瘤胃微生物组成、日粮与饲料消化率及其关系的研究进展

在反刍动物的瘤胃内栖息着复杂、多样、非致病的各种微生物,包括瘤胃细菌、厌氧真菌和瘤胃原虫,还有少数噬菌体.每克瘤胃内容物含有109～1010个细菌和105～106个瘤胃原虫,瘤胃中厌氧真菌的数量较少.经过长期的适应和选择,微生物和宿主之间、微生物与微生物之间处于一种相互依赖、相互制约的动态平衡系统中.反刍动物的种类、日粮都会导致瘤胃微生物的不同,反之不同的瘤胃微生物又会影响瘤胃对饲料的消化率.因此,对瘤胃微生物的研究,希望更多地掌握反刍动物种类、日粮和微生物及饲料消化率相互之间的关系,为合理应用饲料、调控瘤胃微生物从而提高饲料消化率提供科学依据,在很大程度上对生产实践具有一定的经济意义.     

浅谈反刍动物粗饲料中含氮物的利用

随着反刍动物瘤胃生理和生化过程的研究和发展,人们已经揭示出了瘤胃氮代谢的基本规律以及瘤胃微生物(细菌、纤毛虫和原虫)有利用非蛋白氮(NPN)合成细菌蛋白质以供宿主动物利用方面的作用机制和影响因素。在此基础上,已经开始制定出新     

中国农业科学院哈尔滨兽医研究所——基础免疫创新团队

<正>研究内容1.高致病性猪繁殖与呼吸综合征病毒(HP-PRRSV)-细菌混合感染致病力增强分子机制研究:建立HP-PRRSV-细菌混合感染动物或细胞模型,研究病毒-细菌混合感染导致炎症反应增强和动物死亡增加的分子机制。2.利用基因敲除小鼠为模型,研究病毒、细菌及寄生虫感染后逃逸宿主天然免疫的分子机制。3.寻找具有抑制慢病毒复制活性的宿主限制性因子,研究慢病毒和宿主相互作用的分子机制。理解这些限制性因子的抗病毒活性将有助于发现新型的抗病毒分子机制和开发新的抗病毒治疗方法。     

多组学技术在奶牛瘤胃微生物与宿主互作机制中的研究进展

对于反刍动物而言,瘤胃对宿主的新陈代谢、机体免疫调节具有重要的意义。作为瘤胃内强大而丰富的群落,瘤胃微生物结构紊乱不仅会导致生产性能下降,还会引发机体全身炎症反应。近年来,多组学技术分析瘤胃微生物与宿主的关系及其调控机制已逐渐受到研究者的关注。本文从瘤胃微生物的影响因素、多组学技术在瘤胃微生物中的应用及瘤胃微生物紊乱对宿主的影响等多方面进行综述,以通过调节瘤胃微生物来提高奶牛生产性能、增强机体免疫力以及预防疾病等新的视角为奶牛养殖者提供参考。     

瘤胃微生物区系及相互关系的研究进展

反刍动物瘤胃是一个复杂的生态系统,其中瘤胃微生物与宿主之间、微生物与微生物之间处于一种既协同又制约的动态平衡关系.微生物间的关系会直接或间接影响反刍动物对饲料的消化和利用.笔者将微生物的特点、微生物间的关系以及微生物对饲料的降解进行了阐述,并介绍了分子生物学方法在瘤胃微生物研究中的进展.     

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。     

瘤胃微生物的多样性及其研究技术

瘤胃微生物群落具有丰富的多样性,主要包括瘤胃原虫、瘤胃细菌和厌氧真菌,它们各自有其独特的功能,同时又相互联系,共同与宿主之间构成了复杂的微生态系统。为了探明瘤胃微生态系统,一方面要完善传统的微生物分离技术,另一方面要大力发展分子生物学技术,将二者结合起来进行研究。     

