氨氮对拟穴青蟹的急性毒性及对其血清免疫相关酶活力的影响
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摘要
采用静水法研究氨氮对拟穴青蟹(Scylla paramamosain)的急性毒性及在氨氮初始浓度分别为0(C0,对照组)、10(C10组)、20(C20组)、30(C30组)、40(C40组)、50 mg/L(C50组),胁迫时间分别为0、6、24、48、72 h的条件下对其血清中碱性磷酸酶(AKP)、酸性磷酸酶(ACP)、溶菌酶(LZM)、超氧化物歧化酶(SOD)和酚氧化酶(PO)活力的影响。结果显示,总氨氮对拟穴青蟹24 h和48 h的半致死浓度分别为104.793、66.124 mg/L,安全浓度为7.90 mg/L,非离子氨对拟穴青蟹24 h和48 h的半致死浓度分别为8.396、5.298 mg/L,安全浓度为0.63 mg/L。在胁迫6、24、48与72 h时,各实验组的LZM活力均显著低于对照组(P<0.01)。相较于对照组,C10、C20及C40组在24 h的AKP、ACP活力均显著升高(P<0.05)。C20组在24 h的SOD活力则显著低于其他胁迫时间点(P<0.05)。胁迫72 h时,C30、C40及C50组的PO活力显著高于对照组(P<0.05)。该实验条件下,不高于40 mg/L的氨氮可在24 h内使拟穴青蟹血清中的AKP与ACP活力显著升高,而50 mg/L的氨氮则对其具有抑制作用;各实验组浓度氨氮均在72 h内对拟穴青蟹血清中的LZM活力具有显著的抑制作用,对PO活力具有明显的刺激作用,对SOD活力无显著影响。
Using the toxicity method, we studied the acute toxicity of ammonia nitrogen to Scylla paramamosain and its influence on phenol oxidase(PO), alkaline phosphatase(AKP), acid phosphatase(ACP), superoxide dismutase(SOD), and lysozyme(LZM) in the conditions of different ammonia nitrogen concentrations of 0(C0, control group), 10(C10), 20(C20), 30(C30), 40(C40), 50 mg/L(C50) and different times of 0, 6, 24, 48, and 72 h. The results showed that the semi-lethal concentration(LC50) at 24 and 48 h of ammonia-N were 104.793 and 66.124 mg/L respectively, and the safe concentration(SC) was 7.90 mg/L. The LC50 at 24 and 48 h of NH3-N were 8.396 and 5.298 mg/L respectively, and the SC was 0.63 mg/L. The LZM activity of each treatment group was significantly lower than that of C0 group(P<0.01) at 6, 24, 48, and 72 h. AKP and ACP activity of C10, C20, and C40 groups was significantly higher than that of control group at 24 h(P<0.05), while the SOD activity of the C20 group was significantly lower than that of other stress times(P<0.05). At 72 h, the PO activity of C30, C40, and C50 groups were significantly higher than that of C0 group(P<0.05). Under the experimental conditions, ammonia nitrogen that was lower than 40 mg/L could significantly enhance the AKP and ACP activity of S. paramamosain over a 24 h period, while 50 mg/L ammonia nitrogen had inhibitory effects. Ammonia nitrogen in all groups had significant inhibitory effects on LZM activity of S. paramamosain, which had no significant effect on SOD activity, but had an obvious enhancement effect on activity of PO by 72 h.
Using the toxicity method, we studied the acute toxicity of ammonia nitrogen to Scylla paramamosain and its influence on phenol oxidase(PO), alkaline phosphatase(AKP), acid phosphatase(ACP), superoxide dismutase(SOD), and lysozyme(LZM) in the conditions of different ammonia nitrogen concentrations of 0(C0, control group), 10(C10), 20(C20), 30(C30), 40(C40), 50 mg/L(C50) and different times of 0, 6, 24, 48, and 72 h. The results showed that the semi-lethal concentration(LC50) at 24 and 48 h of ammonia-N were 104.793 and 66.124 mg/L respectively, and the safe concentration(SC) was 7.90 mg/L. The LC50 at 24 and 48 h of NH3-N were 8.396 and 5.298 mg/L respectively, and the SC was 0.63 mg/L. The LZM activity of each treatment group was significantly lower than that of C0 group(P<0.01) at 6, 24, 48, and 72 h. AKP and ACP activity of C10, C20, and C40 groups was significantly higher than that of control group at 24 h(P<0.05), while the SOD activity of the C20 group was significantly lower than that of other stress times(P<0.05). At 72 h, the PO activity of C30, C40, and C50 groups were significantly higher than that of C0 group(P<0.05). Under the experimental conditions, ammonia nitrogen that was lower than 40 mg/L could significantly enhance the AKP and ACP activity of S. paramamosain over a 24 h period, while 50 mg/L ammonia nitrogen had inhibitory effects. Ammonia nitrogen in all groups had significant inhibitory effects on LZM activity of S. paramamosain, which had no significant effect on SOD activity, but had an obvious enhancement effect on activity of PO by 72 h.
