研究论文 RESEARCH PAPER

发酵豆粕替代鱼粉时添加不同水平晶体精氨酸对凡纳滨对虾生长及抗应激能力的免疫调控

  • 刘韬 ,
  • 潘韵超 ,
  • 夏燕 ,
  • 杨培江 ,
  • 华雪铭
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  • 1. 上海海洋大学, 农业农村部鱼类营养与环境生态研究中心, 上海 201306;
    2. 上海海洋大学, 农业农村部 淡水水产种质资源重点实验室, 上海 201306;
    3. 上海海洋大学, 水产科学国家级实验教学示范中心, 上海 201306;
    4. 九江大北农水产科技有限公司, 九江 332000;
    5. 上海市奉贤区水产技术推广站, 上海 201499;
    6. 上海市奉贤区奉城镇农业农村服务中心, 上海 201411
刘韬(1993-),男,江西九江人,硕士研究生,从事水产动物营养学研究。E-mail:1071222421@qq.com

收稿日期: 2022-01-08

  网络出版日期: 2022-10-17

基金资助

上海市科技兴农项目[沪农科创字(2019)第3-5号]; 上海市科委高校能力建设项目(14320502000)

Immune Regulation of Different Levels of Crystalline Arginine on Growth and Anti-Stress Ability of Litopenaeus vannamei When Fish Meal Replaced by Fermented Soybean Meal

  • LIU Tao ,
  • PAN Yunchao ,
  • XIA Yan ,
  • YANG Peijiang ,
  • HUA Xueming
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  • 1. Centre for Research on Environmental Ecology and Fish Nutrition of the Ministry of Agriculture, Shanghai Ocean University, Shanghai 201306, China;
    2. Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture, Shanghai Ocean University, Shanghai 201306, China;
    3. National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China;
    4. Jiujiang Dabei Nong Fishery Science and Technology Co., Ltd., Jiujiang 332000, China;
    5. Shanghai Fengxian District Aquatic Technology Promotion Station, Shanghai 201499, China;
    6. Shanghai Fengxian District Fengcheng Town Agricultural and Rural Service Center, Shanghai 201411, China

Received date: 2022-01-08

  Online published: 2022-10-17

摘要

发酵豆粕作为鱼粉替代物已广泛应用于水产动物饲料中,但在替代时会出现氨基酸平衡性的问题,而精氨酸作为一种必需氨基酸,在水产动物的新陈代谢中发挥重要作用。为了探讨发酵豆粕替代鱼粉时精氨酸的重要性,本试验以凡纳滨对虾[初重(0.62±0.10) g]为研究对象,在以鱼粉(22.53%)和发酵豆粕(10.00%)为主要蛋白质源、精氨酸水平为1.98%的基础饲料中,分别添加0(对照, A00组)、0.2%(A02组)、0.4%(A04组)、0.6%(A06组)、0.8%(A08组)的晶体精氨酸。养殖60 d后,评估对虾生长性能、体成分、血清和肝胰腺生化指标、鳃和肝胰腺免疫相关基因表达水平,并进行耐低氧和抗弧菌试验。结果显示: 1)各组间对虾成活率无显著差异(P>0.05), A02组和A08组增重率和特定生长率显著高于除A06组外的其他组(P < 0.05)。2)各组间血清丙二醛(MDA)含量无显著差异(P>0.05),血清谷草转氨酶(AST)、谷丙转氨酶(ALT)活性在A02组最高, A08组血清AST活性显著低于其他组(P < 0.05),血清ALT活性显著高于除A00组外的其他组(P < 0.05);肝胰腺MDA含量在A02组最低。3)耐低氧能力在A02组最强,其次为A04组,且A04组抗弧菌能力最强。4) A04组鳃中的免疫缺陷(IMD)、溶菌酶(LZM)、肝胰腺髓样分化因子88(MyD88)基因相对表达量皆为最低,显著低于A00组(P < 0.05), A00组肝胰腺转录激活因子4(ATF4)基因相对表达量最低且显著低于另外2组(P < 0.05)。综合生长、肝胰腺健康、抗应激能力和基因表达水平,在鱼粉用量为22.5%、发酵豆粕用量为10%时,添加0.4%的晶体L-精氨酸,即饲料精氨酸水平为2.37%能在转录层面上有效调控凡纳滨对虾的免疫系统,增强机体的抗应激能力,保护肝胰腺健康且不影响对虾的生长性能。

