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1、第一编 环境微生物学基础第二章 环境中的微生物第七节 微生物的分类、鉴定和保藏 Classification、Identification And Conservation of Microorganisms,The Introduced Contents,Yeast(for the production of bread,beer and vinegar,etc)Enzyme preparations produced by microbiologyComposting based on microbiologyMetallurgy and production by microbiolog
2、yInformation storage(into the Bacterium),JapanInformation storage(into the Bacterium)ig.Bacillus Subtilis(Bacillus natto,纳豆菌),Human and Fungus,曲霉病(aspergillosis),天花症状,埃波拉病毒,How to recognize the microorganisms?,How to apply the microorganisms to the above sides?How to diagnose the diseases caused by
3、the microorganisms?,Definition：According to the affinity,Classification、Identification And Conservation of Microorganisms,深层地表中的产甲烷菌,tasks：Classification、Identification、Nomenclature,第一节通用分类单元Section General Taxon,种以上的系统分类单元（seven grade）界 Kingdom门 Phylum（或Division）亚门纲 Class 亚纲超目目 Order 亚目 科 Family 亚科
4、 族 亚族 属 Genus 种 Species,Nomenclature属名（Genus name）：拉丁文的名词或用作名词的形容词，单数，首字母大写，表示微生物的主要特征，由微生物构造，形状或由科学家命名。种名（species name）：拉丁文形容词，字首小写，为微生物次要特征，如微生物色素、形状、来源或科学家姓名等。,国际命名法则：林奈的双名法,亚种：subsp.（subspecies）变种：var.(variety),学 名=属 名+种名+命名人姓氏,枯草芽孢杆菌：Bacillus subtilis Cohn,荧光假单胞菌纤维素亚种：Pseudomonas fluorescens su
5、bsp.cellulosae,Ig.大肠埃希氏杆菌 Escherichia coli（Migula）Castellani et Chalmers 1919Ig.金黄色葡萄球菌 Staphylococcus aureus Rosenbach 1884,当泛指某一属微生物，而不特指该属中某一种（或未定种名）时，可在属名后加sp.或ssp.（分别代表species 缩写的单数和复数形式）Ig.Saccharomyces sp.（表示酵母菌属中的一个种。）,菌株名称在种名后面自行加上数字、地名或符号等，Ig.Bacillus subtilis AS1.389 AS=Academia Sinica Ba
6、cillus subtilis BF7658 BF=北纺 Clostridium acetobutylicum ATCC824 丙酮丁醇梭菌ATCC American Type Culture Collection美国模式菌种保藏中心 当文章中前面已出现过某学名时，后面的可将其属名缩写成13个字母。Ig.Escherichia coli 可缩写成 E.coli Staphylococcus aureus可缩写成 S.aureus三名法：用于对亚种的命名，这时在属和种名后加写一个subsp.，然后再附上亚种名称（斜排体）。如：Bacillus thuringiensis subsp.galler
7、ia 苏云金芽孢杆菌腊螟亚种,微生物的名字有俗名和学名两种。Ig.红色面包霉-粗糙脉孢霉 绿脓杆菌-铜绿假单胞菌学名是微生物的科学名称，它是按照有关微生物分类国际委员会拟定的法则命名的。学名由拉丁词、或拉丁化的外来词组成。学名的命名有双名法和三名法两种。,Classification Unit from subspecies,自学内容（self-study content）：亚种（subspecies）、变种（variety）、菌株（strain）、种（species）、模式种（type species）、新种（species nova，sp.nov或nov sp.）。,species and
8、 new species,种是一个基本分类单位，是一大群表型特征高度相似、亲缘关系极其接近、与同属内其他种有明显差异的菌株的总称。新种：sp.nov.或nov.sp.，新被鉴定的种发表时应在其学名后标上sp.nov.的符号，新种发表前应将其模式菌株的培养物存放在一个永久性的保藏机构，并应允许人们从中取得。,subspeciesvarietystrain,Classification Unit from subspecies,Speciestype species,Bacillus halodurans species nova，sp.Nov or nov sp.The Internationa
9、l Journal of Systematic and Evolutionary Microbiology(IJSEM),Domain,三原界系统,Three Domains Theory,BacteriaEukaryaArchaea,General System of Classification,Bacteria：Bergeys Manual of Systematic Bacteriology（vol.7）、The Genus Bacillus（vol.1）Actinomycete：Key to the Families and Genus of Actinomycete(By Micr
