蒙脱石、枯草芽孢杆菌及其互作对蛋鸡血浆激素含量、卵巢繁殖相关基因表达和肠道微生物的影响
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摘要
本试验旨在研究蒙脱石(MMT)、枯草芽孢杆菌(BS)及其互作对蛋鸡血浆激素含量、卵巢繁殖相关基因表达和肠道微生物的影响。采用2×2双因子随机设计,选择360只29周龄健康的罗曼蛋鸡,随机分为4组(每组6个重复,每个重复15只),分别饲喂基础饲粮(对照组)、基础饲粮+0.5 g/kg MMT (MMT组)、基础饲粮+5×108CFU/kg BS (BS组)、基础饲粮+0.5 g/kg M M T+5×108CFU/kg BS(M M T+BS组)。预试期7 d,正试期70 d。结果表明:1)与对照组相比,添加MMT显著降低了蛋鸡血浆皮质醇和促肾上腺皮质激素含量(P<0.05),添加BS显著提高了血浆促卵泡激素含量(P<0.05)。2)与对照组相比,添加MMT显著下调了蛋鸡卵巢催乳素受体和促卵泡激素受体mRNA的表达量(P<0.05),添加BS显著降低了胃饥饿素mRNA的表达量(P<0.05),二者对胃饥饿素mRNA的表达量存在显著互作效应(P<0.05)。3)与对照组相比,添加MMT和BS均显著降低了蛋鸡十二指肠产气荚膜梭菌和沙门氏菌的相对含量(P<0.05),添加M M T显著降低了回肠产气荚膜梭菌的相对含量(P<0.05),二者对小肠产气荚膜梭菌、大肠杆菌和沙门氏菌相对含量存在显著交互作用(P<0.05)。可见,饲粮中单独添加MMT或BS及二者联用均能改善蛋鸡肠道菌群,下调卵巢催乳素受体mRNA的表达量;添加MMT能降低血浆应激激素含量,BS可提高血浆促卵泡激素含量;此外,添加BS及其与MMT互作能够下调卵巢胃饥饿素mRNA的表达量。
This study was conducted to evaluate the effects of montmorillonite(MMT),Bacillus subtilis(BS) and their interaction on hormone contents in plasma,reproduction related genes expression in the ovarian and intestinal microflora of laying hens.The trial used a 2×2 factorial random design.A total of 360 healthy29-week-old Lohmann hens were randomly arranged into 4 groups with 6 replicates per group and 15 hens in each replicate,and the hens in those 4 groups were fed a basal diet(group control),the basal diet+0.5 g/kg MMT(MMT group),the basal diet+5×108 CFU/kg BS(BS group),and the basal diet+0.5 g/kg MMT+5×108 CFU/kg BS(MMT +BS group),respectively.The adjustment period lasted for 7 days,and the experimental period lasted for 70 days.The results showed as follows:1) compared with control group,dietary MMT significantly decreased the contents of cortisol and adrenocorticotropic hormone in the plasma of laying hens(P<0.05);dietary BS significantly increased follicle stimulating hormone content in plasma(P<0.05).2) Compared with control group,dietary MMT significantly down-regulated the mRNA expression levels of prolactin receptor and follicle stimulating hormone receptor in the ovary of laying hens(P<0.05);dietary BS significantly decreased the mRNA expression level of ghrelin in the ovary(P<0.05).There was a significant interaction between MMT and BS on the mRNA expression level of ghrelin(P< 0.05).3) Compared with control group,addition of both MMT and BS significantly decreased the relative contents of Clostridium perfringens and Salmonella in the duodenum of laying hens(P<0.05);dietary MMT significantly decreased the relative content of Clostridium perfringens in the ileum(P<0.05).There was a significant interaction between MMT and BS on the relative contents of Clostridium perfringens,Escherichia coli and Salmonella in the small intestine(P<0.05).In conclusion,single or combined addition of MMT and BS can improve the intestinal microflora and down-regulate the mRNA expression level of prolactin receptor in the ovary;dietary MMT can decrease the stress hormone contents in the plasma,and addition of BS can increase the content of follicle stimulating hormone.In addition,BS,BS and MMT interaction can down-regulate the mRNA expression level of ghrelin in the ovary.
