慢性瘤胃酸中毒对瘤胃发酵功能及乳酸代谢菌的影响
|
摘要
本研究选择6只体重相近,体况良好,并安装有永久性瘤胃瘘管的泌乳期关中奶山羊作为试验对象。试验分四期渐近式进行,并对瘤胃pH值进行24h实时监测,根据瘤胃pH值在5.5～5.8范围内持续的时间来确定SARA的发生。四期试验日粮的精粗比分别为5:5、6:4、7:3及8:2。实时监测结果表明,试验动物在精料比例为80%的水平下被成功诱导发生SARA。
本试验结合传统厌氧培养法、16S rRNA斑点杂交法及实时荧光定量PCR(Real-Time PCR)法来研究SARA状态下瘤胃发酵功能、血气指标发生的相应改变以及瘤胃细菌、乳酸产生菌(牛链球菌Strepococcus bovis、乳酸杆菌Lactobacillus)和乳酸利用菌(反刍兽新月单胞菌Selenomonas ruminantium、埃氏巨型球菌Megasphera elsdenii)的变化情况。研究结果表明:
(1)SARA状态下,丁酸浓度显著升高,且增长幅度最大,是导致瘤胃TVFA浓度升高的主要因素(P<0.05);在前三期试验内,乳酸浓度低于检测阈值;在SARA条件下乳酸浓度稍有增加。
(2)试验初期至SARA发生的整个过程中,细菌总数随着精料比例的提高不断增加,特别是在SARA期,细菌总数出现明显增加(P<0.01)。
(3)随着精料比例的不断提高,乳酸杆菌和牛链球菌的浓度不断增加。乳酸杆菌在70%精料水平时开始出现明显的增幅(P<0.05),而牛链球菌则是在80%精料水平时显著增加(P<0.01)。Real-Time PCR法测定的牛链球菌总体上也呈现略微升高的趋势,各期之间差异不显著(P>0.05)。
(4)SARA条件下,反刍兽新月单胞菌及埃氏巨型球菌的相对数量显著增加(P<0.01),前三期试验之间无明显差异(P>0.05)。
(5)在SARA状态下,瘤胃pH值下降(P>0.05);血浆pCO2分压升高,HCO3-浓度下降,血浆BE完全耗尽,AnGap浓度升高(P<0.05),SARA条件下血清Na+、K+及Cl-浓度变化不大(P>0.05)。
Six GuanZhong dairy goats in lactating period were used that had similar weights and steered with ruminal cannula. The trial was divided into four stages, the ratio of concentrate and roughage of four stages were 5:5, 6:4, 7:3 and 8:2 respectively. Ruminal pH was monitored in 24 hours uninterruptedly. According to the time that ruminal pH remained 5.5～5.8, determined whether SARA happened or not. The results of monitored ruminal pH showed that the goats were induced SARA in the level of 80% concentration. A study on rumen fermentation、blood-gas index、the populations of total bacteria and lactate-metabolizing bacteria (Strepococcus bovis, Lactobacillus, Selenomonas ruminantium and Megasphera elsdenii) using oligonucleotide probes to 16S rRNA、traditional cultivation method and Real-Time PCR was carried out under SARA situation. The results of this research as followed:
1. Extend of increased butyrate was the most in three kinds of VFA. The increase of butyrate leaded to total VFA increased significantly (P<0.05); the concentration of lactate was lower than detection threshold in former three stages; it increased slightly under SARA condition.
2. From the beginning to SARA happened, the concentration of total bacteria increased gradually, the population increased significantly under SARA situation especially (P<0.01).
3. The populations of Lactobacillus and Strepococcus bovis increased following the percentage of concentrate increased. Obvious increase of Lactobacillus occurred until the level of 70% concentrate (P<0.05). Significant increase of Strepococcus bovis occurred until 80% concentrate (P<0.01). The population of Strepococcus bovis showed a slight increase by Real-Time PCR (P>0.05).
4. The populations of Selenomonas ruminantium and Megasphera elsdenii increased significantly under SARA stage (P<0.01). There were no siginificant differences between the former there stages (P>0.05).
5. When SARA happened, the pH of blood decreased (P>0.05), pCO2 of plasma increased , concentration of HCO3-decreased, BE of blood exhausted, AnGap’s concentration increased(P<0.05); The concentrations of Na+, Ka+ and Cl- were influenced slightly with SARA.
