宏基因组学技术在瘤胃微生物研究中的应用研究进展

反刍动物瘤胃中存在大量微生物,这些瘤胃微生物(rumen microorganisms)是影响反刍动物健康和生产性能的关键因素。宏基因组学技术的发展为研究瘤胃微生物开辟了新的途径和方法。利用宏基因组学技术可研究瘤胃微生物的多样性、瘤胃微生物对宿主生理代谢的影响、瘤胃微生物和宿主的相互作用关系,并且还可以挖掘与代谢相关的新功能基因。笔者综述了宏基因组学技术发展背景,以及该技术在研究反刍动物瘤胃微生物方面的主要应用和不足,包括宿主及其生存环境对瘤胃微生物的影响、日粮中精粗比和饲料添加剂对瘤胃微生物的影响、瘤胃微生物中的抗生素耐药基因(antibiotic resistance genes, ARGs)、瘤胃微生物对反刍动物产奶性能的影响,并展望了宏基因组学技术在研究瘤胃微生物方面的应用前景,旨在为反刍动物瘤胃微生物的研究提供参考。     

瘤胃发育研究进展

瘤胃是反刍家畜进行消化、吸收以及代谢的主要器官,瘤胃发育主要表现为瘤胃上皮和瘤胃乳头解剖结构以及生理功能的变化,而饲料组成和营养水平可在不同程度上影响其形态和功能,文章从影响瘤胃发育的要素、瘤胃上皮和乳头等方面对瘤胃发育进行概述。     

体外法研究瘤胃筛选真菌对崂山奶山羊瘤胃发酵的影响

为了分析评价瘤胃筛选真菌在瘤胃微生物降解利用秸秆等纤维性饲料中的功效。本研究通过模拟瘤胃内环境,运用体外法研究了瘤胃厌氧真菌筛选菌对瘤胃发酵指标的影响,确定筛选真菌A、B、C和D对产气量、NH3-N、菌体蛋白和挥发性脂肪酸(VFA)4种瘤胃发酵指标的影响。旨在初步探讨瘤胃真菌筛选菌株对其瘤胃发酵功能的影响,为研究瘤胃真菌筛选菌株添加到瘤胃内对瘤胃降解粗纤维饲料的可能性提供参考。试验结果表明,按培养底物玉米秸秆添加2%真菌D对改善瘤胃发酵功能的综合作用效果最为理想。     

添加草药改善瘤胃功能

添加草药改善瘤胃功能ＣｈａｒａｎｊｉｔＳＲａｎｄｈａｗａ等著任鹏摘译曹友高校瘤胃内物理、化学及微生物因素，如瘤胃活力、ｐＨ、日粮的品质、瘤胃微生物类型，均能影响瘤胃的发酵。１生物胺会降低瘤胃发酵速率一些具药理活性的物质可间接影响瘤胃发酵，如产生于肠道...     

奶牛氨基酸过瘤胃技术及效果概述

本文综述了过瘤胃氨基酸的概念、分类、保护效果、过瘤胃氨基酸对奶牛的影响和影响过瘤胃氨基酸效果的因素及应用前景。     

牛瘤胃积食的综合防治技术应用

瘤胃积食在畜牧养殖业中比较常见,对于养殖户的影响也比较大,其发生是由于瘤胃内有过多的粗饲料在牛的瘤胃内不能排出导致瘤胃体积增大,胃壁扩张,从而影响瘤胃正常的消化机能。本文主要讨论了牛瘤胃积食的发病原因、发病症状、诊断方法、治疗技术和预防技术。     

反刍动物瘤胃氮代谢及其与瘤胃微生物相关性的研究进展

通过饲料养分调控瘤胃氮代谢是提高反刍动物氮利用效率的一种有效方式,其中能量和蛋白质对瘤胃氮代谢的影响尤为显著。能量和蛋白质通过调控瘤胃中的微生物进而影响瘤胃氮代谢过程,但两者对瘤胃中不同微生物的影响程度存在差异。本文综述了瘤胃中的氮降解过程、尿素氮循环、能氮平衡对瘤胃氮代谢的调控以及能量和蛋白质与瘤胃微生物之间的关系,为提高反刍动物氮利用效率和减少反刍动物氮排放量的研究提供科学依据。     

