JPH04505262A - How to stabilize bacteria - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] How to stabilize bacteria When bacteria complete their growth cycle in a particular medium or environment, these bacteria The cuteria begin to die or survive prolonged starvation, dehydration or extreme temperatures. Start forming spores. Most types of bacteria form environmentally resistant spores Because it cannot, its living ability prevents these deaths, or at least prevents them from dying. This can only be increased by finding conditions or chemicals that reduce the rate of sterilization. Ru. Can protect non-spore-forming bacteria from destruction over time Some examples of chemicals have been previously patented; In 1976, Hor+l1ll　el　! By I; also aqueous azide level In 1987, Wong! + If.

These inhibitory chemicals are associated with bacterial lysis, cell death due to cell wall breakdown. The non-spore-forming bacteria were allowed to survive for a long period of time without any problems. In addition, long-term After storage, recovery of these sulfides and azides from aqueous suspensions is rapid. Ta. This can be done in the presence of a supporting medium consisting of bran or grains, or no additional medium at all. Recovery of living organisms from freeze-drying or air-drying is extremely difficult in situations where This is in contrast to the fact that it is extremely slow.

These chemical systems for rendering bacteria dormant must be maintained and ready for use. It is easy to provide an aqueous suspension in the desired state and is easy to apply to the desired process. Ru. Their main advantage is that the level of bacteria in these suspensions is low and relatively large. volume of water is part of the product component. This is because the product is bulky and heavy. Because of this, storage is difficult, and large volumes of water are difficult to transport by ship because shipping costs are high. occurs.

In addition, as well as bacterial growth during the final stages of food or nutrient consumption, The sporulation process involves the production of extracellular enzymes from the bacterial cytoplasm or secretion. included. The purpose of these enzymes is to produce food for bacteria from specific organic matter. It will be made possible. These enzymes can attack bacterial cell wall components. , its presence is harmful to certain bacteria that grow quickly and proliferate. It is. In the presence of these extracellular enzymes, bacteria that have grown to high levels can be The ability to store without harmful effects is a significant advantage.

Of course, the entire aqueous environment is removed from bacteria via freezing or air drying This condition can be achieved by doing so.

Also, like enzymes, this technology concentrates all salts, allowing the bacterial components of the system to dry out. be done. This results in high levels of bacterial kill during these drying techniques. This is probably one reason why. The present invention explores one improvement.

According to the present invention, a method is provided for stabilizing bacteria present in an aqueous growth medium. be done. This method separates bacteria from the growth medium by centrifugation and creating a bacterial paste and storing the paste in a protected environment. It consists of

This protective environment is known as VitkiBl, Saranl or It may also include an impermeable sheath made of a similar material.

Alternatively, this protective environment may be liquid, preferably non-aqueous, but aqueous. or water. By resuspending the above paste in a liquid, Bacteria become more concentrated than in their original environment. Preferred non-aqueous The liquid is glycerol, glycol or the like.

The above paste, by itself or when resuspended in a liquid, has a gelatin-like , encapsulated in an impermeable but soluble or collapsible membrane. This will cause bugs. A solution is obtained that is free of teratinium and, more importantly, has little overall water, salt or salt content. and bacteria that do not contain enzymes that attack bacterial cells and significantly reduce their vitality. Terrier paste is obtained.

Since bacteria have a wet weight of approximately 0-+2 grams/cell, 1012 cells per gram of paste or 1 kg of paste Contains 1015 cells per cell. This kind of paste is Large volume bacterial suspensions containing bacterial concentrations at the 100 million (108) level For example, one batch of such a paste containing 1015 cells A program would make ten thousand (10') liters of such a suspension. subordinate Therefore, a paste weighing 1 kilogram (2,2 bonds) is mixed with the above aqueous suspension. It can be used as a substitute for a single shipping container carried by a ship.

The cost difference based on ocean freight shipping rates could be several thousand dollars. air cargo The difference in shipping rates would be enormous.

