急性氨氮胁迫对尖吻鲈稚鱼消化酶及抗氧化酶活性的影响
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摘要
【目的】明确氨氮对尖吻鲈(Lates calcarifer)稚鱼消化酶及抗氧化酶活性的影响,为提高尖吻鲈仔、稚鱼养殖存活率及丰富尖吻鲈养殖生态学理论提供依据。【方法】对15日龄的尖吻鲈稚鱼进行急性氨氮胁迫试验,分别于氨氮胁迫0、6、12、24、36、48、72和96 h时测定超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、酸性磷酸酶(ACP)、谷胱甘肽过氧化物酶(GSH-Px)等抗氧化酶及脂肪酶(LPS)、淀粉酶(AMS)、胰蛋白酶(TRYP)等消化酶活性,揭示急性氨氮胁迫下尖吻鲈稚鱼抗氧化酶及消化酶的变化规律。【结果】氨氮胁迫下,5 mg/L处理组尖吻鲈稚鱼SOD活性在胁迫36 h后显著高于对照组(P<0.05,下同),10 mg/L处理组除胁迫72 h外,其他时间点均低于对照组;5 mg/L处理组尖吻鲈稚鱼POD活性随胁迫时间的延长始终高于对照组,10 mg/L处理组在胁迫6～36 h时远高于对照组,但低于5 mg/L处理组;5 mg/L处理组尖吻鲈稚鱼CAT活性在整个试验过程中均显著高于对照组,10 mg/L处理组的CAT活性变化波动明显,整体上呈上升—下降的交替式变化趋势;5 mg/L处理组尖吻鲈稚鱼ACP活性呈先下降后上升的变化趋势,且从试验开始后各时间点均显著低于对照组,10 mg/L处理组整体上呈上升—下降—上升—下降的变化趋势;5和10 mg/L处理组的尖吻鲈稚鱼GSH-Px活性变化趋势基本一致,先保持相对稳定,后随胁迫时间延长呈上升—下降—上升的变化趋势。在消化酶方面,5 mg/L处理组尖吻鲈稚鱼LPS活性从氨氮胁迫36 h起显著低于对照组,10 mg/L处理组从胁迫12 h起呈上升—下降—上升—下降的变化趋势;5和10 mg/L处理组尖吻鲈稚鱼AMS活性整体上呈先下降后上升的变化趋势,而TRYP活性均呈阶段性的上升—下降交替变化趋势。【结论】氨氮胁迫对尖吻鲈稚鱼消化酶及抗氧化酶具有明显影响,在短时间(<6 h)内尖吻鲈稚鱼对氨氮有一定耐受性,SOD、CAT等部分抗氧化酶在氨氮浓度的剧烈变化下其活性诱导增强以消除体内过多的自由基,消化酶活性呈不同程度的升高则是为了补充机体能量消耗。
【Objective】The effect of ammonia nitrogen on the digestive enzyme activities and antioxidant enzyme activities of larval Lates calcarifer was clarified,providing basis for improving the survival rate of larval L. calcarifer and enriching the ecological theory of breeding.【Method】The acute stress of ammonia nitrogen on larval L. calcarifer(15 days post hatching)was conducted. Antioxidant enzyme activities such as superoxide dismutase(SOD),peroxidase(POD),catalase(CAT),acid phosphatase(ACP),glutathione peroxidase(GSH-Px)and digestive enzyme activities such as lipase(LPS),amylase(AMS),trypsin(TRYP)were measured in the stress time of 0,6,12,24,36,48,72 and 96 h to reveal the change regulation of the activities of antioxidant enzymes and digestive enzymes in larval L. calcarifer under acute ammonia stress.【Result】Under stress of ammonia nitrogen,SOD activity of larval L. calcarifer from the 5 mg/Lexperimental group after 36 h was significantly higher than the control group(P<0.05,the same as below). SOD activity of the individuals in 10 mg/L experimental group was significantly lower than that in the control group at all other time points except for 72 h. POD activity of larval L. calcarifer from 5 mg/L experimental group was always higher than the control group with the extension of stress time,and POD activity of 10 mg/L group were greatly higher than the control group but lower than 5 mg/L group from 6 h to 36 h of stress. CAT activity of the fish from experimental group was significantly higher than the control group in the whole process. CAT activity of 10 mg/L treatment group fluctuated greatly,showing an alternating trend of up-down. In the 5 mg/L treatment group,ACP activity of larval L. calcarifer decreased firstly and then increased,and was significantly lower than the control group after the start of experiment at all time points,but in the treatment group of 10 mg/L, the overall trend of increase-decrease-increase-decrease was presented.GSH-Px activity of the fish in the experimental groups of 5 and 10 mg/L were basically consistent,which remained stable firstly,and then exhibited the trend of rise-fall-rise again during the stress process. In terms of digestive enzymes,LPS activity of larval L. calcarifer from 5 mg/L experimental group was significantly lower than the control group after 36 h of stress,but the group of 10 mg/L exhibited the trend of increase-decrease-increase-decrease since 12 h of stress. AMS activity of larval L. calcarifer in experimental groups of 5 and 10 mg/L exhibited the trend of fall-rise. TRYP activity of the fish from experimental groups of 5 and 10 mg/L exhibited rise and fall alternately.【Conclusion】Ammonia nitrogen stress greatly affects the digestive enzyme and antioxidant enzyme of larval L. calcarifer. Larval L. calcarifer exhibits a certain tolerance to ammonia nitrogen within short time(<6 h). Some antioxidant enzymes of larval L. calcarifer such as SOD,CAT induces under the dramatic changes of ammonia nitrogen to eliminate excessive free radicals. Digestive enzyme activities exhibits different rising levels in order to replenish the energy consumption.
