T-RFLP分析技术在肉鸡肠道微生物研究中的应用

本文阐述了末端限制性片段长度多态性(T-RFLP)分析技术的基本原理和技术路线,讨论了应用T-RFLP分析技术相对于应用传统方法研究肉鸡肠道微生物的优势,总结了应用T-RFLP分析技术研究肉鸡肠道微生物的发展历程,分析了应用T-RFLP分析技术研究肉鸡肠道微生物存在的问题及解决方法,并对其应用前景进行了展望.     

芽孢乳杆菌对肉仔鸡生产性能、肠道发育和微生物菌群的影响

本试验旨在研究不同水平芽孢乳杆菌对内仔鸡生产性能、肠道发育及微生物菌群的影响.选取1日龄爱拔益加(AA)肉仔鸡2 400只,随机分为4组(每组6个重复,每个重复100只):对照组饲喂基础饲粮,试验组饲喂分别添加200、100和50 mg/kg芽孢乳杆菌的试验饲粮,试验期42 d.结果表明:1)饲粮添加200 mg/kg...     

Significance of earthworms in stimulating soil microbial activity

 The stimulatory effect of earthworms (Lumbricus terrestris L.) on soil microbial activity was studied under microcosm-controlled conditions. The hypothesis was tested that microbial stimulation observed in the presence of a soil invertebrate would be due to the utilization of additional nutritive substances (secretion and excretion products) that it provides. Changes in microbial activity were monitored by measuring simultaneously CO₂ release and protozoan population density. The increase in CO₂ released in the presence of earthworms was found to result from both earthworm respiration and enhanced microbial respiration. The stimulation of microbial activity was confirmed by a significant increase in protozoan population density, which was 3–19 times greater in the presence of earthworms. The respiratory rate of L. terrestris was estimated to be 53 μl O₂ g^–1 h^–1. Earthworm respiration significantly correlated with individual earthworm weight, but there was no correlation between the increase in microbial respiration and earthworm weight. This finding does not support the hypothesis given above that enhanced microbial respiration is due to utilization of earthworm excreta. A new hypothesis that relationships between microbial activity and earthworms are not based on trophic links alone but also on catalytic mechanisms is proposed and discussed. Received: 26 August 1997     

黑土长期定位试验原状土搬迁对土壤微生物性质的影响

通过测定黑土长期定位试验原状土整体搬迁前后四种处理(CK、NPK、M、MNPK)土壤可培养微生物数量、土壤微生物量含量及土壤酶活性变化,探讨搬迁的特定条件对土壤微生物性质的影响。结果表明,1搬迁前后5年各处理三大类微生物数量、微生物量碳、氮含量和四种土壤酶活性在0~20 cm土层中均高于20~40 cm土层;2每年在两个土层中MNPK处理的三大类微生物数量和微生物量碳、氮含量均最高,CK处理均最低,而每年在两个土层中土壤过氧化氢酶和转化酶活性CK和M处理均明显高于NPK和MNPK处理,而磷酸酶活性CK和M处理则低于NPK和MNPK处理,脲酶活性在4个处理中差异不大;3比较搬迁前后5年间各项微生物指标变化发现,各项指标年际间虽存在变化但程度不大,搬迁后2011年和2012年相对较高,同为小麦茬的搬迁前2010年和搬迁后2013年各项指标较为接近。研究说明,土壤深度和施肥对土壤各项生物指标影响较大,年际间变化也受到轮作方式的影响,搬迁对土壤微生物扰动影响远小于施肥和耕作方式等的影响。     

