脱氧雪腐镰刀菌烯醇对断奶仔猪生长性能、血清细胞因子及肠道免疫水平的影响
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摘要
[目的]本试验旨在通过脱氧雪腐镰刀菌烯醇(DON)体内试验,根据仔猪血清细胞因子水平、肠黏膜sIgA水平及肠道免疫球蛋白mRNA相对表达量的变化,研究DON对断奶仔猪免疫机制的影响。[方法]选取30头21日龄"杜×长×大"三元杂交断奶仔猪,随机分为对照组、DON低剂量组和DON高剂量组,对照组饲喂基础饲粮,低剂量组和高剂量组分别饲喂含1和2 mg·kg-1DON的基础饲粮。试验期60 d。在试验开始后的0、14、28、42和60 d,所有仔猪经前腔静脉采血,检测血清细胞因子IFN-γ、IL-2、IL-4和IL-6水平;试验结束时,每组随机选取5头仔猪剖杀,取十二指肠、空肠和回肠,检测肠道黏膜sIgA水平及肠组织IgA、IgM、IgG mRNA相对表达量。[结果]DON能显著或极显著降低仔猪平均日采食量与平均日增重,增加料重比(P<0.05,P<0.01);显著降低血清IFN-γ、IL-2、IL-4、IL-6水平和小肠黏膜sIgA水平及小肠组织IgA、IgM、IgG mRNA相对表达量(P<0.05),其中DON高剂量组的作用效果比低剂量组更加明显。[结论]DON可以降低仔猪的生长性能、血清细胞因子及肠道免疫水平,对断奶仔猪具有一定的免疫毒性作用。
[Objectives]An experiment was conducted to investigate the immunotoxicity of piglets in vivo exposed by deoxynivalenol(DON),conferring to the levels of serum cytokines,sIgA and mRNA relative expression of intestinal immunoglobulin in weanling piglets.[Methods]Thirty crossbred piglets "Duroc × Landrace×Yorkshire"(21 days old) were selected and randomly divided into three groups of ten piglets each.Piglets in control group were fed with the basal diet,low and high dosage of DON groups were fed with 1 and 2 mg·kg-1 DON diet.The trial was conducted for 60 days.Blood samples were collected from each piglet anterior vena cava for measuring serum IFN-γ,IL-2,I L-4,IL-6 levels on 0,14,21,28,42 and 60 d.At the end of experiment,five piglets from each group were sacrificed,and the intestinal mucosa was collected to detect the levels of sIgA.Each segment of the intestine was collected to determine the relative expression of IgA,IgM,IgG mRNA.[Results]Compared with the control group,DON decreased average daily gain(ADG) and average daily feed intake(ADFI) of piglets significantly(P<0.05 or P<0.01) and increased the ratio of feed to gain significantly(P<0.05 or P<0.01);DON reduced the above four kinds of serum cytokines,and decreased the sIgA levels in intestinal mucosa and IgA,IgM and IgG mRNA relative expression in small intestinal tissues significantly(P<0.05).The negative effect of high dosage of DON group was potent.[Conclusions]DON could reduce growth performance,serum cytokine and intestinal immune level in weanling piglets.These results demonstrate that DON has immunotoxicity to the piglets.
