JP2015172040A - Radioactive substance recovery agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a recovery agent and a recovery method for effectively recovering radioactive substances taken in a living body.SOLUTION: The present invention relates to a radioactive substances recovery agent that comprises a genus Bifidobacterium microorganism as an active ingredient, where the microorganism is contained in a capsule, and the target radioactive substances to be recovered are radioactive cesium, radioactive sodium and radioactive strontium. Further, in order to remove radioactive substances from non-human animals, the animals are made to ingest the radioactive substance recovery agent.

Description

  The present invention relates to a radioactive substance recovery agent.

  The radioactive material released into the atmosphere by the explosion accident at the Tokyo Electric Power Fukushima Daiichi Nuclear Power Station following the Great East Japan Earthquake that occurred in March 2011 caused environmental pollution in a wide area mainly in Fukushima Prefecture. Food contamination by radioactive cesium has become a major problem.

  For example, as a decontaminant for radioactive substances in soil or water, a mixture of a plurality of microorganisms such as lactic acid bacteria, Bacillus bacteria, cyanobacteria, yeast, and humic substances (Patent Document 1), and in a liquid or gas It has been reported that single substance and activated carbon to which oligosaccharides are attached are used for the recovery of radioactive substances (Patent Document 2).

  In addition, the radioactive cesium taken into the animal body was detected at a higher level from the feces excreted from the animal than the organ of the animal, and the radioactive substance was taken up by the bacteria constituting the intestinal flora (Non-Patent Document 1).

JP 2013-174557 A JP 2012-112769 A

Abstracts of Annual Meeting 2013 Agricultural Chemical Society of Japan, 3B21p11

  The objective of this invention is providing the collection | recovery agent and collection | recovery method for collect | recovering effectively the radioactive substance taken in in the living body.

  The present invention provides a radioactive substance recovery agent containing a Bifidobacterium microorganism as an active ingredient.

  In one embodiment, the Bifidobacterium microorganism is Bifidobacterium addresscentis, Bifidobacterium angratam, Bifidobacterium animalis subsp. Animalis, Bifidobacterium animalis subsp. Lactis, Bifidobacterium asteroides, Bifidobacterium bifidum, Bifidobacterium boum, Bifidobacterium breve, Bifidobacterium catenuratam, Bifidobacterium kerinum, Bifidobacterium coryne Foam, Bifidobacterium kunikuri, Bifidobacterium denticorens, Bifidobacterium denthium, Bifidobacterium galcumum, Bifidobacterium galinara , Bifidobacterium globosum, Bifidobacterium indicam, Bifidobacterium infantis, Bifidobacterium inopinatum, Bifidobacterium lactis, Bifidobacterium longum, Bifidobacterium Magnum, Bifidobacterium merlicum, Bifidobacterium minimum, Bifidobacterium perbroram, Bifidobacterium pseudocatenuratam, Bifidobacterium pseudolongum subspice globosum, Bifidobacterium Um pseudolongum subspice pseudolongum, Bifidobacterium prolum, Bifidobacterium luminal, Bifidobacterium luminantium, Bifidobacterium sec Any one selected from the group consisting of R., Bifidobacterium Scurbi, Bifidobacterium subtil, Bifidobacterium Switzerland, Bifidobacterium thermacid film, and Bifidobacterium therm film There are two microorganisms.

  In one embodiment, the Bifidobacterium microorganism is any one microorganism selected from the group consisting of Bifidobacterium longum strains JBL01 and JBL05.

  In one embodiment, the Bifidobacterium microorganism is a radioactive substance recovery agent contained in a capsule.

  In one embodiment, the capsule is either a seamless capsule, a soft capsule, or a hard capsule.

  In one embodiment, the recovered radioactive material is any one or more of radioactive cesium, radioactive sodium, or radioactive strontium.

The present invention is also a method of removing radioactive material from a non-human animal, the method comprising:
A step of ingesting the radioactive substance recovery agent to the animal.

  According to the present invention, for example, it is possible to provide a recovery agent and a recovery method for effectively recovering a radioactive substance taken into a living body. According to such a recovery agent and recovery method, it is possible to remove the radioactive substance from the living body of the animal safely and effectively.

  The “radioactive substance recovery agent” of the present invention refers to an agent that recovers from a region containing the radioactive substance by incorporating the radioactive substance into a microorganism belonging to the genus Bifidobacterium. Here, “recovery” refers to taking a radioactive substance into a microorganism belonging to the genus Bifidobacterium and moving or removing it from a region containing the radioactive substance. Such regions include, for example, in vivo living animals (for example, livestock and poultry such as cows, pigs and chickens; and pets such as dogs and cats; humans) or the surrounding environment (for example, soil, Rivers, lakes, etc.).

  According to the “radioactive substance recovery agent” of the present invention, the radioactive substance can be recovered by bringing the radioactive substance recovery agent into contact with a region containing the radioactive substance. For example, for a radioactive substance taken into an animal body, the radioactive substance can be recovered by ingesting the animal with a radioactive substance recovery agent.