本文引用格式

刘韬 , 潘韵超 , 夏燕 , 杨培江 , 华雪铭 . 发酵豆粕替代鱼粉时添加不同水平晶体精氨酸对凡纳滨对虾生长及抗应激能力的免疫调控[J]. 动物营养学报, 2022 , 34(10) : 6643 -6653 . DOI: 10.3969/j.issn.1006-267x.2022.10.057

Abstract

Fermented soybean meal as a substitute for fish meal has been widely used in aqua-feed, but there is usually poor amino acid balance when substitution occurs. Arginine as one of the essential amino acids, it plays an important role in metabolism of aquatic organisms. To investigate the importance of substituting arginine with fermented soybean meal for fish meal, this experiment took Litopenaeus vannamei [initial weight (0.62±0.10) g] as the research object, and supplemented with 0 (control, A00 group), 0.2% (A02 group), 0.4% (A04 group), 0.6% (A06 group), 0.8% (A08 group) of crystalline arginine in a basal diet, in which fish meal (22.53%) and fermented soybean meal (10.00%) were the main protein sources and the arginine level was 1.98%. After 60 days of feeding, the growth performance, body composition, serum and hepatopancreatic biochemical indexes, related gene expression in gill and hepatopancreas were determined, and hypoxic tolerance test and vibrio resistance test were carried out. The results showed as follows: 1) there was no significant difference in the survival rate of shrimp among groups (P>0.05), the weight gain rate and the specific growth rate in the A02 group and A08 group were significantly higher than those in the other groups (expect for A06 group) (P < 0.05). 2) No significant difference on serum malondialdehyde (MDA) content was found among groups (P>0.05), but the activities of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were the highest in the A02 group, and the activity of serum AST was significantly lower than that in other groups (P < 0.05), and serum ALT activity was significantly higher than that in other groups except A00 group (P < 0.05). The MDA content in hepatopancreas was the lowest in the A02 group. 3) The resistance to hypoxia was the strongest in A02 group, followed by A04 group with the strongest resistance to vibrio. 4) The relative expression levels of immunodeficiency (IMD), lysozyme (LZM) genes in gill and myeloid differentiation factor 88 (MyD88) gene in hepatopancreas in A04 group were the lowest and significantly lower than those in A00 group (P < 0.05), and the relative expression level of transcriptional activator factor 4 (ATF4) gene in hepatopancreas in A00 group were the lowest and significantly lower than that in the other two groups (P < 0.05). To consider the growth, hepatopancreas health, stress resistance and gene expression levels, it was concluded that 0.4% of additive crystalline L-arginine (total 2.37% arginine) in the diet including 22.5% fish meal and 10% fermented soybean meal can effectively modulate immune system at transcriptional level and enhance the anti-stress ability, protect the hepatopancreas health and keep normal growth of shrimp.

