部分扣除Lys、Met及Thr对6～9月龄荷斯坦生长母牛瘤胃内环境的影响
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摘要
研究旨在探究扣除Lys、Met及Thr对6～9月龄生长母牛瘤胃内环境的影响,确定扣除3种过瘤胃氨基酸是否会通过改变MCP产量影响小肠氨基酸的吸收。选取72头5.5月龄左右,体重200 kg左右的荷斯坦母牛,分为4组,每组18头,分别为理论氨基酸平衡组(PC)、扣除30%的Lys组(PD-Lys)、扣除30%的Met组(PD-Met)及扣除30%的Thr组(PD-Thr)。试验期为105 d,预饲15 d,正试期90 d。基础日粮均为同一玉米-豆粕-苜蓿型全混合日粮,所选用的氨基酸添加剂均为过瘤胃氨基酸。试验期内,每30 d采集一次瘤胃液,测定相应的瘤胃发酵参数和微生物区系。结果表明:扣除Lys和Thr对生长母牛瘤胃pH值、NH3-N浓度及MCP含量均没有显著影响(P>0.05),但PD-Met组MCP含量有较PC组降低的趋势。此外,PD-Met组的丙酸摩尔比例较PC组显著升高(P<0.05),从而导致该组乙丙比显著低于PC组(P<0.05)。PD-Met组的瘤胃微生物Ace及Chao指数较PC组显著下降(P<0.05),Prevotella_7丰度显著升高(P<0.05),表明扣除Met影响了瘤胃微生物的生长和物种组成;扣除Lys及Thr均未对瘤胃微生物区系及组成产生显著影响(P>0.05)。本试验中扣除30%的过瘤胃Lys及Thr未影响生长母牛瘤胃发酵参数和瘤胃微生物生长,到达小肠的氨基酸水平不受MCP影响,主要受由日粮所提供的氨基酸水平限制;但扣除Met导致瘤胃微生物丰富度下降,Prevotella_7丰度增加,丙酸比例升高,从而导致MCP合成量有降低趋势。
In order to determine whether deleting Lys, Met and Thr would affect the absorption of amino acids in small intestine by altering the production of microbial protein(MCP),the experiment was conducted for 90 days with 72 Holstein heifers [average(5.5±0.5) months old;(200±9.0) kg of body weight]. Following an initial 15 d adaptation period, heifers were allocated to one of four groups: theoretical amino acid, positive control group(PC); 30% Lys deleted group(PD-Lys); 30% Met deleted group(PD-Met), and 30% Thr deleted group(PD-Thr). Experimental animals were fed cornsoybean-alfalfa based total mixed rations(TMR)supplemented with Lys, Met, and Thr(ruminal bypass). Rumen fluid was collected at 30 days interval to determineruminal fermentation parameters and microflora. The results showed as follows: no significant differences of pH value,NH_3-N concentration, MCP yields were found(P>0.05) among groups, but the MCP yields in PD-Met group tended to be lower than that in PC group. In addition, the molar ratio of propionic acid was higher(P<0.05) in PD-Met group than that in PC group, resulting in a lower ratio of acetate/propionate(P<0.05) in PD-Met group compared to PC group. The values of Chao and Ace of microbial diversity were reduced(P<0.05) as Met level decreased, but the abundance of Prevotella_7 was significantly increased(P<0.05) in PD-Met group. No significant effects on rumen microflora and composition was observed(P>0.05) after the deletion of Lys and Thr. The results indicated that deleting 30% of rumen bypass Lys and Thr did not affect rumen fermentation parameters and rumen microbial growth of growing heifers, so the aminoacid level of small intestine was not affected by MCP, mainly limited by the amino acid level in diet. However, deleting Met led to decrease of microbial abundance in the rumen, as well as the increases of Prevotella_7 abundance and propionic acid ratio, which may resulted in the decreasing trend of MCP.
