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为评估3种不同添加物对凡纳滨对虾(Litopenaeus vannamei)生长性能、非特异免疫反应及抗病力的影响，在基础饲料中分别添加了丁酸梭菌CBG01(Clostridium butyricum CBG01)活菌(CB组)、3%聚β-羟基丁酸酯(PHB组)和1%丁酸钠(BS组)来投喂对虾，对照组投喂基础饲料，养殖6周后测量对虾生长情况、检测血清非特异免疫指标和肝胰腺免疫相关基因表达水平，进行副溶血弧菌(Vibrio parahaemolyticus)攻毒实验。研究表明，CB组和PHB组对虾末体质量和特定生长率显著高于对照组(P<0.05),CB组最高，BS组与对照组差异不显著(P>0.05)。3个处理组对虾成活率和饲料效率均显著高于对照组(P<0.05)。3个处理组对虾血清中溶菌酶、过氧化物酶、超氧化物歧化酶活性以及总抗氧化能力显著高于对照组(P<0.05),CB组对虾血清中碱性磷酸酶、酸性磷酸酶、酚氧化酶活性显著高于对照组(P<0.05),PHB组对虾血清中碱性磷酸酶、酸性磷酸酶、总一氧化氮合酶活性显著高于对照组(P<0.05),BS组对虾血清中酸性磷酸酶活性显著高于对照组(P<0.05)。与对照组相比，CB组对虾肝胰腺中所有免疫相关基因(SOD、LZM、proPO、LGBP、HSP70、Imd、Toll、Relish、TOR、4E-BP、eIF4E1α、eIF4E2)相对表达量均显著上调(P<0.05),而PHB组的Toll基因以及BS组的Imd、Toll、Relish、eIF4E2基因相对表达量与对照组无显著差异(P>0.05)。副溶血弧菌攻毒实验表明，3个处理组对虾的累积死亡率均显著低于对照组(P<0.05)。研究结果表明，饲料中添加丁酸梭菌、PHB和丁酸钠均可不同程度地提高凡纳滨对虾的生长性能和非特异免疫能力，能够显著提高对虾对副溶血弧菌感染的抵抗力。总体比较，添加丁酸梭菌和PHB的作用效果基本相当，在一定程度上要优于添加丁酸钠。在饲料生产加工过程中，在难以添加丁酸梭菌活菌的情况下，建议适当添加聚β-羟基丁酸酯来替代丁酸梭菌。
Abstract：An experiment was conducted to study the effects of three additives on the growth performance, non-specific immunity and disease resistance of Litopenaeus vannamei. Shrimp were fed with live cells of Clostridium butyricum CBG01(CB group), 3% poly-β-hydroxybutyrate(PHB group) and 1% sodium butyrate(BS group) in the basal diets, respectively. The basal diet without additives was used as the control group and the feeding trial lasted for 6 weeks. After feeding trial, the final bodyweight, the non-specific immune parameters in the serum and the relative expression levels of immune-related genes in the hepatopancreas of shrimp were measured. Meanwhile, a Vibrio parahaemolyticus challenge experiment was carried out. Results indicated that the final weight and specific growth rate of shrimp in the CB and PHB groups were significantly improved as compared to the control(P<0.05), especially in the CB group. There was no significant difference between the BS and control groups(P>0.05). The survival rate and feed efficiency of three treatment groups were significantly higher than those of the control group(P<0.05). The activities of lysozyme(LZM), peroxidase(POD), superoxide dismutase(SOD) and total antioxidant capacity(T-AOC) in serum of shrimp of three treatment groups were significantly higher than those of the control group(P<0.05). The activities of alkaline phosphatase(AKP), acid phosphatase(ACP) and phenol oxidase(PO) in serum of the CB group, the activities of AKP, ACP and total nitric oxide synthase(T-NOS) in serum of the PHB group and ACP activity in serum of the BS group were significantly higher than those of the control group(P<0.05). Compared with the control group, the relative expression levels of all immune-related genes(SOD, LZM, proPO, LGBP, HSP70, Imd, Toll, Relish, TOR, 4E-BP, eIF4E1α, eIF4E2) in the hepatopancreas of shrimp of the CB group were significantly higher than those of the control group(P<0.05). However, the relative expression levels of Toll gene in the PHB group and Imd, Toll, Relish, eIF4E2 genes in the BS group were not significantly different from those in the control group(P>0.05). The challenge test of V. parahaemolyticus showed that the cumulative mortalities of shrimp in three treatment groups were significantly lower than that in the control group(P<0.05). The results showed that dietary addition of C. butyricum, PHB and sodium butyrate improved the growth performance and non-specific immunity capacity to varying degrees and significantly enhanced the resistance of L. vannamei against V. parahaemolyticus. By comprehensive comparisons, the effects of adding C. butyricum and PHB were basically comparable and to some extent superior to the addition of sodium butyrate. When it is difficult to add live cells of C. butyricum in feed production and processing, it is suggested that appropriately added poly-β-hydroxybutyrate may substitute C. butyricum.