参考文献

[1] SHIMENO S,KUMON M,ANDON H.The growth performance and body composition of young yellowtail fed with diets containing defatted soybean for a long period[J].Nippon Suisan Gakkaishi,1993,59(5):821-825.  
[2] 周歧存,麦康森,刘永坚,等.动植物蛋白源替代鱼粉研究进展[J].水产学报,2005,29(3):404-410. ZHOU Q C,MAI K S,LIU Y J,et al.Advances in animal and plant protein sources in place of fish meal[J].Journal of Fisheries of China,2005,29(3):404-410.(in Chinese)
[3] WANG P H,LIANG J P,GU Z H,et al.Molecular cloning,characterization and expression analysis of two novel Tolls (LvToll2 and LvToll3) and three putative Spätzle-like Toll ligands (LvSpz1-3) from Litopenaeus vannamei[J].Developmental and Comparative Immunology,2012,36(2):359-371.  
[4] WEST N E,QIAN H,GUZIK T J,et al.Nitric oxide synthase (nNOS) gene transfer modifies venous bypass graft remodeling:effects on vascular smooth muscle cell differentiation and superoxide production[J].Circulation,2001,104(13):1526-1532.  
[5] CHEN G F,LIU Y,JIANG J,et al.Effect of dietary arginine on the immune response and gene expression in head kidney and spleen following infection of Jian carp with Aeromonas hydrophila[J].Fish&Shellfish Immunology,2015,44(1):195-202.  
[6] COSTAS B,CONCEIÇÃO L E C,DIAS J,et al.Dietary arginine and repeated handling increase disease resistance and modulate innate immune mechanisms of Senegalese sole (Solea senegalensis Kaup,1858)[J].Fish&Shellfish Immunology,2011,31(6):838-847.  
[7] COSTAS B,RÂGO P C N P,CONCEIÇÃO L E C,et al.Dietary arginine supplementation decreases plasma cortisol levels and modulates immune mechanisms in chronically stressed turbot (Scophthalmus maximus)[J].Aquaculture Nutrition,2013,19(S1):25-38.
[8] JIANG J,SHI D,ZHOU X Q,et al.In vitro and in vivo protective effect of arginine against lipopolysaccharide induced inflammatory response in the intestine of juvenile Jian carp (Cyprinus carpio var. Jian)[J].Fish&Shellfish Immunology,2015,42(2):457-464.  
[9] LEMAITRE B,HOFFMANN J.The host defense of Drosophila melanogaster[J].Annual Review of Immunology,2007,25:697-743.
[10] 陈义方.不同规格凡纳滨对虾对蛋白质和蛋氨酸需要量研究[D].硕士学位论文.上海:上海海洋大学,2012. CHEN Y F.Dietary protein and methionine requirements of three sizes of Litopenaeus vannamei[D].Master's Thesis.Shanghai:Shanghai Ocean University,2012.(in Chinese)
[11] 郑昌区.两种规格凡纳滨对虾对蛋白质和脂肪需要量的研究[D].硕士学位论文.湛江:广东海洋大学,2012. ZHENG C Q.Study on the requirements of the dietary protein and lipid in two sizes of pacific white shrimp[D].Master's Thesis.Zhenjiang:Guangdong Ocean University,2012.(in Chinese)
[12] ZHANG S P,LI J F,WU X C,et al.Effects of different dietary lipid level on the growth,survival and immune-relating genes expression in Pacific white shrimp,Litopenaeus vannamei[J].Fish&Shellfish Immunology,2013,34(5):1131-1138.  
[13] 胡毅,谭北平,麦康森,等.不同碳水化合物水平饲料对凡纳滨对虾生长及部分生理生化指标的影响[J].水生生物学报,2009,33(2):289-295. HU Y,TAN B P,MAI K S,et al.Influence of dietary carbohydrate levels on growth and some physiological-biochemical index in juvenile Litopenaeus vannamei[J].Acta Hydrobiologica Sinica,2009,33(2):289-295.(in Chinese)