  The radioactive substance recovery agent of the present invention contains a Bifidobacterium microorganism as an active ingredient. For example, the radioactive substance recovery agent consists essentially of a Bifidobacterium microorganism as an active ingredient, or optionally contains a component as described below (also referred to as “optional component”). To do.

  The Bifidobacterium microorganism used in the present invention may be any microorganism belonging to the genus Bifidobacterium. Examples of the microorganism belonging to the genus Bifidobacterium include the following: Bifidobacterium adolescentis, Bifidobacterium angulatum, Bifidobacterium animalis subsp. Bifidobacterium animalis subsp. Animalis), Bifidobacterium animalis subsp. Lactis, Bifidobacterium Bifidobacterium Bifidobacteria Bifidobacterium Bifidobacterium bifium, Bifidobacterium boum, Bifidobacterium breve, Bifidobacterium bibium, Bifidobacterium bif Bifidobacterium coryneform, Bifidobacterium cuniculi, Bifidobacterium denticolens, Bifidobacteria Bifidobacterium dentium, Bifidobacterium galcumum, Bifidobacterium gallinarum, Bifidobacterium galbium, Bifidobacterium globium indicum, Bifidobacterium infantis, Bifidobacterium inopinatum, Bifidobacterium lac is), Bifidobacterium longum, Bifidobacterium magnum, Bifidobacterium mericicum, Bifidobacterium minfum Bifidobacterium parvulorum, Bifidobacterium pseudocatenatum, Bifidobacterium pseudolongum subspice globosum (Bifidobacterium pseudoglobulum) ubsp. globosum), Bifidobacterium pseudopulmonum, Bifidobacterium pullulum, Bifidobacterium pullulum, Bifidobacterium pullulum, Bifidobacterium pullulum Um luminumium (Bifidobacterium ruminantium), Bifidobacterium saeculare, Bifidobacterium sarcobii, Bifidobacterium suf. dobacterium subtile), Bifidobacterium Switzerland (Bifidobacterium suis), Bifidobacterium Therm A Sid film (Bifidobacterium thermacidophilum), and Bifidobacterium Therm film (Bifidobacterium thermophilum) and the like.

  Bifidobacterium microorganisms used in the present invention are, for example, Bifidobacterium longum strains JBL01 and JBL05. Bifidobacterium longum strain JBL01 is a strain isolated from the intestines of healthy individuals, and Bifidobacterium longum strain JBL05 is NITE BP-82, an independent administrative agency, Product Evaluation Technology Infrastructure Organization, Patent Microorganism It can be obtained from the Deposit Center (NPMD). Bifidobacterium longum strain JBL05 is preferred.

  The microorganism belonging to the genus Bifidobacterium may be cultured and recovered by a method commonly used by those skilled in the art and used as it is as a recovery agent. Alternatively, it may be dried to prepare a powder (bacterial powder). Drying can be performed by a treatment method that enables growth when exposed to growth conditions such as intestinal environment or culture medium by performing low-temperature treatment such as freeze-drying and low-temperature drying.

  As a radioactive substance collect | recovered by the radioactive substance collection | recovery agent of this invention, although arbitrary radioactive substances are object, radioactive cesium, radioactive sodium, and radioactive strontium are mentioned, for example. The collected radioactive substance may be one type or a combination of plural types, and any of them may be a target.

  The radioactive substance recovery agent can be used for food, feed and pharmaceutical applications, but is not limited to these applications. Specifically, the radioactive substance recovery agent is not limited to these. For example, it should be prepared and used as a powder, granule, tablet, capsule, paste, cream, jelly, or liquid. Can do. Examples of optional components include inorganic substances and organic substances such as excipients, binders, disintegrating agents, lubricants, flavoring agents, solubilizing agents, suspending agents, and coating agents.

  The radioactive substance recovery agent can be used as a food composition, a feed composition, or a pharmaceutical composition for oral ingestion, and a pharmaceutical product for oral administration, liquid food, food for patients, infant food, nutritional functional food, It can be added to foods for specified health use. The composition for oral use of the radioactive substance recovery agent can also be blended in ordinary food and drink or animal feed. Examples of the food and drink include various beverages such as milk, soft drinks, and jelly beverages, various confections such as candy, gummi, chocolate, biscuits, and crackers, and various foods such as rice, bread, udon, and dressings. However, it is not limited to these. Examples of animal feed include feed for livestock and poultry such as cattle, pigs and chickens, and pet feed such as dogs and cats. Milk or the like can be fermented using a composition containing viable bacteria of the genus Bifidobacterium in the radioactive substance recovery agent and ingested as yogurt or fermented milk.

The amount of the microorganism belonging to the genus Bifidobacterium in the radioactive substance recovery agent can be arbitrarily determined according to its purpose, application, form, dosage form, symptom, body weight and the like. The amount of Bifidobacterium microorganisms in the radioactive substance recovery agent is, for example, in the case of oral ingestion or administration, the number of bacteria per day, for example, 1 × 10 ⁶ to 1 × 10 ¹² , more preferably, It is preferable to be determined so that 1 × 10 ⁸ to 1 × 10 ¹¹ can be ingested.