[1] 孟明翔.南美白对虾养殖策略与关键技术探究[J].南方农机，2022,53(20):51-53.Meng M X.Study on culture strategies and key technologies of Litopenaeus vannamei[J].China Southern Agricultural Machinery,2022,53(20):51-53.
[2] Yin C L,Shen X J,Wang Y,et al.Comparative study of five anti-lipopolysaccharide factor genes in Litopenaeus vannamei[J].Developmental and Comparative Immunology,2023,139:104557.
[3] El-Saadony M T,Alagawany M,Patra A K,et al.The functionality of probiotics in aquaculture:An overview[J].Fish and Shellfish Immunology,2021,117:36-52.
[4] Kuebutornye F K,Abarike E D,Lu Y S.A review on the application of Bacillus as probiotics in aquaculture[J].Fish and Shellfish Immunology,2019,87:820-828.
[5] 覃美兰，何雨卓，谭筱钰，等.丁酸梭菌对吉富罗非鱼幼鱼生长性能、血清生化指标及肝脏抗氧化指标的影响[J].饲料研究，2022(24):47-51.Qin M L,He Y Z,Tan X Y,et al.Effect of Clostridium butyricum on growth performance,serum biochemical indexes and liver antioxidant indexes of juvenile tilapia[J].Feed Research,2022(24):47-51.
[6] Meng X L,Wu S K,Hu W P,et al.Clostridium butyricum improves immune responses and remodels the intestinal microbiota of common carp (Cyprinus carpio L.)[J].Aquaculture,2021,530:735753.
[7] Gao Q X,Xiao Y P,Sun P,et al.In vitro protective efficacy of Clostridium butyricum against fish pathogen infections[J].Indian Journal of Microbiology,2013,53(4):453-459.
[8] Aalamifar H,Soltanian S,Vazirzadeh A,et al.Dietary butyric acid improved growth,digestive enzyme activities and humoral immune parameters in barramundi (Lates calcarifer)[J].Aquaculture Nutrition,2020,26:156-164.
[9] 朱滔，王国英，王根虎.丁酸在动物体内的代谢、作用机理及生理作用[J].饲料研究，2022,45(10):146-150.Zhu T,Wang G Y,Wang G H.Metabolism,mechanism and physiological action of butyric acid in animals[J].Feed Research,2022,45(10):146-150.
[10] 刘艳莉，胡毅，钟蕾，等.丁酸对动物肠道健康的影响及水产应用前景[J].水产科学，2019,38(2):276-281.Liu Y L,Hu Y,Zhong L,et al.A Review:Research progress on effects of butyric acid on intestinal health in animals and its application in aquaculture[J].Fisheries Science,2019,38(2):276-281.
