枯草芽孢杆菌铜对先天性缺铜大鼠生长发育、器官指数、铜沉积量、血清生化和抗氧化指标的影响
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摘要
本试验旨在研究枯草芽孢杆菌铜对先天性缺铜大鼠生长发育、器官指数、铜沉积量、血清生化和抗氧化指标的影响。试验分为模型期和干预试验期。模型期选取怀孕10 d的SD大鼠72只,平均分为模型组和对照组,每组6个重复,每个重复6只。模型组大鼠孕期饲喂缺铜饲粮(铜含量为0.33 mg/kg),对照组大鼠孕期饲喂正常饲粮(在缺铜饲粮基础上添加6.0 mg/kg五水硫酸铜),持续到哺乳期结束,验证建模是否成功。干预试验期选择24日建模成功的缺铜幼龄大鼠108只,随机分成6个试验组,Ⅰ～Ⅲ组为硫酸铜组(缺铜饲粮基础上分别添加3.0、6.0和9.0 mg/kg五水硫酸铜),Ⅳ～Ⅵ组为枯草芽孢杆菌铜组(缺铜饲粮基础上分别添加3.0、6.0和9.0 mg/kg枯草芽孢杆菌铜),每个组3个重复,每个重复6只。试验期35 d。结果表明:1)在铜添加量相同条件下,Ⅳ组体重和血清谷丙转氨酶(ALT)、谷胱甘肽过氧化物酶(GSH-Px)活性显著或极显著高于Ⅰ组(P<0.05或P<0.01),Ⅳ组血清谷草转氨酶/谷丙转氨酶(AST/ALT)显著低于Ⅰ组(P<0.05)。Ⅴ组血清铜蓝蛋白(CP)、碱性磷酸酶(AKP)活性显著或极显高于Ⅱ组(P<0.05或P<0.01),Ⅴ组血清丙二醛(MDA)含量显著低于Ⅱ组(P<0.05)。Ⅵ组血清AKP活性和肾脏中铜沉积量显著或极显高于Ⅲ组(P<0.05或P<0.01)。2)在硫酸铜组中,Ⅱ、Ⅲ组体重、体长显著高于Ⅰ组(P<0.05),Ⅲ组血清CP活性显著或极显著高于Ⅰ、Ⅱ组(P<0.05或P<0.01),Ⅲ组血清AKP活性显著高于Ⅱ组(P <0.05),Ⅲ组血清谷草转氨酶(AST)、GSH-Px和铜锌超氧化物歧化酶(CuZn-SOD)活性及肾脏中铜沉积量显著或极显著高于Ⅰ组(P<0.05或P<0.01),Ⅲ组血清M DA含量极显著低于Ⅰ组(P <0.01)。在枯草芽孢杆菌铜组中,Ⅵ组体重、体长,血清AST、GSH-Px和CuZn-SOD活性,心脏、肝脏、肾脏中铜沉积量显著或极显著高于Ⅳ组(P<0.05或P<0.01),Ⅵ组血清CP和AKP活性显著或极显著高于Ⅳ、Ⅴ组(P <0.05或P <0.01)。由此可见,枯草芽孢杆菌铜能提高先天性缺铜大鼠体重,血清CP、AKP、ALT活性,提高机体抗氧化能力,增加肝脏、肾脏中铜沉积量,效果优于硫酸铜,且降低了饲粮中铜添加量。
This experiment was conducted to study the influences of Bacillus subtilis copper on growth and development,organ indices,copper deposition and serum biochemical and antioxidant parameters of congenital copper deficiency rats. The experiment was divided into model period and intervention period. In the model pe-riod,seventy-two pregnant SD rats were divided into 2 groups( model group and control group) with 6 replicates per group and 6 replicates per replicate. The pregnancy rats in the model group were fed a low copper diet( copper content was 0.33 mg/kg),and the others in the control group were fed a normal diet( low copper diets supplemented with 6.0 mg/kg copper sulfate pentahydrate),continue until the end of the lactation period and to verify that the model was successful. In the intervention period,a total of 108 successfully modeled copper deficiency young rats were randomly divided into 6 groups,groups Ⅰ to Ⅲ were copper sulfate groups( low copper diets supplemented with 3.0,6.0 and 9.0 mg/kg copper sulfate pentahydrate,respectively),and groups Ⅳ to Ⅵ were Bacillus subtilis copper groups( low copper diets supplemented with 3. 0,6. 0 and9.0 mg/kg Bacillus subtilis copper,respectively),each group had 3 replicates and each replicate had 6 rates.The experiment lasted for 35 days. The results showed as follows: 1) under the same copper addition condition,the body weight and activities of glutamic-pyruvic transaminase( ALT) and glutathione peroxidase( GSH-Px) in serum of group Ⅳ were significantly higher than those of group Ⅰ( P<0.05 or P<0.01),and the serum glutamic-oxaloacetic transaminase/glutamic-pyruvic transaminase( AST/ALT) of group Ⅳ was significantly lower than that of group Ⅰ( P<0.05). The activities of ceruloplasmin( CP) and alkaline phosphatase( AKP) in serum of group Ⅴ were significantly higher than those of group Ⅱ( P<0.05 or P<0.01),and the serum malonaldehyde( MDA) content of group Ⅴ was significantly lower than that of group Ⅱ( P <0.05). The serum AKP activity and kidney copper deposition of group Ⅵ were significantly higher than those of group Ⅲ( P<0.05 or P<0.01). 