不同物候期牧草对放牧牦牛瘤胃内环境参数及瘤胃微生物多样性的影响
|
摘要
本试验在测定不同物候期牧草营养价值的基础上,研究不同物候期牧草对放牧牦牛瘤胃内环境参数及瘤胃微生物多样性的影响。分别于返青期、青草期和枯草期采集牧草样品,测定牧草常规营养成分含量。选取平均体重为(108.06±2.86) kg的10只3岁全年放牧的健康青藏高原牦牛(公母各5只)进行序号标记,分别在返青期、青草期和枯草期清晨放牧前空腹经口腔采集瘤胃液,用于分析瘤胃内环境参数和瘤胃微生物多样性。结果显示:牧草中粗蛋白质含量在青草期显著高于返青期和枯草期(P<0.05),中性洗涤纤维含量在枯草期高于青草期和返青期(P>0.05)。瘤胃液氨态氮、乙酸、异丁酸、丁酸、总挥发性脂肪酸含量均在青草期最高;瘤胃液总挥发性脂肪酸含量在枯草期显著低于青草期和返青期(P<0.05)。不同物候期牧草对瘤胃微生物多样性有影响,青草期瘤胃微生物多样性及丰富度最低。拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)及普雷沃氏菌属1(Prevotella_1)的相对丰度在3个物候期均较高;青草期厚壁菌门的相对丰度显著高于返青期和枯草期(P<0.05);在种水平上,瘤胃微生物以纤维素降解菌为主。由此得出,不同物候期牧草常规营养成分含量存在差异,不同物候期牧草常规营养成分含量的变化引起了牦牛瘤胃内环境参数和瘤胃微生物多样性的变化;青草期瘤胃液氨态氮及总挥发性脂肪酸含量最高,在枯草期瘤胃液总挥发性性脂肪酸含量下降,能量源减少,因此在枯草期应对放牧牦牛进行补饲才能保证牦牛生长。
This experiment was aimed to study the effects of forage grass in different phenological periods on ruminal environment parameters and rumen microbial diversity of free-range yak(Bos grunniens) based on the measurement of nutritive value of forage grass in different phenological periods.The forage grass samples were collected during the regreen stage,grassy stage and withering stage,and the common nutrient contents were determined.Ten healthy year-round free-range Qinghai and Tibetan plateau yak(5 males and females) with the body weight of(108.06±2.86) kg and at the age of 3 years were marked,and the rumen fluids of them were collected via oral cavity before free-range with empty stomach in regreen stage,grassy stage and withering stage to measured the ruminal environment parameters and rumen microbial diversity,respectively.The results showed as follows:the crude protein content of forage grass in grassy stage was significantly higher than that in regreen stage and withering stage(P<0.05),and the neutral detergent fiber content of forage grass in withering stage was higher than that in grassy stage and regreen stage(P>0.05).The contents of ammonia nitrogen,acetic acid,isobutyric acid,butyric acid,total volatile acid reached the highest values in the grassy stage,and the content of total volatile fatty acids in withering stage was significantly lower than that in grassy stage and regreen stage(P<0.05).Different phenological periods had effects on rumen microbial diversity.Rumen microbial richness and diversity were the lowest in grassy stage.The relative abundances of Bacteroidetes,Firmicutes and Prevotella_1 were all higher in the three phenological periods.The relative abundance of Firmicutes in grassy stage was significantly higher than that in regreen stage and withering stage(P<0.05).The rumen microbe mainly on cellulose degradation bacteria at species level.The bacteria detected were mainly cellulose degradation bacteria.Thus,the common nutrient contents of forage grass are different in different phenological periods,and the changes of common nutrient contents of forage grass in different phenological periods are the factors that resulted in the changes in rumen environment parameters and rumen microbial diversity of yak,the contents of ammonia nitrogen and total volatile fatty acids in rumen fluid are the highest in grassy stage,but the content of total volatile fatty acids in rumen fluid has decreased in withering stage,therefore it is necessary for yak to have supplementary feeding in withering stage to maintain their growth cause of the reducing energy source.