[Objectives]An experiment was conducted to investigate the immunotoxicity of piglets in vivo exposed by deoxynivalenol(DON),conferring to the levels of serum cytokines,sIgA and mRNA relative expression of intestinal immunoglobulin in weanling piglets.[Methods]Thirty crossbred piglets "Duroc × Landrace×Yorkshire"(21 days old) were selected and randomly divided into three groups of ten piglets each.Piglets in control group were fed with the basal diet,low and high dosage of DON groups were fed with 1 and 2 mg·kg-1 DON diet.The trial was conducted for 60 days.Blood samples were collected from each piglet anterior vena cava for measuring serum IFN-γ,IL-2,I L-4,IL-6 levels on 0,14,21,28,42 and 60 d.At the end of experiment,five piglets from each group were sacrificed,and the intestinal mucosa was collected to detect the levels of sIgA.Each segment of the intestine was collected to determine the relative expression of IgA,IgM,IgG mRNA.[Results]Compared with the control group,DON decreased average daily gain(ADG) and average daily feed intake(ADFI) of piglets significantly(P<0.05 or P<0.01) and increased the ratio of feed to gain significantly(P<0.05 or P<0.01);DON reduced the above four kinds of serum cytokines,and decreased the sIgA levels in intestinal mucosa and IgA,IgM and IgG mRNA relative expression in small intestinal tissues significantly(P<0.05).The negative effect of high dosage of DON group was potent.[Conclusions]DON could reduce growth performance,serum cytokine and intestinal immune level in weanling piglets.These results demonstrate that DON has immunotoxicity to the piglets.
引文
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[8]许伟,范梦雪,孟婷婷,等.脱氧雪腐镰刀菌烯醇对PC12细胞凋亡的影响[J].南京农业大学学报,2016,29(5):838-844.DOI:10.7685/jnau.201508040.Xu W,Fan M X,Meng T T,et al.Effect of deoxynivalenol on apoptosis of PC12 cells[J].Journal of Nanjing Agricultural University,2016,29(5):838-844(in Chinese with English abstract).
[9]Danicke S,Brosig B,Klunker L R,et al.Systemic and local effects of the Fusarium toxin deoxynivalenol(DON)are not alleviated by dietary supplementation of humic substances(HS)[J].Food and Chemical Toxicology,2012,50(3):979-988.
[10]Ren Z H,Wang Y C,Deng H D,et al.Deoxynivalenol-induced cytokines and related genes in concanavalin A-stimulated primary chicken splenic lymphocytes[J].Toxicology in Vitro,2015,29(3):558-563.
[11]张婷,朱风华,高晨,等.脱氧雪腐镰刀菌烯醇对小鼠免疫功能的影响[J].中国兽医杂志,2014,50(9):28-30.Zhang T,Zhu F H,Gao C,et al.Effects of DON on the immune function in mice[J].Chinese Journal of Veterinary Medicine,2014,50(9):28-30(in Chinese with English abstract).
[12]姜云晶,许伟,陈晓芳,等.脱氧雪腐镰刀菌烯醇提取、纯化及含量检测[J].中国兽医学报,2017,37(9):1771-1777.Jiang Y J,Xu W,Chen X F,et al.Extraction,purification and concentration determination of deoxynivalenol[J].Chinese Journal of Veterinary Science,2017,37(9):1771-1777(in Chinese with English abstract).
[13]耿芳芳,许伟,范梦雪,等.脱氧雪腐镰刀烯醇对雏鸡脂质过氧化反应及脑部形态结构的影响[J].南京农业大学学报,2016,39(3):460-466.DOI:10.7685/jnau.201510024.Geng F F,Xu W,Fan M X,et al.Effects of deoxynivalenol on lipid peroxidation reaction and brain morphology in chicken[J].Journal of Nanjing Agricultural University,2016,39(3):460-466(in Chinese with English abstract).
[14]D9ll S,Dnicke S.The fusarium toxins deoxynivalenol(DON)and zearalenone(ZEN)in animal feeding[J].Preventive Veterinary Medicine,2011,102(2):132-145.
[15]蒋竹英,范觉鑫,陈明洪,等.脱氧雪腐镰刀菌烯醇污染饲粮对断奶仔猪血液生理生化指标的影响及竹炭和竹醋液的干预作用[J].动物营养学报,2012,24(12):2459-2468.Jiang Z Y,Fan J X,Chen M H,et al.Effects of diets contaminated by deoxynivalenol on blood physiological and biochemical indexes and intervention effects of bamboo-carbon and bamboo vinegar in weaner piglets[J].Chinese Journal of Animal Nutrition,2012,24(12):2459-2468(in Chinese with English abstract).