【Objective】The effect of ammonia nitrogen on the digestive enzyme activities and antioxidant enzyme activities of larval Lates calcarifer was clarified,providing basis for improving the survival rate of larval L. calcarifer and enriching the ecological theory of breeding.【Method】The acute stress of ammonia nitrogen on larval L. calcarifer(15 days post hatching)was conducted. Antioxidant enzyme activities such as superoxide dismutase(SOD),peroxidase(POD),catalase(CAT),acid phosphatase(ACP),glutathione peroxidase(GSH-Px)and digestive enzyme activities such as lipase(LPS),amylase(AMS),trypsin(TRYP)were measured in the stress time of 0,6,12,24,36,48,72 and 96 h to reveal the change regulation of the activities of antioxidant enzymes and digestive enzymes in larval L. calcarifer under acute ammonia stress.【Result】Under stress of ammonia nitrogen,SOD activity of larval L. calcarifer from the 5 mg/Lexperimental group after 36 h was significantly higher than the control group(P<0.05,the same as below). SOD activity of the individuals in 10 mg/L experimental group was significantly lower than that in the control group at all other time points except for 72 h. POD activity of larval L. calcarifer from 5 mg/L experimental group was always higher than the control group with the extension of stress time,and POD activity of 10 mg/L group were greatly higher than the control group but lower than 5 mg/L group from 6 h to 36 h of stress. CAT activity of the fish from experimental group was significantly higher than the control group in the whole process. CAT activity of 10 mg/L treatment group fluctuated greatly,showing an alternating trend of up-down. In the 5 mg/L treatment group,ACP activity of larval L. calcarifer decreased firstly and then increased,and was significantly lower than the control group after the start of experiment at all time points,but in the treatment group of 10 mg/L, the overall trend of increase-decrease-increase-decrease was presented.GSH-Px activity of the fish in the experimental groups of 5 and 10 mg/L were basically consistent,which remained stable firstly,and then exhibited the trend of rise-fall-rise again during the stress process. In terms of digestive enzymes,LPS activity of larval L. calcarifer from 5 mg/L experimental group was significantly lower than the control group after 36 h of stress,but the group of 10 mg/L exhibited the trend of increase-decrease-increase-decrease since 12 h of stress. AMS activity of larval L. calcarifer in experimental groups of 5 and 10 mg/L exhibited the trend of fall-rise. TRYP activity of the fish from experimental groups of 5 and 10 mg/L exhibited rise and fall alternately.【Conclusion】Ammonia nitrogen stress greatly affects the digestive enzyme and antioxidant enzyme of larval L. calcarifer. Larval L. calcarifer exhibits a certain tolerance to ammonia nitrogen within short time(<6 h). Some antioxidant enzymes of larval L. calcarifer such as SOD,CAT induces under the dramatic changes of ammonia nitrogen to eliminate excessive free radicals. Digestive enzyme activities exhibits different rising levels in order to replenish the energy consumption.
引文
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Newton J R,De Santis C,Jerry D R.2012.The gene expression response of the catadromous perciform barramundi Lates calcarifer to an acute heat stress[J].Journal of Fish Biology,81(1):81-93.
Paust L O,Foss A,Imsland A K.2011.Effects of chronic and periodic exposure to ammonia on growth,food conversion efficiency and blood physiology in juvenile Atlantic halibut(Hippoglossus hippoglossus L.)[J].Aquaculture,315(3):400-406.
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Sun H J,LüK,MINTER E J A,Chen Y,Yang Z,Montagnes D J S.2012.Combined effects of ammonia and microcystin on survival,growth,antioxidant responses,and lipid peroxidation of bighead carp Hypophthalmythys nobilis larvae[J].Journal of Harzardous Materials,221-222:213-219.
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