One embodiment of the radioactive substance recovery agent as an oral composition is a form in which a Bifidobacterium microorganism is contained in a capsule, and this form is also referred to as a capsule preparation. Capsules include seamless capsules, soft capsules, and hard capsules. When the form of the radioactive substance recovery agent is a capsule preparation, the number of Bifidobacterium microorganisms is, for example, 1 × 10 ^{8 to} 5 × 10 ¹⁰ cfu, preferably 5 × 10 ^{8 to} 3 per 1 g of the capsule. It can be prepared to contain x10 ⁹ cfu. Moreover, it can be prepared such that the amount of cells per capsule is, for example, 1 × 10 ^{7 to} 5 × 10 ⁸ cfu, preferably 5 × 10 ^{7 to} 3 × 10 ⁸ cfu.

  Preferably, the capsule film is an acid resistant film. In order to express the function of taking up radioactive substances in the animal body, the microorganisms pass through the stomach and enter the intestines so that ion channels existing in the cell membrane of Bifidobacterium microorganisms can be used in the animal body. It is desirable to be able to reach and grow there. In order for the Bifidobacterium microorganisms to reach the intestine of the animal while remaining alive and to take up radioactive substances in the intestine, it is preferable to prevent the microorganisms from being affected by gastric acid as much as possible.

  The configuration, shape, etc. of the capsule preparation with acid-resistant film (also referred to as “acid-resistant capsule preparation”) are not particularly limited as long as the film is resistant to gastric acid. That is, it is desirable that gastric acid penetrates into the capsule and does not come into contact with microorganisms. The capsule film may be a film that does not dissolve at pH 4 or less, preferably at pH 1 to 3. There is no particular limitation on the encapsulation method.

  The production of the acid-resistant capsule preparation will be described below.

(Seamless capsule formulation)
The shape of the capsule for imparting resistance to gastric acid is preferably a seamless capsule. The “seamless capsule” is a kind of soft capsule and refers to a capsule in which the contents are enclosed with a seamless film. The seamless capsule can have a multilayer structure of two or more layers, and preferably has a multilayer structure of three or more layers. Usually, the innermost layer may contain contents (in the case of the present invention, microorganisms), and the outer layer (or outermost layer) may be a film. In other words, it is a form in which microorganisms are included by the film.

  The preparation of a seamless capsule preparation having a three-layer structure will be described below. It is a schematic cross section of a seamless capsule preparation with a three-layer structure. The three-layer structure is composed of an innermost layer, an endothelial layer that covers the innermost layer, and an outer layer that covers the innermost layer.

  The innermost layer is composed of the microorganism and a non-aqueous solvent or a solid component for suspending / mixing the microorganism (hereinafter, this component is referred to as an innermost layer substance). The innermost layer material is not particularly limited. Examples include various fats and oils, fatty acids, fatty acid esters of sugar, aliphatic hydrocarbons, aromatic hydrocarbons, chain ethers, higher fatty acid esters, higher alcohols, and terpenes. Specifically, soybean oil, sesame oil, palm oil, palm kernel oil, corn oil, cottonseed oil, coconut oil, rapeseed oil, cacao butter, beef tallow, lard, horse oil, whale oil, the above-mentioned natural having a melting point of 40 ° C. or less Examples include, but are not limited to, hydrogenated fats and oils, margarine, shortening, glycerin fatty acid ester, sucrose fatty acid ester, camphor oil, thin cargo oil, α-pinene, and D-limonene. These innermost layer materials can be used alone or in admixture of two or more.

  As the material for the endothelial layer, among the above innermost layer substances, for example, a substance having a melting point of 20 ° C. to 50 ° C. and different from the innermost layer is used. Preferably, a substance that is solid at room temperature is used. As the material for the innermost layer and the material for the endothelial layer, the same kind of fats and oils that have been hydrogenated so as to have different melting points can also be used. For example, hydrogenated palm kernel oil having a melting point of 34 ° C. is used as the innermost layer substance, and hydrogenated palm kernel oil having a melting point of 43 ° C. is used as the material of the endothelial layer, and this endothelial layer suppresses the permeation of moisture and oxygen. And can act to prevent contact with stomach acid. The substance to be selected can be determined in consideration of the storage period of the capsule.

  Materials for the outer layer (the outermost layer in the case of a three-layer structure or more) include a mixture of protein and water-soluble polyhydric alcohol, a mixture of protein, water-soluble polyhydric alcohol and polysaccharide, and a polysaccharide and water-soluble polyhydric alcohol. And a mixture thereof. Examples of proteins include gelatin and collagen. Examples of the water-soluble polyhydric alcohol include sorbitol, mannitol, glycerin, propylene glycol, and polyethylene glycol. Polysaccharides include agar, gellan gum, xanthan gum, locust bean gum, pectin, alginate, carrageenan, gum arabic, dextrin, modified dextrin, starch, modified starch, pullulan, pectin, and carboxymethylcellulose salt. When using pectin, alginate, gellan gum, or carrageenan, an alkali metal salt or an alkaline earth metal salt may be added as appropriate.

