微生物的培養(yǎng)特征(微生物,特征,接種,液體培養(yǎng)基)
發(fā)布時(shí)間：
2022-12-27
作者：
微生物的培養(yǎng)特征(微生物,特征,接種,液體培養(yǎng)基)
一、目的要求
1．了解不同微生物培養(yǎng)在斜面上和液體、半固體培養(yǎng)基中的特征。
2．進(jìn)一步熟練和掌握微生物的無菌操作接種技術(shù)。
二、基本原理
微生物的培養(yǎng)特征是指微生物培養(yǎng)在培養(yǎng)基上所表現(xiàn)出的群體形態(tài)和生長情況。一般可用斜面、液體和半固體培養(yǎng)基來檢驗(yàn)不同微生物的培養(yǎng)特征。它們培養(yǎng)在斜面培養(yǎng)基上，可以呈絲線狀、刺毛狀、串珠狀、疏展?fàn)睢渲罨蚣俑鶢睿▓DⅦ-6）。生長在液體培養(yǎng)基內(nèi)，可以呈混濁、絮狀、粘液狀、形成菌膜、上層清晰而底部顯沉淀狀。穿刺培養(yǎng)在半固體培養(yǎng)基中，可以沿接種線向四周蔓延；或僅沿線生長；也可上層生長得好，甚至連成一片，底部很少生長；或底部長得好，上層甚至不生長。利用微生物的培養(yǎng)特征，可以作為它們的種類鑒定和識別純培養(yǎng)是否污染的參考。
檢驗(yàn)微生物的培養(yǎng)特征，或進(jìn)行其他微生物學(xué)實(shí)驗(yàn)時(shí)，接種過程必須保證不被其他微生物所污染，為此，除工作環(huán)境要求盡可能地避免或減少雜菌污染外，熟練地掌握各種無菌操作接種技術(shù)是很重要的。
三、器材
枯草芽孢桿菌，大腸桿菌(Escherichia coli)，金黃色葡萄球菌，白地霉（Geo-trichum candidum），蕈狀芽孢桿菌（Bacillusmycoides），粘質(zhì)沙雷氏菌（粘質(zhì)賽氏桿菌， Serratia marcescens）；
肉膏蛋白胨斜面培養(yǎng)基，肉膏蛋白胨液體培養(yǎng)基，半固體肉膏蛋白胨培養(yǎng)基，接種環(huán)，接種針，無菌吸管，酒精燈等。
四、操作步驟
1．斜面接種
（1）在肉膏蛋白胨斜面試管上，用記號筆寫上將接種的菌名、日期和接種者。
（2）點(diǎn)燃酒精燈或煤氣燈。
（3）將菌種試管和待接種的斜面試管，用大拇指和食指、中指、無名指握在左手中，并將中指夾在兩試管之間，使斜面向上，成水平狀態(tài)，如圖Ⅶ-7。在火焰邊用右手松動試管塞，以利于接種時(shí)拔出。
（4）右手拿接種環(huán)通過火焰燒灼滅菌（參照實(shí)驗(yàn)一），在火焰邊用右手的手掌邊緣和小指，小指和無名指分別夾持棉塞（或試管帽），將其取出，并迅速燒灼管口。
（5）將滅菌的接種環(huán)伸入菌種試管內(nèi)，先將環(huán)接觸試管內(nèi)壁或未長菌的培養(yǎng)基，達(dá)到冷卻的目的，然后再挑取少許菌苔。將接種環(huán)退出菌種試管，迅速伸入待接種的斜面試管，用環(huán)在斜面上自試管底部向上端輕輕地劃直線，勿將培養(yǎng)基劃破，也不要使環(huán)接觸管壁或管口。
（6）接種環(huán)退出斜面試管，再用火焰燒管口，并在火焰邊將試管塞塞上。將接種環(huán)逐漸接近火焰，再燒灼。如果接種環(huán)上沾的菌體較多時(shí)，應(yīng)先將環(huán)在火焰邊烤干，然后燒灼，以免未燒死的菌種飛濺出污染環(huán)境，接種病原菌時(shí)更要注意此點(diǎn)。
2．液體培養(yǎng)基接種
向肉膏蛋白胨液體培養(yǎng)基中接種少量菌體時(shí)，其操作步驟基本與斜面接種時(shí)相同，不同之處是挑取菌體的接種環(huán)放入液體培養(yǎng)基后，應(yīng)在液體表面處的管內(nèi)壁上輕輕磨擦，使菌體從環(huán)上脫落，混進(jìn)液體培養(yǎng)基，塞好試管塞后，搖動液體，使菌體在液體中分布均勻，或用試管振蕩器混勻。
向液體培養(yǎng)基中接種量大或要求定量接種時(shí)，可將無菌水或液體培養(yǎng)基注入菌種試管，用接種環(huán)將菌苔刮下，再將菌種懸液以無菌吸管定量吸出加入，或直接倒入液體培養(yǎng)基。如果菌種為液體培養(yǎng)物，則可用無菌吸管定量吸出加入或直接倒入液體培養(yǎng)基。整個(gè)接種過程都要求無菌操作。
3．穿刺接種
用接種針挑取菌種（針必須挺直），自培養(yǎng)基的中心垂直地刺入半固體培養(yǎng)基中，直至接近管底，但不要穿透，然后沿原穿刺線將針拔出，塞上試管塞，燒灼接種針，如圖Ⅶ-9所示。
上述幾種接種法的無菌操作，凡未敘述的均按實(shí)驗(yàn)一操作。試驗(yàn)者應(yīng)反復(fù)練習(xí)無菌操作接種技術(shù)，直至較熟練地掌握。
4．將已接種的斜面、液體和半固體培養(yǎng)基放置28—30℃溫箱，培養(yǎng)2—3天后取出觀察結(jié)果。
日水培養(yǎng)基qdrishui.cn
Culture characteristics of microorganisms (microorganism, characteristics, inoculation, liquid medium)
I. purpose requirements
1. To understand the characteristics of different microbial cultures on inclined surface and in liquid and semi-solid medium.