This experiment was aimed to study the effects of forage grass in different phenological periods on ruminal environment parameters and rumen microbial diversity of free-range yak(Bos grunniens) based on the measurement of nutritive value of forage grass in different phenological periods.The forage grass samples were collected during the regreen stage,grassy stage and withering stage,and the common nutrient contents were determined.Ten healthy year-round free-range Qinghai and Tibetan plateau yak(5 males and females) with the body weight of(108.06±2.86) kg and at the age of 3 years were marked,and the rumen fluids of them were collected via oral cavity before free-range with empty stomach in regreen stage,grassy stage and withering stage to measured the ruminal environment parameters and rumen microbial diversity,respectively.The results showed as follows:the crude protein content of forage grass in grassy stage was significantly higher than that in regreen stage and withering stage(P<0.05),and the neutral detergent fiber content of forage grass in withering stage was higher than that in grassy stage and regreen stage(P>0.05).The contents of ammonia nitrogen,acetic acid,isobutyric acid,butyric acid,total volatile acid reached the highest values in the grassy stage,and the content of total volatile fatty acids in withering stage was significantly lower than that in grassy stage and regreen stage(P<0.05).Different phenological periods had effects on rumen microbial diversity.Rumen microbial richness and diversity were the lowest in grassy stage.The relative abundances of Bacteroidetes,Firmicutes and Prevotella_1 were all higher in the three phenological periods.The relative abundance of Firmicutes in grassy stage was significantly higher than that in regreen stage and withering stage(P<0.05).The rumen microbe mainly on cellulose degradation bacteria at species level.The bacteria detected were mainly cellulose degradation bacteria.Thus,the common nutrient contents of forage grass are different in different phenological periods,and the changes of common nutrient contents of forage grass in different phenological periods are the factors that resulted in the changes in rumen environment parameters and rumen microbial diversity of yak,the contents of ammonia nitrogen and total volatile fatty acids in rumen fluid are the highest in grassy stage,but the content of total volatile fatty acids in rumen fluid has decreased in withering stage,therefore it is necessary for yak to have supplementary feeding in withering stage to maintain their growth cause of the reducing energy source.
引文
[1]LESLIE D M,Jr,SCHALLER G B.Bos grunniens and Bos mutus(Artiodactyla:Bovidae)[J].Mammalian Species,2009,836(4):1-17.
[2]徐琴.饲粮和品种对山羊瘤胃微生物多样性的影响[D].硕士学位论文.雅安:四川农业大学,2015.
[3]淡瑞芳,张海涛,龙瑞军,等.藏系绵羊瘤胃细菌数量及其放牧地牧草养分随季节变化的研究[J].草业学报,2009,18(1):100-104.
[4]赵辛,韩正康.灌注和日粮添喂挥发性脂肪酸及其盐类对山羊瘤胃代谢及自由采食量的影响[J].南京农业大学学报,1986,9(1):74-83.
[5]王月,王志新,李金泉,等.季节和地区对内蒙古白绒山羊瘤胃发酵特性的影响[J].中国畜牧杂志,2015,51(7):65-69.
[6]张丽英.饲料分析及饲料质量检测技术[M].3版.北京:中国农业大学出版社,2007:48-50.
[7]AOAC.Official methods of analysis of AOAC[S].15th ed.Arlington:Association of Official Analytical Chemists,1990.
[8]VAN SOEST P J,ROBERTSON J B,LEWIS B A.Methods for dietary fiber,neutral detergent fiber,and nonstarch polysaccharides in relation to animal nutrition[J].Journal of Dairy Science,1991,74(10):3583-3597.
[9]BRODERICK G A,KANG J H.Automated simultaneous determination of ammonia and total amino acids in ruminal fluid and in vitro media[J].Journal of Dairy Science,1980,63(1):64-75.
[10]BUXTON D R.Quality-related characteristics of forages as influenced by plant environment and agronomic factors[J].Animal Feed Science and Technology,1996,59(1/2/3):37-49.
