亚硝态氮对中华鳑鲏的急性毒性及短期影响
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摘要
静水试验条件下研究了亚硝态氮对体质量(0.636±0.09) g的中华鳑鲏急性毒性及短期胁迫下对其体内两种酶活性的影响。在水温25℃、溶解氧6.3 mg/L、pH 7.4时,测定亚硝态氮对中华鳑鲏的24 h半致死质量浓度,并设置高、中、低3个亚硝态氮质量浓度组[1.28、0.64、0.43 mg/L(1/10×24 h半致死质量浓度、1/20×24 h半致死质量浓度、1/30×24 h半致死质量浓度)],于0、24、48、72 h采集血样,测定血清中过氧化氢酶和碱性磷酸酶的活性。试验结果表明,亚硝态氮对中华鳑鲏的24 h半致死质量浓度为12.76 mg/L。在亚硝态氮胁迫作用下,中华鳑鲏血清中过氧化氢酶活性先显著上升,48 h后开始下降;碱性磷酸酶活性变化则是先显著下降,在48 h恢复,然后再下降。碱性磷酸酶活性在24 h和72 h均受到明显抑制。亚硝态氮质量浓度和暴露时间两者之间存在显著交互作用,碱性磷酸酶可以作为亚硝态氮毒性检测的生物标记物。
Both acute toxicity of nitrite nitrogen(6, 10, 11, 14, 16.6 mg/L and 18 mg/L) to fish Rhodens sinensis [body weight(0.636±0.09) g]and short term effects of nitrite nitrogen on serum activities of catalase(CAT) and alkaline phosphatase(AKP) were investigated at pH 7.4, dissolved oxygen(DO) of 6.3 mg/L and 25 ℃ in a static test. The activities of CAT and AKP were determined in serum of the fishes exposed to 0.43 mg/L(1/30×24 h LC_(50)), 0.64 mg/L(1/20×24 h LC_(50)) and 1.28 mg/L(1/10×24 h LC_(50)) NaNO_(2 )concentrations at 0, 24, 48 h, and 72 h. The 24 h LC_(50) of nitrite was showed to be 12.76 mg/L, and CAT activity was increased drastically at first and subsequently decreased after 48 h, while the activity of AKP decreased at 24 h, recovered at 48 h, and then decreased again. AKP activity was inhibited significantly at 24 h, and 72 h. There was significant interaction between nitrite concentration and exposure time, indicating that AKP could be used as a biochemical marker for nitrite stress.
Both acute toxicity of nitrite nitrogen(6, 10, 11, 14, 16.6 mg/L and 18 mg/L) to fish Rhodens sinensis [body weight(0.636±0.09) g]and short term effects of nitrite nitrogen on serum activities of catalase(CAT) and alkaline phosphatase(AKP) were investigated at pH 7.4, dissolved oxygen(DO) of 6.3 mg/L and 25 ℃ in a static test. The activities of CAT and AKP were determined in serum of the fishes exposed to 0.43 mg/L(1/30×24 h LC_(50)), 0.64 mg/L(1/20×24 h LC_(50)) and 1.28 mg/L(1/10×24 h LC_(50)) NaNO_(2 )concentrations at 0, 24, 48 h, and 72 h. The 24 h LC_(50) of nitrite was showed to be 12.76 mg/L, and CAT activity was increased drastically at first and subsequently decreased after 48 h, while the activity of AKP decreased at 24 h, recovered at 48 h, and then decreased again. AKP activity was inhibited significantly at 24 h, and 72 h. There was significant interaction between nitrite concentration and exposure time, indicating that AKP could be used as a biochemical marker for nitrite stress.
引文
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[12] 王建国,王权,黄爱军,等.高锰酸钾和三氯异氰尿酸对中华鳑鲏的急性毒性作用[J].江苏农业科学,2013,41(1):239-241.