新型脲酶抑制剂对绵羊瘤胃酶活性和细菌总数的影响

为了研究新型脲酶抑制剂对绵羊瘤胃酶活性和细菌总数的影响 ,进行了两个实验。实验一 ,采用反转设计 ,用4头装有瘤胃瘘管的绵羊 ,分别测定了瘤胃液中脲酶、纤维素酶、蛋白水解酶、总脱氢酶的活性和瘤胃细菌总数。结果表明 ,在瘤胃内灌注50mg脲酶抑制剂 (氢醌 ,HQ )可以使脲酶的活性降低65 % (P<0.01) ,使纤维素酶的活性提高28 % (P<0.05) ,对蛋白水解酶、总脱氢酶的活性和瘤胃细菌总数没有影响(P>0.05)。实验二 ,采用滤纸片法观察了氢醌在10、20、40和80mg/L浓度范围内对瘤胃微生物的抑制作用。测定结果表明 ,除了80mg/L氢醌对瘤胃微生物有微弱的抑制作用外 ,其它浓度的氢醌对瘤胃微生物的生长没有影响。上述结果表明 ,HQ是瘤胃微生物脲酶的专一性抑制剂 ,对瘤胃微生物的繁殖和代谢活性无不良影响。HQ在较低浓度范围内使用 ,就可以非常有效地减缓瘤胃中尿素的释放速度 ,对瘤胃微生物和宿主动物安全 ,无毒副作用 ,在反刍动物尿素利用方面具有非常大的开发潜力。     

中国农业科学院哈尔滨兽医研究所——基础免疫创新团队

<正>研究内容1.高致病性猪繁殖与呼吸综合征病毒(HP-PRRSV)-细菌混合感染致病力增强分子机制研究:建立HP-PRRSV-细菌混合感染动物或细胞模型,研究病毒-细菌混合感染导致炎症反应增强和动物死亡增加的分子机制。2.利用基因敲除小鼠为模型,研究病毒、细菌及寄生虫感染后逃逸宿主天然免疫的分子机制。     

PCR-DGGE技术研究青贮桑叶对肉牛瘤胃细菌区系的影响

研究饲料中添加青贮桑叶对肉牛瘤胃细菌区系的影响.选用18月龄,体质量约350 kg的各4头本地黄牛公牛和利木赞×福州牛杂交母牛,每种牛分别饲喂青贮桑叶比例为0(对照组)、7.5%、15%和22.5%的日粮,饲养14周后屠宰取样,提取样品总DNA,利用聚合酶链式反应和变性梯度凝胶电泳相结合的(PCR-DGGE)技术对瘤胃细菌区系进行相似性和多样性分析.结果表明:牛的品种影响瘤胃细菌区系结构及多样性;青贮桑叶添加水平影响瘤胃细菌区系结构,对细菌多样性的影响不显著.     

甘露寡糖添加方式对哺乳期犊牛瘤胃细菌菌群结构的影响

为利用16S rDNA技术研究甘露寡糖不同添加方式对哺乳期犊牛瘤胃细菌菌群结构影响,本研究选用出生日龄一致、体重接近、健康状况良好的荷斯坦公犊牛20头,随机分为4组。CR组为对照组,饮用乳及开食料中均不添加甘露寡糖;ORa组在饮用乳中添加5 g甘露寡糖,开食料中不添加甘露寡糖;ORb组在开食料中添加5 g甘露寡糖,饮用乳中不添加甘露寡糖;ORc组在饮用乳及开食料中各添加2.5 g甘露寡糖(混合添加)。应用16S rDNA测序技术,研究了甘露寡糖添加方式对犊牛瘤胃细菌构建的影响。结果表明:甘露寡糖添加方式会影响哺乳期犊牛瘤胃细菌总数,但对哺乳期犊牛瘤胃细菌菌群多样性未产生显著影响(P>0.05)。在门水平上,与对照组相比,ORb组颗粒料中添加甘露寡糖显著降低了厚壁菌门与放线菌门的丰度(P<0.05),极显著提高了变形菌门的丰度(P<0.01),优势菌门增加为3种,分别为拟杆菌门、厚壁菌门与变形菌门。在属水平上,甘露寡糖不同添加方式导致哺乳期犊牛瘤胃细菌菌群结发生了改变,其中甘露寡糖不同添加方式对小杆菌属和脱硫弧菌属在瘤胃内丰度均产生了影响,但对ORa组与ORc组犊牛瘤胃细菌影响不及ORb组;且ORb组犊牛瘤胃细菌功能性菌属普雷沃氏-7属、氨基酸球菌属、优杆菌属和琥珀酸弧菌等均发生显著变化。基于以上结果,得出如下结论:本研究条件下,甘露寡糖添加方式对哺乳期犊牛瘤胃细菌菌群多样性未产生显著影响(P>0.05),但开食料中寡糖添加方式对菌群结构产生部分影响,其中发挥瘤胃蛋白降解作用和淀粉降解作用的琥珀酸弧菌科-UCG-001属、利用乳酸的新月形单胞菌属丰度极显著提高(P<0.01),而利用乳糖的优杆菌属丰度显著降低,降解半纤维素的小杆菌属丰度极显著降低(P<0.01)。     