引文
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Mugnier C,Zipper E,Goarant C,et al.Combined effect of exposure to ammonia and hypoxia on the blue shrimp Litopenaeus stylirostris survival and physiological response in relation to molt stage.Aquaculture,2008,274(2-4):398-407
Qian JH,Li ZM,Liu JY,et al.Acute toxicity of ammonia-N and its effect on the immune parameters of Fenneropenaeus merguiensis.Journal of Applied Oceanography,2016,35(2):211-216[钱佳慧,栗志民,刘建勇,等.氨氮对墨吉明对虾的急性毒性及对其免疫因子的影响.应用海洋学学报,2016,35(2):211-216]
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Racotta IS,Hernándezherrera R.Metabolic responses of the white shrimp,Penaeus vannamei,to ambient ammonia.Comparative Biochemistry&Physiology Part A:Molecular&Integrative Physiology,2000,125(4):437-443
Ruyet PL,Chartois H,Quemener L.Comparative acute ammonia toxicity in marine fish and plasma ammonia response.Aquaculture,1995,136(1-2):181-194
Shao XB,LüYL,Li K,et al.The acute toxicity of ammonia nitrogen to Scylla serrata,and the effects of temperature to the rate of oxygen consumption and suffocation point.Journal of Zhejiang Ocean University(Natural Science),2011,30(6):511-514[邵鑫斌,吕永林,李凯,等.氨氮对锯缘青蟹的急性毒性和温度对其耗氧率、窒息点的影响.浙江海洋学院学报(自然科学版),2011,30(6):511-514]
Wang WN,Wang AL,Zhang YJ,et al.Effects of nitrite on lethal and immune response of Macrobrachium nipponense.Aquaculture,2004,232(1):679-686
Wang Y,Hu YB,Jiang NC.Effect of ammonia nitrogen and nitrite nitrogen on immune enzymes of Macrobrachium rosenbergii.Journal of Zhejiang University(Science Edition),2005,32(6):698-705[王玥,胡义波,姜乃澄.氨态氮、亚硝态氮对罗氏沼虾免疫相关酶类的影响.浙江大学学报(理学版),2006,32(6):698-705]
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Ashida M,S?derh?ll K.The prophenoloxidase activating system in crayfish.Comparative Biochemistry and Physiology,1984,77B(1):21-26
Ashida M.Purification and characterization of prophenoloxidase from hemolymph of the silkworm Bombyx mori.Archives of Biochemistry&Biophysics,1971,144(2):749-762
Cheng W,Chen JC.The virulence of Enterococcus to freshwater prawn Macrobrachium rosenbergii and its immune resistance under ammonia stress.Fish&Shellfish Immunology,2002,12(2):97-109
Deng KY,Meng XH,Kong J,et al.Effects of poly-β-hydroxybutyrate on non-specific immunity and expression of immune gene under the ammonia stress of Litopenaeus vannamei.Progress in Fishery Sciences,2015,36(6):71-78[邓康裕,孟宪红,孔杰,等.聚β-羟基丁酸酯对凡纳滨对虾(Litopenaeus vannamei)非特异性免疫力及氨氮胁迫后免疫基因表达量的影响.渔业科学进展,2015,36(6):71-78]
Dong YB,Wang K,Wang LT.Research progress in toxicity of ammonia nitrogen to aquatic organisms.Tianjin Fisheries,2011(Z1):8-13[董玉波,王轲,王林同.氨氮对水生生物毒性的研究进展.天津水产,2011(Z1):8-13]
Grinde B,Jollés J,Jollés P.Purification and characterization of two lysozymes from rainbow trout(Salmo gairdneri).European Journal of Biochemistry,1988,173:269-273
Han CY,Zheng QM,Chen GD,et al.Effect of ammonia-N stress on non-specific immunity of tilapia(Oreochromis niloticus×O.areus).South China Fisheries Science,2014(3):47-52[韩春艳,郑清梅,陈桂丹,等.氨氮胁迫对奥尼罗非鱼非特异性免疫的影响.南方水产科学,2014(3):47-52]
He HQ,Sun F.Research on acidic and alkaline phosphatase characteristics of Penaeus chinensis.Oceanologia et Limnologia Sinica,1992,23(5):555-560[何海琪,孙凤.中国对虾酸性和碱性磷酸酶的特性研究.海洋与湖沼,1992,23(5):555-560]
Hong M,Chen L,Sun X,et al.Metabolic and immune responses in Chinese mitten-handed crab(Eriocheir sinensis)juveniles exposed to elevated ambient ammonia.Comparative Biochemistry&Physiology,Part C:Toxicology&Pharmacology,2007,145(3):363-369