10、oorganism institute of China)Fungus：Smith、Alexopoulos、Ainsworth system,Classification And Identification,.Classic Indicator of Inditification,Morphological Characteristics,八叠球菌,四联球菌,葡萄球菌,梭状芽孢杆菌,短杆菌,长杆菌,螺旋菌,弧菌,螺旋体,Special Morphological Characteristics,Abnomal Morphological Characteristics,结核杆菌的异常形态,结
11、核杆菌的正常形态,Simple Staining,Gram Staining,Colony Characteristics,lawn,Culture Charateristics,反映微生物与氧气的关系,.Identification Technology（mini、simple、fast and automatic）,API System,菌种,基本培养基（液体）,菌悬液,检测、编码、查表、鉴定,适用于API鉴定细菌有700多种,API（API system）（法国生产，可以同时测定20多种生化指标；有20多个塑料小管）,“Enterotube”System,Biolog System(au
12、tomatic and fast),每个孔中含有不同的底物,菌悬液或无菌水,Physiological and Biochemical Characteristics,ig,.The Modern Identification Method,Three Levels,Nucleotide Acid Analysis,、Nucleotide Acid Analysis,GC ratio(guanine plus cytosine base mole percent)G+Cmol%=（G+C）/（A+T+G+C）*100%,Every organsism has its own value of
13、GC ratio.Ig.GC%=25-75%(Bacteria),GC ratio,the Difference of GC Ratio2%,without significance；=2.5-4.0%,the same species；5%,different species；10%,different genus,、Nucleotide Acid Analysis,（2）DNA-DNA HybridizationHomology60%,the same species 70%,the same subspecies=20-60%,the same genus,Effect on DNA b
14、y Different Temperatures,Effect on DNA by Different Temperatures,DNA-DNA Homologous Hybidization,（3）16/18S rRNA,Advantages of 16/18S rRNA Sequece Analysis,a.Common for the Eucaryotes and the procaryotesb.Constent for Physiological function c.High for the content of cellcell d.Stable for the rRNA gen
15、ee.Conservative for some rRNA genef.Moderate for Molecular Weight,(3)16S/18S rRNA sequence Analysis,PrincipleMethod,PCR仪,Evolutionary three,、Nucleotide Acid Analysis,（4）Genome sequeceIg.Bacillus halodurans C-125，E.coli，,、Analysis of the Cell Chemical Components,（1）Cell wall（2）Sugar Type of the Cell
16、Hydrolysate（3）Phosphoric Acid Lipid（4）Mycolic Acid（5）Quinone（6）Fatty Acid and Alcohols(by Gas blaketing Chromatographic Technique),1 Chemical Components of Cell wall,。,2 Sugar Type of the Cell Hydrolysate,5醌类的分析,在细菌和放线菌中所含有甲基萘醌（VK）和泛醌（辅酶Q）。在放线菌分类鉴定上有重要价值。,、numerical taxonomy,Computerig NCBINational
17、Center for Biotechnology Information,一、数值分类法,即统计分类法：在200年前M.Adanson（1727-1806，法国植物学家）发表的分类原理基础上结合现代计算机技术发展而来的。定义：通过广泛比较分类单位的性状特征，然后计算它们之间的相似性，再根据相似性的数值划分类群的一种分类方法。,数值分类的基本步骤,1、计算两菌株间的相关系数2、列出相似度矩阵3、将矩阵图转换成树状,Conservation of Microorganisms,Factors Affecting the Stability of the Microorganisms,Variati
18、on;Pollution;Death,The purposes of the conservation,Non-variation;non-pollution;non-death,The Existing Questions During Conservation,Degradation,Conservation of Microorganisms,Alive、invarable and intortus for the Strains,Basic Requirements,asic ethods,LivingDomancy,Passage by medium,Passage by host,