[11]余苗,钟荣珍,周道玮,等.不同生育期虎尾草的体外发酵产气特性[J].草业科学,2014,31(5):956-964.
[12]LI C Q,ALATENGDALAI,XUE S Y,et al.Estimation of herbage intake and digestibility of grazing sheep in Zhenglan banner of inner Mongolia by using n-alkanes[J].Animal Nutrition,2015,1(4):324-328.
[13]张盼盼,薛树媛,金海,等.不同物候期牧草对放牧绵羊瘤胃发酵指标的影响[J].中国畜牧兽医,2018,45(3):698-704.
[14]杜雪燕,柴沙驼,王迅,等.河南县高山嵩草草地牧草营养价值与载畜量研究[J].河南农业科学,2015,44(11):141-146.
[15]红敏.不同生长阶段天然牧草品质评定比较研究[J].饲料研究,2012(12):79-81.
[16]SHARP W M,JOHNSON R R,OWENS F N.Ruminal VFA production with steers fed whole or ground corn grain[J].Journal of Animal Science,1982,55(6):1505-1514.
[17]淡瑞芳,张海涛,龙瑞军,等.季节变化对放牧藏系绵羊瘤胃发酵特性及产甲烷菌的影响[J].西北农业学报,2013,22(2):1-6.
[18]魏德泳,朱伟云,毛胜勇.日粮不同NFC/NDF比对山羊瘤胃发酵与瘤胃微生物区系结构的影响[J].中国农业科学,2012,45(7):1392-1398.
[19]李小娜,高艳霞,李秋凤,等.盛花期甘农3号苜蓿干草营养成分及瘤胃降解特性研究[J].中国畜牧杂志,2012,48(12):54-58.
[20]MCDONALD P,EDWARDS R A,GREENHALGH JF D.Animal nutrition[M].4th ed.New York:Longman,1988.
[21]凌虹,陈红跃,官继承.日粮碳水化合物结构对瘤胃发酵及消化代谢影响的研究进展[J].中国畜牧兽医,2012,39(9):134-137.
[22]常维毅,郑雪,王晓旭,等.不同季节日粮组成对狍瘤胃内环境的影响[J].中国兽医学报,2012,32(11):1712-1715.
[23]郭春华,张均,王康宁,等.高寒草地生物量及牧草养分含量年度动态研究[J].中国草地学报,2007,29(1):1-5.
[24]韩正康,陈伟华,陆天水.三种饲养制度下山羊瘤胃内pH、氨氮和蛋白质含量的变化[J].南京农业大学学报,1982,5(3):118-124.
[25]申军士.日粮能氮释放同步性对奶牛瘤胃代谢、生产效率与性能的影响研究[D].博士学位论文.杭州:浙江大学,2013.
[26]魏园,赵晨旭,李心慰,等.不同粗纤维源日粮对梅花鹿瘤胃微生物区系的影响[J].中国兽医学报,2018,38(1):217-221,229.
[27]QIN J J,LI R Q,RAES J,et al.A human gut microbial gene catalogue established by metagenomic sequencing[J].Nature,2010,464(7285):59-65.
[28]LEY R E,LOZUPONE C A,HAMADY M,et al.Worlds within worlds:evolution of the vertebrate gut microbiota[J].Nature Reviews Microbiology,2008,6(10):776-788.
[29]EVANS N J,BROWN J M,MURRAY R D,et al.Characterization of novel bovine gastrointestinal tract Treponema isolates and comparison with bovine digital dermatitis treponemes[J].Applied and Environmental Microbiology,2011,77(1):138-147.
[30]MATSUI H,OGATA K,TAJIMA K,et al.Phenotypic characterization of polysaccharidases produced by four Prevotella type strains[J].Current Microbiology,2000,41(1):45-49.
[31]MEYER M,STENZEL U,HOFREITER M.Parallel tagged sequencing on the 454 platform[J].Nature Protocols,2008,3(2):267-278.