[11] Abdel-Tawwab M,Shukry M,Farrag F A,et al.Dietary sodium butyrate nanoparticles enhanced growth,digestive enzyme activities,intestinal histomorphometry,and transcription of growth-related genes in Nile tilapia juveniles[J].Aquaculture,2021,536:736467.
[12] Franke A,Clemmesen C,De Schryver P,et al.Immunostimulatory effects of dietary poly-β-hydroxybutyrate in European sea bass postlarvae[J].Aquaculture Research,2017,48(12):5707-5717.
[13] 杨静，白璐，马红丽，等.聚β-羟基丁酸酯来源及其在水产养殖中的应用[J].水产科学，2020,39(2):292-298.Yang J,Bai L,Ma H L,et al.Sources and applications of Poly-β-hydroxybutyrate in aquaculture[J].Fisheries Science,2020,39(2):292-298.
[14] Li H D,Tian X L,Dong S L.Growth performance,non-specific immunity,intestinal histology and disease resistance of Litopenaeus vannamei fed on a diet supplemented with live cells of Clostridium butyricum[J].Aquaculture,2019,498:470-481.
[15] Duan Y F,Zhang Y,Dong H B,et al.Effect of dietary poly-β-hydroxybutyrate (PHB) on growth performance,intestinal health status and body composition of Pacific white shrimp Litopenaeus vannamei (Boone,1931)[J].Fish and Shellfish Immunology,2017,60:520-528.
[16] 张月.聚β-羟基丁酸酯对凡纳滨对虾生长性能和肠道健康的改善作用研究[D].上海：上海海洋大学，2017.Zhang Y.Effect of Poly-β-hydroxybutyrate on Growth Performance and Intestine Health of Litopenaeus vannamei[D].Shanghai:Shanghai Ocean University,2017.
[17] 张晓晓，汪多，田相利，等.包膜丁酸钠对凡纳滨对虾生长和血清非特异性免疫酶活性的影响[J].中国海洋大学学报(自然科学版),2017,47(S1):27-34.Zhang X X,Wang D,Tian X L,et al.Effects of coated sodium butyrate on growth performance and serum non-specific immunity enzymes of Litopenaeus vannamei[J].Periodical of Ocean University of China,2017,47(S1):27-34.
[18] 刘龙镇，田相利，王明阳，等.不同复合微生态制剂添加方式对凡纳滨对虾生长、非特异性免疫及抗病力的影响[J].中国海洋大学学报(自然科学版),2018,48(12):23-31.Liu L Z,Tian X L,Wang M Y,et al.Effects of additive patterns of omnibiotics on the growth performance,non-specific immunity and disease resistance of Litopenaeus vannamei[J].Periodical of Ocean University of China,2018,48(12):23-31.
[19] Luo K,Tian X L,Wang B,et al.Evaluation of paraprobiotic applicability of Clostridium butyricum CBG01 in improving the growth performance,immune responses and disease resistance in Pacific white shrimp,Penaeus vannamei[J].Aquaculture,2021,544:737041.
[20] 王苓，田相利，董双林，等.两株芽孢杆菌对凡纳滨对虾生长和血清非特异性免疫指标的影响研究[J].中国海洋大学学报(自然科学版),2017,47(4):14-21.Wang L,Tian X L,Dong S L,et al.Effects of two Bacillus on growth performance and serum non-specific immunity of Litopenaeus vannamei[J].Periodical of Ocean University of China,2017,47(4):14-21.
[21] 王明阳，田相利，刘龙镇，等.饲料中添加凝结芽孢杆菌对凡纳滨对虾生长、血清非特异性免疫指标及抗病力的影响[J].中国海洋大学学报(自然科学版),2018,48(S1):8-15.Wang M Y,Tian X L,Liu L Z,et al.Effects of Bacillus coagulans added to feed on the growth performance,serum non-specific immunity and disease resistance of Litopenaeus vannamei[J].Periodical of Ocean University of China,2018,48(S1):8-15.