瘤胃挥发性脂肪酸的作用及影响因素

瘤胃挥发性脂肪酸与反刍动物瘤胃营养以及瘤胃微生物有着密切的关系。反刍动物瘤胃对养分的消化吸收以及微生物动力学可产生大量的挥发性脂肪酸,其主要作用为供能和维持瘤胃环境。本文综述了瘤胃挥发性脂肪酸(VFA)的作用、影响其产生和组成的因素以及其对反刍动物的影响。     

过瘤胃氨基酸在奶牛生产中的应用

本文综述了过瘤胃氨基酸的概念、稳定性、保护效果的测定方法及过瘤胃氨基酸对奶牛生产性能的影响和影响过瘤胃氨基酸效果的因素。     

绵羊瘤胃水平衡、瘤胃发酵与微生物蛋白质合成

瘤胃原虫对细菌的吞食作用是降低瘤胃微生物蛋白质合成量的重要因素。提高瘤胃内容物的稀释率可以减少原虫对细菌的吞食作用及微生物的维持能量需要,因而瘤胃水平衡对于瘤胃微生物区系、瘤胃发酵及微生物蛋白质的合成量有重要影响。本研究进行了以下几方面的工作:(1)研究瘤胃内水的来源与去路及影响水平衡的因素,探讨根据瘤胃水平衡测定绵羊唾液分泌方法;(2)研究瘤胃内灌注人工唾液对瘤胃液体平衡、瘤胃发酵及微生物蛋白质合成的影响;瘤胃液稀释率与微生物蛋白质合成之间的关系。(3)探讨在生产实际中,调控瘤胃水平衡     

Comparison of rumen bacteria distribution in original rumen digesta,rumen liquid and solid fractions in lactating Holstein cows

Background: Original rumen digesta, rumen liquid and solid fractions have been frequently used to assess the rumen bacterial community. However, bacterial profiles in rumen original digesta, liquid and solid fractions vary from each other and need to be better established.Methods: To compare bacterial profiles in each fraction, samples of rumen digesta from six cows fed either a high fiber diet(HFD) or a high energy diet(HED) were collected via rumen fistulas. Rumen digesta was then squeezed through four layers of cheesecloth to separate liquid and solid fractions. The bacterial profiles of rumen original digesta, liquid and solid fractions were analyzed with High-throughput sequencing technique.Results: Rumen bacterial diversity was mainly affected by diet and individual cow(P 0.05) rather than rumen fraction. Bias distributed bacteria were observed in solid and liquid fractions of rumen content using Venn diagram and LEf Se analysis. Fifteen out of 16 detected biomarkers(using LEf Se analysis) were found in liquid fraction, and these 15 biomarkers contributed the most to the bacterial differences among rumen content fractions.Conclusions: Similar results were found when using samples of original rumen digesta, rumen liquid or solid fractions to assess diversity of rumen bacteria; however, more attention should be draw onto bias distributed bacteria in different ruminal fractions, especially when liquid fraction has been used as a representative sample for rumen bacterial study.     

