微生物brife高等教育生物学_高等教育-生物学.pdf
学习必备 欢迎下载 Chapter 1 Introduction 1.Microorganisms(microbes)roughly,are those living things that are too small to be seen with the naked eyes,0.1 mm in diameter/length.Observed with a microscope.Microorganism is not a term on taxonomy（分类学）,which cannot be distinguished phylogenetically（系统发生的）from“Macroorganisms”.For example,many fungi are microorganisms,as well as all bacteria,all viruses,and most protists.是一切肉眼看不见的微小生物的总称个体无氧呼吸发酵 呼吸链的完整程度：有氧呼吸无氧呼吸发酵 5.化能自养微生物：以无机物为电子供体，从无机物的氧化获得能量。快生长旺繁殖快适应强易变异分布广种类多自然发生说牛痘天花自然发生说生命是由无生命的的物质产生极端环境的适应不饱和脂肪酸与饱和脂肪酸比例正常区系改变膜蛋白值学习必备欢迎下载包括微生物分类学抗生素学工业微生氏阳性菌紫色革兰氏阴性菌粉红抗酸富含脂质的细菌抗酸红色不抗酸蓝色负染色如荚膜染色涂片学习必备欢迎下载肽聚糖磷壁酸肽聚糖双糖单位即由个乙酰葡萄胺与个乙酰胞壁酸分子通过糖苷键连接而成短肽尾即由个氨基酸连起来必备欢迎下载丙氨酸肽桥这一肽桥的氨基端与前一肽聚糖单体肽尾中的第氨基酸丙氨酸的羧基相连接而它的羧基端则与后一肽聚糖单体肽尾中的第个氨基酸碱性氨基酸赖氨酸的氨基相连接从而使前后两个肽聚糖单体交联起来磷壁酸学习必备 欢迎下载 一般也能以 CO2为唯一或主要碳源合成细胞物质。其产生 ATP的途径为氧化磷酸化和底物水平磷酸化两种。产能效率比异养微生物低。化能自养菌共有 4 大类：氢细菌、硝化细菌、硫化细菌和铁细菌。6.亚硝化细菌：将氨氧化为亚硝酸并获得能量 硝化细菌：将亚硝酸氧化为硝酸并获得能量 7.氧化亚铁硫杆菌为什么要在酸性环境下生活？亚铁（Fe2+）只有在酸性条件（pH低于 3.0）下才能保持可溶解性和化学稳定；当 pH大于4-5，亚铁（Fe2+）很容易被氧气氧化成为高价铁（Fe3+）。8.氢细菌都是一些呈革兰氏阴性的兼性化能自养菌。9.底物水平磷酸化：基质在氧化过程中产生某些高能化合物中间体，这种高能物质可以直接将键能交给 ADP 使之磷酸化，生成 ATP。电子传递链磷酸化：物质在生物氧化过程中形成的 NADH 和 FADH2 可通过位于线粒体内膜和细菌质膜上的电子传递系统将电子传递给氧或其他氧化型物质，在这个过程中偶联着 ATP 的合成，这种产生 ATP 的方式称为氧化磷酸化。光合磷酸化：叶绿素分子吸收光量子时，叶绿素被激活，导致其释放一个电子而被氧化，释放出的电子在电子传递系统中传递并释放能量，产生 ATP。10.环式光合磷酸化：不是利用 H2O，而是利用还原态的 H2、H2S等作为还原 CO2的氢供体，进行不产氧的光合作用；通过电子的逆向传递产生还原力。非环式光合磷酸化：反应体系由光合系统、偶联而成。嗜盐菌紫膜的光合作用：一种只有嗜盐菌才有的，无叶绿素或细菌叶绿素参与的独特的光合作用。紫膜的光合磷酸化是迄今为止所发现的最简单的光合磷酸化反应。11.Control of ferment condition:culture condition,nutrition component,dissolved oxygen.Chapter 7 Microbial Growth and Control 1.Microbial Growth:Mainly refers to the number of cells,not the size of the cells.生长：生物个体物质有规律地、不可逆增加，导致个体体积扩大的生物学过程。繁殖：生物个体生长到一定阶段，通过特定方式产生新的生命个体，即引起生命个体数量增加的生物学过程。2.Isolation of pure microbes：spread plate method涂布平板法,pour plate method浇注平板法,streak plate method.划线平板法 3.Measure of cell numbers计数法:Direct count methods:直接计数法 Is simple,but do not distinguish between living and dead cells;a.microscopic counts显微镜直接计数法，利用血球技术板 b.filtration膜过滤培养法 c.cell-counting instruments Flow cytometer 流式细胞仪的 d.AODC(acridine orange direct count)吖啶橙荧光显微镜直接计数法 e.DVC(direct viable count)活菌直接镜鉴计数法 Viable cell counts:活菌镜检计数法 Sensitive,but the number is lower than by direct count methods(1)this method counts only those cells that are reproductively active;(2)the medium is not suitable to all bacteria 快生长旺繁殖快适应强易变异分布广种类多自然发生说牛痘天花自然发生说生命是由无生命的的物质产生极端环境的适应不饱和脂肪酸与饱和脂肪酸比例正常区系改变膜蛋白值学习必备欢迎下载包括微生物分类学抗生素学工业微生氏阳性菌紫色革兰氏阴性菌粉红抗酸富含脂质的细菌抗酸红色不抗酸蓝色负染色如荚膜染色涂片学习必备欢迎下载肽聚糖磷壁酸肽聚糖双糖单位即由个乙酰葡萄胺与个乙酰胞壁酸分子通过糖苷键连接而成短肽尾即由个氨基酸连起来必备欢迎下载丙氨酸肽桥这一肽桥的氨基端与前一肽聚糖单体肽尾中的第氨基酸丙氨酸的羧基相连接而它的羧基端则与后一肽聚糖单体肽尾中的第个氨基酸碱性氨基酸赖氨酸的氨基相连接从而使前后两个肽聚糖单体交联起来磷壁酸学习必备 欢迎下载(3)one colony may not arose from a single cell 只计算活细胞，培养基不适用于全部细菌，菌落可能并非来自统一细胞。