JP2006320658A - Artificial insemination method, frozen buffer, and artificial pollination method - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for raising the success rate of artificial insemination or artificial pollination, and to provide a frozen buffer. <P>SOLUTION: A deposit and a clear supernatant liquid are separated from the mixed liquid of the fine particles of fossil seashell being the fossil of chlamys islandica erythrocomata with water (104). The clear supernatant liquid is somatidcontaining water and mixed with semen containing sperm to be the mixed semen (106), and brought into contact with ova so as to perform fertilization (108). <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to an artificial insemination method, a freezing buffer, and an artificial pollination method using somatid, which is a supermicroorganism living in a living body such as a human body.

  As described in Non-patent Document 1, for example, somatides in the human body, mainly blood, as shown in FIG. 11, from the primitive phase somatid 1 to spores 2, double spores 3, and bacterial forms 4, double bacterial form 5, rod-like form 6, bacterial form 7 with double spores, bacterial form 8 with granular double spores, spherical bacterial forms 9, 10, bursting state 11, yeast form 12, offspring cyst It is said that a so-called somatoid cycle that returns to the first primitive phase somatid 1 after repeating the child form 13, the baby cub form 14, the mycelium form 15, and the mycelium rupture state 16 is repeated.

  When the environment is not good, the baby worm form 13 has a resistant mycelium form indicated by reference numerals 17A and 17B. Further, the mycelium form 15 is a fibrous leaf-like body indicated by reference numeral 18 when the environment is not good.

  Somatid is almost a primitive phase, especially in the blood of people with good health and high immunity. From such a state, for example, when a person takes a strong drug or drinks alcohol, the primitive phase somatid 1 enters the inside of platelets or red blood cells. In other words, as a result of the adverse effects on the environment for Somatid, it is considered that they are temporarily evacuated into platelets and red blood cells. Furthermore, it will be in the state after bacteria form 4.

  When the primitive phase somatid 1 is in the blood, it can freely enter any part of the living body. Specifically, it passes through the cell membrane to reach the internal mitochondria, where it reaches the mitochondria. Is believed to be triggering the citric acid cycle.

  In general, when the primitive phase somatide is present in a large amount in the blood, the immunity of the living body is considerably improved due to the relationship between the primitive phase somatide and the mitochondria as described above.

  However, as described above, due to slight changes in blood, somatid may show evacuation behavior, or in some cases, forms a shell itself and evacuates in large quantities inside it, so for example extracted from the natural world Even when a large amount of the primitive phase somatid is ingested, the original function of somatid cannot be satisfied due to slight changes in the environment.

  In the case of fertilization of animals and pollination of plants, the present inventors are concerned with the somatide as described above, and when the number of somatides contained in this semen or pollen is small or the activity is low We found that the success rate of fertilization and pollination decreases.

Completely healed by Gaston Nessan (Publisher: Tokuma Shoten)

  The present inventor found that ancient somatids exist in shell fossils from 2000 to 25 million years ago, and as a result of the research, the ancient somatids are resistant to environmental changes in the living body, and the blood that humans receive in daily life It was found that evacuation behavior was not exhibited by environmental changes such as alcohol consumption and drug intake.

  In addition, the present inventors have found that the success rate of fertilization or pollination increases when ancient somatide is mixed with semen for artificial insemination, or mixed with pollen for artificial pollination.

  In addition, in the case of artificial insemination of livestock, for example, cattle, sperm collected in advance and frozen with liquid nitrogen is injected into the cow, but ancient somatid is used as a freezing buffer for freezing this sperm. It has been found that the success rate of fertilization increases when mixed.

  As a result of diligent research by the present inventors, the present invention uses an ancient somatid extracted from a shell fossil to increase the success rate of artificial insemination and artificial pollination, and in artificial insemination using frozen sperm, We have found a freezing buffer that can increase the success rate of fertilization.

  That is, the above object can be achieved by the following artificial insemination method, frozen buffer, and artificial pollination method.

  (1) An artificial insemination method for animals characterized in that ancient somatid is mixed with semen containing sperm to make mixed semen, and this mixed semen is fertilized by contacting with an egg.

  (2) The method for artificial insemination of animals according to (1), wherein the semen is frozen together with a freezing buffer containing ancient somatide and thawed at the time of insemination.

