朱旭枫,邓秋霞,郭慧,李广丽,朱春华

• 1:广东海洋大学水产学院
• 2:广东省名特优鱼类生殖调控与繁育工程技术研究中心
• 3:广东省海水养殖生物育种工程实验室
• 4:广东省水产经济动物病原生物学及流行病学重点实验室

摘要(Abstract)：

【目的】研究水解单宁饲养的凡纳滨对虾(Litopenaeus vannamei)在副溶血弧菌(Vibrio parahaemolyticu)胁迫24 h后的生理响应,初步探究水解单宁对凡纳滨对虾血液和血细胞的影响。【方法】在基础饲料中分别添加质量分数为0%(对照组)、0.05%、0.10%、0.15%和0.20%的水解单宁(HTs),饲喂凡纳滨对虾60 d。对对虾注射0.2 mL的副溶血弧菌(2.5×10~8 CFU·mL~(-1)),感染24 h后,分析血细胞凋亡率、活性氧含量(ROS)、一氧化氮含量(NO)、酯酶活性、Toll样受体(TLR)和热休克蛋白70(HSP70)基因的表达水平;测定血清的碱性磷酸酶(AKP)、酸性磷酸酶(ACP)、超氧化物歧化酶(SOD),总抗氧化能力(T-AOC)和溶菌酶(LZM)活性。【结果】在感染副溶血弧菌24 h后,实验组对虾的血细胞凋亡率,ROS、NO水平显著低于对照组,酯酶活性显著高于对照组(P <0.05)。实验组凡纳滨对虾血清的AKP、ACP、SOD活性,T-AOC和LZM活性均显著高于对照组(P <0.05)。实验组对虾血细胞TLR和HSP70基因的表达水平显著上调(P <0.05)。【结论】饲料中添加水解单宁可提高凡纳滨对虾的非特异性免疫能力和抗氧化功能,阻止氧化损伤的发生,清除机体受到刺激产生的ROS和NO,增强酯酶活性,减少细胞凋亡,从而提高凡纳滨对虾抗副溶血弧菌感染的能力。

关键词(KeyWords)： 凡纳滨对虾;水解单宁;副溶血弧菌;非特异性免疫;抗氧化;血细胞

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(31600321);; 广东省现代农业产业技术体系创新团队建设专项资金(2019KJ149)