2. Further proficiency and mastery of sterilization and inoculation technology of microorganism.
2. Basic principles
Microbial culture characteristics refer to the population morphology and growth of microbial culture on the medium. Culture characteristics of different microorganisms can be tested by slant, liquid and semi-solid media. They cultivate in slant medium, can be a wire line, hairy, beaded, hydrophobic exhibition, dendritic or rhizoid (figure Ⅶ - 6). Growth in liquid medium, can be cloudy, flocculent, mucous, forming bacterial film, upper clear and bottom precipitation. Puncture culture can spread around the inoculation line in semi-solid medium. Or just growing along the line; It can also grow well on the top, or even into a piece, with little growth on the bottom. Or the bottom grows well, and the top doesn't even grow. The characteristics of microbial culture can be used as a reference for the identification and identification of contamination of pure culture.
Microbial cultivation characteristics test, or other microbiology experiment, inoculation process must ensure that no contamination by other microorganisms, to that end, in addition to the work environment requirements as much as possible to avoid or reduce the bacterial pollution, skillfully master all kinds of asepsis inoculation technology is very important.
3, equipment
Bacillus subtilis, escandicherichia coli, staphylococcus aureus, geo-trichum dum, Bacillusmycoides, and myxanthomycoides.
Meat paste peptone inclined surface medium, meat paste peptone liquid medium, semi-solid meat paste peptone medium, inoculation ring, inoculation needle, sterile straw, alcohol lamp, etc.
4. Operation steps
1. Cant vaccination
(1) write the name, date and inoculant of the bacteria to be inoculated on the test tube with peptone inclined surface.
(2) light alcohol or gas lamps.
(3) with the bevel of the bacteria in vitro and inoculation test tubes, with their thumb and forefinger, middle finger, ring finger grip in the left hand, and the middle finger between tube and ramp up, into a state level, as shown in figure Ⅶ - 7. Loosen the tube plug with your right hand at the edge of the flame to allow it to be pulled out during inoculation.
(4) right hand inoculation loops through the flame burning sterilization (with reference to the experiment one), at the edge of the flame edge and the little finger of the palm, with the right hand little finger and ring finger, respectively, gripping cotton plug (or tube cap), removed, and rapid burning nozzle.
(5) insert the sterilized inoculation ring into the test tube of bacteria species, first contact the ring with the inner wall of the test tube or the medium of ungrown bacteria to achieve the purpose of cooling, and then pick up a little fungus moss. Will ring out strains in vitro, quickly into the vaccination cant tube, with a ring on the slope to the top from the bottom of the tube gently lines, not medium cut, will also make ring contact surface or nozzle.
(6) exit the beveled tube with the inoculation ring, then burn the nozzle with the flame, and plug the tube into the flame side. Bring the inoculation ring gradually close to the flame and then burn. If there is a large amount of bacteria on the inoculation ring, the ring should be baked dry at the edge of the flame and then burned, so as to prevent unburned bacteria from flying out of the environment and contaminating the environment. More attention should be paid to this when inoculation of pathogenic bacteria.
2. Liquid culture medium inoculation
To meat extract peptone liquid medium vaccination when a small amount of bacteria, the basic steps when inoculated with slope is the same, the difference is the pick of the bacteria after inoculation loops in a liquid medium, should be in the liquid surface of the tube wall friction, gently fell off the ring to bacteria, in liquid medium, after good tube plug plug, shake the liquid, the bacteria in a liquid uniform distribution, or use the test tube oscillator blending, as shown in figure Ⅶ - 8.
To the liquid medium large amount or require quantitative inoculated, the strains of sterile water or liquid medium can be injected into a test tube, the bacteria with inoculation loops moss under, then strain suspension quantitative suck out to join in sterile pipette, or directly in a liquid medium. If the strain is a liquid culture, a sterile straw can be used to absorb it quantitatively and add it directly into the liquid culture medium. Sterilization is required throughout the inoculation process.
3. Targeted vaccination
With vaccination needle pick strains (needle must be straight), since the culture medium at the center of the vertically Pierce semisolid medium until close to the end of tube, but not to penetrate, and then pull the needle out along original piercing line, and inserting tube plug, burning vaccination needle, as shown in figure Ⅶ - 9.
The aseptic operations of the above inoculation methods shall be carried out in accordance with experiment 1. The experimenter should practice the sterile inoculation technique repeatedly until he/she is familiar with it.
4. Cant will have been vaccinated, liquid and semisolid medium placed 28 to 30 ℃ temperature box, cultivate 2-3 days to take out the observations.