[14] 夏明宏.凡纳滨对虾幼虾对生物素、烟酸、叶酸和胆碱需要量的研究[D].硕士学位论文.宁波:宁波大学,2014. XIA M H.Study on the requirements of biotin,niacin,folic acid and choline for juvenile Pacific white shrimp,Litopenaeus vannamei[D].Master's Thesis.Ningbo:Ningbo University,2014.(in Chinese)
[15] WILLIAMS K,BARLOW C,RODGERS L.Efficacy of crystalline and protein-bound amino acids for amino acid enrichment of diets for barramundi/Asian seabass (Lates calcarifer Bloch)[J].Aquaculture Research,2002,32(S1):415-429.
[16] 郭旭,戴习林,华雪铭,等.发酵豆粕部分替代鱼粉后凡纳滨对虾对赖氨酸的需求量[J].水产学报,2021,45(6):910-919. GUO X,DAI X L,HUA X M,et al.Optimum lysine requirement of juvenile Pacific white shrimp,Litopenaeus vannamei when fermented soybean meal partially replacing fish meal in the feed[J].Journal of Fisheries of China,2021,45(6):910-919.(in Chinese)
[17] 曾雯娉.凡纳滨对虾幼虾对赖氨酸、蛋氨酸、精氨酸和苯丙氨酸需要量的研究[D].硕士学位论文.湛江:广东海洋大学,2012. ZENG W P.Study on the requirements of lysine,methionine,arginine and phenylalanine for juvenile pacific white shrimp,Litopenaeus vannamei[D].Master's Thesis.Zhenjiang:Guangdong Ocean University,2012.(in Chinese)
[18] 黄凯,王武,李春华.南美白对虾必需氨基酸的需要量[J].水产学报,2003,27(5):456-461. HUANG K,WANG W,LI C H.Requirement of essential amino acids for Penaeus vannamei[J].Journal of Fisheries of China,2003,27(5):456-461.(in Chinese)
[19] 刘韬,黄旭雄,苏美英,等.发酵豆粕替代鱼粉对凡纳滨对虾生长、免疫相关酶及免疫相关基因表达的影响[J].水产学报,2018,42(9):1417-1427. LIU T,HUANG X X,SU M Y,et al.Effects of fermented soybean meal replacing fish meal on the growth performance,immune-related enzymes and gene expression of Litopenaeus vannamei[J].Journal of Fisheries of China,2018,42(9):1417-1427.(in Chinese)
[20] AOAC.Official methods of analysis[M].14th ed.Arlington:AOAC,1984.
[21] SUNYER J O,TORT L.Natural hemolytic and bactericidal activities of sea bream Sparus aurata serum are effected by the alternative complement pathway[J].Veterinary Immunology and Immunopathology,1995,45(3/4):333-345.
[22] 闫伟.L-精氨酸对免疫应激鹅生长性能、器官发育及血液生化指标的影响[D].硕士学位论文.扬州:扬州大学,2010. YAN W.Effect of L-arginine on growth performance,organ development and blood biochemical measurements of Yangzhou geese after immunological stress[D].Master's Thesis.Yangzhou:Yangzhou University,2010.(in Chinese)
[23] LIEBOLDT M A,HALLE I,FRAHM J,et al.Effects of graded dietary L-arginine supply on organ growth in four genetically diverse layer lines during rearing period[J].The Journal of Poultry Science,2016,53(2):136-148.  
[24] KHAN M A,ABIDI S F.Dietary arginine requirement of Heteropneustes fossilis fry (Bloch) based on growth,nutrient retention and haematological parameters[J].Aquaculture Nutrition,2011,17(4):418-428.  
[25] 陈娇娇.精氨酸对草鱼幼鱼生长、肠道结构调控及机制研究[D].硕士学位论文.武汉:华中农业大学,2017. CHEN J J.Studies on the regulation and the related mechanism of arginine on growth and intestinal structure of juvenile grass carp (Ctenopharyngodon idellus)[D].Master's Thesis.Wuhan:Huazhong Agricultural University,2017.(in Chinese)
[26] GAO Y J,YANG H J,GUO D Q,et al.Dietary arginine requirement of juvenile grass carp Ctenopharyngodon idella(Valenciennes in Cuvier&Valenciennes,1844)[J].Aquaculture Research,2015,46(12):3070-3078.  