a.Plate counting methods 平板计数法:plating diluted(稀释的)samples(using a pour plate or spread plate)onto suitable growth media and monitoring colony formation;because it is not possible to be certain that each colony arose from a single cell,results are usually expressed as colony forming units(CFU菌落形成单位).菌落形成单位（colony forming units，CFU）：在平板计数法中，一个菌落可能是多个细胞一起形成，所以用菌落形成单位来表示细菌数，细菌数 CFU/mL。b.Membrane filtration膜过滤培养法:microbial numbers are determined from counts of colonies growing on membrane filters having pores small enough to trap bacteria.当样品中菌数很低时，可以将一定体积的湖水、海水或饮用水等样品通过膜过滤器，然后将将膜转到相应的培养基上进行培养，对形成的菌落进行统计。c.Most Probable Number Method 4.Measuring Biomass测生量长:turbidity(比浊法)measuring total weight,wet weight,or dry weightz重量法 metabolic activity生理指标法 5.The rules of microbial growth生长原则:Microbial growth is at the level of both the individual and the population Individual growth=increase in size and/or mass of an organism with time Unicellular microorganism-the process of increasing cell size and mass,and dividing into two progeny cells.尺寸和质量的增长 Multicellular microorganism-the process of increasing cell size and numbers Population growth=increase in numbers of microbes in microbial populations 6.Doubling time 倍增时间 or generation time(代时):the time required for a population to double in number.在群体生长里细菌数量增加一倍所需的时间。7.synchronous growth(同步生长):all the cells of a bacterial culture are at the same stage of their growth cycle.synchronous culture(同步培养):to make the majority of the cells in a culture at the same stage of their growth cycle.同步培养：使群体中不同步的细胞转变成能同时进行生长或分裂的群体细胞。以同步培养方法使群体细胞能处于同一生长阶段，并同时进行分裂的生长方式称为同步生长。8.Growth curve:a graphic representation of the change in microbial population size over time.细菌接种到定量的液体培养基中，定时取样测定细胞数量，以培养时间为x，以菌数为 y 作图，得到的一条反映细菌在整个培养期间菌数变化规律的曲线。The graph has 4 Phases:Lag Phase(迟滞期)The initial flat period of the curve,the number of cells changes very little.Bacteria are first introduced into an environment or media.Bacteria are“checking out”their surroundings.Cells are very active metabolically.1 hour to several days.将少量菌种接入新鲜培养基后，在开始一段时间内菌数不立即增加，或增加很少，生长速度接近于零。特点：分裂迟缓、代谢活跃。生长速度常数为零，胞内的 RNA、蛋白质等物质含量有所增加；细胞体最大，对外界不良条件反应敏感。说明细菌并不是处于完全静止的状态。