不同脆化阶段草鱼肠道菌群动态变化、血清酶指标及生长性能

本研究旨在探讨草鱼(Ctenopharyngodon idellus)不同脆化阶段的肠道菌群组成结构、血清酶活性及生长性能,进而衡量脆肉鲩形成过程中的生理生化变化,以期为脆肉鲩的健康养殖提供科学数据.脆化不同阶段(30、60、90和120 d)的草鱼,在脆化结束后转投喂人工配合饲料30 d(第150天),鉴定整个过程中草鱼肠道菌群组成、血清酶活性和生长性能的变化.结果表明,60～150 d蚕豆组草鱼体重均低于对照组(P＜0.05,P＜0.01),90～120 d蚕豆组草鱼头肾体指数(head kidney somatic index,HKBI)和肝体指数(hepatosomatic index,HSI)均低于对照组(P＜0.05或P＜0.01),150 d时蚕豆组草鱼肝体指数与对照组无显著差异(P＞0.05);蚕豆组草鱼血清碱性磷酸酶、溶菌酶和补体C3(第30天除外)活性均低于对照组的,在转投喂饲料后,3种血清酶活性均出现回升趋势;与对照组相比,蚕豆组草鱼肠道菌群多样性降低,芽胞杆菌属(Bacillus)和鲸杆菌属(Cetobacterium)等有益菌数量减少,维氏气单胞菌(Aeromonas veronii)、温和气单胞菌(Aeromonas sobria)和鳗弧菌(Vibrio anguillarum)等条件致病菌数量增多.两组草鱼肠道核心菌群主要为变形菌门(Proteobacteria)、厚壁菌门(Firmicutes)、拟杆菌门(Bacteroidetes)和梭杆菌门(Fusobacteria).本研究结果提示,随着脆化时间的增加,摄食蚕豆改变草鱼肠道菌群组成、抑制血清酶活性和生长性能,在转投饲料后其肠道菌群、血清酶活性及生长性能均有所改善.本研究结果对脆肉鲩的健康养殖具有重要的科学指导意义.     

The Application of Microbiome Culturomics in Veterinary Medicine

Under laboratory conditions, the number of cultured microorganisms accounts for about 1% of the total number of microorganisms in nature, which limits people's understanding and utilization of 99% of the unknown microorganisms. However, relevant researches show that those "uncultured microorganisms" can be developed and utilized, and the uncultured microorganisms are the main body of the unknown microorganisms. The microbial culturomics explored the application of multiple culture conditions and long-term culture, it was combined with Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF-MS) and 16S ribosomal RNA (rRNA) sequencing to identify all kinds of microorganisms on a large scale. At the same time, whole-genome sequencing (WGS) and Metagenomics sequencing technology were used to analyze unknown microorganisms in depth. In this paper, the latest progress of culturomics in the ruminant gastrointestinal tract, poultry cecum, and livestock nasal microflora in recent years was reviewed, and the feasibility of applying the method of microflora culturomics in animal disease prevention and control was discussed. As a new research idea, culturomics has some immature aspects, but its development prospect is very broad. The complementary of microflora culturomics and other research methods have gradually become a breakthrough in the development of veterinary microbiology.     

多菌种复合益生素对三黄鸡的生产性能及肠道主要菌群的影响

选用1 560只1日龄的广西岑溪三黄肉仔鸡,随机分成2组,每组780只,对照组饲喂基础日粮,试验组日粮由基础日粮加0.1%的益生素组成,饲养时间6周,研究多菌种复合菌益生素对三黄鸡的生产性能和肠道主要菌群的影响。试验结果表明:全期试验组的平均日增重比对照组高12.58%(P<0.01),料重比比对照组低9.23%(P<0.05);试验结束时,雏鸡空肠和盲肠大肠杆菌数量分别比对照组低27.61%(P<0.05)和25.57%(P<0.05),乳酸菌数量分别比对照组高16.12%(P>0.05)和11.28%(P>0.05)。     