This study was conducted to evaluate the effects of montmorillonite(MMT),Bacillus subtilis(BS) and their interaction on hormone contents in plasma,reproduction related genes expression in the ovarian and intestinal microflora of laying hens.The trial used a 2×2 factorial random design.A total of 360 healthy29-week-old Lohmann hens were randomly arranged into 4 groups with 6 replicates per group and 15 hens in each replicate,and the hens in those 4 groups were fed a basal diet(group control),the basal diet+0.5 g/kg MMT(MMT group),the basal diet+5×108 CFU/kg BS(BS group),and the basal diet+0.5 g/kg MMT+5×108 CFU/kg BS(MMT +BS group),respectively.The adjustment period lasted for 7 days,and the experimental period lasted for 70 days.The results showed as follows:1) compared with control group,dietary MMT significantly decreased the contents of cortisol and adrenocorticotropic hormone in the plasma of laying hens(P<0.05);dietary BS significantly increased follicle stimulating hormone content in plasma(P<0.05).2) Compared with control group,dietary MMT significantly down-regulated the mRNA expression levels of prolactin receptor and follicle stimulating hormone receptor in the ovary of laying hens(P<0.05);dietary BS significantly decreased the mRNA expression level of ghrelin in the ovary(P<0.05).There was a significant interaction between MMT and BS on the mRNA expression level of ghrelin(P< 0.05).3) Compared with control group,addition of both MMT and BS significantly decreased the relative contents of Clostridium perfringens and Salmonella in the duodenum of laying hens(P<0.05);dietary MMT significantly decreased the relative content of Clostridium perfringens in the ileum(P<0.05).There was a significant interaction between MMT and BS on the relative contents of Clostridium perfringens,Escherichia coli and Salmonella in the small intestine(P<0.05).In conclusion,single or combined addition of MMT and BS can improve the intestinal microflora and down-regulate the mRNA expression level of prolactin receptor in the ovary;dietary MMT can decrease the stress hormone contents in the plasma,and addition of BS can increase the content of follicle stimulating hormone.In addition,BS,BS and MMT interaction can down-regulate the mRNA expression level of ghrelin in the ovary.
引文
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[2]ROZENBOIMI,TAKO E,GAL-GARBER O,et al.The effect of heat stress on ovarian function of laying hens[J].Poultry Science,2007,86(8):1760-1765.
[3]SAHIN K,KUCUK O.A simple way to reduce heat stress in laying hens as judged by egg laying,body weight gain and biochemical parameters[J].Acta Veterinaria Hungarica,2001,49(4):421-430.
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[5]雷凯.地衣芽孢杆菌对蛋鸡生产性能的影响及机理研究[D].博士学位论文.杭州:浙江大学,2015:55-57.
[6]李天杰.益生菌对蛋鸡生产性能及肠道微生物变化与基因表达研究[D].硕士学位论文.雅安:四川农业大学,2016:54-57.
[7]陈继发,朱瑾,康克浪,等.枯草芽孢杆菌、蒙脱石及其互作对产蛋鸡生产性能、养分表观利用率和肠黏膜养分转运载体基因表达的影响[J]动物营养学报,2018,30(9):3693-3701.
[8]CAO S T,WANG L,JIAO L F,et al.Effects of diosmectite-Lactobacillus acidophilus on grow th performance,intestine microbiota,mucosal architecture of weaned pigs[J].Animal Feed Science and Technology,2016,220:180-186.
[9]CHEN X L,WANG J K,WU Y M,et al.Effects of chemical treatments of rice strawon rumen fermentation characteristics,fibrolytic enzyme activities and populations of liquid-and solid-associated ruminal microbes in vitro[J].Animal Feed Science and Technology,2008,141(1/2):1-14.
[10]杜恩存.百里香酚和香芹酚对肉仔鸡肠上皮屏障和免疫功能的调节作用[D].博士学位论文.北京:中国农业大学,2016:45.
[11]FENG Y N,GONG J S,YU H,et al.Identification of changes in the composition of ileal bacterial microbiota of broiler chickens infected with Clostridium perfringens[J].Veterinary Microbiology,2010,140(1/2):116-121.
[12]PEPE G J,BILLIAR R B,LEAVITT MG,et al.Expression of estrogen receptorsαandβin the baboon fetal ovary[J].Biology of Reproduction,2002,66(4):1054-1060.
[13]GEORGE J W,DILLE E A,HECKERT L L,et al.Current concepts of follicle-stimulating hormone receptor gene regulation[J].Biology of Reproduction,2011,84(1):7-17.
[14]HU Y D,HUANG Q K,ZHU Q,et al.Identification and association of single-nucleotide polymorphisms in gonadotropin-inhibitory hormone(G nIH)gene with egg production traits in Erlang mountainous chickens[J].Genetics and Molecular Research,2015,14(1):294-303.
[15]VIANI I,VOTTERO A,TASSI F,et al.Ghrelin inhibits steroid biosynthesis by cultured granulosa-lutein cells[J].The Journal of Clinical Endocrinology&Metabolism,2008,93(4):1476-1481.