2) In copper sulfate groups,the body weight and body length of groups Ⅱand Ⅲ were significantly higher than those of group Ⅰ( P<0.05),the serum CP activity of group Ⅲ was significantly lower than that of groups Ⅰ and Ⅱ( P<0.05 or P<0.01),the serum AKP activity of group Ⅲ was significantly lower than that of group Ⅱ( P < 0. 05),the activities of glutamic-oxaloacetic transaminase( AST),GSH-Px and Cu-Zn superoxide dismutase( CuZn-SOD) in serum and kidney copper deposition of group Ⅲ were significantly higher than those of group Ⅰ( P<0.05 or P<0.01),and the serum MDA content of group Ⅲ was significantly lower than that of group Ⅰ( P<0.05). In Bacillus subtilis copper groups,the body weight,body length and the activities of AST,GSH-Px and CuZn-SOD in serum and the copper deposition in heart,liver and kidney of group Ⅵ were significantly higher than those of group Ⅳ( P<0.05 or P<0.01),the activities of CP and AKP in serum of group Ⅵ were significantly higher than those of groups Ⅳand Ⅴ( P<0.05 or P<0.01). In conclusion,the Bacillus subtilis copper can increase the body weight and the activities of CP,AKP and ALT in serum of congenital copper deficiency rats,improve the body antioxidant capacity,increase the copper deposition in liver and kidney,the effects are better than copper sulfate,and reduce the copper addition in the diet.[Chinese Journal of Animal Nutrition,2019,31( 5) : 2232-2242]
This experiment was conducted to study the influences of Bacillus subtilis copper on growth and development,organ indices,copper deposition and serum biochemical and antioxidant parameters of congenital copper deficiency rats. The experiment was divided into model period and intervention period. In the model pe-riod,seventy-two pregnant SD rats were divided into 2 groups( model group and control group) with 6 replicates per group and 6 replicates per replicate. The pregnancy rats in the model group were fed a low copper diet( copper content was 0.33 mg/kg),and the others in the control group were fed a normal diet( low copper diets supplemented with 6.0 mg/kg copper sulfate pentahydrate),continue until the end of the lactation period and to verify that the model was successful. In the intervention period,a total of 108 successfully modeled copper deficiency young rats were randomly divided into 6 groups,groups Ⅰ to Ⅲ were copper sulfate groups( low copper diets supplemented with 3.0,6.0 and 9.0 mg/kg copper sulfate pentahydrate,respectively),and groups Ⅳ to Ⅵ were Bacillus subtilis copper groups( low copper diets supplemented with 3. 0,6. 