氨氮胁迫对刺参生理健康指标的影响
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摘要
研究了氨氮胁迫对刺参(Apostichopus japonicas)免疫和抗氧化指标的影响,氨氮染毒梯度设置为:对照组(自然海水)、0.5、1.25mg/L处理组,取样时间为0、2、6、12、24和48h,分别取刺参体腔液和体壁组织,测定体腔液总体腔细胞数量、凝集活性、溶菌和抗菌活力以及体壁、体腔液总抗氧化活力(TAOC)、超氧化物歧化酶(SOD)活力、还原型谷胱甘肽(GSH)含量、还原型/氧化型谷胱甘肽含量比值(GSH/GSSG)。结果表明:氨氮胁迫对刺参免疫和抗氧化指标影响显著(P<0.05),而对照组无明显变化。各处理组总体腔细胞数量、溶菌活力分别在48、12h内呈峰值变化,分别于12、6h达到最小值和最大值,然后恢复至对照组水平,各处理组抗菌活力、凝集活性随胁迫时间呈下降趋势,1.25mg/L染毒组抗菌活力在12h后明显低于对照组水平。各处理组体壁T-AOC、SOD活力和GSH含量、GSH/GSSG分别在48、12h内呈峰值变化,均于6h时达到最大值,分别于48h时和24h后显著低于或恢复至对照组水平;各处理组体腔液T-AOC、SOD活力和GSH含量实验时间内呈下降趋势,分别在6、12和6h后显著低于对照组水平,然后保持稳定,而GSH/GSSG与对照组无明显差异。由此可见,氨氮对刺参免疫和抗氧化指标具有明显的抑制效应,其中体壁、体腔液T-AOC、SOD活力和体腔液GSH含量表现出显著的时间-剂量效应,可作为刺参在氨氮胁迫下生理健康的评价指标。
The study was to evaluate the effects of ammonia on immune and antioxidant parameters of Apostichopusjaponicus.A.japonicus were exposed to three ammonia levels:0(control),0.5,1.25mg/L for 48 hand sampled randomly at 0,2,6,12,24 and 48h,and then measured total coelomocyte counts(TCC),lysozyme activity(LZM),antimicrobial activity,agglutinating activity in coelomic fluid and total antioxidant capacity(T-AOC),superoxide dismutase(SOD)activity,reduced glutathione content(GSH)and GSH/GSSG in body wall and coelomic.The results indicated that ammonia exposure had a significant effect on immune and antioxidant parameters of A.japonicus(P<0.05).TCC and LZM activity of the experimental groups decreased within 12 hor increased within 6hrespectively,then recovered to the control level.Antimicrobial activity of 1.25mg/L treatment group and agglutinating activity of all treatment groups reached to a low level after 12 and 24hrespectively.T-AOC and SOD activity in body wall increased within 6h,and decreased to a lower level than control at 48h(P<0.05).GSH content and GSH/GSSG in body wall reached to the maximum at 6h,and recovered to the control at 24 h,12hrespectively.T-AOC,SOD activity and GSH content in coelomic fluid decreased significantly at 12 h,and then became stable.T-AOC and SOD in body wall and T-AOC,SOD and GSH content in coelomic fluid had a negative correlation with exposure concentration.It showed that ammonia exposure had a significant inhibiting effect on immune and antioxidant parameters of A.japonicus.T-AOC,SOD activity in body wall and coelomic fluid and GSH in coelomic fluid had a significant does-time effect,and could be used as evaluation indexes for physical health during ammonia stress.
The study was to evaluate the effects of ammonia on immune and antioxidant parameters of Apostichopusjaponicus.A.japonicus were exposed to three ammonia levels:0(control),0.5,1.25mg/L for 48 hand sampled randomly at 0,2,6,12,24 and 48h,and then measured total coelomocyte counts(TCC),lysozyme activity(LZM),antimicrobial activity,agglutinating activity in coelomic fluid and total antioxidant capacity(T-AOC),superoxide dismutase(SOD)activity,reduced glutathione content(GSH)and GSH/GSSG in body wall and coelomic.The results indicated that ammonia exposure had a significant effect on immune and antioxidant parameters of A.japonicus(P<0.05).TCC and LZM activity of the experimental groups decreased within 12 hor increased within 6hrespectively,then recovered to the control level.Antimicrobial activity of 1.25mg/L treatment group and agglutinating activity of all treatment groups reached to a low level after 12 and 24hrespectively.T-AOC and SOD activity in body wall increased within 6h,and decreased to a lower level than control at 48h(P<0.05).GSH content and GSH/GSSG in body wall reached to the maximum at 6h,and recovered to the control at 24 h,12hrespectively.T-AOC,SOD activity and GSH content in coelomic fluid decreased significantly at 12 h,and then became stable.T-AOC and SOD in body wall and T-AOC,SOD and GSH content in coelomic fluid had a negative correlation with exposure concentration.It showed that ammonia exposure had a significant inhibiting effect on immune and antioxidant parameters of A.japonicus.T-AOC,SOD activity in body wall and coelomic fluid and GSH in coelomic fluid had a significant does-time effect,and could be used as evaluation indexes for physical health during ammonia stress.
引文
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[19]岳峰,潘鲁青,谢鹏,等.氨氮胁迫对三疣梭子蟹酚氧化酶原系统和免疫指标的影响[J].中国水产科学,2010,17(4):761-770.
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[23]Haug,T.,A.K.Kjuul,O.B.Styrvold,et al.,Antibacterial activity in Strongylocentrotus droebachiensis(Echinoidea),Cucumaria frondosa(Holothuroidea),and Asterias rubens(Asteroidea)[J].Journal of invertebrate pathology,2002,81(2):94-102.