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[20] 马盛兰.氨氮和亚硝酸盐对鲢鳙鱼种的急性毒性研究[J].工程技术:全文版,2016(6):243-245.
[21] 汪家鑫,张钊,胡盼,等.亚硝酸氮对红鳍东方鲀的毒性效应[J].广东海洋大学学报,2013,33(6):52-56.
[22] 索萦.亚硝酸盐和氨氮对暗纹东方鲀免疫相关生理生化指标及基因表达的影响[D].广州:华南师范大学,2014.
[23] 潘坤,王卫民,祝东梅.氨态氮、亚硝态氮、硝态氮对麦穗鱼的急性毒性研究[J].中国农学通报,2012,28(35):96-101.
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[26] Cogun H Y,Firidin G,Aytekin T,et al.Acute toxicity of nitrite on some biochemical,hematological and antioxidant parameters in Nile tilapia (Oreochromis niloticus L,1758)[J].Fresenius Environmental Bulletin,2017,26(2):1712-1719.
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[28] Jiang Q,Zhang W,Tan H,et al.Analysis of gene expression changes,caused by exposure to nitrite,in metabolic and antioxidant enzymes in the red claw crayfish,Cherax quadricarinatus[J].Ecotoxicology & Environmental Safety,2014,104(5):423-428.
[29] Zhang Z,Wu R S,Mok H O,et al.Isolation,characterization and expression analysis of a hypoxia-responsive glucose transporter gene from the grass carp,Ctenopharyngodon idellus[J].European Journal of Biochemistry,2003,270(14):3010-3017.
[30] 许星鸿,阎斌伦,徐加涛,等.亚硝态氮胁迫对日本蟳免疫指标的影响[J].水产科学,2012,31(7):392-397.
[31] 冼健安,张秀霞,郭慧,等.亚硝酸盐胁迫对罗氏沼虾血细胞及其抗氧化酶活力的影响[J].生物安全学报,2016,25(4):300-307.
[32] Zheng J,Yong M,Su Y,et al.Effects of nitrite stress on mrna expression of antioxidant enzymes,immune-related genes and apoptosis-related proteins in Marsupenaeus japonicus[J].Fish & Shellfish Immunology,2016(58):239-252.
[33] Tian J,Ren Q,Yang J X,et al.Effect of nitrite stress on gene expression of antioxidant enzymes,heat shock protein 70,and metabolic enzymes in gill tissue of adult red swamp crayfish,Procambarus clarkii[J].Journal of Crustacean Biology,2014,34(6):754-759.
[34] Xu Z,Li T,Li E,et al.Comparative transcriptome analysis reveals molecular strategies of oriental river prawn Macrobrachium nipponense in response to acute and chronic nitrite stress[J].Fish & Shellfish Immunology,2015(48):254-265.
[35] Du J,Zhu H,Liu P,et al.Immune responses and gene expression in hepatopancreas from Macrobrachium rosenbergii challenged by a novel pathogen Spiroplasma Mr-1008[J].Fish & Shellfish Immunology,2013,34(1):315-323.
[36] 王国江.氨氮和亚硝态氮胁迫对日本沼虾血淋巴的影响[D].保定:河北大学,2008.
[37] Guo H,Xian J A,Wang A L.Analysis of digital gene expression profiling in hemocytes of white shrimp Litopenaeus vannamei under nitrite stress[J].Fish & Shellfish Immunology,2016(56):1-11.
[38] Sun S,Ge X,Xuan F,et al.Nitrite-induced hepatotoxicity in bluntsnout bream (Megalobrama amblycephala):the mechanistic insight from transcriptome to physiology analysis[J].Environmental Toxicology & Pharmacology,2014,37(1):55-65.
[39] Guo H,Ye C X,Wang A L,et al.Trascriptome analysis of the Pacific white shrimp Litopenaeus vannamei exposed to nitrite by rna-seq[J].Fish & Shellfish Immunology,2013,35(6):2008-2016.