19、freezing,drying,Inclined plane/plate,Liquid nitrogen、low temperature refrigerator,Sand-tube、freezing vacuum drying,Pincinple of the conservation,To make the strains to be domancy or semi-domancy in order,Non-variation;non-pollution;non-death,Pay attention to the followting sides：Different strains wi
20、th different conservative methods.To conservate according to the properties of the strains,the features and the existing conditions.To apply various methods to conservate the objective strains.,Basic Methods for Conservation,Dryinglow-temperature,Methods,Suspension method of distilled waterPassage b
21、y inclined planeimmersion in mineral oil Dry and Freezing Freezing Freezing vacuum dryingConservation by host Gene Engineer strainsConservation Institute of strains,Methods,Temporary Passage by inclined planePuncture method,Long-term Liquid paraffin methodGlycerol tubing-methodSand-tube methodSand-s
22、oil tube method Filter paper methodFreezing-dry methodLiquid Nitrogen,Liquid paraffin method,simple,Passage by inclined plane,Thinking:the advantages and the disadvantages of this method,Immersion in mineral oil,Mineral oil160,12h,Mineral oil is higher than the stain culture by 1cm.12年Which carbon w
23、ill be use?,strain,Pucture inclined plane,Dry and by-vector,Freezing,-20(Common)-60 Liquid nitrogen,Freezing vacuum drying,Strain:stable phase(C=1091019个/ml)To suspense the liquid strain with the protetive agent(such as degreasing milk,sucrose,animal serum and glutamate sodium)The water content is 1
24、%5%.To seal and conservate at the room temporature or in the refrigerator10 yearsBut complex,Freezing vacuum drying,Host conservation,For the following sides:Some animals/plants/viruses,Gene Engineer strains,Conservation Institute of strains,In 1979，CCCCM was founded by China.,Conservation Institute
25、 of strains,Chinese institutes,国外著名菌种保藏中心,美国标准菌种收藏所(ATCC)，美国马里兰州，罗克维尔市。冷泉港研究室(CSH)，美国。国立卫生研究院(NIH)，美国，马里兰州，贝塞斯达。美国农业部北方开发利用研究部(NRRL)，美国，皮奥里亚市。威斯康新大学，细菌学系（WB）,美国，威斯康新州马迪孙。国立标准菌种收藏所(NCTC)，英国，伦敦。英联邦真菌研究所(CMI)，英国，丘(园)。荷兰真菌中心收藏所(CBS)，荷兰，巴尔恩市。日本东京大学应用微生物研究所(IAM)，日本，东京。发酵研究所(IFO)，日本，大阪。日本北海道大学农业部(AHU)，日本，北
26、海道扎幌市。科研化学有限公司(KCC)，日本，东京。国立血清研究所(SSI)，丹麦。世界卫生组织(WHO)。,细菌的保藏,?How to prevent the strains from degradating?Perhaps,we can do this according to the followling facets:To decrease the times of the passage;To provide the better living conditions;To separate the pure strains and check the characteristics;
27、To employ the effective conservative methods.,Thinkting 1,设计一个从某河水中分离细菌，并从中鉴定某株产脂酶细菌的实验方案。答题提示：（1）产脂酶的细菌为目标菌；（2）方案表述要有逻辑性、条理性、可操作性；（3）注意着重点评各个关键环节。,Thinkting 1,设计一个从土壤中选择性分离鉴定细菌的实验方案。答题提示：（1）任意设定自己感兴趣的属种（诸如耐高温、耐酸碱、耐盐、产酶（纤维素酶、脂酶、蛋白酶、漆酶）的菌株及降解环境中的塑料、农药、石油、洗涤剂和重金属及类金属等的菌株）为目标菌；（2）方案表述要有逻辑性、条理性、可操作性；（3）注意着重点评各个关键环节。,