本试验结合传统厌氧培养法、16S rRNA斑点杂交法及实时荧光定量PCR(Real-Time PCR)法来研究SARA状态下瘤胃发酵功能、血气指标发生的相应改变以及瘤胃细菌、乳酸产生菌(牛链球菌Strepococcus bovis、乳酸杆菌Lactobacillus)和乳酸利用菌(反刍兽新月单胞菌Selenomonas ruminantium、埃氏巨型球菌Megasphera elsdenii)的变化情况。研究结果表明:
(1)SARA状态下,丁酸浓度显著升高,且增长幅度最大,是导致瘤胃TVFA浓度升高的主要因素(P<0.05);在前三期试验内,乳酸浓度低于检测阈值;在SARA条件下乳酸浓度稍有增加。
(2)试验初期至SARA发生的整个过程中,细菌总数随着精料比例的提高不断增加,特别是在SARA期,细菌总数出现明显增加(P<0.01)。
(3)随着精料比例的不断提高,乳酸杆菌和牛链球菌的浓度不断增加。乳酸杆菌在70%精料水平时开始出现明显的增幅(P<0.05),而牛链球菌则是在80%精料水平时显著增加(P<0.01)。Real-Time PCR法测定的牛链球菌总体上也呈现略微升高的趋势,各期之间差异不显著(P>0.05)。
(4)SARA条件下,反刍兽新月单胞菌及埃氏巨型球菌的相对数量显著增加(P<0.01),前三期试验之间无明显差异(P>0.05)。
(5)在SARA状态下,瘤胃pH值下降(P>0.05);血浆pCO2分压升高,HCO3-浓度下降,血浆BE完全耗尽,AnGap浓度升高(P<0.05),SARA条件下血清Na+、K+及Cl-浓度变化不大(P>0.05)。
Six GuanZhong dairy goats in lactating period were used that had similar weights and steered with ruminal cannula. The trial was divided into four stages, the ratio of concentrate and roughage of four stages were 5:5, 6:4, 7:3 and 8:2 respectively. Ruminal pH was monitored in 24 hours uninterruptedly. According to the time that ruminal pH remained 5.5～5.8, determined whether SARA happened or not. The results of monitored ruminal pH showed that the goats were induced SARA in the level of 80% concentration. A study on rumen fermentation、blood-gas index、the populations of total bacteria and lactate-metabolizing bacteria (Strepococcus bovis, Lactobacillus, Selenomonas ruminantium and Megasphera elsdenii) using oligonucleotide probes to 16S rRNA、traditional cultivation method and Real-Time PCR was carried out under SARA situation. The results of this research as followed:
1. Extend of increased butyrate was the most in three kinds of VFA. The increase of butyrate leaded to total VFA increased significantly (P<0.05); the concentration of lactate was lower than detection threshold in former three stages; it increased slightly under SARA condition.
2. From the beginning to SARA happened, the concentration of total bacteria increased gradually, the population increased significantly under SARA situation especially (P<0.01).
3. The populations of Lactobacillus and Strepococcus bovis increased following the percentage of concentrate increased. Obvious increase of Lactobacillus occurred until the level of 70% concentrate (P<0.05). Significant increase of Strepococcus bovis occurred until 80% concentrate (P<0.01). The population of Strepococcus bovis showed a slight increase by Real-Time PCR (P>0.05).
4. The populations of Selenomonas ruminantium and Megasphera elsdenii increased significantly under SARA stage (P<0.01). There were no siginificant differences between the former there stages (P>0.05).
5. When SARA happened, the pH of blood decreased (P>0.05), pCO2 of plasma increased , concentration of HCO3-decreased, BE of blood exhausted, AnGap’s concentration increased(P<0.05); The concentrations of Na+, Ka+ and Cl- were influenced slightly with SARA.
引文
1 Hungate R E. The Rumen and Its Microbes. Academic Press. New York, NY 1966.
2 Mackie R I, Aminov R I, White B A, et al. Molecular ecology and diversity in gut microbial ecosysems.In: Cronje P B ed, Ruminant Physiology: Digestion,Metabolism, Growth and Reproduction,London: CAB Internationl, 2000, 61-77.
3 0zkose E, Thomas B J, Davies D R, et al. Cyllamyces aberensis gen.nov.sp.nov.anew anaerobic gut fungus with branched sporangiophores isolated from cattle.[J]. Can. J. Bot, 2001,79:666-673.
4 Cheng K J, Costerton J W. Adherent rumen bacteria: their role in the digestion of plant material, urea and epithelial cells. In: Ruchebush Y and Thivend P, ed. Digestive Physiology and Metabolism in Ruminants[C]. Lancaster: MTP Press, 1980,9: 227-250.
5 Wcelnerney M J. Syntrophic association of a butyrate-Degrading bacterium and Methanosarcina enriched from bovine rumen fluid [J]. Appl.Environ.Microbiol, 1981,41:826-828.
6 Bryant M P. Cultural methods and some characteristics of some of the more nume- rorous groups of bacteria in the bovine rumen [J]. J.dairy.sci, 1953, 36:205-217.
7 Wcelnerney M J. Syntrophic association of a butyrate-Degrading bacterium and Methanosarcina enriched from bovine rumen fluid [J]. Appl.Environ.Microbiol, 1981, 41: 826-828.
