牛链球菌在瘤胃中产酸的代谢机制及调控

牛链球菌(S.bovis)是瘤胃主要的乳酸产生菌,在饲喂高精料饲粮导致瘤胃乳酸中毒进程中扮演重要角色。已有研究证实S.bovis利用碳水化合物代谢产酸主要受葡萄糖转运方式、酵解产酸途径中酶和中间代谢物调控。另外,研究也发现环境p H、增殖生长阶段及分解代谢控制蛋白(Ccp A)等对其产酸速率和模式也有显著影响。本文对近年来有关S.bovis利用饲料中碳水化合物发酵产酸代谢途径及影响因素研究加以综述,为从微生物代谢角度解析瘤胃乳酸中毒机制提供参考。     

牛支原体能量耦合因子转运蛋白S组件MbBioY摄取外源生物素的功能分析

牛支原体(M.bovis)感染与生存依赖于外界微环境提供的各类营养代谢因子。开展M.bovis外源生物素摄取的机制研究,将对M.bovis防控具有极其重要的意义。通过对M.bovis PG45 MbBioY(MBOVPG45＿0349)基因进行系统发育分析、分子对接分析,筛选M.bovis PG45 MbBioY突变株,比较PG45和PG45ΔMbBioY的生物素敏感性,异源构建MbBioY功能性克隆验证其功能。结果表明,PG45染色体上存在一种崭新的生物素膜转运蛋白,全长996 bp, GC含量30.95%,蛋白长度为332个氨基酸,是由7个跨膜结构域组成的能量偶合因子(ECF)转运蛋白S成分MbBioY。进一步验证后发现,当M.bovis缺失MbBioY会影响其生长,表现出生物素代谢营养缺陷的表型。通过异源敲入生物素代谢营养缺陷大肠杆菌MG1655,表明MbBioY单独存在时,就能够恢复生物素代谢营养缺陷大肠杆菌缺失株的生理功能。初步证明,MbBioY编码的是能量偶合因子(ECF)转运蛋白的S成分具有摄取外源生物素的能力,该研究结果将补充细菌生物素代谢调控的多样性和复杂性,为M.b...     

瘤胃中乳酸的代谢及其调控机制

有关瘤胃酸中毒发生机制研究表明,瘤胃乳酸的累积可能对酸中毒诱导起重要作用,而高精料日粮下瘤胃乳酸累积主要取决于瘤胃乳酸产生菌和乳酸利用菌间的平衡程度。本文综述了瘤胃微生物对乳酸的代谢机制,包括主要乳酸产生菌[溶纤维丁酸弧菌(Butyrivibrio fibrisolvens)、牛链球菌(Streptococcus bovis)、乳酸杆菌(Lactobacillus)]和主要乳酸利用菌[反刍兽新月单胞菌(Selenomonus ruminantium)、埃氏巨型球菌(Megasphaera elsdenii)],并简要概述了酸中毒的调控方法,旨在为反刍动物瘤胃酸中毒的乳酸中毒机制深入解析提供参考。     

饲粮碳水化合物对羔羊瘤胃发育影响的研究进展

瘤胃作为反刍动物特有的消化器官,它的消化代谢和营养吸收功能对反刍动物的生长具有重要意义。羔羊瘤胃的结构和机能尚不完善,其发育受到多种因素的影响,而饲粮碳水化合物是调控瘤胃发育的一个重要因素。本文综述了饲粮碳水化合物对羔羊瘤胃发育影响的研究进展,旨在为通过调整羔羊饲粮碳水化合物结构来改善羔羊瘤胃发育的理论研究与生产实践提供依据。     

奶牛瘤胃原虫对营养物质代谢的影响

奶牛瘤胃内栖居着大量微生物,主要包括细菌、真菌和原虫。文章分析了奶牛瘤胃原虫在碳水化合物代谢(淀粉、可溶性糖和纤维素代谢)、蛋白质代谢和瘤胃内甲烷代谢等过程中的作用,并总结了其对奶牛的营养代谢的利弊影响。     

反刍动物瘤胃酸中毒机制解析及其营养调控措施

反刍动物瘤胃亚急性酸中毒(SARA)是现代集约化生产中常见的营养代谢病.“反刍动物SARA的发生和调控机制”是重要的科学问题,对其的进一步深入研究不仅在学术上有重要意义,而且对降低经济损失具有重要的现实意义.本文分析了国际上反刍动物瘤胃中毒研究的最新成果和进展,对其产生原因和机制以及预防措施进行了系统的阐述.本文深入探讨了引起瘤胃酸中毒的3个学说,“有机酸中毒学说”、“乳酸中毒学说”和“内毒素和组织胺中毒学说”;较为深入地解析了饲粮中碳水化合物组成、瘤胃微生物菌群及其代谢变化对瘤胃酸中毒发生的影响.同时,也对SARA的预防及其营养调控措施进行了论述,并提出通过科学合理的饲养管理,合理设计饲粮的碳水化合物组成,使用益生菌、缓冲剂、有机酸、硫胺素,接种疫苗,调控瘤胃内有机酸的产生和利用的途径来预防反刍动物瘤胃酸中毒的发生,以期为今后我国在预防瘤胃酸中毒方面的研究和生产应用提供参考.     

