What is anaerobic system?
are closely related to human and animal life. With the development of medicine and biotechnology, human research on anaerobic bacteria has become more and more extensive. Due to the anaerobic characteristics of anaerobic bacteria, their culture must be carried out under anaerobic conditions. It is easy and practical to culture anaerobic bacteria by the anaerobic culture method, so the anaerobic system
is an important means to cultivate, test, and research anaerobic bacteria in hospitals, factories, education, and research fields.
The anaerobic system
is a special device that can be used to culture and operate bacteria in an anaerobic environment, which can culture the most difficult to grow anaerobic organisms, and avoid the risk of death of anaerobic organisms that are exposed to oxygen when operating in the atmosphere. Therefore, this device is ideal for anaerobic organism detection and research.
What are anaerobic organisms?
"Anaerobic" is common in the biological world. Anaerobic organisms
are organisms that do not require oxygen to grow. Anaerobic organisms break down organic matter such as sugars and proteins in the absence of oxygen, but the energy released is very little, so anaerobic organisms are generally small in size, and among them are generally bacteria. They can be broadly divided into three types, namely, specialized anaerobic organisms, parthenogenic anaerobic organisms, and oxygen-tolerant anaerobic organisms.Anaerobic bacteria
are a major component of the normal flora and are widely present on the deep mucosal surfaces of the human skin and cavities, and they can cause infections in any tissue and organ of the body. Anaerobic bacterial infections occur in cases of tissue ischemia, necrosis, or infection by aerobic bacteria, resulting in a decrease in the oxygen concentration of local tissues. It causes diseases such as gas gangrene, tetanus, botulism, etc. According to its different degrees of tolerance to oxygen, it can be divided into specialized anaerobic bacteria, microaerobic, and parthenogenic anaerobic bacteria.
A. Specialized anaerobic organisms
Some anaerobic organisms die when exposed to an oxygenated environment. These organisms are called "specialized anaerobes" and they survive by fermentation or anaerobic respiration. In an aerobic environment, specialized anaerobes are deficient in superoxide dismutase and catalase, enzymes that help remove lethal superoxide from the cells of specialized anaerobes.
Bifidobacterium is a Gram-positive, specialized anaerobic bacterium with the typical biology of Gram-positive bacteria. Bifidobacterium is one of the most important physiological bacteria in the human and animal intestines, and it constitutes a microbial community with other physiological bacterial members and a microecological system with the host. It has maintenance of intestinal microecological balance, nutritional and growth-promoting effects, immune enhancement, anti-aging effects, prevention and control of hypertension and atherosclerosis, and anti-tumor effects.
B. Parthenogenic anaerobic organisms
Parthenogenic anaerobic organisms can use oxygen in an aerobic environment for aerobic respiration. However, in the absence of oxygen, some of them ferment and some of them respire anaerobically. The conditions that affect the conversion are the concentration of oxygen and fermentable substances. For example, when fermentable sugar is given to brewer's yeast, its observable oxygen consumption stops immediately, which is called "Pasteur mutation". This is because the energy produced by the comparison is not worth the effort of respiration, until when fermentable substances are available and the brewer's yeast chooses to ferment even though the energy produced by fermentation is much lower than the energy produced by respiration. This change from respiration to fermentation is faster than the reverse process because it is used to grow through fermentation and the mitochondria need time to get started. Examples of parthenogenic anaerobic bacteria are Staphylococcus spp., Corynebacterium spp., and Listeria spp. Yeasts among fungi are also parthenogenic anaerobes.
C. Aerobically tolerant anaerobic organisms
Aerobic tolerant anaerobic organisms can survive in the presence of oxygen, but they do not use oxygen as the ultimate electron acceptor, and all aerobic tolerant anaerobic organisms perform fermentation. Among anaerobic organisms, some can survive under aerobic conditions (called aerobic tolerant organisms), but some cannot (called specialized anaerobic organisms), and the O2- produced by cells can cause harm to cells. Aerobic tolerant organisms have SOD (superoxide dismutase), which can catalyze the reaction "2O2-+2H+=H2O2+O2", and H2O2 is converted into H2O by catalase or peroxidase, which relieves the harm of O2- to the organism; while specialized anaerobic organisms do not have SOD, and whether they have SOD is the main reason why anaerobic organisms have different tolerance to oxygen.