8 Amann R I, Ludwing W, Schleifer K H. Phylogenetic identification and in situ detection of individual microbial cells without cultivation [J]. Appl.Environ.Microbiol, 1995,59:143-169.
9 Roszak D B, Grimes D J, Colwell R R. Viable but nonrecoverable stage of Salmonella enteritidis in aquatic systems [J]. Can. J.Microbiol, 1984,30:334-338.
10 冯仰廉.中国反刍家畜在某些方面的研究进展[M].中国农业大学自编教材,1991.
11 Andrae, J G, Duckett S K, Hunt C W, et al. Effects of feeding high-oil corn to beef steers on carcass characteristics and meat quality [J]. J.Anim.Sci.,2001, 79:582-588.
12 Oetzel G R. Introduction to Ruminal Acidosis in Dairy Cattle. Preconvention Seminar
8:Dairy herd problem investigations. American association of bovine practitioners.
34th Annual Convention.2001, September, 11-12.
13 Allen M S, Beede D. Causes, detection and prevention of ruminal acidosis in dairy cattle examined [J]. Feedstuff.1996,68:13-18.
14 Owens F N, Secrist D S, Hill W J, et al. Acidosis in cattle: A review. J. Anim.Sci.,1998, 76: 275-286.
15 Mutsvangwa T, Wright T. Sub-Acute Ruminal Acidosis (SARA) in Dairy Cows.Ministry ofAgriculture Food and Rural Affairs [J]. Dairy Cattle Nutritionist.2003,30-31.
16 Nocek J E. Bovine acidosis:Implications on laminitis[J]. J. Dairy Sci,1997, 80: 1005-1028.
17 Slyter L L. Influence of acidosis on rumen function [J]. J.Anim.Sci,1976, 43: 910-929.
18 Enemark J M, Peters D G,J?rgensen R J. Rumen acidosis with special emphasis on diagnostic aspects of subclinical rumen acidosis A review[J]. Veterinarijair Zootechnika.2002, 20:16-29.
19 Krehbiel C R, Stock R A, Herold D W, et al. Feeding wet corn gluten feed to reduce subacute acidosis in cattle[J]. J.Anim.Sci.,1995,73: 2931-2939.
20 Mackie R I, Gilchrist F M C. Changes in lactate-producing and lactate-utilizing bacteria in relation to pH in the rumen of sheep during stepwise adaptation to a high-concentrate diet[J]. Appl.Environ.Microbiol.,1979, 38: 422-430.
21 Therion J J, Kistner A, Kornelius J H. Effect of pH on growth rates of rumen amylolytic and lactilytic bacteria[J]. Appl.Environ.Microbiol., 1982,44: 428-434.
22 Hungate R E, Dougherty R W, Bryant M P, et al. Mirobiological and physiological changes associated with acute indigestion in sheep[J]. Cornell Vet.1952, 42:423-426.
23 Dunlop R H, Hammond P B. D-lactic acidosis of ruminants[J]. Ann. NY. Acad. Sci.1965,119:1109-1132.
24 Russell J B, Hino T. Regulation of lactate production in Streptococcus bovis:A spiraling effect that contributes to rumen acidosis[J]. J.Dairy Sci. 1985,68:1712-1721.
25 Cooper R, Klopfenstein T. Effect of Rumensin and feed intake variation on ruminal pH[J]. In: Scientific Update on Rumensin/tylan/Mycoti for the Professional Feedlot Consultant.1996, ppA1~A14. Elanco Animal Health, Indianapolis, IN.
26 Bauer M L, Herold D W, Britton R A et al. Efficacy of laidlomycin propionate to reduce ruminal acidosis in cattle[J]. J.Anim.Sci.,1995,73:3445-3454.
27 Maekawa M, Beauchemin K A, Christensen D A. Effect of concentrate level and feeding management on chewing activities, saliva secretion, and ruminal pH of lactating dairy cows[J]. J.Dairy.Sci., 2002,85:1165-1175.
28 Britton R A, Stock R A. Acidosis,rate of starch digestion and intake. Okla. Agric.Exp. Stn. MP. 1986,121: 125-137.
29 Nordlund K V, Garrett E F. Rumenocentesis: a technique for collecting rumen fluid for the diagnosis of subacute rumen acidosis in dairy herds [J]. Bovine Pract. 1994,28: 109-112.
30 Hibbard B, Peters J P, Chester S T, et al. The effect of slaframine on salivary outputand subacute and acute acidosis in growing beef steers [J]. J.Anim.Sci, 1995, 73: 516-525.
31 Garrett E F, Pereira M N, Nordlund K V et al. Diagnostic methods for the detection of subacute ruminal acidosis in dairy cows [J]. J.Dairy.Sci, 1999,82:1170-1178.
