α-酮戊二酸对氨氮胁迫下草鱼鳃Na~+/K~+-ATP酶活性及血液生化指标的影响
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摘要
试验旨在研究饲粮添加α-酮戊二酸(α-ketoglutarate,α-KG)对氨氮胁迫下草鱼(Ctenopharyngodon idellus)鳃Na~+/K~+-ATP酶活性及血液生化指标的影响。选取初始体重为(24.79±0.11)g的健康草鱼,随机分为3个处理组(对照组,养于曝气后氨氮浓度为1.51 mg/L的自来水中并饲喂基础饲粮;氨氮组,养于氨氮浓度为18.37 mg/L的水中并饲喂基础饲粮;α-KG组,养于氨氮浓度为18.37 mg/L的水中并饲喂添加0.75%α-KG的饲粮)。每处理组设3个重复,每个重复30尾鱼,养殖试验为期42d,分别于第1、第14、第28和第42天采样。结果表明:在饲料中添加α-KG能够有效缓解氨氮胁迫导致的草鱼血浆氨含量(1d)、谷草转氨酶(Aspartate transaminase,AST)活性(14d)、碱性磷酸酶(Alkaline phosphatase,ALP)活性(28d)的显著升高和鳃Na~+/K~+-ATP酶活性(28d)、血浆谷丙转氨酶(Alanine aminotransaminase,ALT)(28d和42d)活性、血浆尿素(UREA)含量(28d)的显著降低,显著增加氨氮胁迫下草鱼血浆球蛋白(Globulin,GLB)含量(28d)。即饲粮α-KG的适量添加能够有效缓解草鱼氨氮胁迫所致的血氨含量升高,维持氨氮胁迫下草鱼鳃Na~+/K~+-ATP酶、血浆谷丙转氨酶、谷草转氨酶、碱性磷酸酶的活性和血浆球蛋白、尿素含量的稳定,从而有利于草鱼缓解氨氮胁迫。
A 42-day feeding trial was conducted to investigate the effects of α-ketoglutarate(α-KG) on gill Na~+/K~+-ATPase activity and blood biochemical parameters of grass carp, Ctenopharyngodon idellus, exposure to ammonia. A total of 270 individuals [initial body weight:(24.79±0.11) g] were randomly assigned to 9 tanks, and these tanks were divided into three groups(Ⅰ: treated with 1.51 mg/L of aerated water and basal diet; Ⅱ: treated with 18.37 mg/L of ammonia solution and basal diet; Ⅲ: treated with 18.37 mg/L of ammonia solution and 0.75% α-KG added diet), with three replicates per each group(n=30). Experimental fish were sampled on 1 st, 14 th, 28 th, and 42 days, respectively.The results showed that:(1) diet α-KG could effectively alleviate a significant increase in the content of plasma ammonia on the 1 st day, the activity of plasma aspartate transaminase on 14 th day and plasma alkaline phosphatase on 28 th day, which were caused as being exposure to ammonia; diet α-KG could effectively alleviate a significant decrease in the activity of gill Na~+/K~+-ATPase on 28 th day, plasma alanine aminotransferase on 28 th and 42 th days, and the content of plasma urea on 28 th day because of being exposure to ammonia. Diet α-KG could significantly increase plasma globulin content on 28 th day exposure to ammonia. In conclusion, dietary α-KG could alleviate the increase of plasma ammonia content exposure to ammonia, and maintain the stabilization in the activity of gill Na~+/K~+-ATPase, plasma alanine transaminase(ALT), plasma aspartate aminotransferase(AST), plasma alkaline phosphatase(ALP), and the content of GLB, as well as urea in grass carp exposure to ammonia.