不同瘤胃细菌培养物添加水平对奶牛瘤胃发酵及BCP含量的影响

本文旨在研究不同剂量的瘤胃细菌培养物对荷斯坦奶牛瘤胃发酵及菌体蛋白(BCP)含量的影响。试验采用单因素试验设计,选用4头装有永久性瘤胃瘘管,体况相近的荷斯坦奶牛,取其瘤胃液混匀,添加不同剂量的瘤胃细菌培养物进行体外培养,分别为:对照组(0 CUF/g),试验Ⅰ组(添加量为3%,4.4×108CUF/g),试验Ⅱ组(添加量为5%,7.8×108CUF/g),试验Ⅲ组(添加量为10%,1.2×109CUF/g),每组分别设2、4、8、12、24、36、48 h 7个指标测定时间点,每个时间点3个重复。结果表明:(1)添加瘤胃细菌培养物使瘤胃液pH有所降低,但始终在6.5～7.0,对瘤胃内环境无不良影响;(2)添加3%的瘤胃细菌培养物显著降低氨态氮(NH3-N)的含量(P<0.05),显著增加了培养液BCP的浓度(P<0.05);(3)添加3%的瘤胃细菌培养物可一定程度增加总挥发性脂肪酸(TVFA)和乙酸浓度,但对其他各挥发性脂肪酸及乙酸/丙酸比值无显著影响。由此可见,添加3%剂量的瘤胃细菌培养物可改变瘤胃的发酵环境,提高BCP浓度。     

Forestomach epithelial receptor activation by rumen fluids from sheep given intraruminal infusions of volatile fatty acids

Forty-four reticuloruminal epithelial receptors were tested with rumen fluids obtained from 12 sheep before they were intraruminally infused with 4.0M acetic acid (8 sheep) or 4.0M butyric acid (4 sheep; preinfusion rumen fluid) and with rumen fluids obtained at the onset of ruminal stasis (abolition rumen fluid). The preinfusion rumen fluids from the 8 acetic acid-infused sheep (mean pH, 6.55) contained 1.7 mM nondissociated volatile fatty acids (VFA)/L and excited none of the 25 receptors tested. Preinfusion rumen fluids from the 4 butyric acid-infused sheep (mean pH, 6.98) contained 0.3 mM nondissociated VFA/L and also did not evoke responses in any of the 19 receptors tested. Abolition rumen fluids from sheep treated with acetic acid excited 17 of the 25 receptors tested and contained 89.4 mM nondissociated VFA/L, of which nondissociated acetic acid comprised 85.0 mM/L. Abolition rumen fluids from sheep treated with butyric acid activated 14 of the 19 receptors tested and contained 61.1 mM nondissociated VFA/L, of which 38.7 mM/L was nondissociated butyric acid. Preinfusion rumen fluids whose pH values were adjusted to that of abolition rumen fluids with HCl contained nondissociated VFA levels ranging from 16.3 mM/L (acetic acid-treated sheep) to 20.6 mM/L (butyric acid-treated sheep) and elicited responses in 4 of 30 receptors tested. Preinfusion rumen fluids whose pH values were adjusted to the pH value of abolition rumen fluid with acetic acid contained 29.5 mM nondissociated VFA/L and excited 7 of 13 tested receptors.(ABSTRACT TRUNCATED AT 250 WORDS)     

Relationship between the volatile fatty acids (VFA) in the rumen and nitrogen secretion into isolated sheep's rumen]

Two sheep with a ruminal fistula and an isolated small rumen were studied for the secretion of ammonia nitrogen, urea nitrogen, and amino nitrogen into the isolated rumen at different levels of volatile fatty acids (VFA) (50, 133-97, and 97-66 M Mol 1(-1)) in the rumen. The VFA level in the rumen was found to exert a great influence on the quantitative secretion of endogenous nitrogen from the blood through the rumen wall into rumen content. When the VFA level in the rumen was increased by administration of a single dose of acetic, propionic, and butyric acid, the secretion of ammonia nitrogen and amino nitrogen abruptly dropped and the secretion of urea into the isolated rumen slightly increased. The over-all amount of nitrogen (NH3-N + urea-N + amino-N) that had passed into the isolated rumen in the course of an hour showed a highly significant correlation with the passage of nitrogen in the form of ammonia and amino nitrogen and was greatest before the application of VFA to the rumen, i.e. at the level of 50 m mol 1-1. Of the metabolites under study, which were passing to the isolated rumen, amino nitrogen shared the greatest proportion (45.38-46.54%). When the VFA level in the rumen was raised, the proportion of ammonia secreted to the isolated rumen decreased and the proportion of urea in the total amount of nitrogen increased.     