32 Burrin D G, Britton R A. Response to monensin in cattle during subacute acidosis [J]. J.Anim. Sci.1986, 63: 888-893.
33 Harmon D L, Britton R A, Prior R L et al. Net portal absorption of lactate and volatile fatty acids in steers experiencing glucose-induced acidosis or fed a 70% concentrate diet ad libitum[J]. J.Anim.Sci, 1985,60: 560-569.
34 Brossard L, Martin C, Michalet-Doreau B. Ruminal fermentative parameters and blood acido-basic balance changes during onset and. recovery of induced latent acidosis in sheep[J]. Anim.Res, 2003,52: 513-530.
35 Armentano L E, Pereira M. Measuring the effectiveness of fiber by animal response trials [J]. J.Dairy.Sci,1997, 80:1416-1425.
36 Grant R J. Interactions among forages and nonforage fiber sources [J]. J.Dairy. Sci,1997,80:1438-1446.
37 肖训军.毛茛科苦木提取物对饲喂高碳水化合物饲粮反刍动物瘤胃发酵的影响[D].硕士学位论文,中国农业大学,2001
38 Keunen J E, Plaizier J C, Kyriazakis L,et al. Effects of a subacute ruminal acidosis model on the diet selection of dairy cows[J]. J.Dairy.Sci. 2002,85(12):3304-3313
39 张树金.奶牛亚急性瘤胃酸中毒(SARA)[J]. 中国动物保健,2004,8:23-24.
40 Aschenbach J R, Gabel G. Effect and absorption of histamine in sheep rumen: significance of acidosis epithelial damage [J]. J.Anim.Sci, 2000,78(2):464-470.
41 汪恩强,黄会岭,李占雷.奶山羊瘤胃酸中毒主要特性的试验研究[J]. 河北农业大学学报,2000,23(3):83-85.
42 Mutsvangwa T. Wright T. Sub-Acute Ruminal Acidosis (SARA) in Dairy Cows.Ministry of Agriculture Food and Rural Affairs[J]. Dairy Cattle Nutritionist.2003,30-31.
43 Stock R A. Nutritional benefits of specialty grain hybrids in beef feedlot diets. J.Anim.Sci., 1999,77:208-212.
44 Krehbiel C R, Stock R A, Herold D W et al. Feeding wet corn gluten feed to reduce subacute acidosis in cattle[J]. J.Anim.Sci, 1995,73:2931-2939.
45 Oetzel G R. Clinical aspects of ruminal acidosis in dairy cattle. Proceedings of the
33rd Annual Convention of the Americn Association of Bovine Practitioner [J].2000, 46-53.
46 Bertocchi L. Acidosis-promotiong factors [J]. Attidella Societa Italianadi Buiatria.1998,30:423-426.
47 Monteils V, Jurjanz S, Colin Schoellen O, et al. Kinetics of ruminal degradation of wheat and potato starches in total mixed rations[J]. Journal of Animal Science, 2002, 80(1):235-241.
48 Beauchemin K A, Yang W Z, Rode L M. Effects of particle size of alfalfa-based dairy cow diets on chewing acting,ruminal fermentation and milk production[J]. Journal of Dariy Science, 2003, 86(2):630-643.
49 Andrighetto J, Berzaghi P, Cozzi G, et al. Longer maize, feeding costs reduced [J]. Informatore Agrario, 1999, 55(31):35-38.
50 Nik Khah A, Fathy MHN, Nehzati G. Effect of ammoniated and roasted barley on rumen pH, milk yield and milk composition of Holstan cow[J]. Iranian Journal of Agriculture Science, 1998, 29(1):147-160.
51 Limin K J. Direct fed microbials for lactating dairy cows proceedings [M]. 12th Annual Florid Ruminant Nutrition Symposium, 2001, 22-28.
52 Ghorbani G R, Morgavi D P, Beauchemin K A, et al. Effects of bacterial direct-fed microbials on ruminal fermentation, blood variables, and the microbial populations of feedlot cattle[J]. J. Anim. Sci.2002, 80:1977-1986.
53 Beauchemin K A, Yang W Z, Rode L M. Effects of Particle Size of Alfalfa-Based Dairy Cow Diets on Chewing Activity, Ruminal Fermentation, and Milk Production [J]. J.Dairy.Sci, 2003, 86:630-643.
54 Shu Q, Gill H S,et al. Immunologicalcross reactivity between the vaccine and other isolates of Streptococcus bovis and Lactobacillus. FEMS Immun and Med Microbiol, 1999, 26:153-158.
55 Jeff D E, Scott A, Martin S A. Factors affecting lactate and malate utilization by Selenomonas ruminantium[J]. Appl.Environ.Microbiol, 1997, 63(12):4853-4858.