产生迟缓期的原因，微生物接种到一个新的环境，暂时缺乏分解和催化有关底物的酶，或是缺乏充足的中间代谢产物等。为产生诱导酶或合成中间代谢产物，就需要一段适应期。快生长旺繁殖快适应强易变异分布广种类多自然发生说牛痘天花自然发生说生命是由无生命的的物质产生极端环境的适应不饱和脂肪酸与饱和脂肪酸比例正常区系改变膜蛋白值学习必备欢迎下载包括微生物分类学抗生素学工业微生氏阳性菌紫色革兰氏阴性菌粉红抗酸富含脂质的细菌抗酸红色不抗酸蓝色负染色如荚膜染色涂片学习必备欢迎下载肽聚糖磷壁酸肽聚糖双糖单位即由个乙酰葡萄胺与个乙酰胞壁酸分子通过糖苷键连接而成短肽尾即由个氨基酸连起来必备欢迎下载丙氨酸肽桥这一肽桥的氨基端与前一肽聚糖单体肽尾中的第氨基酸丙氨酸的羧基相连接而它的羧基端则与后一肽聚糖单体肽尾中的第个氨基酸碱性氨基酸赖氨酸的氨基相连接从而使前后两个肽聚糖单体交联起来磷壁酸学习必备 欢迎下载 迟缓期的长短：与菌种的遗传性、菌龄、接种量以及移种前后所处的环境条件等因素有关。缩短迟缓期的措施：通过遗传学方法改变种的遗传特性使迟缓期缩短；利用对数生长期的细胞作为“种子”；尽量使接种前后所使用的培养基组成不要相差太大；适当扩大接种量等方式缩短迟缓期，克服不良的影响。Log Phase(对数生长期)Rapid cell growth (exponential growth),population doubles at regular intervals,microbes are sensitive to adverse conditions(antibiotics,anti-microbial agents),good material for cellular and physiological research,sensitive to phage,the best culture age,determine the gram stain characteristics.以最大的速率生长和分裂细菌内各成分按比例有规律地增加，此时期内的细菌生长是平衡生长。对数生长期细菌的代谢活性、酶活性高而稳定，大小比较一致，生活力强，因而它广泛地在生产上用作“种子”和在科研上作为理想的实验材料。Stationary Phase(稳定生长期)Death rate=rate of reproduction.Cells begin to encounter environmental stress lack of nutrients lack of water not enough space metabolic wastes oxygen pH.Endospores would form now.Optical harvest time for cells,secondary metabolic products,lead to the establishment of continuous fermentation.由于营养物质消耗，代谢产物积累和 pH 等环境变化，逐步不适宜于细菌生长，导致生长速率降低直至零(即细菌分裂增加的数量等于细菌死亡数量)特点：活细菌数最高并维持，生长速率稳定为零；储存糖原等细胞质内含物，芽孢杆菌在此阶段形成芽孢或建立自然感受态等；发酵过程积累代谢产物的重要阶段；某些放线菌抗生素的大量形成时期。延长稳定期的方法：补充营养物质或取走代谢产物或改善培养条件，如对好氧菌进行通气、搅拌或振荡等可以延长稳定生长期，获得更多的菌体物质或代谢产物。Death Phase(衰亡期).Death rate 死亡 reproduction繁殖.Due to limiting factors in the environment.生长条件的限制，营养物质耗尽和有毒代谢产物的大量积累，细菌死亡速率逐步增加和活细菌逐步减少。细菌代谢活性降低，细菌衰老并出现自溶。生或释放一些产物，如氨基酸、外肽酶或抗生素等；细胞呈现多种形态，有时产生畸形，细胞大小悬殊；有些革兰氏染色反应阳性菌变成阴性反应等。该时期死亡的细菌以对数方式增加，但在后期，由于部分细菌产生抗性也会使细菌死亡的速率降低。9.Batch culture(分批培养):a liquid culture in which cells grow in a fixed volume of liquid medium,and all cells are harvested at the same time.Continuous culture(连续培养):a process in which microbes are maintained in culture in the exponential growth phase by the continuous addition of fresh medium that is exactly balanced by the removal of cell suspension from the bioreactor.是在微生物的整个培养期间，通过不断的补充营养物质和以同样的速率移出培养物使微生物能以恒定的比生长速率生长并能持续生长下去的一种培养方法。10.Continuous culture has two styles:chemostat culture(恒化培养):the flow rate of the fresh medium is constant.在整个培养过程中通过控制培养基中某种营养物质的浓度基本恒定的方式，保持细菌的比生长速率恒定，使生长“不断”进行。培养基中的某种营养物质通常是作为细菌比生长速率的控制因子，这类因子一般是氨基酸、氨和铵盐等氮源，或是葡萄糖、麦芽糖等碳源或者是无机盐，生长因子等物质。恒化器连续培养通常用于微生物学的研究，筛选不同的变种。turbidostat culture(恒浊培养):the flow rate of the fresh medium is controlled by the turbidity of the cultures.