0 and9.0 mg/kg Bacillus subtilis copper,respectively),each group had 3 replicates and each replicate had 6 rates.The experiment lasted for 35 days. The results showed as follows: 1) under the same copper addition condition,the body weight and activities of glutamic-pyruvic transaminase( ALT) and glutathione peroxidase( GSH-Px) in serum of group Ⅳ were significantly higher than those of group Ⅰ( P<0.05 or P<0.01),and the serum glutamic-oxaloacetic transaminase/glutamic-pyruvic transaminase( AST/ALT) of group Ⅳ was significantly lower than that of group Ⅰ( P<0.05). The activities of ceruloplasmin( CP) and alkaline phosphatase( AKP) in serum of group Ⅴ were significantly higher than those of group Ⅱ( P<0.05 or P<0.01),and the serum malonaldehyde( MDA) content of group Ⅴ was significantly lower than that of group Ⅱ( P <0.05). The serum AKP activity and kidney copper deposition of group Ⅵ were significantly higher than those of group Ⅲ( P<0.05 or P<0.01). 2) In copper sulfate groups,the body weight and body length of groups Ⅱand Ⅲ were significantly higher than those of group Ⅰ( P<0.05),the serum CP activity of group Ⅲ was significantly lower than that of groups Ⅰ and Ⅱ( P<0.05 or P<0.01),the serum AKP activity of group Ⅲ was significantly lower than that of group Ⅱ( P < 0. 05),the activities of glutamic-oxaloacetic transaminase( AST),GSH-Px and Cu-Zn superoxide dismutase( CuZn-SOD) in serum and kidney copper deposition of group Ⅲ were significantly higher than those of group Ⅰ( P<0.05 or P<0.01),and the serum MDA content of group Ⅲ was significantly lower than that of group Ⅰ( P<0.05). In Bacillus subtilis copper groups,the body weight,body length and the activities of AST,GSH-Px and CuZn-SOD in serum and the copper deposition in heart,liver and kidney of group Ⅵ were significantly higher than those of group Ⅳ( P<0.05 or P<0.01),the activities of CP and AKP in serum of group Ⅵ were significantly higher than those of groups Ⅳand Ⅴ( P<0.05 or P<0.01). In conclusion,the Bacillus subtilis copper can increase the body weight and the activities of CP,AKP and ALT in serum of congenital copper deficiency rats,improve the body antioxidant capacity,increase the copper deposition in liver and kidney,the effects are better than copper sulfate,and reduce the copper addition in the diet.[Chinese Journal of Animal Nutrition,2019,31( 5) : 2232-2242]
引文
[1] GAETKE L M,CHOW C K. Copper toxicity,oxidative stress,and antioxidant nutrients[J]. Toxicology,2003,189(1/2):147-163.
[2] KIM B E,NEVITT T,THIELE D J. Mechanisms for copper acquisition,distribution and regulation[J]. Nature Chemical Biology,2008,4(3):176-185.
[3] JU C F,YU B,ZHU D,et al.The Effect of dietary sulphate copper supplementation on grow th performance,intestinal digestive enzymes and absorptive transporters in w eanling pigs[J]. Journal of Animal Science and Biotechnology,2010(1):35-43.
[4] KUMAR V,KALITA J,BORA H K,et al. Temporal kinetics of organ damage in copper toxicity:a histopathological correlation in rat model[J]. Regulatory Toxicology and Pharmacology,2016,81:372-380.
[5]王聪,董宽虎,刘强,等.蛋氨酸铜对西门塔尔牛日粮养分消化代谢及血液指标的影响[J].中国生态农业学报,2008,16(6):1523-1527.
[6] HUANG Y,YOO J S,KIM H J,et al.The effects of different copper(inorganic and organic)and energy(tallow and glycerol)sources on growth performance,nutrient digestibility,and fecal excretion profiles in grow ing pigs[J].Asian-Australasian Journal of Animal Sciences,2010,23(5):573-579.