56 冯仰廉.中国反刍家畜在某些方面的研究进展[M].中国农业大学自编教材,1991
57 Tajima K, Arai S, Ogata K, et al. Rumen bacterial community transition during adaptation to high-grain diet [J]. Anaerobe, 1999, 6:273-284.
58 Nagarajia T G, Chengappa M M. Liver abscesses in feedlot: a review, J.Anim.Sci, 1998, 76:287-298.
59 Cheng K J. McAllister TA,Popp J D et al. A review of bloat in feedlot cattle [J]. J.Anim.Sci, 1998:76: 299-308.
60 王全军等.瘤胃微生物可作为新式酶源 [J].中国饲料,2000,13:28-29.
61 郑国展.利用瘤胃微生物酶类增进畜牧业的生产能力[J].动物营养学报,1999,11:61-64.
62 陈庆今等.瘤胃微生物在有机废物处理中的应用研究[J].微生物学杂志,2001,21(2): 41-49.
63 张元庆,孟庆翔.反当动物瘤胃细菌素研究进展[J].中国饲料,2004,(18): 6-7.
64 Stahl D A, Flesher B,Mansfield H R. Use of phylogenetically based hybridization probes for studies of ruminal microbial ecology[J]. Appl.Enuivon.Microbiol, 1988, 54:1079-1084.
65 Tajima K, Aminov R I, Nagamine T, et al. Diet-dependent shift in the bacterial population of the rumen revealed with real-time PCR[J].Appllied and Environmental Microbiology,2001,67(6):2766-2774.
66 Sylvester J T,Karnati S K R,Yu Z,et al. Development of an assay to quantify rumen ciliate protozoal biomass in cows using real-time PCR [J]. Journal of Nutrition,2004,134:3378-3384.
67 Lucy C, Skillman, Andrew F, et al. Development and Validation of a Real-Time PCR Method To Quantify Rumen Protozoa and Examination of Variability between Entodinium Populations in Sheep Offered a Hay-Based Diet [J]. 2005, CSIRO Livestock Industries, CSIRO Centre for Environment and Life Sciences, Private Bag 5, Wembley, Western Australia 6913, Australia.
68 Lin C, Raskin L, Stahl D A. Microbial community structure in gastrointestinal tracts of domestic animals:comparative analyses using rRNA-targeted oligonucleotide probes. FEMS Micobial Ecol, 1997, 22:281-294.
69 Luton P E, Wayne J M, Sharp R J,et al. The mcrA gene as an alternative to 16S rRNA in the phylogenetic analysis of methanogen populations in landfill [J]. Microbiology, 2002, 148:3521-3530.
70 Brossard L,Martin C.Protozoa involved in butyric rather than lactic fermentative pattern during latent acidosis in sheep[J].Reprod.Nutr.Dev.2004,44:195-206.
71 常影. 阉牛亚急性瘤胃酸中毒发病机理和饲喂高油玉米籽粒缓解效果的研究[D]. 博士学位论文,中国农业大学, 2006.
72 Olumeyan D B, Nagaraja T G, Miller G W, et al. Rumen microbial changes in cattle fed diets with or without salinomycin[J]. Appl.Environ.Microbiol. 1986, 51(2):340-345.
73 Brown M S,Krehbiel C R,Galyean M L,et al.Evaluation of models of acute and subacute acidosis on dry matter intake,ruminal fermentation,blood chemistry and endocrine profiles of beef steers[J]. J.Anim.Sci.2000, 78:3155-3168.
74 Hungate R E. A roll-tube method for cultivation of strict anaerobes [J]. Methods Microbiol, 1969, 3:117-132.
75 凌代文,东秀珠. 乳酸菌分离鉴定及实验方法[M]. 北京:中国轻工业出版社.
76 Goad D W, Goad C L.Ruminal microbial and fermentative changes associated with experimentally induced subacute acidosis in steers [J].Journal of Animal Science.1998,76(1):234-241.
77 Klieve A V.Establishing populations of Megasphaera elsdenii YE 34 and Butyrivibrio fibrisolvens YE 44 in the rumen of cattle fed high grain diets [J].Journal of Applied Microbiology, 2003, 95(3):621-630.
2 Mackie R I, Aminov R I, White B A, et al. Molecular ecology and diversity in gut microbial ecosysems.In: Cronje P B ed, Ruminant Physiology: Digestion,Metabolism, Growth and Reproduction,London: CAB Internationl, 2000, 61-77.
3 0zkose E, Thomas B J, Davies D R, et al. Cyllamyces aberensis gen.nov.sp.nov.anew anaerobic gut fungus with branched sporangiophores isolated from cattle.[J]. Can. J. Bot, 2001,79:666-673.
4 Cheng K J, Costerton J W. Adherent rumen bacteria: their role in the digestion of plant material, urea and epithelial cells. In: Ruchebush Y and Thivend P, ed. Digestive Physiology and Metabolism in Ruminants[C]. Lancaster: MTP Press, 1980,9: 227-250.