[32]TAJIMA K,AMINOV R I,NAGAMINE T,et al.Dietdependent shifts in the bacterial population of the rumen revealed with real-time PCR[J].Applied and Environmental Microbiology,2001,67(6):2766-2774.
[33]李莉,沈明华,俞红贤,等.含1/4野血犊牦牛血红蛋白、肌红蛋白含量的测定[J].青海大学学报(自然科学版),2005,23(6):42-44.
[34]徐俊.不同牧草来源的NDF在瘤胃中降解特性及其对细菌群落结构的影响[D].博士学位论文.扬州:扬州大学,2014.
[35]LATHAM M J,BROOKER B E,PETTIPHER G L,et al.Adhesion of Bacteroides succinogenes in pure culture and in the presence of Ruminococcus flavefaciens to cell walls in leaves of perennial ryegrass(Lolium perenne)[J].Applied and Environmnetal Microbiology,1978,35(6):1166-1173.
[36]FONDEVILA M,DEHORITY B A.Interactions between Fibrobacter succinogenes,Prevotella ruminicola,and Ruminococcus flavefaciens in the digestion of cellulose from forages[J].Journal of Animal Science,1996,74(3):678-684.
[2]徐琴.饲粮和品种对山羊瘤胃微生物多样性的影响[D].硕士学位论文.雅安:四川农业大学,2015.
[3]淡瑞芳,张海涛,龙瑞军,等.藏系绵羊瘤胃细菌数量及其放牧地牧草养分随季节变化的研究[J].草业学报,2009,18(1):100-104.
[4]赵辛,韩正康.灌注和日粮添喂挥发性脂肪酸及其盐类对山羊瘤胃代谢及自由采食量的影响[J].南京农业大学学报,1986,9(1):74-83.
[5]王月,王志新,李金泉,等.季节和地区对内蒙古白绒山羊瘤胃发酵特性的影响[J].中国畜牧杂志,2015,51(7):65-69.
[6]张丽英.饲料分析及饲料质量检测技术[M].3版.北京:中国农业大学出版社,2007:48-50.
[7]AOAC.Official methods of analysis of AOAC[S].15th ed.Arlington:Association of Official Analytical Chemists,1990.
[8]VAN SOEST P J,ROBERTSON J B,LEWIS B A.Methods for dietary fiber,neutral detergent fiber,and nonstarch polysaccharides in relation to animal nutrition[J].Journal of Dairy Science,1991,74(10):3583-3597.
[9]BRODERICK G A,KANG J H.Automated simultaneous determination of ammonia and total amino acids in ruminal fluid and in vitro media[J].Journal of Dairy Science,1980,63(1):64-75.
[10]BUXTON D R.Quality-related characteristics of forages as influenced by plant environment and agronomic factors[J].Animal Feed Science and Technology,1996,59(1/2/3):37-49.
[11]余苗,钟荣珍,周道玮,等.不同生育期虎尾草的体外发酵产气特性[J].草业科学,2014,31(5):956-964.
[12]LI C Q,ALATENGDALAI,XUE S Y,et al.Estimation of herbage intake and digestibility of grazing sheep in Zhenglan banner of inner Mongolia by using n-alkanes[J].Animal Nutrition,2015,1(4):324-328.
[13]张盼盼,薛树媛,金海,等.不同物候期牧草对放牧绵羊瘤胃发酵指标的影响[J].中国畜牧兽医,2018,45(3):698-704.
[14]杜雪燕,柴沙驼,王迅,等.河南县高山嵩草草地牧草营养价值与载畜量研究[J].河南农业科学,2015,44(11):141-146.
[15]红敏.不同生长阶段天然牧草品质评定比较研究[J].饲料研究,2012(12):79-81.
[16]SHARP W M,JOHNSON R R,OWENS F N.Ruminal VFA production with steers fed whole or ground corn grain[J].Journal of Animal Science,1982,55(6):1505-1514.