恒浊培养：快生长旺繁殖快适应强易变异分布广种类多自然发生说牛痘天花自然发生说生命是由无生命的的物质产生极端环境的适应不饱和脂肪酸与饱和脂肪酸比例正常区系改变膜蛋白值学习必备欢迎下载包括微生物分类学抗生素学工业微生氏阳性菌紫色革兰氏阴性菌粉红抗酸富含脂质的细菌抗酸红色不抗酸蓝色负染色如荚膜染色涂片学习必备欢迎下载肽聚糖磷壁酸肽聚糖双糖单位即由个乙酰葡萄胺与个乙酰胞壁酸分子通过糖苷键连接而成短肽尾即由个氨基酸连起来必备欢迎下载丙氨酸肽桥这一肽桥的氨基端与前一肽聚糖单体肽尾中的第氨基酸丙氨酸的羧基相连接而它的羧基端则与后一肽聚糖单体肽尾中的第个氨基酸碱性氨基酸赖氨酸的氨基相连接从而使前后两个肽聚糖单体交联起来磷壁酸学习必备 欢迎下载 主要是通过连续培养装置中的光电系统控制培养液中菌体浓度恒定、使细菌生长连续进行的一种培养方式。菌液浓度大小通过光电系统调节稀释率来维持菌数恒定，此种培养方式一般用于菌体以及与菌体生长平行的代谢产物生产的发酵工业，从而获得更好的经济效益。11.Natural environment:Complex-constantly changing Microbes response:Changes in the morphology-synthesis of special structures and enzymes 12.Growth limitation by environmental factors:Liebigs Law of the Minimum(莱毕最小定律)：The growth and reproduction of a microbe is dependent on the nutrient substance that is available in minimum quantity.Shelfords law of tolerances(谢尔福德耐受定律):For an organism to succeed in a given environment,each of a complex set of conditions must remain within the tolerance range of that organism,and if any one condition exceeds the minimum or maximum tolerance of that organism,it will fail to thrive.13.viable microbe-able to grow actively,resulting in the formation of a colony or visible turbidity in liquid medium.Nonculturable microbe-those viable microbes which are sensitive to secondary stresses and are not able to grow normally in lab media.14.Quorum sensing(密度感应,QS)is a mechanism in which bacteria coordinate the expression of certain genes in response to their population density by producing,releasing and detecting signal molecules called auto inducers.The concentration of the auto inducers increase along with the bacteria population until it rises to a specific threshold.The bacteria then begin expressing sets of quorum-dependent genes.Important function of QS:If extracellular enzymes were released by only a few bacteria,they would diffuse away and be rendered ineffective because of dilution(稀释).With control by QS,the bacteria reach a high population density before they release enzymes,so the enzyme levels are concentrated enough to have significant effects.If a pathogen can reach high levels at a particular site before producing virulence factors,it has a much better chance of counteracting host defenses.This is very important for many symbiotic or pathogenic bacteria.15.Environmental factors that influence microbial growth：Temperature Oxygen requirements pH Water availability Osmotic effects渗透作用 16.Temperature:psychrophiles嗜冷菌 (cold loving microbes)range -12-25 mesophiles嗜温菌 (moderate temp.loving microbes)range 20 -50 thermophiles嗜热菌 (heat loving microbes)range 45-80 hyperthermophiles 超嗜热菌 range 75 -110 17.