[16]Mingari M C,Gerosa F,Carra G,et al.Human interleukin-2 promotes proliferation of activated B cells via surface receptors similar to those of activated T cells[J].Nature,1984,312(5995):641-643.
[17]Yan D,Zhou H R,Brooks K H,et al.Potential role for IL-5 and IL-6 in enhanced Ig A secretion by Peyer's patch cells isolated from mice acutely exposed to vomitoxin[J].Toxicology,1997,122(1/2):145-158.
[18]蔡国栋,潘顺叶,王凤丽,等.ZEA、DON及其联合染毒对CTLL-2细胞因子分泌的影响[J].中国畜牧兽医,2017,44(5):1533-1540.Cai G D,Pan S Y,Wang F L,et al.Effects of ZEA,DON and their combined effects on secretion of cytokine in CTLL-2 cells[J].China Animal Husbandry&Veterinary Medicine,2017,44(5):1533-1540(in Chinese with English abstract).
[19]李茜,岑雪,潘顺叶,等.脱氧雪腐镰刀菌烯醇诱导CTLL-2细胞凋亡的信号途径研究[J].畜牧与兽医,2016,48(12):12-18.Li Q,Cen X,Pan S Y,et al.Deoxynivalenol-induced pathway of apoptosis in CTLL-2 cells[J].Animal Husbandry&Veterinary Medicine,2016,48(12):12-18(in Chinese with English abstract).
[20]Brandtzaeg P.Role of secretory antibodies in the defence against infections[J].International Journal of Medical Microbiology,2003,293(1):3-15.
[21]Wijburg O L C,Uren T K,Simpfendorfer K,et al.Innate secretory antibodies protect against natural Salmonella typhimurium infection[J].Journal of Experimental Medicine,2006,203(1):21-26.
[22]左涛,律迎春,曹露,等.冰岛参对小鼠肠道黏膜SIgA分泌的影响[J].食品工业科技,2012,33(1):373-375.Zuo T,LüY C,Cao L,et al.Promotion effect of Cucumaria frondosa on the secretion of mice intestinal SIg A[J].Science and Technology of Food Industry,2012,33(1):373-375(in Chinese with English abstract).
[23]宫佳杰,耿芳芳,许伟,等.DON暴露对雏鸡免疫功能的影响[J].中国兽医学报,2016,36(11):1923-1928.Gong J J,Geng F F,Xu W,et al.Effects of DON exposure on immunity function in chicken[J].Chinese Journal of Veterinary Science,2016,36(11):1923-1928(in Chinese with English abstract).
[24]Ren Z H,Zhou R,Deng J L,et al.Effects of the Fusarium toxin zearalenone(ZEA)and/or deoxynivalenol(DON)on the serum Ig A,Ig G and Ig Mlevels in mice[J].Food and Agricultural Immunology,2014,25(4):600-606.
[25]Abbès S,Salah-Abbès J B,Ouanes Z,et al.Preventive role of phyllosilicate clay on the immunological and biochemical toxicity of zearalenone in Balb/c mice[J].International Immunopharmacology,2006,6(8):1251-1258.
[2]Ebrahem M,Kersten S,Valenta H,et al.Effects of feeding deoxynivalenol(DON)-contaminated wheat to laying hens and roosters of different genetic background on the reproductive performance and health of the newly hatched chicks[J].Mycotoxin Research,2014,30(3):131-140.
[3]Ren Z H,Wang Y C,Deng H D,et al.Deoxynivalenol induces apoptosis in chicken splenic lymphocytes via the reactive oxygen species-mediated mitochondrial pathway[J].Environmental Toxicology and Pharmacology,2015,39(1):339-346.