A 42-day feeding trial was conducted to investigate the effects of α-ketoglutarate(α-KG) on gill Na~+/K~+-ATPase activity and blood biochemical parameters of grass carp, Ctenopharyngodon idellus, exposure to ammonia. A total of 270 individuals [initial body weight:(24.79±0.11) g] were randomly assigned to 9 tanks, and these tanks were divided into three groups(Ⅰ: treated with 1.51 mg/L of aerated water and basal diet; Ⅱ: treated with 18.37 mg/L of ammonia solution and basal diet; Ⅲ: treated with 18.37 mg/L of ammonia solution and 0.75% α-KG added diet), with three replicates per each group(n=30). Experimental fish were sampled on 1 st, 14 th, 28 th, and 42 days, respectively.The results showed that:(1) diet α-KG could effectively alleviate a significant increase in the content of plasma ammonia on the 1 st day, the activity of plasma aspartate transaminase on 14 th day and plasma alkaline phosphatase on 28 th day, which were caused as being exposure to ammonia; diet α-KG could effectively alleviate a significant decrease in the activity of gill Na~+/K~+-ATPase on 28 th day, plasma alanine aminotransferase on 28 th and 42 th days, and the content of plasma urea on 28 th day because of being exposure to ammonia. Diet α-KG could significantly increase plasma globulin content on 28 th day exposure to ammonia. In conclusion, dietary α-KG could alleviate the increase of plasma ammonia content exposure to ammonia, and maintain the stabilization in the activity of gill Na~+/K~+-ATPase, plasma alanine transaminase(ALT), plasma aspartate aminotransferase(AST), plasma alkaline phosphatase(ALP), and the content of GLB, as well as urea in grass carp exposure to ammonia.
引文
[1]Chen J C,Liu P C,Lin Y T.Super intensive culture of red-tailed Shrimp Penaeus penicillatus[J].Journal of the World Aquaculture Society,1988,19(3):127-131
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[3]Qian J H,Li Z M,Liu J Y,et al.Acute toxicity of ammonia-N and its effect on the immune parameters of Fenneropenaeus merguiensis[J].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]
[4]Schram E,Roques J A C,Kuijk T V,et al.The impact of elevated water ammonia and nitrate concentrations on physiology,growth and feed intake of pikeperch(Sander lucioperca)[J].Aquaculture,2014,420-421(2):95-104
[5]Junghans P,Derno M,Pierzynowski S,et al.Intraduodenal infusion ofα-ketoglutarate decreases whole body energy expenditure in growing pigs[J].Clinical Nutrition,2006,25(3):489-496
[6]Fu D B,Wang Y W,Hou Y Q,et al.The effects of alphaketoglutarate on the muscle energy metabolism of weaned piglets were stimulated by lipopolysaccharide[J].Chinese Agricultural Science,2011,44(4):814-822[付大波,王友炜,侯永清,等.α-酮戊二酸对脂多糖刺激断奶仔猪肌肉能量代谢的影响.中国农业科学,2011,44(4):814-822]
[7]Wei Y Q.The effects of alpha-ketoglutarate on the metabolism,antioxidant capacity and intestinal mucosal mor-phology and function of pinpo microcarp[D].Thesis for Master of Science.Shanghai Ocean University,Shanghai.2015[魏玉强.α-酮戊二酸对松浦镜鲤氮代谢、抗氧化能力和小肠黏膜形态与功能的影响.硕士学位论文,上海海洋大学,上海.2015]