In order to determine whether deleting Lys, Met and Thr would affect the absorption of amino acids in small intestine by altering the production of microbial protein(MCP),the experiment was conducted for 90 days with 72 Holstein heifers [average(5.5±0.5) months old;(200±9.0) kg of body weight]. Following an initial 15 d adaptation period, heifers were allocated to one of four groups: theoretical amino acid, positive control group(PC); 30% Lys deleted group(PD-Lys); 30% Met deleted group(PD-Met), and 30% Thr deleted group(PD-Thr). Experimental animals were fed cornsoybean-alfalfa based total mixed rations(TMR)supplemented with Lys, Met, and Thr(ruminal bypass). Rumen fluid was collected at 30 days interval to determineruminal fermentation parameters and microflora. The results showed as follows: no significant differences of pH value,NH_3-N concentration, MCP yields were found(P>0.05) among groups, but the MCP yields in PD-Met group tended to be lower than that in PC group. In addition, the molar ratio of propionic acid was higher(P<0.05) in PD-Met group than that in PC group, resulting in a lower ratio of acetate/propionate(P<0.05) in PD-Met group compared to PC group. The values of Chao and Ace of microbial diversity were reduced(P<0.05) as Met level decreased, but the abundance of Prevotella_7 was significantly increased(P<0.05) in PD-Met group. No significant effects on rumen microflora and composition was observed(P>0.05) after the deletion of Lys and Thr. The results indicated that deleting 30% of rumen bypass Lys and Thr did not affect rumen fermentation parameters and rumen microbial growth of growing heifers, so the aminoacid level of small intestine was not affected by MCP, mainly limited by the amino acid level in diet. However, deleting Met led to decrease of microbial abundance in the rumen, as well as the increases of Prevotella_7 abundance and propionic acid ratio, which may resulted in the decreasing trend of MCP.
引文
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[23]刘开朗,王加启,卜登攀.2008-2009年反刍动物营养研究进展Ⅰ.瘤胃微生物多样性与功能[J].中国畜牧兽医,2010,37(2):5-14.
[24]王欢莉,王洪荣,徐爱秋,等.瘤胃微生物理想氨基酸模式的建立[J].中国畜牧杂志,2014,50(9):62-66.
[25]Karnati S K,Sylvester J T,Noftsger S M,et al.Assessment of ruminal bacterial populations and protozoal generation time in cows fed different methionine sources[J].Journal of Dairy Science,2007,90(2):798-809.
[26]Sylvester J T,Karnati S K R,Yu Z,et al.Evaluation of a realtime PCR assay quantifying the ruminal pool size and duodenal flow of protozoal nitrogen[J].Journal of Dairy Science,2005,88(6):2083-2095.
[27]刘文杰,雒秋江,陈勇,等.添喂赖氨酸对小尾寒羊瘤胃微生物区系和代谢的影响[C]//全国动物生理生化第十二次学术交流会论文摘要汇编.2012.
[28]Purushe J,Fouts D E,Morrison M,et al.Comparative Genome Analysis of Prevotella ruminicola and Prevotella bryantii:Insights into Their Environmental Niche[J].Microbial Ecology,2010,60(4):721-729.
[29]外源氨基酸对人工瘤胃中微生物生长及氨基酸组成的影响[D].扬州大学,2009.
[2]王洪荣,徐爱秋,王梦芝,等.氨基酸对体外培养瘤胃微生物生长及发酵的影响[J].畜牧兽医学报,2010,41(9):1109-1116.
[3]Rodriguez C A,Gonzalez J,Alvirl M R,et al.Composition of bacteria harvested from the liquid and solid fractions of the rumen of sheep as influenced by feed intake[J].British Journal of Nutrition,2000,84(3):369-376.
[4]周勃,冯仰廉.不同能量水平日粮对肉牛瘤胃微生物氨基酸组成的影响[J].中国畜牧杂志,2000,36(4):3-5.
[5]王洪荣,王梦芝,曹恒春,等.日粮不同蛋白质来源对山羊瘤胃微生物氨基酸组成的影响[J].动物营养学报,2009,21(6):834-845.
[6]Gedge J.Note on the Anatomical Development of the Ruminant Stomach[J].Journal of Anatomy&Physiology,1868,2(2):323-452.
[7]Zinn R A,Shen Y.An evaluation of ruminally degradable intake protein and metabolizable amino acid requirements of feedlot calves[J].Journal of Animal Science,1998,76(5):1280-1289.
[8]Chung T K,Baker D H.Ideal amino acid pattern for 10-kilogram pigs[L500;L600];pigs;amino acids;growth;nitrogen metabolism;nitrogen balance;diet IND92069907[J].Journal of Animal Science,1992,70(10):3102-3111.