[4]He C H,Fan Y H,Wang Y,et al.The individual and combined effects of deoxynivalenol and aflatoxin b1on primary hepatocytes of Cyprinus carpio[J].International Journal of Molecular Sciences,2010,11(10):3760-3768.
[5]史建荣,刘馨,仇剑波,等.小麦中镰刀菌毒素脱氧雪腐镰刀菌烯醇污染现状与防控研究进展[J].中国农业科学,2014,47(18):3641-3654.Shi J R,Liu X,Qiu J B,et al.Deoxynivalenol contamination in wheat and its management[J].Scientia Agricultura Sinica,2014,47(18):3641-3654(in Chinese with English abstract).
[6]王伟,朱江辉,邵兵,等.中国居民小麦粉与玉米制品中脱氧雪腐镰刀烯醇与玉米赤霉烯酮暴露水平评估[J].中国预防医学杂志,2015,49(3):223-227.Wang W,Zhu J H,Shao B,et al.Probabilistic assessment of dietary exposure to both deoxynivalenol and zearalenone from cerea-based products in Chinese populations[J].Chinese Journal of Preventive Medicine,2015,49(3):223-227(in Chinese with English abstract).
[7]杨雪丽,张格祥,杨勤德,等.新疆市售小麦粉中脱氧雪腐镰刀菌烯醇的污染水平调查[J].中国卫生检验杂志,2016,26(4):578-581.Yang X L,Zhang G X,Yang Q D,et al.Contamination survey of the deoxynivalenols in wheat flour from in the market of Xinjiang[J].Chinese Journal of Health Laboratory Technology,2016,26(4):578-581(in Chinese with English abstract).
[8]许伟,范梦雪,孟婷婷,等.脱氧雪腐镰刀菌烯醇对PC12细胞凋亡的影响[J].南京农业大学学报,2016,29(5):838-844.DOI:10.7685/jnau.201508040.Xu W,Fan M X,Meng T T,et al.Effect of deoxynivalenol on apoptosis of PC12 cells[J].Journal of Nanjing Agricultural University,2016,29(5):838-844(in Chinese with English abstract).
[9]Danicke S,Brosig B,Klunker L R,et al.Systemic and local effects of the Fusarium toxin deoxynivalenol(DON)are not alleviated by dietary supplementation of humic substances(HS)[J].Food and Chemical Toxicology,2012,50(3):979-988.
[10]Ren Z H,Wang Y C,Deng H D,et al.Deoxynivalenol-induced cytokines and related genes in concanavalin A-stimulated primary chicken splenic lymphocytes[J].Toxicology in Vitro,2015,29(3):558-563.
[11]张婷,朱风华,高晨,等.脱氧雪腐镰刀菌烯醇对小鼠免疫功能的影响[J].中国兽医杂志,2014,50(9):28-30.Zhang T,Zhu F H,Gao C,et al.Effects of DON on the immune function in mice[J].Chinese Journal of Veterinary Medicine,2014,50(9):28-30(in Chinese with English abstract).
[12]姜云晶,许伟,陈晓芳,等.脱氧雪腐镰刀菌烯醇提取、纯化及含量检测[J].中国兽医学报,2017,37(9):1771-1777.Jiang Y J,Xu W,Chen X F,et al.Extraction,purification and concentration determination of deoxynivalenol[J].Chinese Journal of Veterinary Science,2017,37(9):1771-1777(in Chinese with English abstract).
[13]耿芳芳,许伟,范梦雪,等.脱氧雪腐镰刀烯醇对雏鸡脂质过氧化反应及脑部形态结构的影响[J].南京农业大学学报,2016,39(3):460-466.DOI:10.7685/jnau.201510024.Geng F F,Xu W,Fan M X,et al.Effects of deoxynivalenol on lipid peroxidation reaction and brain morphology in chicken[J].Journal of Nanjing Agricultural University,2016,39(3):460-466(in Chinese with English abstract).