[8]Hu Q Z,Hou Y Q,Ding B Y,et al.The effects of alphaketoglutarate on the morphology and function of small intestine histology[J].Journal of Animal Nutrition,2008,20(6):662-667[胡泉舟,侯永清,丁斌鹰,等.α-酮戊二酸对仔猪小肠组织学形态与功能的影响.动物营养学报,2008,20(6):662-667]
[9]Ai F,Wang L S,Li J N,et al.Effects ofα-ketoglutarate supplementation in low-phosphorous diets on growth performance,body composition and serum biochemical indexes of Songpu mirror carp[J].Acta Hydrobiologica Sinica,2018,42(3):525-532[艾凤,王连生,李晋南,等.低磷饲料中添加α-酮戊二酸对松浦镜鲤生长性能、体成分和血清生化指标的影响.水生生物学报,2018,42(3):525-532]
[10]Yan L Q,Liao M,Xie J Q,et al.The effects of l-arginine and alpha-ketoglutarate on intestinal absorption,antioxidant capacity and energy metabolism of hot stress chicken[J].China Journal of Animal Husbandry,2016,52(15):33-41[晏利琼,廖满,谢佳倩,等.L-精氨酸和α-酮戊二酸对热应激肉鸡肠道吸收功能、抗氧化能力、能量代谢的影响.中国畜牧杂志,2016,52(15):33-41]
[11]Huang Y,Hu Y,Wen H,et al.Effects of vitamin E on the growth,immunity and ammonia stress of young fish[J].Acta Hydrobiologica Sinica,2013,37(3):507-514[黄云,胡毅,文华,等.维生素E对青鱼幼鱼生长、免疫及抗氨氮胁迫能力的影响.水生生物学报,2013,37(3):507-514]
[12]Wang L,Hou Y Q,Ding W Y,et al.The effects of alphaketoglutarate on the plasma insulin,amino acid content and the m TOR signaling pathway of the gastrocnemius muscle[J].Journal of Animal Nutrition,2010,22(6):1724-1729[王蕾,侯永清,丁斌鹰,等.α-酮戊二酸对脂多糖应激仔猪血浆胰岛素、氨基酸含量及腓肠肌m TOR信号传导途径的影响.动物营养学报,2010,22(6):1724-1729]
[13]Li S F,Liu H X,Zhang Y B,et al.The protective effects of alpha-ketoacids against oxidative stress on rat spermatozoa in vitro[J].Asian Journal of Andrology,2010,12(2):247-256
[14]Liang J.Study on the mitigation effect of alpha-ketoglutarate on the ammonia nitrogen stress of grass carp[D].Thesis for Master of Science.Hunan Agricultural University,Changsha.2014[梁健.α-酮戊二酸对草鱼氨氮胁迫的缓解作用研究.硕士学位论文,湖南农业大学,长沙.2014]
[15]Wang H Q,Zhao Y R,Jin B T,et al.Effects of dietary alpha-ketoglutarate supplementation on growth and serum biochemical parameters of grass carp(Ctenopharyngodon idella)fingerling[J].The Israeli Journal of Aquaculture-Bamidgeh,IJA_68.2016.1239
[16]Zhou Y X,Zhang P Y,Zhou R Z.The acute and subacute toxicity to ammonia on grass carp(Ctenopharyngodon idellus)[J].Acta Hydrobiologica Sinica,1986,10(1):32-38[周永欣,张甫英,周仁珍.氨对草鱼的急性和亚急性毒性.水生生物学报,1986,10(1):32-38]
[17]Teng E J,Liu T L,An H.Conversion method of nonionic ammonia for surface water environmental quality standard[J].Environmental monitoring in china,1995,11(4):9-21[滕恩江,刘廷良,安华.地面水环境质量标准非离子氨换算方法.中国环境监测,1995,11(4):9-21]
[18]Pagliarani A,Bandiera P,Ventrella V,et al.Response of Na+-dependent ATPase activities to the contaminant ammonia nitrogen in tapes philippinarum:possible ATPase involvement in ammonium transport[J].Archives of Environmental Contamination&Toxicology,2008,55(1):49-56
[19]Zhao F,Zhuang P,Zhang L Z,et al.The influence of salinity domestication on the activity of Na+/K+-ATP enzyme activity,serum osmotic pressure and ion concentration[J].Journal of Fisheries,2006,30(4):444-449[赵峰,庄平,章龙珍,等.盐度驯化对史氏鲟鳃Na+/K+-ATP酶活力、血清渗透压及离子浓度的影响.水产学报,2006,30(4):444-449]