反刍动物碳水化合物高效利用的综合调控

碳水化合物是反刍动物的主要能量来源,在维持动物生长发育、机体代谢和生产性能等方面发挥关键作用。碳水化合物降解生成的瘤胃挥发性脂肪酸和小肠葡萄糖是反刍动物的主要供能形式,淀粉在小肠降解生成葡萄糖的供能效率显著优于瘤胃发酵生成挥发性脂肪酸,理论上将部分在瘤胃中降解的淀粉转移到小肠消化可有效改善能量利用效率,并降低瘤胃酸中毒的风险。文章主要以本课题组近年的研究进展为基础,并参考部分相关研究成果,从碳水化合物与瘤胃代谢调控、碳水化合物平衡指数、小肠碳水化合物高效利用、奶牛围产期葡萄糖营养平衡,以及基于CNCPS和CPM-Dairy的中国饲料数据库建设等5个方面论述反刍动物碳水化合物高效利用的综合调控技术,以期为提高反刍动物碳水化合物利用效率、改善动物健康和促进畜牧业高效发展提供理论依据。     

2007～2008年国际反刍动物营养研究进展 Ⅲ. 碳水化合物营养

碳水化合物是反刍动物饲料的主要组成部分,研究碳水化合物的营养对提高反刍动物生产效率具有重要作用。近年来,根据饲料中碳水化合物各组分在瘤胃中的降解情况进行分类的方法;非结构性碳水化合物中淀粉的消化特性;结构性碳水化合物中中性洗涤纤维在饲料中的含量及对消化的影响;饲料的物理特性对瘤胃发酵的影响;碳水化合物需要量的指标;碳水化合物饲料原料的开发和利用方面都有较多研究。作者综述了饲料中碳水化合物的组成,结构性和非结构性碳水化合物的营养,碳水化合物的物理特性与营养,反刍动物的碳水化合物需要量,奶牛、肉牛及其他反刍动物饲料原料的研究进展。今后,将利用分子生物学技术在碳水化合物的生理功能、作用机理、调控机制及体内的代谢途径等方面进行深入的研究,而且也将在饲料资源利用效果方面进行多方面研究。     

皂甙调控反刍动物瘤胃发酵和生产性能的研究进展

皂甙作为一种植物提取物,能够通过影响瘤胃微生物种群数量和活性,调控瘤胃发酵模式,降低甲烷排放、提高反刍动物生产性能。本文结合最新的研究报道,将皂甙对原虫、产甲烷菌、真菌和细菌的影响作为出发点,论述了皂甙对瘤胃内氮代谢和碳水化合物代谢的影响机理,以及皂甙对反刍动物生产性能的影响,以期为皂甙产品作为反刍动物瘤胃调控剂的研究和开发提供参考。     

不同碳水化合物对犊牛消化生理功能的研究进展

碳水化合物是多羟基醛、酮及其衍生物的总称,是反刍动物瘤胃微生物的主要能量来源,在犊牛营养中起着重要的作用.介绍了碳水化合物的分类及在瘤胃中的消化代谢情况,并就近年来不同类型碳水化合物在犊牛开食料中的应用进行综述.     

Analysis of Rumen Microbial Diversity and Functional Prediction of Chinese Simmental Cattle Based on 16S rRNA High-throughput Sequencing Technology

To systematically explore the microbial diversity and function in the rumen of Chinese Simmental cattle,this experiment used 16S rRNA gene high-throughput sequencing technology to detect and analyze the rumen fluid samples of Simmental cattle (about 20 months old,the average weight was 550 kg),and its function was predicted by PICRUSt.The results showed that a total of 66 712 high-quality sequences were obtained from the Illumina Miseq sequencing platform,and 1 797 operational classification units (OTU) were obtained by cluster analysis.The taxonomic identification was divided into 13 phylums,20 classes,22 orders,33 families and 59 genuses;Firmicutes and Bacteroidetes were dominant bacteria,accounting for 62.46% and 22.45%,respectively;Based on the genus composition,[Eubacterium]_coprostanoligenes_group (9.16%),Ruminococcus_2 (7.90%),norank_f__Bacteroidales_RF16_group(6.23%),norank_f__Ruminococcaceae(4.79%),Rikenellaceae_RC9_gut_group(4.72%),norank_f_Bacteroidales_BS11_gut_group(4.49%),Ruminococcus_1(4.43%),etc.The PICRUSt function of the 16S rRNA genome predicted that the rumen flora function was mainly concentrated in carbohydrate transport and metabolism,and might contain a large amount of cellulose and lignin degrading enzyme genes.In summary,the high-throughput sequencing technology based on 16S rRNA gene fully revealed the diversity of rumen bacteria in Simmental cattle,and predicted that it was rich in protein decomposition and lignocellulose degrading enzymes.The basis of microbial cognition provided a reference for further research on rumen microbial functional genes closely related to some important nutritional and physiological functions.     

