The separation and cultivation of anaerobic bacteria

1 Purpose1.1 Understanding the growth characteristics of anaerobic
microbials

1.2 Observing the morphological characteristics of anaerobic microorganisms (Bifidobacteria
1.3 Mastering the roll tube separation of anaerobic microorganisms,
Training
and Counting Technology
2 Principles Currently, simple and effective techniques for cultivating anaerobic microorganisms include: anaerobic tank culture technology, anaerobic tank culture technology, anaerobic bag culture technology, Hengeter anaerobic roller tube technology. Hengate anaerobic roller technology is described here.
Hengate anaerobic roller tube technology is an anaerobic culture technique first proposed by
U.S.
Microbiology and applied to the study of anaerobic microorganisms in the stomach in 1950. Since then, the technology has undergone decades of continuous improvement, which has made Hengert anaerobic technology more interesting, and gradually developed into a complete set of technologies for the study of anaerobic microorganisms. And over the years, practice has proved to be a very effective technique for studying strict, specialized anaerobic bacteria.
Henget anaerobic tube culture technology can be used not only for the separation of anaerobic bacteria such as Bifidobacteria, and live bacteria culture count, but also for the isolation and identification of harmful corrupt bacteria (e.g. tyrosine) or pathogens (e.g. Botox Thyroid spores).
3 Material3.1 Samples
Bifidos yogurt (liquid), Bifidobacteria preparation (solid).
3.2
modified MRS
PTYG culture base for 3.2 years.
3.3 Instruments and appliances
Hengeert anaerobic roller tube device set, anaerobic tube, anaerobic bottle, roller machine, quantitative sampler
4 processcopper column deoxygenation → pre-reduced culture base→ dilution Preparation of →diluted samples→ roll tube→ culture →counting
5 Method 5.1 copper column system deoxygenation
copper column is a hard glass tube with copper wire or copper chips inside. The tube is 40-400mm in size, two sections are processed into
flepper
, the outer wall is surrounded by
heating
bands, and connected to the transformer to control the voltage and stabilize the temperature of the copper column. The copper column is connected to the hose at both ends, the gas cylinders at one end and the trachea port at the other end. Since gases such as N2, CO2 and H2 from gas cylinders usually contain O2, when these gases pass through copper columns at temperatures of about 360 degrees C, trace amounts of O2 in copper and gas are combined to produce CuO, and copper columns change from bright yellow to black.
when H2 is passed into the oxidized copper column, H2 and the oxygen in CuO combine to form H2O, which is reduced to copper and the copper column is bright yellow. This copper column can be used repeatedly and continuously for deoxygenation purposes. Of course, the H2 source can also be produced by a hydrogen generator.
5.2 Preparation of pre-reduced media and dilutions
When making pre-reduced media and dilutions, the prepared media and dilutions are boiled to drive oxygen, and then installed into the screws with a semi-quantitative sampler while hot In oral anaerobic
test tube
, the general
agar
medium is 4.5-5 ml, the dilution is 9 ml, and inserted into a long needle with N2 gas to exclude O2. At this point, it is clear that the redox indicator added to the culture base - the edge of the sky blue to red finally turned colorless, indicating that the test tube has become oxygen-free state, and then cover the nut butene plug and screw cover, sterilization standby.