[20]Wei Y,Xiang F H,Sun H J,et al.Changes in the selected hematological parameters and gill Na+/K+-ATPase activity of juvenile crucian carp Carassius auratus during elevated ammonia exposure and the post-exposure recovery[J].Biochemical Systematics&Ecology,2010,38(4):557-562
[21]Francois N R L,Savoie A,Siikavuopio S I.Gill metabolic and osmoregulatory responses of juvenile Atlantic cod(Gadus morhua)to chronic ammonia exposure[J].Journal of Applied Ichthyology,2008,24(1):112-115
[22]Alam M,Frankel T L.Gill ATPase activities of silverperch,Bidyanus bidyanus(Mitchell),and golden perch Macquaria ambigua(Richardson):Effects of environmental salt and ammonia[J].Aquaculture,2006,251(1)118-133
[23]Wei Y Y,Xu Q Y,Li J N,et al.The effects of alpha-ketoglutarate on the growth performance,body composition and serum biochemical indexes of the pteroscope were added to different protein levels[J].Journal of Animal Nutrition,2013,25(12):2958-2965[位莹莹,徐奇友,李晋南,等.不同蛋白质水平饲料中添加α-酮戊二酸对松浦镜鲤生长性能、体成分和血清生化指标的影响.动物营养学报,2013,25(12):2958-2965]
[24]Zhang L,Fan Q X,Zhao Z G,et al.Effects of chronic congestion stress on the growth of carp and blood biochemical indexes[J].Journal of Dalian Ocean University,2007,22(6):465-469[张磊,樊启学,赵志刚,等.慢性拥挤胁迫对鲤生长及血液生化指标的影响.大连海洋大学学报,2007,22(6):465-469]
[25]Hou Y E,Feng J,Yang Q H,et al.Progress in the study of fish immunoglobulin[J].Heilongjiang Animal Husbandry Veterinarian,2010,(17):37-39[侯月娥,冯娟,杨清华,等.鱼类免疫球蛋白的研究进展.黑龙江畜牧兽医,2010,(17):37-39]
[26]Xu Y,Xiao W,Li D Y,et al.Effects of chronic ammonia-nitrogen stress on the growth and physiological function of nirotifish larvae[J].Journal of Southern Agriculture,2015,46(2):327-331[徐杨,肖炜,李大宇,等.慢性氨氮胁迫对尼罗罗非鱼幼鱼生长及生理功能的影响.南方农业学报,2015,46(2):327-331]
[27]Zhang X Q,Xu D S,Li X Q,et al.The effects of neutral phytase on the growth of the growth and the apparent digestibility,deposition rate and plasma biochemical indexes of the growth and nutrient content of the carbium were added[J].Journal of Animal Nutrition,2016,28(5):1598-1608[张晓清,徐树德,李小勤,等.不同磷酸二氢钙含量饲料中添加中性植酸酶对异育银鲫生长及营养物质表观消化率、沉积率和血浆生化指标的影响.动物营养学报,2016,28(5):1598-1608]
[28]Jayakumar P,Jothivel N,Paul V.Heavy metals induced alterations in the acid phosphatase activity in the edible freshwater mussel Lamellidens marginalis(Lamarck)[J].Internet Journal of Toxicology,2007,5(2):1-8
[2]Braissant O,Mclin V A,Cudalbu C.Ammonia toxicity to the brain[J].Journal of Inherited Metabolic Disease,2013,36(4):595-612
[3]Qian J H,Li Z M,Liu J Y,et al.Acute toxicity of ammonia-N and its effect on the immune parameters of Fenneropenaeus merguiensis[J].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]
[4]Schram E,Roques J A C,Kuijk T V,et al.The impact of elevated water ammonia and nitrate concentrations on physiology,growth and feed intake of pikeperch(Sander lucioperca)[J].Aquaculture,2014,420-421(2):95-104