  The above-mentioned three-layered seamless capsule preparation is prepared by a technique well known to those skilled in the art, for example, a dropping method using a triple nozzle described in Japanese Patent No. 1398836. In this dropping method, the innermost layer substance combined with microorganisms (for example, freeze-dried cells) (preferably into a hydrophobic solvent substance that is non-flowable at 20 ° C. to 50 ° C.) from the innermost nozzle of the concentric triple nozzle. A suspension of microorganisms (preferably freeze-dried cells) from the intermediate nozzle, the substance constituting the endothelial layer (for example, a melt of a substance that is solid at room temperature), and the outer layer (from the outermost nozzle) A three-layer structure in which microorganisms are contained in the innermost layer by simultaneously discharging a solution of a substance to be a film) and dropping it into a carrier liquid (for example, corn oil, rapeseed oil, etc.) flowing under cooling A “seamless” capsule is formed. Thus, the microorganisms are included or contained by a seamless outer layer coating.

  The capsules formed as described above are then dried. As a drying method, for example, normal temperature ventilation drying is performed. The drying is generally performed by, for example, drying at 5 ° C. to 30 ° C. air. The drying time is preferably 2 to 12 hours. A method of further vacuum drying or vacuum freeze drying described above in JP-A-07-0669867 can be suitably used for capsules that have been subjected to normal drying as described above. In vacuum drying, for example, the vacuum can be kept at 0.5 to 0.02 torr. In vacuum freeze-drying, for example, it can be frozen at −20 ° C. or less at the degree of vacuum and dried. The time required for vacuum drying or vacuum freeze drying is not particularly limited, but is generally 5 to 60 hours, preferably 24 to 48 hours. If it is 5 hours or less, the drying is insufficient, and the moisture present in the capsule can adversely affect the contents.

  In the capsule obtained by the method described in JP-A-07-0669867, moisture in the capsule is sufficiently removed by vacuum freeze-drying, the Aw value is 0.20 or less, and the thermal conductivity is 0.16 kcal / mh ° C. It can be: Of course, moisture is reduced by vacuum drying or vacuum freeze drying, and at the same time, the capsules are sufficiently dried and porous, so that the thermal conductivity is also much lower than that obtained by simply drying.

  The Aw value is not the absolute amount of water present in the sample, but the value determined by the presence of water, that is, the degree of freedom of water in the sample. It is an index representing moisture that can be involved, and is measured by an electrical resistance water activity measurement method (for example, Aw meter WA-360, Shibaura Electronics Co., Ltd.). The thermal conductivity is measured by the Fitch method or the like. The Aw value is preferably 0.20 or less, and the thermal conductivity is preferably 0.02 to 0.08 kcal / mh ° C.

  In order to impart acid resistance to the capsule film of the seamless capsule preparation, an acid-resistant outer layer is formed, or the formed seamless capsule film (outermost layer) is treated to be acid-resistant.

  As a method of forming the acid-resistant outer layer, pectin, alginate, gum arabic, etc., for example, 0.01 to 20% by mass, preferably 0.1 to 0.1% with respect to gelatin, agar, carrageenan and the like having gelling ability. The method of adding so that it may become 10 mass% is mentioned.

  Examples of a method for imparting acid resistance to the formed seamless capsule film (outermost layer) include crosslinking treatment of the outer layer (outermost layer) of the seamless capsule and coating treatment of the surface of the seamless capsule. These treatments are preferably performed alone or in combination.

  When the outer layer containing protein is subjected to a crosslinking treatment, first, a seamless capsule is prepared and then thoroughly washed with water. The seamless capsule washed with water is added to an aqueous solution containing a crosslinking agent to crosslink the surface of the outer layer. A conventionally well-known crosslinking agent can be used as a crosslinking agent. Examples of the crosslinking agent include formaldehyde, acetaldehyde, propionaldehyde, glyoxal, glutaraldehyde, cinnamaldehyde, vanillyl aldehyde, acetone, ethyl methyl ketone, ethylene oxide, propylene oxide, potassium alum, and ammonium alum. By adding 1 part by weight of a seamless capsule to 50 to 100 parts by weight of an aqueous solution containing, for example, 0.1 to 2 w / v%, preferably 0.5 to 2 w / v% of a crosslinking agent, and stirring for 10 to 300 seconds, Outer layer processing can be performed. In addition, the usage-amount of a crosslinking agent and the time to act vary with crosslinking agents. After the surface of the outer coating is crosslinked, it is sufficiently washed with water to remove the aqueous solution containing the crosslinking agent and dry the moisture contained in the outer layer.