Hong ML,Chen LQ,Gu SZ,et al.Effects of ammonia exposure on immunity indicators of haemolymph and histological structure of hepatopancreas in Chinese mitten crab(Eriocheir sinensis).Journal of Fishery Sciences of China,2007,14(3):412-418[洪美玲,陈立侨,顾顺樟,等.氨氮胁迫对中华绒螯蟹免疫指标及肝胰腺组织结构的影响.中国水产科学,2007,14(3):412-418]
Hu Y,Huang Y,Zhong L,et al.Effects of ammonia stress on the gill Na+/K+-ATPase,microstructure and some serum physiological-biochemical indices of juvenile black carp(Mylopharyngodon piceus).Journal of Fisheries of China,2012,36(4):538-545[胡毅,黄云,钟蕾,等.氨氮胁迫对青鱼幼鱼鳃丝Na+/K+-ATP酶、组织结构及血清部分生理生化指标的影响.水产学报,2012,36(4):538-545]
Huang HZ,Li Y,Song XH,et al.NH+4-N stress on immune function of Eriocheir sinensis.Oceanologia et Limnologia Sinica,2006,37(3):198-205[黄鹤忠,李义,宋学宏,等.氨氮胁迫对中华绒螯蟹(Eriocheir sinensis)免疫功能的影响.海洋与湖沼,2006,37(3):198-205]
Huang HZ,Liang SR.Toxicity treatment of 3 kinds of drugs on Eriocheir sinensis larvae.Reservoir Fisheries,1998(6):15-16[黄鹤忠,梁守仁.3种药物对河蟹幼体的毒性试验.水生态学杂志,1998(6):15-16]
Kong XH,Wang GZ,Ai CX,et al.Comparative research on total antioxidant capacity and SOD activity in different organs and tissues in Scylla serrata.Journal of Applied Oceanography,2003,22(4):469-474[孔祥会,王桂忠,艾春香,等.锯缘青蟹不同器官组织中总抗氧化能力和SOD活性的比较研究.应用海洋学学报,2003,22(4):469-474]
Le Moullac G,Haffner P.Environmental factors affecting immune responses in Crustacea.Aquaculture,2000,191:121-131
Liu HZ,Zheng FR,Sun XQ,et al.Effect of exposure to ammonia nitrogen stress on immune enzyme of holothurian Apostichopus japonicus.Marine Science,2012,36(8):47-52[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.海洋科学,2012,36(8):47-52]
Mugnier C,Zipper E,Goarant C,et al.Combined effect of exposure to ammonia and hypoxia on the blue shrimp Litopenaeus stylirostris survival and physiological response in relation to molt stage.Aquaculture,2008,274(2-4):398-407
Qian JH,Li ZM,Liu JY,et al.Acute toxicity of ammonia-N and its effect on the immune parameters of Fenneropenaeus merguiensis.Journal of Applied Oceanography,2016,35(2):211-216[钱佳慧,栗志民,刘建勇,等.氨氮对墨吉明对虾的急性毒性及对其免疫因子的影响.应用海洋学学报,2016,35(2):211-216]
Qiu DQ,Zhou XJ,Qiu MS,et al.Research on anti-disease ability of Litopenaeus vannamei and the biological control of Vibrio parahaemolyticus bacteriophage under stresses of ammonia nitrogen.Acta Hydrobiologica Sinica,2008,32(4):455-461[邱德全,周鲜娇,邱明生.氨氮胁迫下凡纳滨对虾抗病力和副溶血弧菌噬菌体防病效果研究.水生生物学报,2008,32(4):455-461]
Racotta IS,Hernándezherrera R.Metabolic responses of the white shrimp,Penaeus vannamei,to ambient ammonia.Comparative Biochemistry&Physiology Part A:Molecular&Integrative Physiology,2000,125(4):437-443
Ruyet PL,Chartois H,Quemener L.Comparative acute ammonia toxicity in marine fish and plasma ammonia response.Aquaculture,1995,136(1-2):181-194
Shao XB,LüYL,Li K,et al.The acute toxicity of ammonia nitrogen to Scylla serrata,and the effects of temperature to the rate of oxygen consumption and suffocation point.Journal of Zhejiang Ocean University(Natural Science),2011,30(6):511-514[邵鑫斌,吕永林,李凯,等.氨氮对锯缘青蟹的急性毒性和温度对其耗氧率、窒息点的影响.浙江海洋学院学报(自然科学版),2011,30(6):511-514]
Wang WN,Wang AL,Zhang YJ,et al.Effects of nitrite on lethal and immune response of Macrobrachium nipponense.Aquaculture,2004,232(1):679-686
Wang Y,Hu YB,Jiang NC.Effect of ammonia nitrogen and nitrite nitrogen on immune enzymes of Macrobrachium rosenbergii.Journal of Zhejiang University(Science Edition),2005,32(6):698-705[王玥,胡义波,姜乃澄.氨态氮、亚硝态氮对罗氏沼虾免疫相关酶类的影响.浙江大学学报(理学版),2006,32(6):698-705]
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