[5]Junghans P,Derno M,Pierzynowski S,et al.Intraduodenal infusion ofα-ketoglutarate decreases whole body energy expenditure in growing pigs[J].Clinical Nutrition,2006,25(3):489-496
[6]Fu D B,Wang Y W,Hou Y Q,et al.The effects of alphaketoglutarate on the muscle energy metabolism of weaned piglets were stimulated by lipopolysaccharide[J].Chinese Agricultural Science,2011,44(4):814-822[付大波,王友炜,侯永清,等.α-酮戊二酸对脂多糖刺激断奶仔猪肌肉能量代谢的影响.中国农业科学,2011,44(4):814-822]
[7]Wei Y Q.The effects of alpha-ketoglutarate on the metabolism,antioxidant capacity and intestinal mucosal mor-phology and function of pinpo microcarp[D].Thesis for Master of Science.Shanghai Ocean University,Shanghai.2015[魏玉强.α-酮戊二酸对松浦镜鲤氮代谢、抗氧化能力和小肠黏膜形态与功能的影响.硕士学位论文,上海海洋大学,上海.2015]
[8]Hu Q Z,Hou Y Q,Ding B Y,et al.The effects of alphaketoglutarate on the morphology and function of small intestine histology[J].Journal of Animal Nutrition,2008,20(6):662-667[胡泉舟,侯永清,丁斌鹰,等.α-酮戊二酸对仔猪小肠组织学形态与功能的影响.动物营养学报,2008,20(6):662-667]
[9]Ai F,Wang L S,Li J N,et al.Effects ofα-ketoglutarate supplementation in low-phosphorous diets on growth performance,body composition and serum biochemical indexes of Songpu mirror carp[J].Acta Hydrobiologica Sinica,2018,42(3):525-532[艾凤,王连生,李晋南,等.低磷饲料中添加α-酮戊二酸对松浦镜鲤生长性能、体成分和血清生化指标的影响.水生生物学报,2018,42(3):525-532]
[10]Yan L Q,Liao M,Xie J Q,et al.The effects of l-arginine and alpha-ketoglutarate on intestinal absorption,antioxidant capacity and energy metabolism of hot stress chicken[J].China Journal of Animal Husbandry,2016,52(15):33-41[晏利琼,廖满,谢佳倩,等.L-精氨酸和α-酮戊二酸对热应激肉鸡肠道吸收功能、抗氧化能力、能量代谢的影响.中国畜牧杂志,2016,52(15):33-41]
[11]Huang Y,Hu Y,Wen H,et al.Effects of vitamin E on the growth,immunity and ammonia stress of young fish[J].Acta Hydrobiologica Sinica,2013,37(3):507-514[黄云,胡毅,文华,等.维生素E对青鱼幼鱼生长、免疫及抗氨氮胁迫能力的影响.水生生物学报,2013,37(3):507-514]
[12]Wang L,Hou Y Q,Ding W Y,et al.The effects of alphaketoglutarate on the plasma insulin,amino acid content and the m TOR signaling pathway of the gastrocnemius muscle[J].Journal of Animal Nutrition,2010,22(6):1724-1729[王蕾,侯永清,丁斌鹰,等.α-酮戊二酸对脂多糖应激仔猪血浆胰岛素、氨基酸含量及腓肠肌m TOR信号传导途径的影响.动物营养学报,2010,22(6):1724-1729]
[13]Li S F,Liu H X,Zhang Y B,et al.The protective effects of alpha-ketoacids against oxidative stress on rat spermatozoa in vitro[J].Asian Journal of Andrology,2010,12(2):247-256
[14]Liang J.Study on the mitigation effect of alpha-ketoglutarate on the ammonia nitrogen stress of grass carp[D].Thesis for Master of Science.Hunan Agricultural University,Changsha.2014[梁健.α-酮戊二酸对草鱼氨氮胁迫的缓解作用研究.硕士学位论文,湖南农业大学,长沙.2014]
[15]Wang H Q,Zhao Y R,Jin B T,et al.Effects of dietary alpha-ketoglutarate supplementation on growth and serum biochemical parameters of grass carp(Ctenopharyngodon idella)fingerling[J].The Israeli Journal of Aquaculture-Bamidgeh,IJA_68.2016.1239
[16]Zhou Y X,Zhang P Y,Zhou R Z.The acute and subacute toxicity to ammonia on grass carp(Ctenopharyngodon idellus)[J].Acta Hydrobiologica Sinica,1986,10(1):32-38[周永欣,张甫英,周仁珍.氨对草鱼的急性和亚急性毒性.水生生物学报,1986,10(1):32-38]