[2] Jia R,Han C,Lei J L,et al.Effects of nitrite exposure on haematological parameters,oxidative stress and apoptosis in juvenile turbot (Scophthalmus maximus)[J].Aquatic Toxicology,2015,169(7):1-9.
[3] Huey D W,Beitinger T L,Wooten M C.Nitrite-induced methemoglobin formation and recovery in channel catfish (Ictalurus punctatus) at three acclimation temperatures[J].Bulletin of Environmental Contamination and Toxicology,1984,32(1):674-681.
[4] Pottinger T G.Modulation of the stress response in wild fish is associated with variation in dissolved nitrate and nitrite [J].Environmental Pollution,2017(225):550-558.
[5] Frances J,Allan G L,Nowak B F.The effects of nitrite on the short-term growth of silver perch (Bidyanus bidyanus)[J].Aquaculture,1998,163(1):63-72.
[6] 王权,王建国,封琦,等.中华鳑鲏的生物学特性及人工养殖技术[J].江苏农业科学,2014,42(5):193-194.
[7] 王建国,王权,封琦,等.一种鳑鲏和沙塘鳢生态立体养殖方法:中国,CN104756911A[P].2015-07-08.
[8] 贺艳辉,袁永明,王红卫,等.一种河蟹、青虾和中华鳑鲏的生态混养方法:中国,CN104054614A[P].2014-09-24.
[9] 熊良伟,王帅兵,王建国,等.一种宽体金线蛭与鳑鲏混养技术:中国,CN105145425A[P].2015-12-16.
[10] 王建国,王权,熊良伟,等.一种鳑鲏和小龙虾生态混合养殖方法:中国,CN104770317A[P].2015-07-15.
[11] 杨建华,宋维彦.3种重金属离子对中华鳑鲏鱼的急性毒性及安全浓度研究[J].安徽农业科学,2010,38(23):12481-12482.
[12] 王建国,王权,黄爱军,等.高锰酸钾和三氯异氰尿酸对中华鳑鲏的急性毒性作用[J].江苏农业科学,2013,41(1):239-241.
[13] 喻倩,王金庆,刘娅琴,等.高密度养殖鳑鲏鱼对水体营养盐浓度和浮游植物群落的影响[J].上海农业学报,2014,30(4):52-56.
[14] 龚学臣.SPSS软件在交互效应方差分析中的应用[J].河北北方学院学报:自然科学版,2015,31(1):19-22.
[15] 邵翠红,高山林,孙素霞.SPSS11.0软件用于方差分析中的数据转换[J].电脑开发与应用,2005,18(2):60.
[16] Svobodová Z,MáChová J,Poleszczuk G,et al.Nitrite poisoning of fish in aquaculture facilities with water-recirculating systems[J].Acta Veterinaria Brno,2005,74(1):9584-9585.
[17] William M,Lewis J.Toxicity of nitrite to fish:a review[J].Transactions of the American Fisheries Society,1986,115(2):183-195.
[18] Eddy F B,Williams E M.Nitrite and freshwater fish[J].Chemistry & Ecology,1987,3(1):1-38.
[19] 陈万光,凡秀丽,王琳丹.亚硝态氮对高体鳑鲏鱼苗的急性毒性[J].江苏农业科学,2012,40(2):188-189.
[20] 马盛兰.氨氮和亚硝酸盐对鲢鳙鱼种的急性毒性研究[J].工程技术:全文版,2016(6):243-245.
[21] 汪家鑫,张钊,胡盼,等.亚硝酸氮对红鳍东方鲀的毒性效应[J].广东海洋大学学报,2013,33(6):52-56.
[22] 索萦.亚硝酸盐和氨氮对暗纹东方鲀免疫相关生理生化指标及基因表达的影响[D].广州:华南师范大学,2014.