[17]淡瑞芳,张海涛,龙瑞军,等.季节变化对放牧藏系绵羊瘤胃发酵特性及产甲烷菌的影响[J].西北农业学报,2013,22(2):1-6.
[18]魏德泳,朱伟云,毛胜勇.日粮不同NFC/NDF比对山羊瘤胃发酵与瘤胃微生物区系结构的影响[J].中国农业科学,2012,45(7):1392-1398.
[19]李小娜,高艳霞,李秋凤,等.盛花期甘农3号苜蓿干草营养成分及瘤胃降解特性研究[J].中国畜牧杂志,2012,48(12):54-58.
[20]MCDONALD P,EDWARDS R A,GREENHALGH JF D.Animal nutrition[M].4th ed.New York:Longman,1988.
[21]凌虹,陈红跃,官继承.日粮碳水化合物结构对瘤胃发酵及消化代谢影响的研究进展[J].中国畜牧兽医,2012,39(9):134-137.
[22]常维毅,郑雪,王晓旭,等.不同季节日粮组成对狍瘤胃内环境的影响[J].中国兽医学报,2012,32(11):1712-1715.
[23]郭春华,张均,王康宁,等.高寒草地生物量及牧草养分含量年度动态研究[J].中国草地学报,2007,29(1):1-5.
[24]韩正康,陈伟华,陆天水.三种饲养制度下山羊瘤胃内pH、氨氮和蛋白质含量的变化[J].南京农业大学学报,1982,5(3):118-124.
[25]申军士.日粮能氮释放同步性对奶牛瘤胃代谢、生产效率与性能的影响研究[D].博士学位论文.杭州:浙江大学,2013.
[26]魏园,赵晨旭,李心慰,等.不同粗纤维源日粮对梅花鹿瘤胃微生物区系的影响[J].中国兽医学报,2018,38(1):217-221,229.
[27]QIN J J,LI R Q,RAES J,et al.A human gut microbial gene catalogue established by metagenomic sequencing[J].Nature,2010,464(7285):59-65.
[28]LEY R E,LOZUPONE C A,HAMADY M,et al.Worlds within worlds:evolution of the vertebrate gut microbiota[J].Nature Reviews Microbiology,2008,6(10):776-788.
[29]EVANS N J,BROWN J M,MURRAY R D,et al.Characterization of novel bovine gastrointestinal tract Treponema isolates and comparison with bovine digital dermatitis treponemes[J].Applied and Environmental Microbiology,2011,77(1):138-147.
[30]MATSUI H,OGATA K,TAJIMA K,et al.Phenotypic characterization of polysaccharidases produced by four Prevotella type strains[J].Current Microbiology,2000,41(1):45-49.
[31]MEYER M,STENZEL U,HOFREITER M.Parallel tagged sequencing on the 454 platform[J].Nature Protocols,2008,3(2):267-278.
[32]TAJIMA K,AMINOV R I,NAGAMINE T,et al.Dietdependent shifts in the bacterial population of the rumen revealed with real-time PCR[J].Applied and Environmental Microbiology,2001,67(6):2766-2774.
[33]李莉,沈明华,俞红贤,等.含1/4野血犊牦牛血红蛋白、肌红蛋白含量的测定[J].青海大学学报(自然科学版),2005,23(6):42-44.
[34]徐俊.不同牧草来源的NDF在瘤胃中降解特性及其对细菌群落结构的影响[D].博士学位论文.扬州:扬州大学,2014.
[35]LATHAM M J,BROOKER B E,PETTIPHER G L,et al.Adhesion of Bacteroides succinogenes in pure culture and in the presence of Ruminococcus flavefaciens to cell walls in leaves of perennial ryegrass(Lolium perenne)[J].Applied and Environmnetal Microbiology,1978,35(6):1166-1173.
[36]FONDEVILA M,DEHORITY B A.Interactions between Fibrobacter succinogenes,Prevotella ruminicola,and Ruminococcus flavefaciens in the digestion of cellulose from forages[J].Journal of Animal Science,1996,74(3):678-684.