  Moreover, you may perform the bridge | crosslinking by the enzyme process which uses transglutaminase as a bridge | crosslinking process of the outer layer containing said protein. In this case, 1 part by mass of the produced seamless capsule is added to 50 to 100 parts by mass of an aqueous solution containing, for example, 0.1 to 10 w / v%, preferably 0.5 to 2 w / v% enzyme, and stirred for 1 to 300 minutes. By doing so, the outer layer can be treated and washed with water and dried as described above.

  When performing the coating treatment, after drying the produced wet seamless capsule, shellac, ethylcellulose, hydroxypropylmethylcellulose, hydroxypropylcellulose, polyvinylpyrrolidone, cellulose TC-5, vinylpyrrolidone-vinyl acetate copolymer, zein, ethylene Based on wax, etc., using conventional methods such as castor oil, rapeseed oil, dibutyl phthalate, polyethylene glycol, glycerin, stearic acid, fatty acid ester, sorbitan palmitate, polyoxyethylene stearate, acetylated monoglyceride as plasticizer Coat seamless capsules according to

  Furthermore, by providing enteric properties to the capsule film, the capsule can be protected from acidic solutions in the stomach (for example, gastric acid) and disintegrated in the intestine, thereby releasing microorganisms inside the capsule in the intestine. It becomes possible. Entericity is generally imparted by those skilled in the art by producing enteric capsules. Further, an enteric coating can be obtained by using a mixture containing gelatin and pectin as the outer layer material of the seamless capsule. Prepared by adding pectin, alginate, gum arabic, etc. to gelatin, agar, carrageenan, etc. having gelling ability so as to be 0.01 to 20% by mass, preferably 0.1 to 10% by mass, for example. The acid-resistant outer layer also has enteric properties.

  The seamless capsule formulation can be spherical in shape due to its manufacturing method. The average particle diameter of the seamless capsule is, for example, 0.3 to 10 mm, preferably 1.5 to 8.0 mm.

  The thus obtained seamless capsule preparation can be stored for 6 months or more while maintaining the activity of microorganisms at room temperature, and when stored at 10 ° C. or lower, it can be stored for a long period of 1 year or longer. is there.

(Soft capsule formulation)
As in the case of the seamless capsule preparation, the soft capsule preparation contains a suspension of microorganisms in a non-aqueous solvent and is contained in a film sheet. The material of the film sheet is the same as the material of the outer layer of the seamless capsule.

  The soft capsule preparation can be prepared by a known technique, for example, a method described in Japanese Patent No. 2999535. For example, a rotary die can be used to encapsulate and encapsulate the coated sheet by heating through a mold while injecting and filling the contents. In order to have a function of releasing microorganisms in the intestine, oil obtained as a release agent is removed from the obtained soft capsules by washing with a polar solvent (for example, methanol, ethanol, propanol, isopropanol). Thereafter, as in the case of the seamless capsule, the crosslinking treatment and the coating treatment may be performed in combination, or any one of the treatments may be performed to make it acid resistant.

  The acid-resistant film sheet is, for example, 0.01 to 20% by mass, preferably 0.1 to 10% by mass of pectin, alginate, gum arabic and the like with respect to gelatin, agar, carrageenan and the like having gelling ability. And can be prepared based on a known method. Alternatively, the film sheet may be combined with a crosslinking treatment and a coating treatment, or any one of the treatments may be performed to make it acid resistant. Using the acid-resistant film sheet thus obtained, for example, the capsule is molded and the contents are introduced into the capsule by a known technique, and then the contents are encapsulated by welding the seam of the capsule. A soft capsule preparation in which microorganisms are contained by an acid-resistant film can be produced.

  Soft capsule formulations can have a spherical, elliptical, or rectangular structure. The soft capsule preferably has a major axis of 3 to 16 mm and a minor axis of 2 to 10 mm, more preferably a major axis of 5 to 7 mm and a minor axis of 2 to 3 mm.

  The soft capsule formulation thus obtained can be stored for 6 months or more while maintaining the activity of microorganisms at room temperature, and when stored at 10 ° C. or less, it can be stored for a long period of 1 year or more. is there.

(Hard capsule formulation)
A hard capsule formulation can be produced by pre-molding a capsule film into a body and a cap, filling the contents into the capsule body, and then combining the capsule cap.

  Examples of the film material for the hard capsule preparation include gelatin, cellulose, pullulan, carrageenan, and cellulose derivatives such as hydroxypropylmethylcellulose. Hard capsules can be molded by methods commonly used by those skilled in the art. The molded capsule may be a commercially available capsule. The contents can be wrapped in a film and sealed.

  The contents can be a mixture of microorganisms with an excipient (for example, anhydrous silicic acid, synthetic aluminum silicate, lactose, corn starch, crystalline cellulose) or a powder containing dried microorganism powder. .

  The capsule film may be coated after the contents are placed in the capsule. The coating can be applied with the materials and methods described in the outer layer of the seamless capsule, thereby imparting acid resistance and preferably disintegration (enteric) in the intestine. This coating may also have the role of encapsulating the capsule film and including the contents.