  (3) The ancient somatid has a fossil shellfish fossil shell fossil inhabited 2000 to 25 million years ago with a particle size of 0.1 to 5 μm, and a ratio of at least 5 cc to 1 g of the fine shellfish fossil. Mixing with water, holding the liquid mixture of the fine particles and water for at least 4 hours, and then containing ancient somatid-containing water consisting of the supernatant separated from the precipitate mainly composed of calcium carbonate, or powdered form extracted therefrom The method for artificial insemination of animals according to (1) or (2), characterized by comprising the ancient somatid.

  (4) To the ancient somatid-containing water, weak acidic water containing at least one kind of weak acid that is harmless to the human body, including acetic acid, dilute hydrochloric acid, citric acid, and malic acid, is added, and the ancient somatid water is converted to PH7.5. The method for artificial insemination of animals according to any one of (1) to (3), characterized in that a weak alkali is used.

  (5) Shell fossil that is a fossil of Kamionisiki shellfish inhabited 2000 to 25 million years ago is made 0.1 to 5 μm in diameter, and mixed with water at a ratio of at least 5 cc per 1 g of this fine shell fossil, Sperm containing an ancient somatid-containing water consisting of a supernatant liquid separated from a precipitate containing calcium carbonate as a main component after holding the mixed liquid of fine particles and water for at least 4 hours, or a powdered somatid extracted therefrom Frozen buffer for cryopreservation.

  (6) To the ancient somatid-containing water, weak acidic water containing at least one kind of weak acid harmless to human body, including acetic acid, dilute hydrochloric acid, citric acid and malic acid, is added, and the ancient somatid water is converted to PH7.5. The frozen buffer for sperm cryopreservation according to (5), characterized by having a weak alkali to the extent.

  (7) Shell fossil which is a fossil of Kamionisiki shellfish inhabited 2000 to 25 million years ago is made into a particle size of 0.1 to 5 μm, and mixed with water at a ratio of at least 5 cc to 1 g of the fine shellfish fossil, After holding the liquid mixture of the fine particles and water for at least 4 hours, water containing ancient somatid consisting of a supernatant separated from a precipitate mainly composed of calcium carbonate, or powdered ancient somatid extracted therefrom, A method for artificially pollinating a plant, comprising applying pollen to the pistil before, after or at the same time, between the root and the tip of the pistil.

  (8) A fossil shellfish fossil that has been inhabited 2000 to 25 million years ago has a particle diameter of 0.1 to 5 μm, and is mixed with water at a ratio of at least 5 cc to 1 g of the fine shellfish fossil, After holding the liquid mixture of the fine particles and water for at least 4 hours, water containing ancient somatid consisting of a supernatant separated from a precipitate mainly composed of calcium carbonate, or powdered ancient somatid extracted therefrom, The artificial pollination method for plants according to (7), wherein the method is mixed with an artificial pollination solution containing pollen and applied to a pistil.

  According to this invention, the success rate of fertilization can be increased by mixing ancient somatid in the semen during artificial insemination. In addition, if an ancient somatid is included in the freezing buffer for storing sperm in a frozen state, the success rate of fertilization can be increased. Furthermore, the success rate of artificial pollination can be increased by mixing ancient somatide with an artificial pollination solution containing pollen to cause pollination.

  The method of artificial insemination of animals according to the best mode is to mix fine particles of shell fossils, which are fossil of Kamionisiki shellfish, and water, for example, negative ion water, to separate into a precipitate and a supernatant. The above-mentioned purpose is achieved by mixing the liquid as ancient somatid-containing water with the semen during artificial insemination and bringing it into contact with the ovum.

  Further, the above object is achieved by mixing the above-mentioned ancient somatid-containing water or the powdered ancient somatid extracted into the artificial pollination solution containing pollen and applying it to the pistil. Furthermore, the said objective is achieved by mixing the ancient somatide containing water and comprising the frozen buffer for sperm cryopreservation.

  Next, an artificial insemination method according to Embodiment 1 of the present invention will be described with reference to FIG. 1 (flow chart).

  In step 101, the fossil of Kamioniki shellfish (hereinafter referred to as “shellfish fossil”) inhabited about 2000 to 25 million years ago is processed into fine particles having a particle diameter of 0.1 to 5 μm.

  Next, in step 102, it is mixed with water at a ratio of at least 5 cc with respect to 1 g of the fine shell fossil.

  In step 103, the mixed liquid of the fine particles and water is held in, for example, a glass container for at least 4 hours.