[23] 潘坤,王卫民,祝东梅.氨态氮、亚硝态氮、硝态氮对麦穗鱼的急性毒性研究[J].中国农学通报,2012,28(35):96-101.
[24] Tomasso J R.Comparative toxicity of nitrite to freshwater fishes[J].Aquatic Toxicology,1986,8(2):129-137.
[25] Kroupová K H,Valentová O,Svobodová Z,et al.Toxic effects of nitrite on freshwater organisms:a review[J].Reviews in Aquaculture,2018,10(3):525-542.
[26] Cogun H Y,Firidin G,Aytekin T,et al.Acute toxicity of nitrite on some biochemical,hematological and antioxidant parameters in Nile tilapia (Oreochromis niloticus L,1758)[J].Fresenius Environmental Bulletin,2017,26(2):1712-1719.
[27] Yildiz H Y,Robaina L,Pirhonen J,et al.Fish welfare in aquaponic systems:its relation to water quality with an emphasis on feed and faeces—a review[J].Water,2017,9(1):1-17.
[28] Jiang Q,Zhang W,Tan H,et al.Analysis of gene expression changes,caused by exposure to nitrite,in metabolic and antioxidant enzymes in the red claw crayfish,Cherax quadricarinatus[J].Ecotoxicology & Environmental Safety,2014,104(5):423-428.
[29] Zhang Z,Wu R S,Mok H O,et al.Isolation,characterization and expression analysis of a hypoxia-responsive glucose transporter gene from the grass carp,Ctenopharyngodon idellus[J].European Journal of Biochemistry,2003,270(14):3010-3017.
[30] 许星鸿,阎斌伦,徐加涛,等.亚硝态氮胁迫对日本蟳免疫指标的影响[J].水产科学,2012,31(7):392-397.
[31] 冼健安,张秀霞,郭慧,等.亚硝酸盐胁迫对罗氏沼虾血细胞及其抗氧化酶活力的影响[J].生物安全学报,2016,25(4):300-307.
[32] Zheng J,Yong M,Su Y,et al.Effects of nitrite stress on mrna expression of antioxidant enzymes,immune-related genes and apoptosis-related proteins in Marsupenaeus japonicus[J].Fish & Shellfish Immunology,2016(58):239-252.
[33] Tian J,Ren Q,Yang J X,et al.Effect of nitrite stress on gene expression of antioxidant enzymes,heat shock protein 70,and metabolic enzymes in gill tissue of adult red swamp crayfish,Procambarus clarkii[J].Journal of Crustacean Biology,2014,34(6):754-759.
[34] Xu Z,Li T,Li E,et al.Comparative transcriptome analysis reveals molecular strategies of oriental river prawn Macrobrachium nipponense in response to acute and chronic nitrite stress[J].Fish & Shellfish Immunology,2015(48):254-265.
[35] Du J,Zhu H,Liu P,et al.Immune responses and gene expression in hepatopancreas from Macrobrachium rosenbergii challenged by a novel pathogen Spiroplasma Mr-1008[J].Fish & Shellfish Immunology,2013,34(1):315-323.
[36] 王国江.氨氮和亚硝态氮胁迫对日本沼虾血淋巴的影响[D].保定:河北大学,2008.
[37] Guo H,Xian J A,Wang A L.Analysis of digital gene expression profiling in hemocytes of white shrimp Litopenaeus vannamei under nitrite stress[J].Fish & Shellfish Immunology,2016(56):1-11.
[38] Sun S,Ge X,Xuan F,et al.Nitrite-induced hepatotoxicity in bluntsnout bream (Megalobrama amblycephala):the mechanistic insight from transcriptome to physiology analysis[J].Environmental Toxicology & Pharmacology,2014,37(1):55-65.
[39] Guo H,Ye C X,Wang A L,et al.Trascriptome analysis of the Pacific white shrimp Litopenaeus vannamei exposed to nitrite by rna-seq[J].Fish & Shellfish Immunology,2013,35(6):2008-2016.