The effects of oral magnesium hydroxide administration on rumen fluid in cattle

This study was conducted to determine the effects of oral magnesium hydroxide administration on rumen fluid in cattle. Six lactating Holstein cows (4-7 years of age) with rumen fistulas were studied. Cattle were randomly assigned to receive boluses of magnesium hydroxide (162 g) or a powdered form (450 g dissolved in 3.5 L of water) PO daily for 3 days. Analysis of rumen fluid, blood gas tensions, and pH and measurement of serum magnesium concentrations were conducted daily. The study was discontinued after 72 hours, or sooner if rumen pH exceeded 8.0. After at least 3 weeks, the study was repeated with each cow receiving the other form of magnesium hydroxide (powder or bolus). Compared with baseline rumen pH (mean +/- SD: 6.22 +/- 0.28), magnesium hydroxide boluses caused a significant increase (P < .05) in rumen pH after 48 (7.27 +/- 0.11) and 72 (8.01 +/- 0.16) hours of administration, whereas the powdered form caused a significant increase (P < .05) in rumen pH after 24 (7.54 +/- 0.19) and 48 (8.43 +/- 0.22) hours of administration. Both the powdered and bolus forms of magnesium hydroxide decreased rumen protozoal numbers and increased methylene blue reduction times compared with baseline values. There was no change in blood pH, bicarbonate, or base excess values. Serum magnesium concentrations were significantly increased (P < .05) in cows that received the magnesium hydroxide powder. The results of this study indicate that magnesium hydroxide has a potent alkalinizing effect on rumen pH and significantly decreases rumen microbial activity.     

Effect of N fertilisation rate, energy supplementation and supplementation strategy on efficiency of N utilisation in the sheep rumen.

The effect of nitrogen (N) fertilisation (200 vs. 400 kg N ha-1 year-1) of pasture cut in the beginning (end of May) and the end (end of August) of the grazing season and of simultaneous or separated feeding of maize and grass (400 kg N ha-1 year-1) on efficiency of N utilisation in the rumen has been studied using four rumen cannulated wethers. Doubling N fertiliser rate increased grass CP production by about 60%, but induced extensive excess of rumen degradable N, reflected in high urinary urea excretion (rpearson = 0.747). The latter was lower (74% at the maximum) when feeding less fertilised or older grass or when supplementing with maize silage. Although simultaneous feeding of maize silage with grass changed patterns of rumen ammonia concentrations, no change in urinary excretion of purine derivatives was observed. Hence, faecal or urinary N excretion was unaffected by the supplementation strategy. Microbial growth efficiency was estimated from urinary excretion of purine derivatives and fermented OM. The latter was calculated from total rumen CH4 production, based on rumen fermentation stoichiometry and taking into account proportional concentrations of individual volatile fatty acids. Higher levels of intake tended to improve rumen microbial growth efficiency slightly (rpearson = 0.406), which, however, could not compensate for the reduced effective rumen DM degradability (rpearson = -0.442). The latter was apparently associated with a partial shift of the fermentation from the rumen to the hindgut, as suggested from the negative correlation (rpearson = -0.745) between faecal RNA concentrations and rumen effective degradability.     

幼龄反刍动物纤维营养需要与影响因素

幼龄反刍动物饲粮的纤维性碳水化合物(FC)来源与组成对调控其生长发育和瘤胃功能建立有重要作用,其来源各异,粒度不同,对正处于生长期的幼龄反刍动物的采食和消化、瘤胃液p H及瘤胃发育等生理参数的影响存在差异。本文综述了近年来关于幼龄反刍动物饲粮中性洗涤纤维(NDF)来源和粒度的研究,分别从生产性能、瘤胃发酵和瘤胃发育3个方面总结了二者对幼龄反刍动物的影响,并对相关机制进行了探讨。     