Preparing such a paste maintains the viability and longevity of bacteria. This is crucial to achieving this goal. The centrifuged paste is free of evaporation loss and oxygen. must be kept cool. To do this, the bacteria must be placed in a cylindrical Sharpless (Sha) is deposited inside the pod so that the sausage is backed Continuous flow centrifuges such as rple+) and Visking or Saran in the centrifuges It is necessary to use an insert made of a material such as aqueous environment, enzymes, nutrients and The described method will A bacterial paste contained inside an impermeable casing that does not change over time. is manufactured. Bacterial food remains in the aqueous concentrate, so bacteria Growth can be arrested and centrifuged at any stage of its development. And bacteria commercially available Waring (W! ng) or Oster (0+lu) machines. Resuspending the bacteria into an aqueous suspension obtained using a simple food mixer such as a It can be held without any environmental influence until it becomes cloudy. far Storage of the so-called bacterial sausages produced in the human body is easily carried out in commercial refrigerators. be exposed. In addition, filling the tube prevents dehydration, so it Within the tube, the contents are not dehydrated beyond that achieved by centrifugation.

The bacteria present in the sausage can be used to grow new culture colonies. To use it as terrier seed, cut a portion out of the sausage and use it. and can be added to any growth medium. Next, if a microbiologist Bacteria growth is initiated using foolproof methods.

The production of such bacterial sausages is, in particular, associated with any of the following industrial processes: It can be used for, but is not limited to, the following purposes.

1) Production of mixed cultures or other types of treatment for general wastewater.

2) Maintenance of bacterial cultures from equipment that is occasionally subjected to toxic or conditional shocks. Possessed. These preserved cultures grow concentrated seed cultures that are introduced into plants. By having bacteria readily available for use, such contingencies can be avoided. A means of immediate on-site relief from I will provide a.

3) Specialized preparations of bacterial mixtures can eliminate native bacteria in existing equipment. Both the bacteria and the bacteria that need to be added to the natural bacteria in the equipment It would be possible to manufacture it using Such sausages may be a conditional problem or a poisonous problem. may function to restart a treatment plant that has been affected several times by Additional species to optimize and improve the operation of the plant on a regular and continuous basis It can function as

(4) The sausage contains a huge number of live bacteria that are concentrated and immediately available. Providing a means of transportation over vast distances from one place to another in the form that is possible. provide

For all intents and purposes, this bacterial suspension in the sausage is Additional growth occurs even though it is immediately available because it does not contain water or water. or if a stable bacterial mixture is available nearby that has virtually eliminated the possibility of mortality. It will be done.

When the bacterial paste is produced as described above, it contains highly concentrated bacteria. The rear mixture is used as a system to provide the required functions and capabilities of the equipment. become available for use. Such systems provide a method for rendering bacteria dormant and Continuing, this results in products produced in non-aqueous media as described below. , can be further improved.

Ability to distribute bacteria in concentrated form in the sediment at the bottom of the water receptor, treatment equipment or aqueous cultivation processes are the primary would provide benefits. Currently available systems, as described above, relies on physically injecting bacteria into the system. Centrifuged Pe Bacteria prepared as yeast are non-aqueous, denser than water, and easily soluble in water. The preservation, distribution and and utilization will be significantly improved. Minimal impact on the environment and overall water and Some of these available liquids supply the requirements of a miscible liquid system. There is an example. An example is glycerol-glycerin ethylene glycol 1.2-Propylene glycol lj-propylene glycol By introducing bacteria into such a medium from a paste, a stable suspension can be achieved. A cloudy liquid is obtained. This is because, although these liquid media are biodegradable, their preparation This is because there is no water or nutrients in the material for bacteria to grow.

The growth of bacteria, or any form of life for that matter, requires four basic components: These are carbon, nitrogen, phosphorus and water. Bacteria are the Centrifuged from most of the nutrients used for growth and the water in which growth takes place , by resuspending it in an anhydrous liquid such as glycol. , the necessary conditions for its growth are not provided. Because nitrogen, phosphorus and water Both contain glycerol, ethylene glycol, or various polylene glycols. This is because it is not part of the sexual body. Bacterial paste added to anhydrous, non-aqueous liquid The bacterial suspension formed from Bacteria lack the nutrients they need to grow and multiply. The stability of this system is given by the growth-limiting conditions of the suspension itself and the anhydrous conditions.

