Penaeus californiensis)[J].Comparative Biochemistry and Physiology Part A:Physiology,1993,106(2):299-303.
• [19]Hernández-López J,Gollas-Galván T,Vargas-Albores F.Activation of the prophenoloxidase system of the brown shrimp(Penaeus californiensis)Holmes[J].Comparative Biochemistry and Physiology Part C:Pharmacology,Toxicology and Endocrinology,1996,113(1):61-66.
• [20]Song Y L,Hsieh Y T.Immunostimplation of tiger shrimp(Penaeus monodon)hemocytes for generation of microbicidal substances:analysis of reactive oxygen species[J].Developmental&Comparative Immunology,1994,18(3):201-209.
• [21]姜令绪,潘鲁青,肖国强.氨氮对凡纳对虾免疫指标的影响[J].中国水产科学,2004,11(6):537-541.Jiang L X,Pan L Q,Xiao G Q.Effects of ammonia-N on immune parameters of white shrimp Litopenaeus vannamei[J].Journal of Fishery Sciences of China,2004,11(6):537-541.
• [22]汪小锋,樊廷俊,丛日山,等.几种免疫促进剂对中国对虾血细胞数量、形态结构以及酚氧化酶产量和活性的影响[J].水产学报,2005,29(1):66-73.Wang X F,Fan T J,Cong R S,et al.Effects of several immunostimulants on the number,ultrastructure of hemocytes and the yield,activities of phenoloxidase in Penaeus chinensis[J].Journal of Fisheries of China,2005,29(1):66-73.
• [23]Burgents J E,Burnett K G,Burnett L E.Disease resistance of Pacific white shrimp,Litopenaeus vannamei,following the dietary administration of a yeast culture food supplement[J].Aquaculture,2004,231(1-4):1-8.
• [24]Wang Y C,Chang P S,Chen H Y.Tissue distribution of prophenoloxidase transcript in the Pacific white shrimp(Litopenaeus vannamei)[J].Fish&Shellfish Immunology,2006,20(3):414-418.
• [25]Dziarski R.Peptidoglycan recognition proteins(PGRPs)[J].Molecular Immunology,2004,40(12):877-886.
• [26]Amparyup P,Charoensapsri W,Tassanakajon A.Two prophenoloxidases are important for the survival of Vibrio harveyi challenged shrimp Penaeus monodon[J].Developmental&Comparative Immunology,2009,33(2):247-256.
• [27]Ai H S,Huang Y C,Li S D,et al.Characterization of a prophenoloxidase from hemocytes of the shrimp Litopenaeus vannamei that is down-regulated by white spot syndrome virus[J].Fish&Shellfish Immunology,2008,25(1-2):28-39.
• [28]Murphy R,DeCoursey T E.Charge compensation during the phagocyte respiratory burst[J].Biochimica et Biophysica Acta(BBA)-Bioenergetics,2006,1757(8):996-1011.
• [29]Parihar M S,Javeri T,Hemnani T,et al.Responses of superoxide dismutase,glutathione peroxidase and reduced glutathione antioxidant defenses in gills of the freshwater catfish(Heteropneustes fossilis)to short-term elevated temperature[J].Journal of Thermal Biology,1997,22(2):151-156.
• [30]Fridovich I.Superoxide dismutases.An adaptation to a paramagnetic gas[J].Biology Chemistry,1989,264(14):7761-7764.
• [31]Jiang G,Yu R,Zhou M.Modulatory effects of ammonia-N on the immune system of Penaeus japonicas to virulence of white spot syndrome virus[J].Aquaculture,2004,241(1):61-75.
• [32]Sinthujaroen P,Tonganunt-Srithaworn M,Eurwilaichitr L,et al.Protection of Litopenaeus vannamei against the white spot syndrome virus using recombinant Pm-fortilin expressed in Pichia pastoris[J].Aquaculture,2015,435:450-457.

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