[22] Tran T T,Bott N,Lam N D,et al.The role of Pseudomonas in heterotrophic nitrification:A case study on shrimp ponds(Litopenaeus vannamei) in Soc Trang province[J].Microorganisms,2019,7(6):155.
[23] Kewcharoen W,Srisapoome P.Probiotic effects of Bacillus spp.from Pacific white shrimp(Litopenaeus vannamei) on water quality and shrimp growth,immune responses,and resistance to Vibrio parahaemolyticus (AHPND strains)[J].Fish and Shellfish Immunology,2019,94:175-189.
[24] Hoseinifar S H,Sun Y Z,Wang A R,et al.Probiotics as means of diseases control in aquaculture,a review of current knowledge and future perspectives[J].Frontiers in Microbiology,2018,9:2429.
[25] Yang G,Tian X L,Dong S L.Bacillus cereus and rhubarb regulate the intestinal microbiota of sea cucumber(Apostichopus japonicus Selenka):Species-species interaction,network,and stability[J].Aquaculture,2019,512:734284.
[26] Duan Y F,Zhang Y,Dong H B,et al.Effect of dietary Clostridium butyricum on growth,intestine health status and resistance to ammonia stress in Pacific white shrimp Litopenaeus vannamei[J].Fish and Shellfish Immunology,2017,65:25-33.
[27] Duan Y F,Zhang Y,Dong H B,et al.Effect of the dietary probiotic Clostridium butyricum on growth,intestine antioxidant capacity and resistance to high temperature stress in kuruma shrimp Marsupenaeus japonicus[J].Journal of Thermal Biology,2017,66:93-100.
[28] Qiao G,Xu C,Sun Q R,et al.Effects of dietary poly-β-hydroxybutyrate supplementation on the growth,immune response and intestinal microbiota of soiny mullet (Liza haematocheila)[J].Fish and Shellfish Immunology,2019,91:251-263.
[29] Sui L,Cai J,Sun H,et al.Effect of poly-β-hydroxybutyrate on Chinese mitten crab,Eriocheir sinensis,larvae challenged with pathogenic Vibrio anguillarum[J].Journal of Fish Diseases,2012,35:359-364.
[30] Najdegerami E,Tran T N,Defoirdt T,et al.Effects of poly-β-hydroxybutyrate(PHB) on Siberian sturgeon(Acipenser baerii) fingerlings performance and its gastrointestinal tract microbial community[J].FEMS Microbiology Ecology,2012,79:25-33.
[31] Zhao H X,Wang G X,Wang H R,et al.Effects of dietary sodium butyrate on growth,digestive enzymes,body composition and nutrient retention-related gene expression of juvenile yellow catfish (Pelteobagrus fulvidraco)[J].Animal Nutrition,2021,7:539-547.
[32] Liu M M,Guo W,Wu F,et al.Dietary supplementation of sodium butyrate may benefit growth performance and intestinal function in juvenile grass carp (Ctenopharyngodon idellus)[J].Aquaculture Research,2017,48:4102-4111.
[33] 李光，樊景凤，林凤翱，等.对虾的免疫机制及其疾病免疫预防的研究进展[J].水产科学，2007,26(1):56-60.Li G,Fan J F,Lin F X,et al.Research progress on immune mechanism and immunoprophylaxis of diseases in shrimp[J].Fisheries Science,2007,26(1):56-60.
[34] Shi Y Z,Suwaree K,Chen Y Y,et al.White shrimp Litopenaeus vannamei hemocytes receiving fucoidan release endogenous molecules that activate and synergize innate immunity in the presence of fucoidan[J].Aquaculture,2020,519:734720.
[35] Tassanakajon A,Somboonwiwat K,Supungul P,et al.Discovery of immune molecules and their crucial functions in shrimp immunity[J].Fish and Shellfish Immunology,2013,34:954-967.