[16]汪峰.鸡PRL、PRLR基因表达及其与繁殖性能的关系[D].硕士学位论文.南京:南京农业大学,2007:26-31.
[17]HU X R,LV G L,CHEN L S,et al.Study on the mechanism of the interaction between montmorillonite and bacterium[J].Acta Pharmaceutica Sinica,2002,37(9):718-720.
[18]HU C H,QIAN Z C,SONG J,et al.Effects of zinc oxide-montmorillonite hybrid on grow th performance,intestinal structure,and function of broiler chicken[J].Poultry Science,2013,92(1):143-150.
[19]XIA MS,HU C H,XU Z R.Effects of copper-bearing montmorillonite on grow th performance,digestive enzyme activities,intestinal microflora and morphology of male broilers[J].Poultry Science,2004,83(11):1868-1875.
[20]孟艳莉.凹凸棒石、蒙脱石及其复合物对断奶仔猪肠道的保护作用研究[D].硕士学位论文.北京:中国农业科学院,2011:24-25.
[21]GUO J R,DONG X F,LIU S,et al.Effects of longterm Bacillus subtilis CGMCC 1.921 supplementation on performance,egg quality,and fecal and cecal microbiota of laying hens[J].Poultry Science,2017,96(5):1280-1289.
[22]GAO Z H,WU H H,SHI L,et al.Study of Bacillus subtilis on grow th performance,nutrition metabolism and intestinal microflora of 1 to 42 d broiler chickens[J].Animal Nutrition,2017,3(2):109-113.
[23]KNAP I,KEHLET A B,BENNEDSEN M,et al.Bacillus subtilis(DSM17299)significantly reduces Salmonella in broilers[J].Poultry Science,2011,90(8):1690-1694.
[24]LA RAGIONE R M,WOODWARD MJ.Competitive exclusion by Bacillus subtilis spores of Salmonella enterica serotype Enteritidis and Clostridium perfringens in young chickens[J].Veterinary Microbiology,2003,94(3):245-256.
[25]LECLRE V,BCHET M,ADAMA,et al.Mycosubtilin overproduction by Bacillus subtilis BBG100 enhances the organism's antagonistic and biocontrol activities[J].Applied and Environmental Microbiology,2005,71(8):4577-4584.
[26]STEIN T.Bacillus subtilis antibiotics:structures,syntheses and specific functions[J].Molecular Microbiology,2005,56(4):845-857.
[27]LI S L,JIANG C X,CHEN X M,et al.Lactobacillus casei immobilized onto montmorillonite:survivability in simulated gastrointestinal conditions,refrigeration and yogurt[J].Food Research International,2014,64:822-830.
[28]ALBENGRES E,URIEN S,TILLEMENT J P,et al.Interactions between smectite,a mucus stabilizer,and acidic and basic drugs:in vitro and in vivo studies[J].European Journal of Clinical Pharmacology,1985,28(5):601-605.
[29]KAMADA N,CHEN G Y,INOHARA N,et al.Control of pathogens and pathobionts by the gut microbiota[J].Nature Immunology,2013,14(7):685-690.
[2]ROZENBOIMI,TAKO E,GAL-GARBER O,et al.The effect of heat stress on ovarian function of laying hens[J].Poultry Science,2007,86(8):1760-1765.
[3]SAHIN K,KUCUK O.A simple way to reduce heat stress in laying hens as judged by egg laying,body weight gain and biochemical parameters[J].Acta Veterinaria Hungarica,2001,49(4):421-430.
[4]LEI K,LI Y,YU D Y,et al.Influence of dietary inclusion of Bacillus licheniformis on laying performance,egg quality,antioxidant enzyme activities,and intestinal barrier function of laying hens[J].Poultry Science,2013,92(9):2389-2395.
[5]雷凯.地衣芽孢杆菌对蛋鸡生产性能的影响及机理研究[D].博士学位论文.杭州:浙江大学,2015:55-57.
[6]李天杰.益生菌对蛋鸡生产性能及肠道微生物变化与基因表达研究[D].硕士学位论文.雅安:四川农业大学,2016:54-57.
[7]陈继发,朱瑾,康克浪,等.枯草芽孢杆菌、蒙脱石及其互作对产蛋鸡生产性能、养分表观利用率和肠黏膜养分转运载体基因表达的影响[J]动物营养学报,2018,30(9):3693-3701.
[8]CAO S T,WANG L,JIAO L F,et al.Effects of diosmectite-Lactobacillus acidophilus on grow th performance,intestine microbiota,mucosal architecture of weaned pigs[J].Animal Feed Science and Technology,2016,220:180-186.