作者(Author): 朱旭枫,邓秋霞,郭慧,李广丽,朱春华

参考文献(References)：

• [1]吴丹.不同生长阶段凡纳滨对虾与罗氏沼虾营养成分的研究[D].上海:上海海洋大学, 2019.
• [2]廖静.见证凡纳滨对虾良种的诞生和崛起[J].海洋与渔业, 2019(12):70-71.
• [3] BONDAD-REANTASO M G, SUBASINGHE R P,ARTHUR J R, et al. Disease and health management in Asian aquaculture[J]. Veterinary Parasitology, 2005,132(3/4):249-272.
• [4] SAULNIER D, HAFFNER P, GOARANT C, et al.Experimental infection models for shrimp vibriosis studies:a review[J]. Aquaculture, 2000, 191(1/2/3):133-144.
• [5] NELAPATI S, NELAPATI K, CHINNAM B. Vibrio parahaemolyticus-An emerging foodborne pathogen[J].Veterinary World, 2012:48.
• [6] THEETHAKAEW C, NAKAMURA S, MOTOOKA D, et al. Plasmid dynamics in Vibrio parahaemolyticus strains related to shrimp Acute Hepatopancreatic Necrosis Syndrome(AHPNS)[J]. Infection, Genetics and Evolution,2017, 51:211-218.
• [7] LIGHTNER D V, REDMAN R M, PANTOJA C R, et al.Historic emergence, impact and current status of shrimp pathogens in the Americas[J]. Journal of Invertebrate Pathology, 2012, 110(2):174-183.
• [8] DAS A, NAKHRO K, CHOWDHURY S, et al. Effects of potential probiotic Bacillus amyloliquifaciens FPTB16 on systemic and cutaneous mucosal immune responses and disease resistance of catla(Catla catla)[J]. Fish and Shellfish Immunology, 2013, 35(5):1547-1553.
• [9]孙晓蛟,金海峰,闫研,等.不同抗生素替代品组合对断奶仔猪生长性能、腹泻及抗氧化能力的影响[J].中国畜牧兽医, 2020, 47(2):460-468.
• [10]刘汉锁,龙沈飞,尚庆辉,等.水解单宁酸的作用机制及其在畜禽生产中的应用进展[J].动物营养学报, 2019,31(9):3991-3999.
• [11] IJI P A, KHUMALO K, SLIPPERS S, et al. Intestinal function and body growth of broiler chickens on maize-based diets supplemented with Mimosa tannins and a microbial enzyme[J]. Journal of the Science of Food and Agriculture, 2004, 84(12):1451-1458.
• [12] EBADI M R, POURREZA J, JAMALIAN J, et al. Amino acid content and availability in low, medium and high tannin Sorghum grain for poultry[J]. International Journal of Poultry Science, 2004, 4(1):27-31.
• [13] MANSOORI B. Absorption capacity of chicken intestine for d-xylose in response to graded concentrations of tannic acid[J]. Animal Feed Science and Technology, 2009,151(1/2):167-171.
• [14] MANSOORI B, ROGIEWICZ A, SLOMINSKI B A. The effect of canola meal tannins on the intestinal absorption capacity of broilers using a D-xylose test[J]. Journal of Animal Physiology and Animal Nutrition, 2015, 99(6):1084-1093.
• [15] PRABU D L, CHANDRASEKAR S, AMBASHANKAR K, et al. Effect of dietary Syzygium cumini leaf powder on growth and non-specific immunity of Litopenaeus vannamei(Boone 1931)and defense against virulent strain of Vibrio parahaemolyticus[J]. Aquaculture, 2018, 489:9-20.
• [16] KIRAN G S, PRIYADHARSHINI S, SAJAYAN A, et al.Dietary administration of gelatinised polyhydroxybutyrate to Penaeus vannamei improved growth performance and enhanced immune response against Vibrio parahaemolyticus[J]. Aquaculture, 2020, 517:734773.
• [17] CHEN S J, ZHUANG Z X, YIN P, et al. Changes in growth performance, haematological parameters,hepatopancreas histopathology and antioxidant status of Pacific white shrimp(Litopenaeus vannamei)fed oxidized fish oil:Regulation by dietary myo-inositol[J]. Fish and Shellfish Immunology, 2019, 88:53-64.
• [18] LIU T, ZHANG G G, FENG Y, et al. Dietary soybean antigen impairs growth and health through stress-induced non-specific immune responses in Pacific white shrimp,Litopenaeus vannamei[J]. Fish and Shellfish Immunology,2019, 84:124-129.
• [19] LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method[J]. Methods, 2001, 25(4):402-408.
• [20] ZHU X F, GUO H, LI G L, et al. Effects of dietary hydrolyzable tannins on growth performance, antioxidant capacity, intestinal microflora and resistance against Vibrio parahaemolyticus of juvenile Pacific white shrimp,Litopenaeus vannamei(Boone, 1931)[J]. Aquaculture Reports, 2021, 19:100601.
• [21]刘泓宇,李正,孙武卫,等.饲料中添加益生菌或植物提取物对凡纳滨对虾生长、免疫力及抗病力的影响[J].饲料工业, 2014, 35(12):21-26.
• [22]袁香丽.低鱼粉饲料添加壳寡糖对凡纳滨对虾生长、免疫和抗副溶血弧菌感染能力的影响[D].厦门:集美大学, 2017.