5 Wcelnerney M J. Syntrophic association of a butyrate-Degrading bacterium and Methanosarcina enriched from bovine rumen fluid [J]. Appl.Environ.Microbiol, 1981,41:826-828.
6 Bryant M P. Cultural methods and some characteristics of some of the more nume- rorous groups of bacteria in the bovine rumen [J]. J.dairy.sci, 1953, 36:205-217.
7 Wcelnerney M J. Syntrophic association of a butyrate-Degrading bacterium and Methanosarcina enriched from bovine rumen fluid [J]. Appl.Environ.Microbiol, 1981, 41: 826-828.
8 Amann R I, Ludwing W, Schleifer K H. Phylogenetic identification and in situ detection of individual microbial cells without cultivation [J]. Appl.Environ.Microbiol, 1995,59:143-169.
9 Roszak D B, Grimes D J, Colwell R R. Viable but nonrecoverable stage of Salmonella enteritidis in aquatic systems [J]. Can. J.Microbiol, 1984,30:334-338.
10 冯仰廉.中国反刍家畜在某些方面的研究进展[M].中国农业大学自编教材,1991.
11 Andrae, J G, Duckett S K, Hunt C W, et al. Effects of feeding high-oil corn to beef steers on carcass characteristics and meat quality [J]. J.Anim.Sci.,2001, 79:582-588.
12 Oetzel G R. Introduction to Ruminal Acidosis in Dairy Cattle. Preconvention Seminar
8:Dairy herd problem investigations. American association of bovine practitioners.
34th Annual Convention.2001, September, 11-12.
13 Allen M S, Beede D. Causes, detection and prevention of ruminal acidosis in dairy cattle examined [J]. Feedstuff.1996,68:13-18.
14 Owens F N, Secrist D S, Hill W J, et al. Acidosis in cattle: A review. J. Anim.Sci.,1998, 76: 275-286.
15 Mutsvangwa T, Wright T. Sub-Acute Ruminal Acidosis (SARA) in Dairy Cows.Ministry ofAgriculture Food and Rural Affairs [J]. Dairy Cattle Nutritionist.2003,30-31.
16 Nocek J E. Bovine acidosis:Implications on laminitis[J]. J. Dairy Sci,1997, 80: 1005-1028.
17 Slyter L L. Influence of acidosis on rumen function [J]. J.Anim.Sci,1976, 43: 910-929.
18 Enemark J M, Peters D G,J?rgensen R J. Rumen acidosis with special emphasis on diagnostic aspects of subclinical rumen acidosis A review[J]. Veterinarijair Zootechnika.2002, 20:16-29.
19 Krehbiel C R, Stock R A, Herold D W, et al. Feeding wet corn gluten feed to reduce subacute acidosis in cattle[J]. J.Anim.Sci.,1995,73: 2931-2939.
20 Mackie R I, Gilchrist F M C. Changes in lactate-producing and lactate-utilizing bacteria in relation to pH in the rumen of sheep during stepwise adaptation to a high-concentrate diet[J]. Appl.Environ.Microbiol.,1979, 38: 422-430.
21 Therion J J, Kistner A, Kornelius J H. Effect of pH on growth rates of rumen amylolytic and lactilytic bacteria[J]. Appl.Environ.Microbiol., 1982,44: 428-434.
22 Hungate R E, Dougherty R W, Bryant M P, et al. Mirobiological and physiological changes associated with acute indigestion in sheep[J]. Cornell Vet.1952, 42:423-426.
23 Dunlop R H, Hammond P B. D-lactic acidosis of ruminants[J]. Ann. NY. Acad. Sci.1965,119:1109-1132.
24 Russell J B, Hino T. Regulation of lactate production in Streptococcus bovis:A spiraling effect that contributes to rumen acidosis[J]. J.Dairy Sci. 1985,68:1712-1721.
25 Cooper R, Klopfenstein T. Effect of Rumensin and feed intake variation on ruminal pH[J]. In: Scientific Update on Rumensin/tylan/Mycoti for the Professional Feedlot Consultant.1996, ppA1~A14. Elanco Animal Health, Indianapolis, IN.
26 Bauer M L, Herold D W, Britton R A et al. Efficacy of laidlomycin propionate to reduce ruminal acidosis in cattle[J]. J.Anim.Sci.,1995,73:3445-3454.
27 Maekawa M, Beauchemin K A, Christensen D A. Effect of concentrate level and feeding management on chewing activities, saliva secretion, and ruminal pH of lactating dairy cows[J]. J.Dairy.Sci., 2002,85:1165-1175.