  The acid-resistant film sheet is, for example, 0.01 to 20% by mass, preferably 0.1 to 10% by mass of pectin, alginate, gum arabic and the like with respect to gelatin, agar, carrageenan and the like having gelling ability. And can be prepared based on a known method. Alternatively, the film sheet may be combined with a crosslinking treatment and a coating treatment, or any one of the treatments may be performed to make it acid resistant. By using the acid-resistant film sheet thus obtained, for example, a hard capsule can be molded by a known technique, and the contents are put inside the molded hard capsule and the seam of the capsule is welded. The contents can be enclosed, and a hard capsule preparation containing microorganisms by an acid-resistant film can be produced.

  The hard capsule formulation thus obtained can be stored for 6 months or more while maintaining the activity of the bacteria at room temperature, and when stored at 10 ° C. or lower, it can be stored for a long period of 1 year or longer. is there.

  The present invention is also a method for removing a radioactive substance from an animal other than a human, and this method includes the step of ingesting the radioactive substance recovery agent into the animal. The radioactive substance recovery agent to be ingested is preferably a capsule preparation as described above, more preferably an acid-resistant capsule preparation as described above.

  EXAMPLES Hereinafter, although an Example demonstrates this invention more concretely, this invention is not limited by these Examples.

(Example 1: Radiocesium uptake of Bifidobacterium)
In this example, viable bacteria of Bifidobacterium longum strains JBL01 (isolated from the intestines of healthy individuals), JBL05 strains and JCM1217 strains (obtained from the microbial material development room (JCM) of RIKEN BioResource Center) were frozen. Bacterial powder obtained by drying was used (after centrifugal concentration, 10% cellulose (Avicel manufactured by Asahi Kasei Co., Ltd.) suspension water for the pellet volume was added and freeze-dried). As the base solution of the radiation source, the surface layer soil contaminated with radioactive cesium was dissolved by heating with nitric acid, etc., and the supernatant was separated by centrifugation, dried once, and then used as an aqueous solution.

10 mg of bacterial powder was suspended in 10 mL of MRS broth medium (Becton, Dickinson and Company) and pre-cultured at 37 ° C. for 18 hours. The viable cell count of the bacterial solution was determined by taking a correlation between OD and viable cell count. Bouillon was prepared from beef meat sampled within 20 km of Fukushima nuclear power plant according to the bacteriological training requirements. To the MRS broth medium, 10% bouillon was added, and 0.5 mL of the above suspension was added to 20 mL of the medium so that the number of life bacteria was 5 × 10 ⁷ cells / mL. Time anaerobic culture was performed. Thereafter, the culture solution was centrifuged at 8,000 × g for 10 minutes at 4 ° C., and the obtained supernatant was collected in a beaker. Distilled water was added to the precipitate after separating and removing the supernatant, and the suspension was sufficiently suspended, followed by further centrifugation at 8,000 × g for 10 minutes at 4 ° C. The obtained supernatant was put into a beaker and made up to 70 mL to obtain a supernatant sample. The resulting bacterial cells were made up to 70 mL with distilled water, 0.7 g of superabsorbent resin CP1 (Kennis Co., Ltd.) was added, and the cells were gelled in a U8 container so that the bacterial cells were dispersed. A bacterial cell sample was used.

  Cesium 137 (Cs-137) with known radioactivity is correlated with the scintillation measurement at the energy level of the substance in a three-point germanium counter, and Cs-137 in the germanium counter in the supernatant sample and cell sample. The radioactivity of Cs-137 in the microorganism and in the culture medium was measured from the scintillation measurement value of energy corresponding to. The radioactivity uptake rate of the microorganism was calculated from the measured values of the remaining medium and the radioactivity in the microbe (the radioactivity uptake rate (%) corresponds to the radioactivity measurement in the microbe relative to the initial amount). The results are shown in Table 1.

  The results shown in Table 1 showed that all Bifidobacterium tested had radiocesium uptake ability (recovery ability), but JBL05 strain had particularly high Cs-137 uptake ability. It was done.

(Preparation Example 1: Preparation of acid-resistant seamless capsule formulation)
As described below, it was prepared using a capsule manufacturing apparatus equipped with a concentric triple nozzle. 400 g of hardened oil (hydrogenated palm kernel oil having a melting point of 34 ° C.) was melted, and 100 g of viable powder of any of JCM1217, JBL01 and JBL05 was dispersed therein. A hardened oil (hydrogenated palm kernel oil having a melting point of 43 ° C.) melted from the inner nozzle of the concentric triple nozzle and the outer intermediate nozzle of this dispersion was further added to a gelatin solution (gelatin 600 g, glycerin 300 g, and Dissolve 100 g of pectin dissolved in 4 kg of purified water at the same time, and drop it into rapeseed oil flowing under cooling at 15 ° C., so that the cells in a three-layered seamless capsule with a diameter of 2.5 mm A formulation including was prepared. The capsule preparation was air-dried at 20 ° C. for 10 hours, and further vacuum-dried at room temperature, so that the water activity in the capsule had an Aw value of 0.20 or less and a thermal conductivity of 0.16 kcal / mh ° C. or less. Reduced to. The amount of bacterial cells per capsule was 1.0 × 10 ⁸ cfu.

