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），粘質沙雷氏菌（粘質賽氏桿菌， Serratia marcescens）；
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.
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.