[7]刘冬,黄守敏,毕璋友,等.益生菌混合培养富集锌条件优化研究[J].安徽农业科学,2015,43(20):259-262.
[8] REEVES P G,NIELSEN F H,FAHEY G C,Jr.AIN-93 purified diets for laboratory rodents:final report of the American institute of nutrition ad hoc w riting committee on the reformulation of the AIN-76A rodent diet[J]. The Journal of Nutrition,1993,123(11):1939-1951.
[9]吕咸坤.家畜缺乏微量元素铜、锌、锰的症状及防治[J].饲料博览,2017(5):58.
[10] ERGAZ Z,SHOSHANI-DROR D,GUILLEMIN C,et al.The effect of copper deficiency on fetal grow th and liver anti-oxidant capacity in the Cohen diabetic rat model[J]. Toxicology and Applied Pharmacology,2012,265(2):209-220.
[11] DZIEZYC K,LITWIN T,SOBA N'SKA A,et al.Symptomatic copper deficiency in three w ilson’s disease patients treated w ith zinc sulphate[J]. Neurologiai Neurochirurgia Polska,2014,48(3):214-218.
[12]胥彩玉,胥彩霞,刘国华,等.有机与无机微量元素比例对肉仔鸡生长和组织生化指标的影响[J].动物营养学报,2015,27(5):1549-1558.
[13]赵春雨,张天芮,刘博,等.日粮铜来源及水平对仔猪生长性能及组织铜含量的影响[J].家畜生态学报,2015,36(9):46-50.
[14]张名爱,杨文娇,张泽楠,等.饲粮添加枯草芽孢杆菌对5~16周龄五龙鹅肠道发育、微生物菌群结构及血清酶活性的影响[J].动物营养学报,2017,29(9):3175-3183.
[15] SMITH M S. Responses of chicks to dietary supplements of copper sulphate[J]. British Poultry Science,1969,10(2):97-108.
[16]田相迪,朱风华,孙金全,等.不同剂量纳米氧化铜对肉仔鸡生长性能的影响[J].饲料研究,2007(7):41-44.
[17]郑鑫,刘国文,王哲,等.铜对猪原代培养肝细胞中铜蓝蛋白酶活性的影响[J].饲料与畜牧,2006(3):13-16.
[18]赵春雨,张天芮,刘博,等.日粮铜来源及水平对猪血液生化指标及含铜酶活性的影响[J].中国畜牧杂志,2015,51(21):39-44.
[19]钱剑,刘国文,王哲,等.铜对软骨细胞碱性磷酸酶活性影响的实验观察[J].中国兽医杂志,2005,41(3):21-22.
[20] BOAL A K,ROSENZWEIG A C. Structural biology of copper trafficking[J]. Chemical Review s,2009,109(10):4760-4779.
[21] YAN Q,XIE S,ZHU X,et al.Dietary methionine requirement for juvenile rockfish,Sebastes schlegeli[J].Aquaculture Nutrition,2007,13(3):163-169.
[22] LIN L,ZENG X L,ZHANG J. Effect of profenofos poisoning on liver lipid peroxidation and liver function in rabbits[J]. Chinese Journal of Clinical Rehabilitation,2004,8(21):4380-4381.
[23]谭志刚,王墙,林滋,等.铜过载对小鼠肝肾的损伤[J].畜牧与兽医,2015,47(7):88-90.
[24]李万立,罗海吉,查龙应.铜过量小鼠肝损伤模型的建立及各项指标的观察研究[J].热带医学杂志,2008,8(12):1210-1212.
[25]冯杰,刘欣,吴新民,等.酪蛋白铜对仔猪生长及血清铜蓝蛋白和SOD活性的影响[J].中国畜牧杂志,2005,41(3):14-17.
[26] PAYNTER D I. The role of dietary copper,manganese,selenium,and vitamin E in lipid peroxidation in tissues of the rat[J]. Biological Trace Element Research,1980,2(2):121-135.