饲粮纤维水平对育肥羔羊瘤胃微生物组成及多样性的影响

试验旨在研究提高饲粮纤维水平对育肥羔羊瘤胃微生物组成及多样性的影响。试验选择健康无病、体重相近的2月龄杜湖杂交公羔,按照体重随机分为2组,每组38只,分别饲喂不同的饲粮,对照组（LW_CK）饲粮为精料+苜蓿,处理组（LW_EG）为精料+苜蓿+全株玉米青贮+花生秧;试验期150 d,其中预试期20 d,正试期130 d,试验结束后,每组随机选择5只羔羊屠宰取瘤胃内容物,通过16S rDNA高通量测序技术分析其微生物多样性及结构变化。结果表明,处理组羔羊瘤胃中细菌多样性显著高于对照组（P<0.05）。在门水平上,处理组羔羊瘤胃中优势菌门为拟杆菌门（Bacteroidetes）和厚壁菌门（Firmicutes）,对照组优势菌门为拟杆菌门和变形菌门（Proteobacteria）。提高纤维水平后显著增加了育肥羔羊瘤胃中厚壁菌门的相对丰度（P<0.05）;显著降低了变形菌门的相对丰度（P<0.05）。在属水平上,对照组与处理组羔羊瘤胃菌群共鉴定出195个属,处理组优势菌属为普雷沃氏菌属（Prevotella_1）和解琥珀酸菌属（Succiniclasticum）,对照组优势菌属为琥珀酸弧菌属（Succinivibrionaceae）和普雷沃氏菌属（Prevotella_7）。提高饲粮纤维水平,增加了育肥羔羊瘤胃中解琥珀酸弧菌属、密螺旋体属（Treponema_2）、疣微菌属（Ruminococcaceae）、月形单胞菌属（Selenomonas_1）的相对丰度,显著降低了琥珀酸弧菌属相对丰度（P<0.05）。综上所述,适量提高饲粮中纤维水平对育肥羔羊瘤胃微生物菌群结构有一定影响,能够促进部分纤维降解菌的增殖,有利于育肥羔羊的瘤胃发酵和养分消化利用。     

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.     

Viral networks and detection of potential zoonotic viruses in bats and rodents: A worldwide analysis

Bats and rodents are recognized to host a great diversity of viruses and several important viral zoonoses, but how this viral diversity is structured and how viruses are connected, shared and distributed among host networks is not well understood. To address this gap in knowledge, we compared the associative capacity of the host–virus networks in rodents and bats with the identification of those viruses with zoonotic potential. A virus database, detected by molecular methods, was constructed in the two taxonomic groups. We compiled 5,484 records: 825 in rodents and 4,659 in bats. We identified a total of 173 and 166 viruses, of which 53 and 40 are zoonotic viruses, in rodents and bats, respectively. Based on a network theory, a non‐directed bipartite host–virus network was built for each group. Subsequently, the networks were collapsed to represent the connections among hosts and viruses. We identified both discrete and connected communities. We observed a greater degree of connectivity in bat viruses and more discrete communities in rodents. The Coronaviridae recorded in bats have the highest values of degree, betweenness and closeness centralities. In rodents, higher degree positions were distributed homogeneously between viruses and hosts. At least in our database, a higher proportion of rodent viruses were zoonotic. Rodents should thus not be underestimated as important reservoirs of zoonotic disease. We found that viruses were more frequently shared among bats than in rodents. Network theory can reveal some macroecological patterns and identify risks that were previously unrecognized. For example, we found that parvovirus in megabats and Gbagroube virus in rodents may represent a zoonotic risk due to the proximity to humans and other zoonotic viruses. We propose that epidemiological surveillance programmes should consider the connectivity of network actors as a measure of the risks of dispersion and transmission.     

酵母发酵饲料对瘤胃菌群数量及多样性的影响

本试验研究日粮中添加酵母发酵饲料对蒙古绵羊瘤胃菌群数量及多样性的影响.选用10只安装永久性瘤胃瘘管且体重约40 kg的14月龄蒙古羯羊,分为对照组和试验组,每组各5只.对照组饲喂基础日粮,试验组饲喂基础日粮+酵母发酵饲料(13.37％).预试期15 d,正试期5 d.于正试期晨饲后0、3、6、9、12 h依次采集瘤胃液...     