(Preparation Example 2: Preparation of acid-resistant soft capsule preparation)
A soft capsule content solution was prepared by suspending 50 g of the bacterial powder of any of JCM1217, JBL01 and JBL05 in 300 g of rapeseed oil. 400 g of gelatin and 100 g of glycerin were added to 200 g of distilled water and dissolved by stirring at 60 ° C., and this was formed into a sheet shape to obtain a gelatin film, which was used as a soft capsule film. This gelatin film was encapsulated by feeding it between a pair of rotating cylindrical molds and ejecting the content liquid between the gelatin films with a pump interlocking therewith. 400 g of the obtained encapsulated product was put into a rolling granulator, and a solution prepared by dissolving 10 g of shellac and 1 g of castor oil in 400 g of a methanol-ethyl acetate (1: 1, v / v) mixed solution was applied to the entire surface of the soft capsule. It sprayed so that it might become a coating film thickness 0.3mm. In this manner, 400 g of a soft capsule preparation having a major axis of 4 mm and a minor axis of 3 mm, including bacterial cells and coated with acid resistance was obtained. The amount of bacterial cells per capsule was 1.2 × 10 ⁸ cfu.

(Preparation Example 3: Preparation of acid-resistant hard capsule formulation)
The viable powder of any one of JCM1217, JBL01 and JBL05 was used as the contents of the hard capsule. As the hard capsule film, a commercially available pharmacopoeia No. 5 capsule was used. The contents were encapsulated by filling the capsule body and combining the cap with this. 100 g of the obtained encapsulated product was put in a rolling granulator, and a solution prepared by dissolving 10 g of shellac and 1 g of castor oil in 400 g of a methanol-ethyl acetate (1: 1, v / v) mixed solution was added to the entire surface of the hard capsule. It sprayed so that it might become a coating film thickness of 0.3 mm, and 100 g of hard capsule formulation which included the microbial cell and was acid-resistant coated was obtained. The amount of bacterial cells per capsule was 1.8 × 10 ⁸ cfu.

(Example 2: Behavior of viable bacteria in capsule)
The capsule preparations described in Preparation Examples 1 to 3 were subjected to a disintegration test described in the 16th revision Japanese Pharmacopoeia.

1 g of various capsule preparations prepared as described in Preparation Examples 1 to 3 were immersed in the assumed intragastric test solution (a) for 2 hours using a disintegration tester NT-60H (manufactured by Toyama Sangyo Co., Ltd.). Then, it was immersed for 2 hours in the assumed small intestine test solution (b) below.
(A) Presumed test solution in stomach: First solution (pH 1.2) (2.0 g of sodium chloride dissolved in 100 mL of water, 7.0 mL of 6M hydrochloric acid added, and made up to 1,000 mL)
(B) Presumed test solution in the small intestine: Second solution (pH 6.8) (250 mL of 0.2 mol / L potassium dihydrogen phosphate aqueous solution, 250 mL of 0.2 mol / L potassium dihydrogen phosphate aqueous solution was mixed and 1,000 mL) )

  Various capsules were immersed in the initial stage of the test, after immersion in the assumed test liquid in the stomach (a) (that is, after the first liquid treatment), and after immersion in the assumed test liquid in the stomach (a) and in the assumed small intestine test liquid (b) ( That is, after treatment with the second solution), homogenization was performed in 10 mL of 1% sodium ascorbate-added physiological saline, and the number of bacteria was measured using an MRS agar medium under anaerobic conditions. Table 2 shows the results of changes in the number of bacteria when the number of bacteria measured at the initial stage of the test for various capsules is taken as 100.

  As shown in Table 2, in any of the capsule preparations (seamless capsules, soft capsules, and hard capsules) described in Preparation Examples 1 to 3, almost none of the Bifidobacterium tested It was suggested that it was transported to the intestines.

(Example 3: Difference in recoverability of radioactive cesium due to life and death of Bifidobacterium)
Regarding cesium 137 (Cs-137) with known radioactivity, the recovery ability of each of live and dead bacteria of JCM1217, JBL01 and JBL05 was examined.

  For each of the viable bacteria of JCM1217, JBL01 and JBL05 that had been cultured on the MRS agar medium, colony 2 platinum ears were inoculated on the MRS agar medium and cultured at 37 ° C. for 48 hours under anaerobic conditions. Next, colonies that grew on the MRS agar medium were recovered while washing with phosphate buffered saline (PBS), and boiled at 100 ° C. for 10 minutes to inactivate the cells. After boiling, the cell suspension was centrifuged at 5,000 × g for 10 minutes at 4 ° C., PBS was added to the precipitate and suspended sufficiently, and further 10 ° C. at 5,000 × g at 4 ° C. Centrifugation was performed for a minute, and the precipitate was used as inactivated cells (dead cells). As live bacteria, cells grown on MRS agar medium (not boiled) were used.