芽孢杆菌制剂对艾维茵肉鸡胴体性能和肠道菌群的影响

试验选用7日龄艾维茵肉仔鸡480只,随机分为8组,每组3个重复,每个重复20只;选择适宜肉鸡饲喂的3种芽孢杆菌制剂,分别为枯草芽孢杆菌BS3(Bacillus subtilis 3)、产蛋白酶活力高的蜡样芽孢杆菌BC1(Bacillus cereus 1)和产纤维素酶活力高的蜡样芽孢杆菌BC2(B.cereus 2),将其单一或复合的添加到肉鸡日粮中,研究芽孢杆菌制剂对肉鸡胴体性能、免疫器官指数及肠道菌群的影响;8个试验组分别为对照组和BC1、BC2、BS3单一供给组及BC1+BC2、BC1+BS3、BC2+BS3、BC1+BC2+BS3 4个混合供给组,饲料中芽孢杆菌的添加量为3×105 CFU.g-1,混合组的配比为1∶1和1∶1∶1。结果表明:与对照组相比,添加BC1+BC2对提高肉鸡日增质量和降低料质量比效果显著(P<0.05),其中日增质量提高了9.97%,料质量比降低了8.46%;添加BC1和BC1+BC2组肉鸡的腿肌率分别提高了8.46%和7.95%(P<0.05),BC1、BC1+BC2和BC1+BC2+BS3组的腹脂率较对照组显著降低,分别降低了26.6%、30.2%和25.8%;日粮中添加BC1+BC2和BC1+BC2+BS3显著提高了脾脏指数(P<0.05),添加BC1+BC2、BC1+BS3和BC1+BC2+BS3组的法氏囊指数显著提高,添加BC1+BC2、BC1+BS3和BC1+BC2+BS3显著降低了肉鸡盲肠道内的大肠杆菌数(P<0.05)。本试验结果表明适用肉鸡的最佳芽孢杆菌制剂是BC1+BC2。     

低温沼气功能菌群的选育与试验

通过低温定向培养,从高纬、高寒野外厌氧环境样品和冬季沼气池底泥样品中分别选育出野生低温功能菌群和沼气池低温菌群。在13℃和10℃低温条件下,野生低温菌群产气率达到0.14 m3/d.m3和0.10 m3/d.m3,但所产沼气的甲烷含量较低,只有51.0%和48.0%。同样温度下,沼气池低温菌群的产气率只有0.06 m3/d.m3和0.04 m3/d.m3,但所产沼气的甲烷含量较高,达到62%和56%。两类菌群混合发酵可显著改善产气性能,相同温度下,产气率可达到0.15 m3/d.m3和0.11 m3/d.m3,高于两类菌群单独发酵,是常温菌群的3倍以上;混合低温菌群所产沼气的甲烷含量为57.0%和52.0%,介于野生低温菌群和沼气池低温菌群之间,比常温菌群提高19.0%和16.0%。研究结果表明,混合低温菌群具有较好的低温产气性能和广阔的应用前景。     

复合微生态制剂对异育银鲫肠道菌群和消化机能的影响

为了研究复合微生态制剂对异育银鲫肠道菌群和消化机能的影响,在饲料中添加不同水平的酿酒酵母(0.04%、0.06%、0.08%)、嗜酸乳酸菌(0.01%、0.02%、0.03%)以及枯草芽孢杆菌(0.01%、0.03%、0.05%)3种益生菌制剂,采用L9(34)正交设计法配制成9种复合微生态制剂。结果表明,酿酒酵母对肠道乳酸菌和芽孢杆菌含量的影响达到极显著水平(P<0.01),枯草芽孢杆菌对肠道乳酸菌含量的影响达到显著水平(P<0.05)。3种益生菌对大肠杆菌和气单胞菌含量的影响均显著(P<0.05)。3种试验益生菌对异育银鲫前、中、后肠段的淀粉酶、脂肪酶及蛋白酶均有不同程度的影响,但差异不显著(P>0.05)。酿酒酵母对肠道消化酶和营养物质表观消化率的影响最大,对磷表观消化率影响的极显著(P<0.01),对干物质表观消化率影响显著(P<0.05)。酿酒酵母、嗜酸乳杆菌和枯草芽孢杆菌3菌种适当搭配可改善异育银鲫肠道微生态环境,有益于消化酶的分泌和饲料养分的消化,较优的组合为酿酒酵母0.08%、嗜酸乳杆菌0.03%、枯草芽孢杆菌0.01%。