[27] O’DONOHUE J W,REID M A,VARGHESE A,et al.M icronodular cirrhosis and acute liver failure due to chronic copper self-intoxication[J]. European Journal of Gastroenterology&Hepatology,1993,5(7):561-562.
[28] ARAYA M,OLIVARES M,PIZARRO F,et al. Copper exposure and potential biomarkers of copper metabolism[J].Biometals,2003,16(1):199-204.
[29] CHOLEWI N'SKA E,JU S'KIEWICZ J,OGNIK K.Comparison of the effect of dietary copper nanoparticles and one copper(Ⅱ)salt on the metabolic and immune status in a rat model[J].Journal of Trace Elements in M edicine and Biology,2018,48:111-117.
[30] OZKUL H,KIRKPINAR F,MERT S,et al.Effects of high levels of dietary copper sulfate and copper proteinate on grow th performance,retention for copper and zinc of rats[J]. Journal of Animal and Veterinary Advances,2011,10(11):1373-1377.
[31] ENGLE T E,SPEARS J W.Effects of dietary copper concentration and source on performance and copper status of grow ing and finishing steers[J]. Journal of Animal Science,2000,78(9):2446-2451.
[32]潘娜,朱风华,王友令,等.日粮添加高水平硫酸锌和纳米氧化铜对鸡脏器组织锌铜沉积的影响[J].中国农业科学,2011,44(23):4874-4881.
[2] KIM B E,NEVITT T,THIELE D J. Mechanisms for copper acquisition,distribution and regulation[J]. Nature Chemical Biology,2008,4(3):176-185.
[3] JU C F,YU B,ZHU D,et al.The Effect of dietary sulphate copper supplementation on grow th performance,intestinal digestive enzymes and absorptive transporters in w eanling pigs[J]. Journal of Animal Science and Biotechnology,2010(1):35-43.
[4] KUMAR V,KALITA J,BORA H K,et al. Temporal kinetics of organ damage in copper toxicity:a histopathological correlation in rat model[J]. Regulatory Toxicology and Pharmacology,2016,81:372-380.
[5]王聪,董宽虎,刘强,等.蛋氨酸铜对西门塔尔牛日粮养分消化代谢及血液指标的影响[J].中国生态农业学报,2008,16(6):1523-1527.
[6] HUANG Y,YOO J S,KIM H J,et al.The effects of different copper(inorganic and organic)and energy(tallow and glycerol)sources on growth performance,nutrient digestibility,and fecal excretion profiles in grow ing pigs[J].Asian-Australasian Journal of Animal Sciences,2010,23(5):573-579.
[7]刘冬,黄守敏,毕璋友,等.益生菌混合培养富集锌条件优化研究[J].安徽农业科学,2015,43(20):259-262.
[8] REEVES P G,NIELSEN F H,FAHEY G C,Jr.AIN-93 purified diets for laboratory rodents:final report of the American institute of nutrition ad hoc w riting committee on the reformulation of the AIN-76A rodent diet[J]. The Journal of Nutrition,1993,123(11):1939-1951.
[9]吕咸坤.家畜缺乏微量元素铜、锌、锰的症状及防治[J].饲料博览,2017(5):58.
[10] ERGAZ Z,SHOSHANI-DROR D,GUILLEMIN C,et al.The effect of copper deficiency on fetal grow th and liver anti-oxidant capacity in the Cohen diabetic rat model[J]. Toxicology and Applied Pharmacology,2012,265(2):209-220.
[11] DZIEZYC K,LITWIN T,SOBA N'SKA A,et al.Symptomatic copper deficiency in three w ilson’s disease patients treated w ith zinc sulphate[J]. Neurologiai Neurochirurgia Polska,2014,48(3):214-218.
[12]胥彩玉,胥彩霞,刘国华,等.有机与无机微量元素比例对肉仔鸡生长和组织生化指标的影响[J].动物营养学报,2015,27(5):1549-1558.