[36] Amparyup P,Charoensapsri W,Tassanakajon A.Prophenoloxidase system and its role in shrimp immune responses against major pathogens[J].Fish and Shellfish Immunology,2013,34:990-1001.
[37] Han M M,Gao T H,Liu G X,et al.The effect of a polystyrene nanoplastic on the intestinal microbes and oxidative stress defense of the freshwater crayfish,Procambarus clarkii[J].Science of the Total Environment,2022,833:155722.
[38] Chen S J,Yu Y Y,Gao Y J,et al.Exposure to acute ammonia stress influences survival,immune response and antioxidant status of pacific white shrimp (Litopenaeus vannamei) pretreated with diverse levels of inositol[J].Fish and Shellfish Immunology,2019,89:248-256.
[39] Pan C H,Chien Y H,Hunter B.The resistance to ammonia stress of Penaeus monodon Fabricius juvenile fed diets supplemented with astaxanthin[J].Journal of Experimental Marine Biology and Ecology,2003,297:107-118.
[40] 姜国建，于仁诚，周名江.活性氮中间体和一氧化氮合成酶系统在水产养殖生物病害防御中的作用[J].海洋科学，2006,3(3):90-93.Jiang G J,Yu R C,Zhou M J.Role of reactive nitrogen intermediates(RNIs) and nitric oxide synthase in disease resistance of maricultured organisms[J].Marine Sciences,2006,3(3):90-93.
[41] Wu C H,Siva V,Song Y L.An evolutionarily ancient NO synthase(NOS) in shrimp[J].Fish and Shellfish Immunology,2013,35:1483-1500.
[42] 李维康，李立贤，刘泓宇，等.低鱼粉饲料添加丁酸梭菌对凡纳滨对虾生长、抗氧化能力和非特异性免疫的影响[J].广东海洋大学学报，2022,42(2):29-37.Li W K,Li L X,Liu H Y,et al.Effects of Clostridium butyricum on growth,antioxidant capacity and non-specific immunology of Litopenaeus vannamei fed with concentrated cottonseed protein replacement of fishmeal[J].Journal of Guangdong Ocean University,2022,42(2):29-37.
[43] Liu L F,Wang Y X,Ren J P,et al.Effect of dietary supplementation with sodium butyrate and tributyrin on the growth performance and intestinal microbiota of Pacific white shrimp (Litopenaeus vannamei)[J].Aquaculture International,2022,30:2477-2489.
[44] 袁伟，戴习林，戈潘缘元，等.RNA干扰不同类型TLR基因对罗氏沼虾免疫相关基因表达的影响[J].上海海洋大学学报，2021,30(4):590-600.Yuan W,Dai X L,Ge P Y Y,et al.Effects of RNA interferences with different Toll-like receptors on the expression of immune-related genes in Macrobrachium rosenbergii[J].Journal of Shanghai Ocean University,2021,30(4):590-600.
[45] Yan P Y,Lin C,He M,et al.Immune regulation mediated by JAK/STAT signaling pathway in hemocytes of Pacific white shrimps,Litopenaeus vannamei stimulated by lipopolysaccharide[J].Fish and Shellfish Immunology,2022,130:141-154.
[46] Li F H,Xiang J H.Signaling pathways regulating innate immune responses in shrimp[J].Fish and Shellfish Immunology,2013,34:973-980.
[47] Zhuang Y,Huang H,Liu X L,et al.Effect of bovine lactoferricin on the growth performance,digestive capacity,immune responses and disease resistance in Pacific white shrimp,Penaeus vannamei[J].Fish and Shellfish Immunology,2022,123:282-289.
[48] Huang Y,Li T T,Jin M,et al.Newly identified PcToll4 regulates antimicrobial peptide expression in intestine of red swamp crayfish Procambarus clarkii[J].Gene,2017,610:140-147.