  About 20 mg of viable cells and dead cells were added to brain heart infusion (BHI) agar medium (Becton, Dickinson and Company) to which 10% of bouillon prepared in the same manner as in Example 1 was added. The cells were cultured at 37 ° C. for 48 hours under anaerobic conditions.

  Then, each microbial cell was collect | recovered, wash | cleaning using PBS. The collected cell suspension was centrifuged at 5,000 × g for 10 minutes at 4 ° C. to separate the precipitate and the supernatant. Add distilled water to the precipitate and suspend sufficiently, then make up to 70 mL with distilled water, add 0.5 g of superabsorbent resin CP1 (Kennis Co., Ltd.), and disperse the bacterial cells in a U8 container. Was gelled to obtain a sample for measuring Cs-137 in microorganisms. On the other hand, the supernatant was combined with the medium after collecting the cells, and the volume was made up to 70 mL to obtain a sample for measuring the remaining medium Cs-137.

  The measurement of the radioactivity of Cs-137 remaining in the cells and in the medium, and the calculation of the radioactivity uptake rate of the microorganisms from the measured values of the remaining medium and in the microorganisms were carried out in the same manner as in Example 1 above. The results are shown in Table 3.

As shown in Table 3, for all Bifidobacterium species tested, radioactive cesium uptake was very low in dead bacteria, whereas radioactive cesium uptake (recovery ability) was seen in live bacteria. It was. This is because there is a process in which K ⁺ is taken into the cells in the Na ⁺ / K ⁺ pump of the cell membrane of the living cells by Na ⁺ / K ⁺ ATPase, and Cs ⁺ is also taken into the cells at the same time. ⁺ / K ⁺ ATPase activity causes Cs ^{+ to} be taken into the cells, but dead cells do not work because the ATPase is deactivated and Cs ⁺ is no longer taken into the cells. It is thought that was seen.

  From the results of Examples 2 and 3 as described above, it is suggested that by transferring live bacteria in the intestine, live Bifidobacterium takes the radioactive substance from the body and removes the radioactive substance from the living body of the animal. It was done.

  ADVANTAGE OF THE INVENTION According to this invention, the collection | recovery agent and collection | recovery method for collect | recovering the radioactive substance taken in in the living body effectively can be provided. According to such a recovery agent and recovery method, it is possible to remove the radioactive substance from the living body of the animal safely and effectively.

Claims (7)

  A radioactive substance recovery agent containing a Bifidobacterium microorganism as an active ingredient.   The microorganism belonging to the genus Bifidobacterium is Bifidobacterium addresscentis, Bifidobacterium angratam, Bifidobacterium animalis subspice animalis, Bifidobacterium animalis subspice lactis, Bifidobacterium・ Asteroides, Bifidobacterium bifidum, Bifidobacterium boum, Bifidobacterium breve, Bifidobacterium catenuratam, Bifidobacterium kerinum, Bifidobacterium coryneform, Bifidobacterium・ Kunikuri, Bifidobacterium denticorens, Bifidobacterium denticium, Bifidobacterium gallicum, Bifidobacterium galinarum, Bifidobacterium Globosum, Bifidobacterium indicum, Bifidobacterium infantis, Bifidobacterium inopinatum, Bifidobacterium lactis, Bifidobacterium longum, Bifidobacterium magnum, Bifidobacterium Phytobacterium merisicum, Bifidobacterium minimum, Bifidobacterium perbroram, Bifidobacterium pseudocatenuratam, Bifidobacterium pseudolongum subspice globosum, Bifidobacterium pseudoron Gum subspice pseudoron gum, Bifidobacterium prolum, Bifidobacterium luminous, Bifidobacterium luminous, Bifidobacterium sexual, Bifidobacte Um Squardbi, Bifidobacterium subtil, Bifidobacterium Switzerland, Bifidobacterium thermacid film, and Bifidobacterium thermum film, any one microorganism The radioactive substance recovery agent according to claim 1.   The radioactive substance recovery agent according to claim 1 or 2, wherein the Bifidobacterium microorganism is any one microorganism selected from the group consisting of Bifidobacterium longum strains JBL01 and JBL05.   The radioactive substance recovery agent according to any one of claims 1 to 3, wherein the Bifidobacterium microorganism is contained in a capsule.   The radioactive substance recovery agent according to claim 4, wherein the capsule is any one of a seamless capsule, a soft capsule, and a hard capsule.   The radioactive substance collection | recovery agent in any one of Claim 1 to 5 whose radioactive substance collect | recovered is any one or more of radioactive cesium, radioactive sodium, or radioactive strontium. A method for removing radioactive material from animals other than humans,
A method comprising the step of ingesting the radioactive substance recovery agent according to any one of claims 1 to 6 to the animal.