These liquids are completely miscible with water, so bacteria made with these materials The rear suspension immediately disperses once introduced into the aqueous environment. like phosphorus and nitrogen Growth is guaranteed if the necessary nutrients are available. Then, preparation of suspension The original organic liquid used in the process is either diluted beyond practical use or consumed. as a food source to initiate bacterial growth in the new aqueous burial. Function. For this reason, once Bacteria growth maintenance media are created, they They continue to grow and multiply again to provide their metabolic, cleaning function. this The technology keeps bacteria growing but inactive until they are needed. fully activated by the simplest procedure of direct introduction into growth maintenance media. keep it in a state where it is possible to change it.

The properties of the four liquid temperature-sensitive chemicals selected here as examples are as follows.

Compound Density Melting point Boiling point Solubility Glycerol 1.269 20 290 Infinite! 120 ethylene 1.108 8 -11.5 199 Infinity + 120 Glycol 1.2-Propylene 1. ．． 0361-189 Mugen B20 Glycol 1.3-Propylene 1.0597-213.5 Infinite 1120 Glycol These properties provide suspensions with several important capabilities.

1) Do not freeze at 0℃. In other words, it does not freeze even at -20°C.

21. Does not evaporate or boil at 100℃.

3) Does not cause any harm to the people handling it. In some cases, injection (injectcd) may not cause any harm.

4) Does not inhibit bacterial growth at all.

5) Provides long-term storage ability.

Once the bacterial suspension described above is made, A liquid system with a higher density than any body of water is available. It becomes something close. Introduction of these bacterial suspensions into the organic sludge layer at the bottom of the water basin There may be times when it would be advantageous to allow this. To achieve this, This bacterial suspension is dissolved by and washed away with the water. A medium must be present to prevent this. In fact, this is due to the slow pace of bacteria. Allowing the introduction of these bacterial suspensions into systems where release is deemed necessary It would be useful to do so. without an appropriate shell to prevent dissolution of the suspended liquid , such a process is not possible. Something denser than fresh water or seawater The bacteria suspended in the medium are incorporated into a form that provides a system for slow distribution. How to do so is described here.

By encapsulating the manufactured suspension using a substance such as gelatin, the suspension can be The necessary system is provided to enable the isolation and use of like gelatin substances that retain bacteria over a long period of time after the encapsulated liquid sinks to the bottom of the puddle. Allows for slow release of turbid liquid. Of course, the release of the suspension contained inside Solid or semi-solid coatings that allow some time to develop are also sufficient. It's a minute. One can imagine a number of different coatings to achieve this goal. You will be able to do that. The basic requirement for this coating is that it is soluble in water. Solubility and insolubility in the liquid used to make the bacterial suspension to be coated Limited to gender.

Such systems are readily available commercially and can be used to make tablets such as vitamins. Since it is used in construction, we will not discuss it here. For example, in the case of glycerol , capsules similar to those sold as vitamin E are quite sufficient.

international search report international search report PCT/GO90101093

Claims (15)

[Claims] 1. A method of stabilizing bacteria present in an aqueous growth medium, which involves centrifugation. A non-aqueous bacterial paste is prepared by isolating bacteria from a growth medium; A method consisting of storing a host in a protective environment. 2. 7. The method of claim 1, wherein said protective environment comprises an impermeable sheath. 3. 2. The method of claim 1, wherein said sheath is comprised of a visking material. 4. 2. The method of claim 1, wherein said sheath is made of Saran material. 5. Claims wherein said protective environment is a liquid environment in which said paste is resuspended. The method described in paragraph 1. 6. 6. The method of claim 5, wherein the liquid environment is aqueous. 7. 7. The method of claim 6, wherein said liquid is water. 8. 6. The method of claim 5, wherein the liquid environment is non-aqueous. 9. Claim 8, wherein the liquid is glycerol, glycol, or the like. How to put it on. 10. The liquid containing the paste is non-permeable but dissolves or disintegrates. Any one of claims 5 to 9, which is encapsulated in a degradable membrane that can be destroyed. The method described in. 11. 11. The method according to claim 10, wherein the membrane is gelatin or the like. 12. Non-aqueous bacterial paste obtained by centrifugation and housed in an impervious sheath A bacterial preparation consisting of 13. Non-aqueous bacterial paste obtained by centrifugation and placed in a non-aqueous liquid A bacterial preparation consisting of 14. From a non-aqueous bacterial paste obtained by centrifugation and resuspended in water. bacterial preparations. 15. The bag according to claim 13 or 14 housed inside a capsule. Terrier preparation.