• [23]马寨璞,张繁霜,佟霁坤.甲壳动物免疫系统概述及其图论表示[J].安徽农业科学, 2011, 39(1):291-294.
• [24]赵柳兰.组织胺对新糠虾和中华绒螯蟹的消化功能、免疫力以及卵巢发育的影响[D].上海:上海海洋大学,2012.
• [25]陈晓瑛,曹俊明,黄燕华,等.饲料中添加低聚木糖对凡纳滨对虾幼虾生长性能、非特异性免疫力、抗氧化功能及抗对虾白斑综合征病毒能力的影响[J].动物营养学报, 2014, 26(8):2397-2407.
• [26] YANG C J, ZHANG J Q, LI F H, et al. A Toll receptor from Chinese shrimp Fenneropenaeus chinensis is responsive to Vibrio anguillarum infection[J]. Fish and Shellfish Immunology, 2008, 24(5):564-574.
• [27]李晶晶,李云,刘红,等.感染溶藻弧菌及白斑综合症病毒后凡纳滨对虾不同组织的Toll样受体基因表达变化研究[J].海洋与湖沼, 2016, 47(2):476-483.
• [28]张红波,万亚涛,王莲哲,等.在细胞应激反应中HSP70的保护与调节作用[J].中国实用医药, 2009,4(14):228-229.
• [29] GULLO C A, TEOH G. Heat shock proteins:to present or not, that is the question[J]. Immunology Letters, 2004,94(1/2):1-10.
• [30] JOHANSSON M W, KEYSER P, SRITUNYALUCKSANA K, et al. Crustacean haemocytes and haematopoiesis[J].Aquaculture, 2000, 191(1/2/3):45-52.
• [31] LORENZON S, DE GUARRINI S, SMITH V J, et al.Effects of LPS injection on circulating haemocytes in crustaceansin vivo[J]. Fish and Shellfish Immunology,1999, 9(1):31-50.
• [32]段亚飞,张家松,董宏标,等.副溶血弧菌对斑节对虾非特异性免疫酶活性的影响[J].水产学报, 2014, 38(9):1557-1564.
• [33] GUO H, XIAN J N, LI B, et al. Gene expression of apoptosis-related genes, stress protein and antioxidant enzymes in hemocytes of white shrimp Litopenaeus vannamei under nitrite stress[J]. Comparative Biochemistry and Physiology Part C:Toxicology and Pharmacology, 2013, 157(4):366-371.
• [34]郭慧,冼健安,王安利.亚硝酸盐对凡纳滨对虾血细胞毒性及p53基因表达的影响[J].水生态学杂志, 2015,36(2):61-67.
• [35]姚翠鸾,王志勇,相建海.甲壳动物血细胞及其在免疫防御中的功能[J].动物学研究, 2006, 27(5):549-557.
• [36]汪蕾,张秀霞,冼健安.硫化物胁迫对凡纳滨对虾血细胞的毒性影响[J].水生态学杂志, 2017, 38(3):97-102.
• [37]李玲,吴方舟,梁金荣,等. Cd对凡纳滨对虾血细胞活性氧含量(ROS)、细胞凋亡率及应激反应基因表达的影响[J].中国水产科学, 2021, 28(2):167-175.
• [38]卢芷程,许銮佳,卢文宇,等.溶藻弧菌对凡纳滨对虾血细胞毒性及细胞凋亡和免疫相关基因的影响[J].南方农业学报, 2018, 49(12):2559-2565.
• [39]朱孟凯,姚翠鸾.温度胁迫对凡纳滨对虾肝胰腺氧代谢及能量代谢的影响[J].水产学报, 2015, 39(5):669-678.
• [40]刘文珍,邱德全.溶藻弧菌对凡纳滨对虾血清中一氧化氮及氧自由基的影响[J].广东海洋大学学报, 2007,27(3):60-63.
• [41] BROWN G C. Nitric oxide and neuronal death[J]. Nitric Oxide, 2010, 23(3):153-165.
• [42] SIMON H U, HAJ-YEHIA A, LEVI-SCHAFFER F. Role of reactive oxygen species(ROS)in apoptosis induction[J].Apoptosis, 2000, 5(5):415-418.
• [43] XIAN J N, ZHANG X X, GUO H, et al. Cellular responses of the tiger shrimp Penaeus monodon haemocytes after lipopolysaccharide injection[J]. Fish and Shellfish Immunology, 2016, 54:385-390.
• [44] ZVEREVA E, SEREBROV V, GLUPOV V, et al. Activity and heavy metal resistance of non-specific esterases in leaf beetle Chrysomela lapponica from polluted and unpolluted habitats[J]. Comparative Biochemistry and Physiology Part C:Toxicology and Pharmacology, 2003,135(4):383-391.
• [45] XIAN J N, LI B, GUO H, et al. Haemocyte apoptosis of the tiger shrimp Penaeus monodon exposed to cadmium[J].Bulletin of Environmental Contamination and Toxicology,2014, 92(5):525-528.
• [46]郭慧,朱晓闻,欧荣华,等. Cu2+胁迫对罗氏沼虾血细胞毒性及caspase-3、α-2M基因表达的影响[J].四川动物, 2017, 36(3):293-299.
• [47] ALEXANDER T C, HAN F X, ARSLAN Z, et al.Toxicity of As in Crassostrea virginica(Gmelin, 1791)from the Northern Gulf of Mexico at the presence of Zn and its antioxidant defense mechanisms[J]. Ecotoxicology and Environmental Safety, 2019, 172:514-522.
• [48]庞树坤.单宁酸对育肥猪的抗氧化能力和预防腹泻病的效果研究[J].今日畜牧兽医, 2015(7):31-33.
• [49] LIU H S, HU J X, MAHFUZ S, et al. Effects of hydrolysable tannins as zinc oxide substitutes on antioxidant status, immune function, intestinal morphology, and digestive enzyme activities in weaned piglets[J]. Animals, 2020, 10(5):757.
• [50] STAR?EVI?K, KRSTULOVI?L, BROZI?D, et al.Production performance, meat composition and oxidative susceptibility in broiler chicken fed with different phenolic compounds[J]. Journal of the Science of Food and Agriculture, 2015, 95(6):1172-1178.
• [51]陈赛娟,刘亚娟,郅永伟,等.单宁酸对生长獭兔毛皮品质、免疫器官发育及抗氧化能力的影响[J].动物营养学报, 2019, 31(11):5151-5156.
• [52]刘华伟.栗树单宁的抗氧化能力及其对肉兔生产性能的影响研究[D].北京:中国农业科学院, 2010.