  In the next step 104, the precipitate mainly composed of calcium carbonate and the supernatant liquid are separated from the mixed liquid after at least 4 hours by, for example, a filter, and put in a container in step 105, and the supernatant liquid is separated. Let the liquid be ancient somatid-containing water.

  This ancient somatid-containing water contains the primitive somatid of the primitive phase, and can evaporate the water, for example, by heating or leaving it. There is a risk of changing to other forms other than somatid. Therefore, the ancient somatid should be retained in the ancient somatid-containing water.

  The present inventor is a bivalve that resembles a modern scallop shell, and the shell fossil shell is made using a shell substrate and is in a dormant state. It was found by.

  This ancient somatid was active in the flesh of the Kamioniki shellfish before it became a fossil, and when the symbiotic Kamioniki shellfish died, the shell portion was searched for a place to evacuate and sleep. It is estimated to be.

  As described above, the meaning of making the fossil shellfish fine particles having a particle size of 0.1 to 5 μm means that if the particle size exceeds 5 μm, a plurality of somatids may expose the surface of the shell inside. Of course, some shell surfaces appear on the surface of the fine particles even at 6 μm or more, but in this case, the yield of collecting ancient somatids from the shells is greatly reduced. . When the particle size of the fine particles is 5 μm, the shell in which the somatid is dormant can be exposed to the surface of the fine particles almost certainly, so that the recovery rate is high.

  Moreover, the reason for making the shell fossil fine particles having a particle size of 0.1 μm or more is that the pulverized fine particle operation of 0.1 μ or less of the shell fossils mainly composed of calcium carbonate is very expensive, In addition, the particle size cannot be reduced unless a special apparatus is used.

  The process of holding the mixture of fine particles and water for at least the fourth order is ideally 24 hours, but if held for 4 hours, the ancient somatid of the primitive phase appears in the shell with a considerable yield. Because.

  In Step 106, for example, the ancient somatid-containing water is mixed with semen containing sperm collected from a bull to obtain mixed semen.

  In Step 107, for example, at least one of a weak acid that is harmless to a living body or sperm including acetic acid, dilute hydrochloric acid, citric acid, malic acid is mixed with the mixed semen, and the pH is weakly alkaline (about 7.5). Adjust to.

  This is because shellfish fossil is mainly composed of calcium carbonate, but for a long time, it may be denatured into calcium oxide (CaO) by heat or pressure. In this case, when CaO is brought into contact with water, strong alkalinity is obtained. As shown, it is necessary to adjust to weak alkalinity optimal for sperm survival.

  In step 108, as described above, the mixed semen adjusted in pH is brought into contact with the ovum and fertilized. In step 106, the pH may be adjusted by adding a weak acid to the ancient somatide-containing water before making the mixed semen. In this way, sperm damage can be prevented when the ancient somatid-containing water is strong alkali.

  Here, ideally, the water mixed with the fine particles is preferably negative ion water. The negative ion water may have a negative ion number of about 80000 to 150,000 in 1 cc.

  In the case of negative ion water, electrons contained in a large amount in the negative ion water serve as food for the ancient Somatid, so that the negative ion water is a good environment for the ancient Somatid. For this reason, ancient somatids easily break into shells and jump out into negative ion water.

  Further, the negative ion water is specifically prepared by bringing a fossil containing silicon as a main component, called black silica, into particles having a particle diameter of 5 to 10 μm and contacting the particles with water for at least 2 hours. It is good to form. Since the black silica fine particles emit a large amount of electrons, the water in contact with the fine particles quickly becomes negative ion water.

  The negative ion water may be formed by using, for example, a commercially available alkaline ion water conditioner. However, according to the experiment by the present inventors, the negative ion water formed by the particles of black silica is the most. Gave good results.

  In addition, at least one of weak acids that are harmless to the human body, including acetic acid, dilute hydrochloric acid, citric acid, and malic acid, may be dissolved in water mixed with the fine particles of the fossil shell.

  According to the inventor's experiment, mixing these weak acid-dissolved water and shell fossil microparticles, but not as much as negative ion water with black silica, can efficiently collect ancient somatids of the primitive phase. did it.

  Next, an artificial insemination method for animals according to Example 2 of the present invention will be described with reference to FIG.

  In step 201, the artificial insemination method mixes ancient somatide-containing water with a freezing buffer to form an ancient somatid-containing freezing buffer, and in the next step 202, the ancient somatid-containing freezing buffer is combined with the semen. In step 203, the frozen semen is thawed and the process proceeds to step 108 in FIG.