甘露寡糖、果寡糖和大豆寡糖组合对锦江黄牛瘤胃液细菌多样性的影响

本试验旨在研究功能性寡糖(甘露寡糖、果寡糖和大豆寡糖)组合对锦江黄牛的瘤胃液细菌多样性的影响.选择3头健康且体重相近的安装有永久性瘤胃瘘管的锦江黄牛,采用自身对照方法,试验分2期进行,第1期:对照期,饲喂基础饲粮,不添加任何寡糖;第2期:添加期,饲喂基础饲粮+功能性寡糖(0.8％甘露寡糖+1.0％果寡糖+0.8％大豆寡糖),每期持续14 d.利用PCR-变性梯度凝胶电泳(DGGE)方法研究分析牛瘤胃液细菌多样性.结果表明:添加功能性寡糖组合后,牛瘤胃液细菌PCR-DGGE图谱条带数明显增加,3头牛增加了5个共同条带.经测序,其中2个是产琥珀酸丝状杆菌属,3个是未鉴定细菌.饲粮中添加功能性寡糖组合可使瘤胃液细菌菌群种类明显增加.结果提示,饲粮添加功能性寡糖组合能够影响瘤胃菌群区系,增强其稳定性.     

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.     

动物肠道菌群与病原微生物感染关系的研究进展

动物肠道内寄居着大量微生物,通常被称为共生菌群。它们对动物的生长、代谢和免疫状态至关重要,还与许多疾病的发生密切相关。病毒、细菌和寄生虫感染都会使机体的肠道菌群发生紊乱,表现在益生菌丰度减少而有害菌丰度增加。其机制包括引发宿主的炎症反应和抑制机体的免疫细胞两方面。同样肠道菌群也会调控病原菌的感染,如肠道菌群对不同的病毒会产生颉颃或促进作用,对细菌和寄生虫分别产生抑制和促进作用。肠道菌群抑制病原菌的机制包括与病原菌竞争代谢产物和诱导宿主的免疫反应。肠道菌群促进病毒感染的机制包括3点,分别为提高病毒的稳定性及其与靶细胞的黏附作用、抑制机体免疫系统和刺激靶细胞的增殖。肠道菌群促进寄生虫感染的可能机制包括降低Th2细胞因子(如IL-4和IL-13)并提高调节性T细胞的表达频率。肠道菌群、病原微生物和宿主不断相互作用,形成一个动态的平衡关系,并在感染过程中不断进化。作者主要综述了病毒、细菌和寄生虫感染对动物肠道菌群的组成和丰度的影响,动物肠道菌群如何影响病毒、细菌和寄生虫的感染进程并分析相关机制,以期了解疾病的发病机理,为疫苗佐剂的研发及制定更有效的预防和治疗策略提供新视角和理论依据。     

猪肠道细菌培养组学研究进展

猪肠道菌群是由所有定殖在猪肠道里的大量细菌、病毒、真菌和古菌等构成的集合.已有研究表明,很多疾病以及猪重要经济性状都与肠道菌群有关.目前,肠道菌群研究使用较多的技术是16S rRNA基因测序和宏基因组测序,但这些技术并不能了解具体菌株的实际功能和生理特性.肠道细菌的分离培养具有特殊重要的意义.近几年来,肠道细菌的培养取...     

Diversity Analysis of Rumen Microbes in Yunnan Gayals (Bos frontalis)

In order to study the rumen microbial diversity in Yunnan gayals (Bos frontalis), we analyzed the diversity of microbes using largely assembled sequence data obtained by metagenomic sequencing DNA isolated from rumen samples of eight adult galyals. The genome primary sequence (total metagenome size was >3 Gb) was assembled with short reads.Based on the gayal rumen microbial metagenomic sequencing, we received a total of 13 546 196 EGTs. Because of the relatively large amount, 10 387 566 EGTs (76.68% of all EGTs ) were randomly selected to analyze microbial communities by BLAST search of the NCBI-NR database. Microbial species annotations analysis revealed that microbial communities from gayals rumen were mainly composed of the bacterial domain, eukarya domain, archaea domain and unknown microbial composition. The majority microbes mainly belonged to ten phylums, and 94% EGTs belonged to Bacteroidetes, Firmicutes, Proteobacteria and Actinobacteria, of which the highest abundance phylums were Bacteroidete and Firmicutes, accounting for 48% and 42%, respectively. In addition, the main cellulolytic bacteria in rumen of gayals were Ruminococcus flavefacie, Butyrivibrio fibrisolvens, Ruminococcus albus and Fibrobacter succinogenes. They accounted for 1.60%, 1.20%, 0.86% and 0.52% of the total bacteria, respectively. These results further indicated that EGTs data could be used to analyze and explain environmental microbial community structure and function. There was great significance to study the composition of microorganisms in special environments.