[27] CHEN G F,FENG L,KUANG S Y,et al.Effect of dietary arginine on growth,intestinal enzyme activities and gene expression in muscle,hepatopancreas and intestine of juvenile Jian carp (Cyprinus carpio var.Jian)[J].The British Journal of Nutrition,2012,108(2):195-207.  
[28] LIANG H L,REN M C,HABTE-TSION H M,et al.Dietary arginine affects growth performance,plasma amino acid contents and gene expressions of the TOR signaling pathway in juvenile blunt snout bream,Megalobrama amblycephala[J].Aquaculture,2016,461:1-8.
[29] KAUSHIK S J,FAUCONNEAU B,TERRIER L,et al.Arginine requirement and status assessed by different biochemical indices in rainbow trout (Salmo gairdneri R.)[J].Aquaculture,1988,70(1/2):75-95.
[30] 万军利,麦康森,艾庆辉.鱼类精氨酸营养生理研究进展[J].中国水产科学,2006,13(4):679-685. WAN J L,MAI K S,AI Q H.The recent advance on arginine nutritional physiology in fish[J].Journal of Fishery Sciences of China,2006,13(4):679-685.(in Chinese)
[31] 袁超.精氨酸对蛋鸡采食及组织蛋白质代谢调控的机理研究[D].博士学位论文.杭州:浙江大学,2016. YUAN C.Research on the mechanism of l-arginine on the regulation of feed intake and tissue protein metabolism in laying hens[D].Ph.D.Thesis.Hangzhou:Zhejiang University,2016.(in Chinese)
[32] 管晓娟.甲壳动物体液免疫相关酶及免疫因子研究概况[J].生命科学仪器,2009,7(6):3-7. GUAN X J.Progress on the researches of humoral immunity of crustacean[J].Life Science Instruments,2009,7(6):3-7.(in Chinese)
[33] YUE Y R,ZOU Z Y,ZHU J L,et al.Effects of dietary arginine on growth performance,feed utilization,haematological parameters and non-specific immune responses of juvenile Nile tilapia (Oreochromis niloticus L.)[J].Aquaculture Research,2015,46(8):1801-1809.  
[34] 朱春华,李广丽,吴天力,等.L-精氨酸对凡纳滨对虾体液免疫因子的影响[J].海洋科学,2009,33(2):55-59. ZHU C H,LI G L,WU T L,et al.Effects of L-arginine on the humoral immuning factors of Litopenaeus vannamei[J].Marine Sciences,2009,33(2):55-59.(in Chinese)
[35] 冀培丰.一氧化氮合酶及其相关因子在凡纳滨对虾免疫反应中特性的研究[D].硕士学位论文.厦门:集美大学,2010. JI P F.The roles and characterizations of NOS and related immune factors in Litopenaeus vannamei immune response[D].Master's Thesis.Xiamen:Jimei University,2010.(in Chinese)
[36] ZAPOL W M,RIMAR S,GILLIS N,et al.Nitric oxide and the lung[J].American Journal of Respiratory and Critical Care Medicine,1994,149(5):1375-1380.  
[37] HOLEN E,ESPE M,ANDERSEN S M,et al.A co culture approach show that polyamine turnover is affected during inflammation in Atlantic salmon immune and liver cells and that arginine and LPS exerts opposite effects on p38MAPK signaling[J].Fish&Shellfish Immunology,2014,37(2):286-298.  
[38] LI F H,XIANG J H.Signaling pathways regulating innate immune responses in shrimp[J].Fish&Shellfish Immunology,2013,34(4):973-980.  
[39] WANG P H,GU Z H,HUANG X D,et al.An immune deficiency homolog from the white shrimp,Litopenaeus vannamei,activates antimicrobial peptide genes[J].Molecular Immunology,2009,46(8/9):1897-1904.
[40] KILBERG M S,SHAN J X,SU N.ATF4-dependent transcription mediates signaling of amino acid limitation[J].Trends in Endocrinology and Metabolism,2009,20(9):436-443.  
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