饲粮不同NFC/NDF对奶山羊瘤胃溶纤维丁酸弧菌、牛链球菌及埃氏巨型球菌含量变化的影响

试验旨在探讨在饲粮不同非纤维性碳水化合物/中性洗涤纤维条件下,奶山羊发生亚急性瘤胃酸中毒（subacute rumen acidosis,SARA）过程中瘤胃溶纤维丁酸弧菌、牛链球菌及埃氏巨型球菌数量的变化。选用6只安装有永久性瘤胃瘘管的泌乳期关中奶山羊为试验动物,采用动物自身前后对照法,试验分4期进行,每期10d,依次饲喂NFC/NDF比分别为1.02（Ⅰ期）、1.24（Ⅱ期）、1.63（Ⅲ期）、2.58（Ⅳ期）的4种饲粮,以逐渐增加饲粮精料含量的方式诱导奶山羊发生SARA,并采用实时定量PCR技术对瘤胃内溶纤维丁酸弧菌、牛链球菌及埃氏巨型球菌的数量变化进行定量分析。结果表明：①随着饲粮NFC/NDF比的逐步升高,溶纤维丁酸弧菌和埃氏巨型球菌的数量均增加,但当饲粮NFC/NDF比升至2.58时,与其他试验期相比,埃氏巨型球菌的数量极显著增加（P〈0.01）,而溶纤维丁酸弧菌的数量却极显著降低（P〈0.01）;②牛链球菌的数量随着饲粮NFC/NDF比的逐步增加呈显著下降趋势（P〈0.05）,但到第Ⅳ期又恢复到第Ⅰ期的数量。结果提示,当奶山羊发生SARA时,瘤胃牛链球菌的数量无明显变化,对低pH值敏感的溶纤维丁酸弧菌数量急剧下降,而耐酸性的埃氏巨型球菌数量表现为大幅增加。     

In vitro metabolism of tyrosine by rumen bacteria, protozoa and their mixture

An anaerobic in vitro study of tyrosine (Tyr) metabolism by rumen bacteria (B), protozoa (P) and their mixture (BP) was conducted using an HPLC analytical method to obtain some basic and systematic information on aromatic amino acid metabolism in the rumen. Most of the Tyr (> 96%) disappeared after 12 h incubation in B and BP, but only 58% in P. Tyr was converted mainly to p‐hydroxyphenylacetic acid (HPA) in all microbial suspensions. About 45% of disappeared Tyr in B and P, and about 35% in BP were converted to HPA. An appreciable amount of phenylalanine (Phe), 13 and 3% of disappeared Tyr, and a small amount of tryptophan (Trp), 8 and 1% of disappeared Tyr, were also produced from Tyr by rumen bacteria and protozoa, respectively. Small amounts of p‐hydroxyphenylpyruvic acid (about 4 and 6% of disappeared Tyr) were produced from Tyr in B and P, respectively. A moderately large amount of phenylacetic acid (14% of disappeared Tyr) was produced from Tyr in P which was 1.9 times higher than that in B. Phenylpropionic acid and p‐hydroxybenzoic acid were produced only in B and BP. It was concluded that the Tyr degradation ability of rumen bacteria was about 1.5 times higher than that of rumen protozoa. Degraded Tyr mainly produces HPA and then two other aromatic amino acids, Phe and Trp, which are considered essential amino acids for ruminants. Therefore, it is speculated that the requirement for Phe and Trp in ruminants may be partially fulfilled if Tyr is sufficiently supplied in rations.     

A net carbohydrate and protein system for evaluating cattle diets: I. Ruminal fermentation.