Structure of the anaerobic system
The anaerobic system is generally composed of four major parts: anaerobic chamber, transfer chamber, bare hand cuff operation hole, air circuit, and circuit control system.
Anaerobic energy system examples
A. Anaerobic chamber
Inner cavity mechanical forced convection and inner cavity positive pressure, to achieve constant temperature, humidity control, deoxygenation, biological detoxification of the four aspects of state stability and uniformity, and can quickly recover, operation, and breed in the same room.
B. Transfer chamber
Adopt a compact cylinder design to achieve easy transfer of samples by one person and one hand.
C. Bare hand cuff operation hole
No need to carry out the vacuum/nitrogen filling replacement process, both hands can go straight into and out of the inner chamber anaerobic.
D. Control system
All-round real-time status self-test alarm function to ensure normal operation of the equipment.
Use of anaerobic system
a. Place the necessary accessories and appliances according to the requirements and put two non-toxic plastic bags into the operation room.
b. Mixing gas bottle, nitrogen bottle input pressure adjustment, adjust the pressure reducing valve so that the input pressure is 0.1Mpa.
c. After opening the equipment rear match, power switch, and press the total power button on the control board to energize the equipment.
d. Put 1000g palladium pellets (sealed) in the operation chamber, dehumidify by the condensing system and put in the anaerobic indicator.
e. Close the outer door of the sampling room tightly, and vacuum calibration. (Press the vacuum pump key on the control surface plate, press number 1 to open, then number 1 to close).
f. In the operation chamber for the first replacement (nitrogen replacement).
a) Insert a rubber tube into the air inlet of the operation chamber first, and another into the plastic bag.
b) Connected to the nitrogen inlet, open the operating room nitrogen control valve, so that the two plastic bags after sufficient nitrogen, close the solenoid valve, and then tighten the bag mouth.
c) Put latex gloves on the observation plate flange ring and tie it tightly.
d) Put the nitrogen in the plastic bag gradually in the operation room, for all the prevention and removal.
g. Perform a second replacement (nitrogen replacement) in the operation chamber. Repeat the nitrogen flushing process once. The sampling chamber is first evacuated, and pay attention to the exhaust valve to close the exhaust at any time.
h. In the operation room for the third replacement (mixed gas replacement).
a) Switching gas circuit to open the operating room mixed gas valve into the gas, flush gas sampling chamber the first vacuum, and pay attention to the exhaust valve to close the exhaust at any time.
b) After the mixture is loaded into the plastic bag, close the solenoid valve. Open the mixture restrictor valve to allow the mixture to pass through the flow stabilizer and adjust the flow meter to a flow rate of approximately 10 ml per minute.
c) The mixed gas in the plastic bag is gradually discharged into the operation room.
d) After three air changes, the oxygen content of the gas in the operation room is a very small amount.
i. Turn on the palladium grain deaerator in the operation room, and turn on the power of the deaerating catalyst for catalytic deaeration. And a little drop of clean water on the anaerobic indicator bar reaction area, 1-2 hours later, observe its color change, the reaction part of the dark blue into light blue for the operation room to reach the anaerobic environment.
j. UV sterilization lamp, indoor sterilization treatment, and sterilization time are self-defined.
Strain placement and culture
a. Check the door of the sampling room and close it tightly.
b. Open the door outside the sampling room, and put the strain into the sampling room that is closed after the door.
c. The sampling chamber is filled with a nitrogen gas replacement process three times. First, evacuate 500 mmHg (66 Kpa) or more, then open the sampling chamber oxygen valve into the air. Make the pointer back to zero and close the valve.
d. If the selected vacuum is low, it is necessary to increase the number of replacements.
e. After the door outside the sampling room is opened and closed tightly, then pump a low vacuum of 100 mmHg (13Kpa) to test.
f. Anaerobic incubator needs long-term continuous use conditions.
a) Put an indicator strip in the operation room daily to observe. If it does not work properly, it must be re-gassed.
b) To continuously input a trace amount of gas mixture for a long time. So that the replenished nitrogen can be combined with a trace amount of nitrogen through catalytic absorption, to ensure the anaerobic state of the room, replenished into the mixture of gas flow selected for about 10 ml per minute.
c) It is necessary to replace the deaerator and desiccant once every running day. (The replaced deaerator and desiccant can be put in a drying oven at 200℃ for 2-3 hours to recover).
g. The temperature in the incubator of the operation room can be selected and controlled at will.
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