[9]CHEN X L,WANG J K,WU Y M,et al.Effects of chemical treatments of rice strawon rumen fermentation characteristics,fibrolytic enzyme activities and populations of liquid-and solid-associated ruminal microbes in vitro[J].Animal Feed Science and Technology,2008,141(1/2):1-14.
[10]杜恩存.百里香酚和香芹酚对肉仔鸡肠上皮屏障和免疫功能的调节作用[D].博士学位论文.北京:中国农业大学,2016:45.
[11]FENG Y N,GONG J S,YU H,et al.Identification of changes in the composition of ileal bacterial microbiota of broiler chickens infected with Clostridium perfringens[J].Veterinary Microbiology,2010,140(1/2):116-121.
[12]PEPE G J,BILLIAR R B,LEAVITT MG,et al.Expression of estrogen receptorsαandβin the baboon fetal ovary[J].Biology of Reproduction,2002,66(4):1054-1060.
[13]GEORGE J W,DILLE E A,HECKERT L L,et al.Current concepts of follicle-stimulating hormone receptor gene regulation[J].Biology of Reproduction,2011,84(1):7-17.
[14]HU Y D,HUANG Q K,ZHU Q,et al.Identification and association of single-nucleotide polymorphisms in gonadotropin-inhibitory hormone(G nIH)gene with egg production traits in Erlang mountainous chickens[J].Genetics and Molecular Research,2015,14(1):294-303.
[15]VIANI I,VOTTERO A,TASSI F,et al.Ghrelin inhibits steroid biosynthesis by cultured granulosa-lutein cells[J].The Journal of Clinical Endocrinology&Metabolism,2008,93(4):1476-1481.
[16]汪峰.鸡PRL、PRLR基因表达及其与繁殖性能的关系[D].硕士学位论文.南京:南京农业大学,2007:26-31.
[17]HU X R,LV G L,CHEN L S,et al.Study on the mechanism of the interaction between montmorillonite and bacterium[J].Acta Pharmaceutica Sinica,2002,37(9):718-720.
[18]HU C H,QIAN Z C,SONG J,et al.Effects of zinc oxide-montmorillonite hybrid on grow th performance,intestinal structure,and function of broiler chicken[J].Poultry Science,2013,92(1):143-150.
[19]XIA MS,HU C H,XU Z R.Effects of copper-bearing montmorillonite on grow th performance,digestive enzyme activities,intestinal microflora and morphology of male broilers[J].Poultry Science,2004,83(11):1868-1875.
[20]孟艳莉.凹凸棒石、蒙脱石及其复合物对断奶仔猪肠道的保护作用研究[D].硕士学位论文.北京:中国农业科学院,2011:24-25.
[21]GUO J R,DONG X F,LIU S,et al.Effects of longterm Bacillus subtilis CGMCC 1.921 supplementation on performance,egg quality,and fecal and cecal microbiota of laying hens[J].Poultry Science,2017,96(5):1280-1289.
[22]GAO Z H,WU H H,SHI L,et al.Study of Bacillus subtilis on grow th performance,nutrition metabolism and intestinal microflora of 1 to 42 d broiler chickens[J].Animal Nutrition,2017,3(2):109-113.
[23]KNAP I,KEHLET A B,BENNEDSEN M,et al.Bacillus subtilis(DSM17299)significantly reduces Salmonella in broilers[J].Poultry Science,2011,90(8):1690-1694.
[24]LA RAGIONE R M,WOODWARD MJ.Competitive exclusion by Bacillus subtilis spores of Salmonella enterica serotype Enteritidis and Clostridium perfringens in young chickens[J].Veterinary Microbiology,2003,94(3):245-256.
[25]LECLRE V,BCHET M,ADAMA,et al.Mycosubtilin overproduction by Bacillus subtilis BBG100 enhances the organism's antagonistic and biocontrol activities[J].Applied and Environmental Microbiology,2005,71(8):4577-4584.
[26]STEIN T.Bacillus subtilis antibiotics:structures,syntheses and specific functions[J].Molecular Microbiology,2005,56(4):845-857.
[27]LI S L,JIANG C X,CHEN X M,et al.Lactobacillus casei immobilized onto montmorillonite:survivability in simulated gastrointestinal conditions,refrigeration and yogurt[J].Food Research International,2014,64:822-830.
[28]ALBENGRES E,URIEN S,TILLEMENT J P,et al.Interactions between smectite,a mucus stabilizer,and acidic and basic drugs:in vitro and in vivo studies[J].European Journal of Clinical Pharmacology,1985,28(5):601-605.
[29]KAMADA N,CHEN G Y,INOHARA N,et al.Control of pathogens and pathobionts by the gut microbiota[J].Nature Immunology,2013,14(7):685-690.