[14]D9ll S,Dnicke S.The fusarium toxins deoxynivalenol(DON)and zearalenone(ZEN)in animal feeding[J].Preventive Veterinary Medicine,2011,102(2):132-145.
[15]蒋竹英,范觉鑫,陈明洪,等.脱氧雪腐镰刀菌烯醇污染饲粮对断奶仔猪血液生理生化指标的影响及竹炭和竹醋液的干预作用[J].动物营养学报,2012,24(12):2459-2468.Jiang Z Y,Fan J X,Chen M H,et al.Effects of diets contaminated by deoxynivalenol on blood physiological and biochemical indexes and intervention effects of bamboo-carbon and bamboo vinegar in weaner piglets[J].Chinese Journal of Animal Nutrition,2012,24(12):2459-2468(in Chinese with English abstract).
[16]Mingari M C,Gerosa F,Carra G,et al.Human interleukin-2 promotes proliferation of activated B cells via surface receptors similar to those of activated T cells[J].Nature,1984,312(5995):641-643.
[17]Yan D,Zhou H R,Brooks K H,et al.Potential role for IL-5 and IL-6 in enhanced Ig A secretion by Peyer's patch cells isolated from mice acutely exposed to vomitoxin[J].Toxicology,1997,122(1/2):145-158.
[18]蔡国栋,潘顺叶,王凤丽,等.ZEA、DON及其联合染毒对CTLL-2细胞因子分泌的影响[J].中国畜牧兽医,2017,44(5):1533-1540.Cai G D,Pan S Y,Wang F L,et al.Effects of ZEA,DON and their combined effects on secretion of cytokine in CTLL-2 cells[J].China Animal Husbandry&Veterinary Medicine,2017,44(5):1533-1540(in Chinese with English abstract).
[19]李茜,岑雪,潘顺叶,等.脱氧雪腐镰刀菌烯醇诱导CTLL-2细胞凋亡的信号途径研究[J].畜牧与兽医,2016,48(12):12-18.Li Q,Cen X,Pan S Y,et al.Deoxynivalenol-induced pathway of apoptosis in CTLL-2 cells[J].Animal Husbandry&Veterinary Medicine,2016,48(12):12-18(in Chinese with English abstract).
[20]Brandtzaeg P.Role of secretory antibodies in the defence against infections[J].International Journal of Medical Microbiology,2003,293(1):3-15.
[21]Wijburg O L C,Uren T K,Simpfendorfer K,et al.Innate secretory antibodies protect against natural Salmonella typhimurium infection[J].Journal of Experimental Medicine,2006,203(1):21-26.
[22]左涛,律迎春,曹露,等.冰岛参对小鼠肠道黏膜SIgA分泌的影响[J].食品工业科技,2012,33(1):373-375.Zuo T,LüY C,Cao L,et al.Promotion effect of Cucumaria frondosa on the secretion of mice intestinal SIg A[J].Science and Technology of Food Industry,2012,33(1):373-375(in Chinese with English abstract).
[23]宫佳杰,耿芳芳,许伟,等.DON暴露对雏鸡免疫功能的影响[J].中国兽医学报,2016,36(11):1923-1928.Gong J J,Geng F F,Xu W,et al.Effects of DON exposure on immunity function in chicken[J].Chinese Journal of Veterinary Science,2016,36(11):1923-1928(in Chinese with English abstract).
[24]Ren Z H,Zhou R,Deng J L,et al.Effects of the Fusarium toxin zearalenone(ZEA)and/or deoxynivalenol(DON)on the serum Ig A,Ig G and Ig Mlevels in mice[J].Food and Agricultural Immunology,2014,25(4):600-606.
[25]Abbès S,Salah-Abbès J B,Ouanes Z,et al.Preventive role of phyllosilicate clay on the immunological and biochemical toxicity of zearalenone in Balb/c mice[J].International Immunopharmacology,2006,6(8):1251-1258.