28 Britton R A, Stock R A. Acidosis,rate of starch digestion and intake. Okla. Agric.Exp. Stn. MP. 1986,121: 125-137.
29 Nordlund K V, Garrett E F. Rumenocentesis: a technique for collecting rumen fluid for the diagnosis of subacute rumen acidosis in dairy herds [J]. Bovine Pract. 1994,28: 109-112.
30 Hibbard B, Peters J P, Chester S T, et al. The effect of slaframine on salivary outputand subacute and acute acidosis in growing beef steers [J]. J.Anim.Sci, 1995, 73: 516-525.
31 Garrett E F, Pereira M N, Nordlund K V et al. Diagnostic methods for the detection of subacute ruminal acidosis in dairy cows [J]. J.Dairy.Sci, 1999,82:1170-1178.
32 Burrin D G, Britton R A. Response to monensin in cattle during subacute acidosis [J]. J.Anim. Sci.1986, 63: 888-893.
33 Harmon D L, Britton R A, Prior R L et al. Net portal absorption of lactate and volatile fatty acids in steers experiencing glucose-induced acidosis or fed a 70% concentrate diet ad libitum[J]. J.Anim.Sci, 1985,60: 560-569.
34 Brossard L, Martin C, Michalet-Doreau B. Ruminal fermentative parameters and blood acido-basic balance changes during onset and. recovery of induced latent acidosis in sheep[J]. Anim.Res, 2003,52: 513-530.
35 Armentano L E, Pereira M. Measuring the effectiveness of fiber by animal response trials [J]. J.Dairy.Sci,1997, 80:1416-1425.
36 Grant R J. Interactions among forages and nonforage fiber sources [J]. J.Dairy. Sci,1997,80:1438-1446.
37 肖训军.毛茛科苦木提取物对饲喂高碳水化合物饲粮反刍动物瘤胃发酵的影响[D].硕士学位论文,中国农业大学,2001
38 Keunen J E, Plaizier J C, Kyriazakis L,et al. Effects of a subacute ruminal acidosis model on the diet selection of dairy cows[J]. J.Dairy.Sci. 2002,85(12):3304-3313
39 张树金.奶牛亚急性瘤胃酸中毒(SARA)[J]. 中国动物保健,2004,8:23-24.
40 Aschenbach J R, Gabel G. Effect and absorption of histamine in sheep rumen: significance of acidosis epithelial damage [J]. J.Anim.Sci, 2000,78(2):464-470.
41 汪恩强,黄会岭,李占雷.奶山羊瘤胃酸中毒主要特性的试验研究[J]. 河北农业大学学报,2000,23(3):83-85.
42 Mutsvangwa T. Wright T. Sub-Acute Ruminal Acidosis (SARA) in Dairy Cows.Ministry of Agriculture Food and Rural Affairs[J]. Dairy Cattle Nutritionist.2003,30-31.
43 Stock R A. Nutritional benefits of specialty grain hybrids in beef feedlot diets. J.Anim.Sci., 1999,77:208-212.
44 Krehbiel C R, Stock R A, Herold D W et al. Feeding wet corn gluten feed to reduce subacute acidosis in cattle[J]. J.Anim.Sci, 1995,73:2931-2939.
45 Oetzel G R. Clinical aspects of ruminal acidosis in dairy cattle. Proceedings of the
33rd Annual Convention of the Americn Association of Bovine Practitioner [J].2000, 46-53.
46 Bertocchi L. Acidosis-promotiong factors [J]. Attidella Societa Italianadi Buiatria.1998,30:423-426.
47 Monteils V, Jurjanz S, Colin Schoellen O, et al. Kinetics of ruminal degradation of wheat and potato starches in total mixed rations[J]. Journal of Animal Science, 2002, 80(1):235-241.
48 Beauchemin K A, Yang W Z, Rode L M. Effects of particle size of alfalfa-based dairy cow diets on chewing acting,ruminal fermentation and milk production[J]. Journal of Dariy Science, 2003, 86(2):630-643.
49 Andrighetto J, Berzaghi P, Cozzi G, et al. Longer maize, feeding costs reduced [J]. Informatore Agrario, 1999, 55(31):35-38.
50 Nik Khah A, Fathy MHN, Nehzati G. Effect of ammoniated and roasted barley on rumen pH, milk yield and milk composition of Holstan cow[J]. Iranian Journal of Agriculture Science, 1998, 29(1):147-160.
51 Limin K J. Direct fed microbials for lactating dairy cows proceedings [M]. 12th Annual Florid Ruminant Nutrition Symposium, 2001, 22-28.
52 Ghorbani G R, Morgavi D P, Beauchemin K A, et al. Effects of bacterial direct-fed microbials on ruminal fermentation, blood variables, and the microbial populations of feedlot cattle[J]. J. Anim. Sci.2002, 80:1977-1986.