[49] 梁芳梅，朱鹏，邱春桃，等.长毛明对虾核转录因子NF-κB家族基因的克隆及在细菌侵染过程中的表达变化[J].中国水产科学，2022,29(11):1551-1563.Liang F M,Zhu P,Qiu C T,et al.Cloning and expression analysis of NF-κB family genes under bacterial infection of Fenneropenaeus penicillatus[J].Journal of Fishery Sciences of China,2022,29(11):1551-1563.
[50] Takahashi S,Shibutani S,Iwata H.Nuclear-targeted 4E-BP1 is dephosphorylated,induces nuclear translocation of eIF4E,and alters mRNA translation[J].Experimental Cell Research,2022,418:113246.
[51] Evsikov A,Marín de Evsikova C.Evolutionary origin and phylogenetic analysis of the novel oocyte-specific eukaryotic translation initiation factor 4E in Tetrapoda[J].Development Genes and Evolution,2009,219:111-118.
[52] Livingstone M,Larsson O,Sukarieh R,et al.A Chemical genetic screen for mTOR pathway inhibitors based on 4E-BP-dependent nuclear accumulation of eIF4E[J].Chemistry and Biology,2009,16(12):1240-1249.
[53] Bondad-Reantaso M G,Subasinghe R P,Arthur J R,et al.Disease and health management in Asian aquaculture[J].Veterinary Parasitology,2005,132:249-272.
[54] Nair A,Leo Antony M,Praveen N K,et al.Evaluation of in vitro and in vivo potential of Bacillus subtilis MBTDCMFRI Ba37 as a candidate probiont in fish health management[J].Microbial Pathogenesis,2021,152:104610.
[55] Karnjana K,Soowannayan C,Wongprasert K.Ethanolic extract of red seaweed Gracilaria fisheri and furanone eradicate Vibrio harveyi and Vibrio parahaemolyticus biofilms and ameliorate the bacterial infection in shrimp[J].Fish and Shellfish Immunology,2019,88:91-101.
[56] Gu Q Q,Wang G H,Li N Q,et al.Evaluation of the efficacy of a novel Vibrio vulnificus vaccine based on antibacterial peptide inactivation in turbot,Scophthalmus maximus[J].Fish and Shellfish Immunology,2021,118:197-204.
[57] Liang F Y,Sun C B,Li S D,et al.Therapeutic effect and immune mechanism of chitosan-gentamicin conjugate on Pacific white shrimp(Litopenaeus vannamei) infected with Vibrio parahaemolyticus[J].Carbohydrate Polymers,2021,269:118334.
[58] Luo M,Feng G Q and Ke H.Role of Clostridium butyricum,Bacillus subtilis,and algae-sourced β-1,3 glucan on health in grass turtle[J].Fish and Shellfish Immunology,2022,131:244-256.
[59] Situmorang M L,Schryver P D,Dierckens K,et al.Effect of poly-β-hydroxybutyrate on growth and disease resistance of Nile tilapia Oreochromis niloticus juveniles[J].Veterinary Microbiology,2016,182:44-49.
[60] Xiao C Y,Zhang Y F,Zhu F.Effect of dietary sodium butyrate on the innate immune response of Procambarus clarkii and disease resistance against white spot syndrome virus[J].Aquaculture,2021,541:736784.
基本信息:
DOI：10.16441/j.cnki.hdxb.20230014
中图分类号:S968.22
引用信息:
[1]田相利,秦光彩,罗凯,等.3种不同添加物对凡纳滨对虾生长、非特异免疫和抗病力的影响[J].中国海洋大学学报(自然科学版),2024,54(02):33-43.DOI:10.16441/j.cnki.hdxb.20230014.
基金信息:
国家重点研究发展计划项目（2020YFD0900201,2019YFD0900403）; 青岛市生物制造行业智库联合基金项目（QDSWZK202111）资助~~
2024-01-16
2024-01-16