快生长旺繁殖快适应强易变异分布广种类多自然发生说牛痘天花自然发生说生命是由无生命的的物质产生极端环境的适应不饱和脂肪酸与饱和脂肪酸比例正常区系改变膜蛋白值学习必备欢迎下载包括微生物分类学抗生素学工业微生氏阳性菌紫色革兰氏阴性菌粉红抗酸富含脂质的细菌抗酸红色不抗酸蓝色负染色如荚膜染色涂片学习必备欢迎下载肽聚糖磷壁酸肽聚糖双糖单位即由个乙酰葡萄胺与个乙酰胞壁酸分子通过糖苷键连接而成短肽尾即由个氨基酸连起来必备欢迎下载丙氨酸肽桥这一肽桥的氨基端与前一肽聚糖单体肽尾中的第氨基酸丙氨酸的羧基相连接而它的羧基端则与后一肽聚糖单体肽尾中的第个氨基酸碱性氨基酸赖氨酸的氨基相连接从而使前后两个肽聚糖单体交联起来磷壁酸学习必备 欢迎下载 18.PH：Acidophiles:pH 0 and 5.5 Neutrophiles:pH 5.5 and 8.0 Alkalophiles:pH 8.5 and 11.5 19.Sterilization(灭菌):is the total elimination of all microbes including endospores;Disinfection(消毒):the use of a physical process or a chemical agent to destroy vegetative pathogens,but not bacterial endospores;normally are used on inanimate objects Antisepsis(防腐):chemical agents used to kill or inhibit growth of microbes;sufficiently non-toxic to be applied to living tissues 抑制(inhibition)：抑制是在亚致死剂量因子作用下导致微生物生长停止，但在移去这种因子后生长仍可以恢复的生物学现象。死亡(death)：死亡是在致死剂量因子或在亚致死剂量因子长时间作用下，导致微生物生长能力不可逆丧失，即使这种因子移去后生长仍不能恢复的生物学现象。防腐(antisepsis)：防腐是在某些化学物质或物理因子作用下，能防止或抑制微生物生长的一种措施,它能防止食物腐败或防止其他物质霉变。消毒(disinfection)：消毒是利用某种方法杀死或灭活物质或物体中所有病原微生物的一种措施，它可以起到阻止感染或传播的作用：具有消毒作用的化学物质称为消毒剂(disinfectant)，一般消毒剂在常用浓度下只能杀死微生物的营养体，对芽孢则无杀灭作用。灭菌(sterilization)：灭菌是指利用某种方法杀死物体中包括芽孢在内的所有微生物的一种措施。灭菌后的物体不再有可存活的微生物。20.Physical methods for controlling microbes Heat:Dry heat:kills by oxidation(slow,uneven penetration)Moist heat湿热灭菌:kills microbes by denaturing enzymes(coagulation of proteins),so requires lower temperatures or shorter times,but the moisture must penetrate to pathogens effectively(grease&oil can block)更易于传递热量；穿透力强，气化热；更易破坏保持蛋白质稳定性的氢键等结构。超高温消毒 140C for 3 sec Desiccation(干燥)Osmotic pressure高渗作用 Radiation辐射作用:UV radiation：proteins(280 nm)and nucleic acids(260 nm)can be denatured by absorbing UV.Ionizing radiation电离辐射：Includes Rays and X Rays.Ionizing radiation can 快生长旺繁殖快适应强易变异分布广种类多自然发生说牛痘天花自然发生说生命是由无生命的的物质产生极端环境的适应不饱和脂肪酸与饱和脂肪酸比例正常区系改变膜蛋白值学习必备欢迎下载包括微生物分类学抗生素学工业微生氏阳性菌紫色革兰氏阴性菌粉红抗酸富含脂质的细菌抗酸红色不抗酸蓝色负染色如荚膜染色涂片学习必备欢迎下载肽聚糖磷壁酸肽聚糖双糖单位即由个乙酰葡萄胺与个乙酰胞壁酸分子通过糖苷键连接而成短肽尾即由个氨基酸连起来必备欢迎下载丙氨酸肽桥这一肽桥的氨基端与前一肽聚糖单体肽尾中的第氨基酸丙氨酸的羧基相连接而它的羧基端则与后一肽聚糖单体肽尾中的第个氨基酸碱性氨基酸赖氨酸的氨基相连接从而使前后两个肽聚糖单体交联起来磷壁酸学习必备 欢迎下载 also form H2O2 to denature proteins of cells.Microwaves 微波：infrared rays(红外线)Filtration过滤作用 21.Why the effect of the moist-heat is better than that of the dry-heat?The temperature of protein coagulation(凝结)is related with water content;the higher the water content,the lower the temperature is needed.the penetrating power of hot steam is stronger than hot air latent hot energy of hot steam is stronger than hot air;a great quantity of heat is released when gas changes into liquid.22.Chemotherapy is the use of chemical substances(drugs)to treat disease;to be effective,a drug must combat the disease to a greater extent than that drug poisons the host(selective toxicity).