[13]赵春雨,张天芮,刘博,等.日粮铜来源及水平对仔猪生长性能及组织铜含量的影响[J].家畜生态学报,2015,36(9):46-50.
[14]张名爱,杨文娇,张泽楠,等.饲粮添加枯草芽孢杆菌对5~16周龄五龙鹅肠道发育、微生物菌群结构及血清酶活性的影响[J].动物营养学报,2017,29(9):3175-3183.
[15] SMITH M S. Responses of chicks to dietary supplements of copper sulphate[J]. British Poultry Science,1969,10(2):97-108.
[16]田相迪,朱风华,孙金全,等.不同剂量纳米氧化铜对肉仔鸡生长性能的影响[J].饲料研究,2007(7):41-44.
[17]郑鑫,刘国文,王哲,等.铜对猪原代培养肝细胞中铜蓝蛋白酶活性的影响[J].饲料与畜牧,2006(3):13-16.
[18]赵春雨,张天芮,刘博,等.日粮铜来源及水平对猪血液生化指标及含铜酶活性的影响[J].中国畜牧杂志,2015,51(21):39-44.
[19]钱剑,刘国文,王哲,等.铜对软骨细胞碱性磷酸酶活性影响的实验观察[J].中国兽医杂志,2005,41(3):21-22.
[20] BOAL A K,ROSENZWEIG A C. Structural biology of copper trafficking[J]. Chemical Review s,2009,109(10):4760-4779.
[21] YAN Q,XIE S,ZHU X,et al.Dietary methionine requirement for juvenile rockfish,Sebastes schlegeli[J].Aquaculture Nutrition,2007,13(3):163-169.
[22] LIN L,ZENG X L,ZHANG J. Effect of profenofos poisoning on liver lipid peroxidation and liver function in rabbits[J]. Chinese Journal of Clinical Rehabilitation,2004,8(21):4380-4381.
[23]谭志刚,王墙,林滋,等.铜过载对小鼠肝肾的损伤[J].畜牧与兽医,2015,47(7):88-90.
[24]李万立,罗海吉,查龙应.铜过量小鼠肝损伤模型的建立及各项指标的观察研究[J].热带医学杂志,2008,8(12):1210-1212.
[25]冯杰,刘欣,吴新民,等.酪蛋白铜对仔猪生长及血清铜蓝蛋白和SOD活性的影响[J].中国畜牧杂志,2005,41(3):14-17.
[26] PAYNTER D I. The role of dietary copper,manganese,selenium,and vitamin E in lipid peroxidation in tissues of the rat[J]. Biological Trace Element Research,1980,2(2):121-135.
[27] O’DONOHUE J W,REID M A,VARGHESE A,et al.M icronodular cirrhosis and acute liver failure due to chronic copper self-intoxication[J]. European Journal of Gastroenterology&Hepatology,1993,5(7):561-562.
[28] ARAYA M,OLIVARES M,PIZARRO F,et al. Copper exposure and potential biomarkers of copper metabolism[J].Biometals,2003,16(1):199-204.
[29] CHOLEWI N'SKA E,JU S'KIEWICZ J,OGNIK K.Comparison of the effect of dietary copper nanoparticles and one copper(Ⅱ)salt on the metabolic and immune status in a rat model[J].Journal of Trace Elements in M edicine and Biology,2018,48:111-117.
[30] OZKUL H,KIRKPINAR F,MERT S,et al.Effects of high levels of dietary copper sulfate and copper proteinate on grow th performance,retention for copper and zinc of rats[J]. Journal of Animal and Veterinary Advances,2011,10(11):1373-1377.
[31] ENGLE T E,SPEARS J W.Effects of dietary copper concentration and source on performance and copper status of grow ing and finishing steers[J]. Journal of Animal Science,2000,78(9):2446-2451.
[32]潘娜,朱风华,王友令,等.日粮添加高水平硫酸锌和纳米氧化铜对鸡脏器组织锌铜沉积的影响[J].中国农业科学,2011,44(23):4874-4881.