53 Beauchemin K A, Yang W Z, Rode L M. Effects of Particle Size of Alfalfa-Based Dairy Cow Diets on Chewing Activity, Ruminal Fermentation, and Milk Production [J]. J.Dairy.Sci, 2003, 86:630-643.
54 Shu Q, Gill H S,et al. Immunologicalcross reactivity between the vaccine and other isolates of Streptococcus bovis and Lactobacillus. FEMS Immun and Med Microbiol, 1999, 26:153-158.
55 Jeff D E, Scott A, Martin S A. Factors affecting lactate and malate utilization by Selenomonas ruminantium[J]. Appl.Environ.Microbiol, 1997, 63(12):4853-4858.
56 冯仰廉.中国反刍家畜在某些方面的研究进展[M].中国农业大学自编教材,1991
57 Tajima K, Arai S, Ogata K, et al. Rumen bacterial community transition during adaptation to high-grain diet [J]. Anaerobe, 1999, 6:273-284.
58 Nagarajia T G, Chengappa M M. Liver abscesses in feedlot: a review, J.Anim.Sci, 1998, 76:287-298.
59 Cheng K J. McAllister TA,Popp J D et al. A review of bloat in feedlot cattle [J]. J.Anim.Sci, 1998:76: 299-308.
60 王全军等.瘤胃微生物可作为新式酶源 [J].中国饲料,2000,13:28-29.
61 郑国展.利用瘤胃微生物酶类增进畜牧业的生产能力[J].动物营养学报,1999,11:61-64.
62 陈庆今等.瘤胃微生物在有机废物处理中的应用研究[J].微生物学杂志,2001,21(2): 41-49.
63 张元庆,孟庆翔.反当动物瘤胃细菌素研究进展[J].中国饲料,2004,(18): 6-7.
64 Stahl D A, Flesher B,Mansfield H R. Use of phylogenetically based hybridization probes for studies of ruminal microbial ecology[J]. Appl.Enuivon.Microbiol, 1988, 54:1079-1084.
65 Tajima K, Aminov R I, Nagamine T, et al. Diet-dependent shift in the bacterial population of the rumen revealed with real-time PCR[J].Appllied and Environmental Microbiology,2001,67(6):2766-2774.
66 Sylvester J T,Karnati S K R,Yu Z,et al. Development of an assay to quantify rumen ciliate protozoal biomass in cows using real-time PCR [J]. Journal of Nutrition,2004,134:3378-3384.
67 Lucy C, Skillman, Andrew F, et al. Development and Validation of a Real-Time PCR Method To Quantify Rumen Protozoa and Examination of Variability between Entodinium Populations in Sheep Offered a Hay-Based Diet [J]. 2005, CSIRO Livestock Industries, CSIRO Centre for Environment and Life Sciences, Private Bag 5, Wembley, Western Australia 6913, Australia.
68 Lin C, Raskin L, Stahl D A. Microbial community structure in gastrointestinal tracts of domestic animals:comparative analyses using rRNA-targeted oligonucleotide probes. FEMS Micobial Ecol, 1997, 22:281-294.
69 Luton P E, Wayne J M, Sharp R J,et al. The mcrA gene as an alternative to 16S rRNA in the phylogenetic analysis of methanogen populations in landfill [J]. Microbiology, 2002, 148:3521-3530.
70 Brossard L,Martin C.Protozoa involved in butyric rather than lactic fermentative pattern during latent acidosis in sheep[J].Reprod.Nutr.Dev.2004,44:195-206.
71 常影. 阉牛亚急性瘤胃酸中毒发病机理和饲喂高油玉米籽粒缓解效果的研究[D]. 博士学位论文,中国农业大学, 2006.
72 Olumeyan D B, Nagaraja T G, Miller G W, et al. Rumen microbial changes in cattle fed diets with or without salinomycin[J]. Appl.Environ.Microbiol. 1986, 51(2):340-345.
73 Brown M S,Krehbiel C R,Galyean M L,et al.Evaluation of models of acute and subacute acidosis on dry matter intake,ruminal fermentation,blood chemistry and endocrine profiles of beef steers[J]. J.Anim.Sci.2000, 78:3155-3168.
74 Hungate R E. A roll-tube method for cultivation of strict anaerobes [J]. Methods Microbiol, 1969, 3:117-132.
75 凌代文,东秀珠. 乳酸菌分离鉴定及实验方法[M]. 北京:中国轻工业出版社.
76 Goad D W, Goad C L.Ruminal microbial and fermentative changes associated with experimentally induced subacute acidosis in steers [J].Journal of Animal Science.1998,76(1):234-241.
77 Klieve A V.Establishing populations of Megasphaera elsdenii YE 34 and Butyrivibrio fibrisolvens YE 44 in the rumen of cattle fed high grain diets [J].Journal of Applied Microbiology, 2003, 95(3):621-630.