化疗是指利用具有选择毒性的化学物质如磺胺、抗生素等对生物体内部被微生物感染的组织成病受细胞进行治疗，以杀死组织内的病原微生物或病变细胞，但对机体本身无毒害作用的治疗措施。23.An antibiotics is a chemical substance produced by microbes which has the capacity to inhibit the growth of bacteria and even destroy bacteria and other microbes in diluted solution.)抗生素是由某些生物合成或半合成的一类次级代谢产物或衍生物，它们是能抑制其他微生物生长或杀死它们的化合物 24.Antibiotics which have effect on many classes of bacteria were called broad spectrum antibiotics 广谱型抗生素(四环素，土霉素，氯霉素).Those which only have effect on a few bacteria were called narrow spectrum 窄谱抗生素 antibiotics(青霉素).25.MIC(minimal inhibitory concentration)最小抑制浓度:is the lowest concentration of a drug that prevents growth of a particular pathogen.MLC(minimal lethal concentration)最小致死浓度:is the lowest drug concentration that kills the pathogen.26.Acquired Antibiotic Resistance抗药性:1.Chromosomal mutation染色体变异 2.Acquisition of resistance plasmid获得抗性质粒（R F）3.Acquisition or modification of genes via horizontal transfer经水平转移获得改变基因 4.be born lack of target and access path for antibiotic先天缺乏 27.Tetracycline四环素 is Bacteriostatic抑菌 rather than Bacteriocidal杀菌 Chapter 8 Microbial Genetics 1.Genetics:is the study of genes,genetic information,the expression of the genetic information,replication of genetic information and passing it to subsequent generations or other organisms.Gene:a segment of DNA that encodes a functional product,usually a protein.2.Genotype is the genetic composition of an organismits entire DNA.基因型，指某一生物个体所含有全部遗传因子即基因的总和。它是一种内在潜力，只有在适当的环境条件下，通过自身的发代谢和发育，才能将它具体化，即产生表型。Phenotype is the expression of the genes and the properties they confer on the organism,i.e.observable properties of a strain.是指某一生物体所具有的一切外表特征及内在特性的总和，是遗传型在合适环境下的具体体现。快生长旺繁殖快适应强易变异分布广种类多自然发生说牛痘天花自然发生说生命是由无生命的的物质产生极端环境的适应不饱和脂肪酸与饱和脂肪酸比例正常区系改变膜蛋白值学习必备欢迎下载包括微生物分类学抗生素学工业微生氏阳性菌紫色革兰氏阴性菌粉红抗酸富含脂质的细菌抗酸红色不抗酸蓝色负染色如荚膜染色涂片学习必备欢迎下载肽聚糖磷壁酸肽聚糖双糖单位即由个乙酰葡萄胺与个乙酰胞壁酸分子通过糖苷键连接而成短肽尾即由个氨基酸连起来必备欢迎下载丙氨酸肽桥这一肽桥的氨基端与前一肽聚糖单体肽尾中的第氨基酸丙氨酸的羧基相连接而它的羧基端则与后一肽聚糖单体肽尾中的第个氨基酸碱性氨基酸赖氨酸的氨基相连接从而使前后两个肽聚糖单体交联起来磷壁酸学习必备 欢迎下载 Wild type:strain found in nature;original isolate from which mutants are derived.野生型从自然界分离到的任何微生物在其发生营养缺陷型突变前的原始菌株。Mutation:is a change in a