[9]Calsamiglia S,Ferret A,Devant M.Effects of pH and pH Fluctuations on Microbial Fermentation and Nutrient Flow from a DualFlow Continuous Culture System[J].Journal of Dairy Science,2002,85(3):574-579.
[10]杨维仁.瘤胃保护性氨基酸对肉牛消化代谢影响及适宜供给量的研究[D].中国农业大学,2004.
[11]Campbell C G,Tigemeyer E C,Cocharan R C,et al.Free amino acid supplementation to steers:effects on ruminal fermentation and performance[J].Journal of Animal Science,1997,75(4):1167-1178.
[12]Satter L D,Slyter L L.Effect of ammonia concentration of rumen microbial protein production in vitro[J].British Journal of Nutrition,1974,32(2):199-208.
[13]韩继福,吴其宏.过瘤胃氨基酸对乳牛瘤胃发酵、产乳量及乳成分的影响[J].安徽农业大学学报,2003,30(2):139-143.
[14]燕磊.瘤胃保护性蛋氨酸对小尾寒羊氨基酸代谢影响的研究[D].山东农业大学,2005.
[15]Windschitlp M,Stern M D.Influence of methionine derivatives on effluent flow of methionine from continuous culture of ruminal bacteria[J].Journal of Animal Science,1988,66(11):2937.
[16]Firkins J L,Fowler C M,Devillard E,et al.Kinetics of microbial methionine metabolism in continuous cultures administered different methionine sources[J].Journal of Dairy Science,2015,98(2):1178-1194.
[17]Vazquez A M,Cassidy T,Mccullough P,et al.Effects of alimet on nutrient digestibility,bacterial protein synthesis,and ruminal disappearance during continuous culture[J].Journal of Dairy Science,2001,84(1):159-166.
[18]甄玉国,马宁,卢德勋.反刍动物小肠氨基酸平衡与调控[J].中国饲料,2002(3):29-30.
[19]Maiga H A.Evaluation of corn distillers grains and ruminally protected lysine and methionine for lactating dairy cows[J].Journal of Dairy Science,1998,81(2):482-491.
[20]Noftsger S,Pierre N R,Sylvester J T.Determination of Rumen Degradability and Ruminal Effects of Three Sources of Methionine in Lactating Cows*[J].Journal of Dairy Science,2005,88(1):223-237.
[21]Robinson P H,Chalupa W,Sniffen C J,et al.Influence of abomasal infusion of high levels of lysine or methionine,or both,on ruminal fermentation,eating behavior,and performance of lactating dairy cows[J].Journal of Animal Science,2000,78(4):1067-1077.
[22]Noftsger S M,Pierre N R,Karnati S K,et al.Effects of 2-Hydroxy-4-(methylthio)Butanoic Acid(HMB)on Microbial Growth in Continuous Culture[J].Journal of Dairy Science,2003,86(8):2629-2636.
[23]刘开朗,王加启,卜登攀.2008-2009年反刍动物营养研究进展Ⅰ.瘤胃微生物多样性与功能[J].中国畜牧兽医,2010,37(2):5-14.
[24]王欢莉,王洪荣,徐爱秋,等.瘤胃微生物理想氨基酸模式的建立[J].中国畜牧杂志,2014,50(9):62-66.
[25]Karnati S K,Sylvester J T,Noftsger S M,et al.Assessment of ruminal bacterial populations and protozoal generation time in cows fed different methionine sources[J].Journal of Dairy Science,2007,90(2):798-809.
[26]Sylvester J T,Karnati S K R,Yu Z,et al.Evaluation of a realtime PCR assay quantifying the ruminal pool size and duodenal flow of protozoal nitrogen[J].Journal of Dairy Science,2005,88(6):2083-2095.
[27]刘文杰,雒秋江,陈勇,等.添喂赖氨酸对小尾寒羊瘤胃微生物区系和代谢的影响[C]//全国动物生理生化第十二次学术交流会论文摘要汇编.2012.
[28]Purushe J,Fouts D E,Morrison M,et al.Comparative Genome Analysis of Prevotella ruminicola and Prevotella bryantii:Insights into Their Environmental Niche[J].Microbial Ecology,2010,60(4):721-729.
[29]外源氨基酸对人工瘤胃中微生物生长及氨基酸组成的影响[D].扬州大学,2009.