[24]Xing,J.,and F.S.Chia,Opsonin-like molecule found in coelomic fluid of a sea cucumber,Holothuria leucospilota,Marine Biology[J].2000,136(6):979-986.
[25]Grundy,J.E.,and K.B.Storey,Antioxidant defenses and lipid peroxidation damage in estivating toads,Scaphiopus couchii[J].Journal of Comparative Biochemistry and Physiology B,1998,168:132-142.
[26]Winston,G.W.,Oxidants and antioxidants in aquatic animals[J].Journal of ComparativeBiochemistry and Physiology C,1991,100:173-176.
[2]Colt,J.E.,and D.A.Armstrong,Nitrogen toxicity to crustaceans,fish[M].mollusks:Bioengineering Symposium for Fish Culture,1981.
[3]Motnikar,S.,R.Marsan,and F.Tetreault,Conditioning green sea urchins in tanks:water quality tolerance limits[J].Bulletin-Aquaculture Association of Cannada,1998,98:96-98.
[4]Wickins,J.F.,and D.O.C.Lee,Crustacean Farming Ranching and Culture[J].Second edition.Blackwell Science,2002:446.
[5]Livingstone,D.R.,Oxidative stress in aquatic organisms in relation to pollution and aquaculture[J].Revue de Medecine Veterinaire.2003,154(6):427-430.
[6]Smith,V.J.,The echinoderms[J].Invertebrate blood cells,1981,2:513-561.
[7]刘洪展,郑风荣,孙修勤,等.氨氮胁迫对刺参几种免疫酶活性的影响[J].海洋科学,2012,36(8):47-52.
[8]臧元奇,田相利,董双林,等.氨氮慢性胁迫对刺参免疫酶活性及热休克蛋白表达的影响[J].中国海洋大学学报(自然科学版),2012,36(8):47-52.
[9]Siikavuopio,S.I.,T.,Dale,A.Foss,et al.,Effects of chronic ammonia exposure on gonad growth and survival in green sea urchin Strongylocentrotus droebachiensis[J].Aquaculture,2004,242(1):313-320.
[10]孙兴涛,王连华.黄河三角洲地区刺参健康养殖技术[J].河北渔业,2010,(2):24-26.
[11]刘峰,王芳,董双林,等.投饵与不投饵海参养殖池塘水质变化的初步研究[J].中国海洋大学学报(自然科学版),2009,39:369-374.
[12]戴聪杰,陈寅山.日本对虾血清和肌肉提取液凝集活力初步研究[J].福建师范大学学报(自然科学版),2002,18(4):81-85.
[13]Luo,T.,H.Yang,F.Li,et al.,Purification,characterization and cDNA cloning of a novel lipopolysaccharide-binding lectin from the shrimp Penaeus monodon[J].Developmental and Comparative Immunology,2006,30(7):607-617.
[14]Hultmark,D.,H.Steiner,T.Rasmuson,et al.,Insect immunity,Purification and properties of three inducible bactericidal proteins from hemolymph of immunized pupae of Hyalophora cecropia[J].European Journal of Biochemistry,1980,106:7-16.
[15]邹国林,桂兴芬,钟晓凌.一种SOD的测活方法-邻苯三酚自氧化法的改进[J].生物化学与生物物理进展,1986,4(4):71-73.
[16]Anderson,M.E.,Determination of glutathione and glutathione disulfide in biological samples[J].Methods Enzymology,1985,113:548-555.
[17]Bradford,M.A.,A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of proteindye binding[J].Analytical Biochemistry,1976,72(1):248-254.
[18]黄鹤忠,李义,宋学宏,等.氨氮胁迫对中华绒螯蟹免疫功能的影响[J].海洋与湖沼,2006,7(3):198-205.
[19]岳峰,潘鲁青,谢鹏,等.氨氮胁迫对三疣梭子蟹酚氧化酶原系统和免疫指标的影响[J].中国水产科学,2010,17(4):761-770.
[20]Canicatti,C.,and G.D'Ancona,Cellular aspects of Holothuria polii immune response[J].Journal of Invertebrate Pathology,1989,53(2):152-158.
[21]Canicattí,C.,Lysosomal enzyme pattern in Holothuria polii coelomocytes[J].Journal of Invertebrate Pathology,1990,56(1):70-74.
[22]Canicatti,C.,and N.Parrinello,Hemagglutinin and hemolysin levels in the coelomic fluid from Holothuria polii(Echinodermata)following sheep erythrocyte injection[J].The Biological Bulletin,1985,168(1):175-182.
[23]Haug,T.,A.K.Kjuul,O.B.Styrvold,et al.,Antibacterial activity in Strongylocentrotus droebachiensis(Echinoidea),Cucumaria frondosa(Holothuroidea),and Asterias rubens(Asteroidea)[J].Journal of invertebrate pathology,2002,81(2):94-102.
[24]Xing,J.,and F.S.Chia,Opsonin-like molecule found in coelomic fluid of a sea cucumber,Holothuria leucospilota,Marine Biology[J].2000,136(6):979-986.
[25]Grundy,J.E.,and K.B.Storey,Antioxidant defenses and lipid peroxidation damage in estivating toads,Scaphiopus couchii[J].Journal of Comparative Biochemistry and Physiology B,1998,168:132-142.
[26]Winston,G.W.,Oxidants and antioxidants in aquatic animals[J].Journal of ComparativeBiochemistry and Physiology C,1991,100:173-176.