新型饲料添加剂对稻草对牛瘤胃降解特性影响的研究

以3头装有瘤胃瘘管的湖北本地黄牛为实验动物 ,研究了在以稻草为主的日粮条件下 ,补喂SL添加剂和B(SL)舔块对稻草干物质(DM)和中性洗涤纤维(NDF)瘤胃降解率的影响 ,同时测定了瘤胃液的 pH和氨氮浓度。结果表明 :添加SL后 ,加快了稻草的消化速度 ;饲喂B(SL)后 ,显著加快了稻草的消化速度(P<0.01) ,明显提高了稻草DM和NDF降解率(P<0.05) ;添加SL或B(SL)舔块后 ,瘤胃液pH有下降趋势 ,但差异不显著 ;2种舔加剂均显著降低采食4h内的瘤胃液氨氮浓度(P<0.01) ,并使氨氮浓度曲线趋于平缓 ,说明有效提高了瘤胃微生物对降解氮的转化能力。     

小麦面筋蛋白对山羊瘤胃运动的影响

采用活体瘤胃运动测定装置，测定山羊日粮中添加小麦面筋蛋白条件下瘤胃运动的变化。结果表明；4只山羊试验期与对照期相比，瘤胃平滑肌收缩力平均提高31．2％，差异极显著（P〈0．05）。15min内，瘤胃平均蠕动持续时间延长34．1％，差异显著（P〈0．05）。瘤胃蠕动频率下降25．1％，差异显著（P〈0．05）。建立的瘤胃运动检测装置能够灵敏地检测山羊瘤胃运动变化规律。同时利用家兔小肠体外培养装置检测到小麦面筋蛋白通过瘤胃微生物降解作用生成的阿片肽活性，为反刍动物瘤胃代谢调控的研究提供理论依据。     

Effects of protozoa on methane production in rumen and hindgut of calves around time of weaning.

Effects of the presence or absence of ciliate protozoa on methanogenesis in the rumen and hindgut were investigated in young calves during a 7-week period. Ten Holstein calves, aged 7 days, were divided in two groups (n = 5) and fed an increasing amount of a commercial milk replacer and small amounts of a calves starter. One group was inoculated with ciliate fauna on two occasions, week 5 and 6, while the second remained ciliate-free. The absence of protozoa in the rumen decreased rumen empty weight (-23%, P < 0.01), and rumen pool size of N (-36%, P < 0.01) and crude fat (-37%, P < 0.05). Rumen bacteria of non-faunated calves contained a higher proportion of total amino acid-N per 16 g N (+3%, P < 0.01) and D-alanine-N per 16 g N (+13%, P < 0.05) compared to faunated calves. Further results contain a reference for a higher bacterial mass in the ciliate-free rumen with an increased number of bacteria adherent to rumen mucosa. The CH4 production in the rumen increased exponentially with the increase in protozoa population size (R2 = 0.68). In presence of 46 x 10(4) protozoa per ml rumen fluid, the in vitro CH4 production of rumen fluid per mol total VFA was about 34% higher in faunated than in non-faunated calves (P < 0.001). Hydrogen (2H) recovery of rumen fermentation was positively correlated (R2 = 0.55) to the CH4 production rate. Methanogens were attached on rumen mucosa. Methanogenesis, induced by rumen mucosa attached bacteria, was stimulated by ruminal protozoa. In the absence of protozoa in the rumen, the acetate-propionate ratio and butyrate proportion of VFA were reduced. In vivo, in the absence of protozoa not only the whole animal CH4 production (-30%, P < 0.05) but also the digestibility of carbohydrates (-4%, P < 0.05) was reduced. Thereby no difference was observed in the intake of ME per kg DM between the groups. In conclusion, the methanogenesis in the rumen, but not in hindgut, is associated with the development of the ruminal protozoa population. The level of methanogenesis (mol/mol VFA) in the hindgut amounts to 20% of the ruminal methanogenesis.