The Cornell Net Carbohydrate and Protein System (CNCPS) has a kinetic submodel that predicts ruminal fermentation. The ruminal microbial population is divided into bacteria that ferment structural carbohydrate (SC) and those that ferment nonstructural carbohydrate (NSC). Protozoa are accommodated by a decrease in the theoretical maximum growth yield (.50 vs .40 g of cells per gram of carbohydrate fermented), and the yields are adjusted for maintenance requirements (.05 vs .150 g of cell dry weight per gram of carbohydrate fermented per hour for SC and NSC bacteria, respectively). Bacterial yield is decreased when forage NDF is < 20% (2.5% for every 1% decrease in NDF). The SC bacteria utilize only ammonia as a N source, but the NSC bacteria can utilize either ammonia or peptides. The yield of NSC bacteria is enhanced by as much as 18.7% when proteins or peptides are available. The NSC bacteria produce less ammonia when the carbohydrate fermentation (growth) rate is rapid, but 34% of the ammonia production is insensitive to the rate of carbohydrate fermentation. Ammonia production rates are moderated by the rate of peptide and amino acid uptake (.07 g of peptide per gram of cells per hour), and peptides and amino acids can pass out of the rumen if the rate of proteolysis is faster than the rate of peptide utilization. The protein-sparing effect of ionophores is accommodated by decreasing the rate of peptide uptake by 34%. Validation with published data of microbial flow from the rumen gave a regression with a slope of .94 and an r2 of .88.     

Preliminary observations of interaction between bacteriophages and Streptococcus bovis bacteria on ruminal epithelium primoculture.

Five Streptococcus bovis strains (47/3, 59/2, 4/1, 46/2 and 44/9) isolated from calf ruminal fluid samples were examined for the adherence to cultured ruminal epithelium cells. Four strains (47/3, 59/2, 4/1 and 46/2) were able to attach to the cultured epithelial cells. However, S. bovis 47/3 strain attached to the target cells in significantly greater numbers than the other strains. Strain 44/9 did not adhere to cells of ruminal epithelium. The adherent bacteria were observed on the surface of differentiated (mainly keratinized) cells of ruminal epithelium primoculture only. The different effect of F4, F5 and F6 bacteriophages was ascertained on S. bovis bacteria adhering to rumen epithelial primoculture. A significant decrease in the number of adherent bacteria was shown after cultivation of strains 47/3 and 4/1 with F6 bacteriophage and of 47/3 strain with F4 phage. The F5 bacteriophage had no significant effect on these bacteria.     

Influence of redox potential on metabolism of glucose in mixed cultures of rumen microorganisms

The rumen inocula taken from cows and sheep were incubated in pH controlled anaerobic batch cultures and the effect of the elevated redox potential (Eh) on metabolism of glucose was examined. In Eh regulated cultures (Eh = +100 mV) the VFA production and production of methane were decreased with a concomitant increase of production of lactate. In these cultures counts of lactilytic bacteria tended to be lower and counts of lactic acid bacteria higher than in cultures with a normal Eh. The elevated Eh also exerted an inhibitory effect on the rumen protozoa. The relevance of these changes to the rumen metabolism is discussed.     

The redox potential of growing cultures of Streptococcus bovis Orla-Jensen compared with other facultative anaerobes.

The changes of redox potential were measured in growing cultures of three strains of Streptococcus bovis, together with three strains of Staphylococcus aureus and one strain of each of Lactobacillus plantaram, Lactobacillus casei, and Eschericia coli. It was found that both S. aureus and E. coli could reduce the redox potential of the growth medium to very low values (between —400 mv and —600 mv), whereas the streptococci and lactobacilli were able to cause only slight or insignificant changes of the redox potential. Respirometric measurements confirmed that the capacity of oxygen consumption of S. bovis was very small compared to that of E. coli and S. aureus. On this basis the authors conclude that S. bovis in all probability is unable to contribute significantly to maintenance of the low redox potential of its natural habitat, the rumen. This function must be carried out by other bacteria, such as enterobacteria or staphylococci, which are capable of performing a true, aerobic respiration.     

Immunisation against lactic acidosis in cattle.

The present study was designed to investigate the efficacy of control of lactic acidosis by immunisation against lactic acid-producing bacteria, Streptococcus bovis and Lactobacillus. Ten steers were allocated to two treatment groups. One group was immunised with a vaccine containing S. bovis (strain Sb-5) and Lactobacillus (LB-27) cells, and the other was a non-immunised control group. The vaccine, using Freund's complete adjuvant for primary immunisation and Freund's incomplete adjuvant for boosters, was administered intramuscularly. After primary immunisation, boosters were given at 2 to 4 week intervals. Both anti- S. bovis and anti- Lactobacillus IgG levels in saliva increased significantly (P < 0.01) after the 1st booster which were lower (P < 0.05) than the IgG levels after the 2nd and 3rd boosters, but were not significantly different (P > 0.05) from the IgG levels prior to a grain challenge (after the 4th booster). There were positive correlations between the anti- S.bovis and anti- Lactobacillus IgG in serum and saliva. Compared with the control group, steers in the immunised group had higher (P < 0.05) feed intakes, lower (P < 0.05) rumen concentrations of lactate and lower numbers of S. bovis and Lactobacillus. Three of the control animals were withdrawn from the grain challenge due to their rumen pH persisting below 5.2, while only one animal in the immunised group was withdrawn. These results suggest that the risk of lactic acidosis can be reduced by immunisation against S. bovis and Lactobacillus.