[17]Teng E J,Liu T L,An H.Conversion method of nonionic ammonia for surface water environmental quality standard[J].Environmental monitoring in china,1995,11(4):9-21[滕恩江,刘廷良,安华.地面水环境质量标准非离子氨换算方法.中国环境监测,1995,11(4):9-21]
[18]Pagliarani A,Bandiera P,Ventrella V,et al.Response of Na+-dependent ATPase activities to the contaminant ammonia nitrogen in tapes philippinarum:possible ATPase involvement in ammonium transport[J].Archives of Environmental Contamination&Toxicology,2008,55(1):49-56
[19]Zhao F,Zhuang P,Zhang L Z,et al.The influence of salinity domestication on the activity of Na+/K+-ATP enzyme activity,serum osmotic pressure and ion concentration[J].Journal of Fisheries,2006,30(4):444-449[赵峰,庄平,章龙珍,等.盐度驯化对史氏鲟鳃Na+/K+-ATP酶活力、血清渗透压及离子浓度的影响.水产学报,2006,30(4):444-449]
[20]Wei Y,Xiang F H,Sun H J,et al.Changes in the selected hematological parameters and gill Na+/K+-ATPase activity of juvenile crucian carp Carassius auratus during elevated ammonia exposure and the post-exposure recovery[J].Biochemical Systematics&Ecology,2010,38(4):557-562
[21]Francois N R L,Savoie A,Siikavuopio S I.Gill metabolic and osmoregulatory responses of juvenile Atlantic cod(Gadus morhua)to chronic ammonia exposure[J].Journal of Applied Ichthyology,2008,24(1):112-115
[22]Alam M,Frankel T L.Gill ATPase activities of silverperch,Bidyanus bidyanus(Mitchell),and golden perch Macquaria ambigua(Richardson):Effects of environmental salt and ammonia[J].Aquaculture,2006,251(1)118-133
[23]Wei Y Y,Xu Q Y,Li J N,et al.The effects of alpha-ketoglutarate on the growth performance,body composition and serum biochemical indexes of the pteroscope were added to different protein levels[J].Journal of Animal Nutrition,2013,25(12):2958-2965[位莹莹,徐奇友,李晋南,等.不同蛋白质水平饲料中添加α-酮戊二酸对松浦镜鲤生长性能、体成分和血清生化指标的影响.动物营养学报,2013,25(12):2958-2965]
[24]Zhang L,Fan Q X,Zhao Z G,et al.Effects of chronic congestion stress on the growth of carp and blood biochemical indexes[J].Journal of Dalian Ocean University,2007,22(6):465-469[张磊,樊启学,赵志刚,等.慢性拥挤胁迫对鲤生长及血液生化指标的影响.大连海洋大学学报,2007,22(6):465-469]
[25]Hou Y E,Feng J,Yang Q H,et al.Progress in the study of fish immunoglobulin[J].Heilongjiang Animal Husbandry Veterinarian,2010,(17):37-39[侯月娥,冯娟,杨清华,等.鱼类免疫球蛋白的研究进展.黑龙江畜牧兽医,2010,(17):37-39]
[26]Xu Y,Xiao W,Li D Y,et al.Effects of chronic ammonia-nitrogen stress on the growth and physiological function of nirotifish larvae[J].Journal of Southern Agriculture,2015,46(2):327-331[徐杨,肖炜,李大宇,等.慢性氨氮胁迫对尼罗罗非鱼幼鱼生长及生理功能的影响.南方农业学报,2015,46(2):327-331]
[27]Zhang X Q,Xu D S,Li X Q,et al.The effects of neutral phytase on the growth of the growth and the apparent digestibility,deposition rate and plasma biochemical indexes of the growth and nutrient content of the carbium were added[J].Journal of Animal Nutrition,2016,28(5):1598-1608[张晓清,徐树德,李小勤,等.不同磷酸二氢钙含量饲料中添加中性植酸酶对异育银鲫生长及营养物质表观消化率、沉积率和血浆生化指标的影响.动物营养学报,2016,28(5):1598-1608]
[28]Jayakumar P,Jothivel N,Paul V.Heavy metals induced alterations in the acid phosphatase activity in the edible freshwater mussel Lamellidens marginalis(Lamarck)[J].Internet Journal of Toxicology,2007,5(2):1-8