  Here, the frozen buffer is for preserving sperm in a frozen state and maintaining high motility and survival rate of the sperm after thawing, and is mainly composed of glycerin, glucose or the like. .

  Thus, if semen is frozen together with a freezing buffer containing an ancient somatid in advance, the fertilization rate can be improved by the activity of the ancient somatid when thawed and brought into contact with the ovum.

  Next, an artificial pollination method according to Example 3 of the present invention will be described with reference to FIG.

  In step 301, pollen in a state where pollination is possible is collected. At this time, for example, using a vacuum inhaler, the pollen is inhaled in a dry state and separated from the air by a filter and collected.

  Next, in step 302, the ancient somatid-containing water formed as described above is applied or dripped onto the pistil root.

  In the next step 303, the collected pollen is attached to the pistil. This attachment work is made to adhere to the pistil with a fluffy application member (not shown).

  The application or dripping of the ancient somatid-containing water may be after the pollen is attached to the pistil, or may be at a location other than the root of the pistil.

  As described above, when somatid-containing water is applied or dripped onto the pistil root or other locations during pollination, ancient somatid in the somatoid-containing water enters inside through the cell wall of the pistil root and enters the seed part. Reach easily. Here, the pollen also reaches and pollinates.

  The ancient somatid that reaches the seed part promotes the synthesis of the protein here, and this protein is converted into an enzyme for germination.

  In general, plants do not contain protein except for seeds and fruit parts, but these somatides are involved in seeds and fruit parts, and proteins that become enzymes for germination are synthesized. Therefore, if the seed contains a large amount of somatide, the vitality of the seed becomes stronger.

  In addition, although the ancient somatid containing water is used in the said Example, this invention is not limited to this, You may use the powder which contains somatid in large quantities from ancient somatid containing water. When using a powdered ancient somatid, the powdered ancient somatid is attached to the pistil simultaneously with or before the pollen.

  In order to form a powdery ancient somatid from the ancient somatid-containing water, the following procedure may be used.

  First, as shown in FIG. 4, in step 401, the ancient somatide-containing water is dried to form a crystalline powder, and in the next step 402, this crystalline powder is put into distilled water. Glucose and carboxylic acid are added to distilled water containing powder and filtered, and in step 405, a crystalline powder containing somatide is obtained.

[Example]
The present inventor contacted shell fossil fine particles with negative ion water to form ancient somatid-containing water from the supernatant, which was observed with a phase contrast microscope.

  The observation results are shown in FIGS.

  As shown in these figures, the ancient somatid 30 is a primitive phase and has a size of about 150 nm. In any of the states of FIG. 5 to FIG. 6 and FIG. 6 to FIG. It was taken at an interval of about 2 seconds, and it can be seen that the ancient Somacid in the primitive phase is actively moving. In this case, according to the observation by the present inventor, the moving speed is about 10 μm / second in FIGS. 5 to 6 and about 1 μm / second in FIGS. 6 to 7. In addition, the circumference | surroundings of ancient somatid 30 are aggregates of particulate calcium carbonate.

  In addition, the present inventors added the precipitate such as calcium carbonate to water containing ancient somatid that has been confirmed to be a primitive phase, and injected 5 cc of 100% ethyl alcohol with respect to 1 cc of water. However, the ancient Somacid of the primitive phase never stopped its movement. On the other hand, when a similar test was performed on somatid collected from the blood of modern humans, modern somatid stopped its movement at all.

  8 to 10 are photographs of the ancient somatid 32 in the primitive phase with a 2 μm diameter shell taken with a phase contrast microscope at intervals of 2 seconds, from which the ancient somatid moves at 5 μm / sec. You can see that

The flowchart which shows the artificial insemination method which concerns on Example 1 of this invention. Flowchart showing an artificial insemination method according to the second embodiment The flowchart which shows the artificial pollination method which concerns on Example 3. Flow chart illustrating a method for forming an ancient somatid in crystalline form from water containing ancient somatid Photograph showing the observation results of primordial phase ancient somatids by phase contrast microscope A photo of the somacid 2 seconds after the photo A photograph of the somacid two seconds later Photograph showing the results of observation of the ancient somatid with shells by phase contrast microscopy A photo of the somacid 2 seconds after the photo A photograph of the somacid two seconds later Plan view schematically showing the somatid cycle

Claims (8)

  An artificial insemination method for animals characterized in that ancient somatide is mixed with semen containing sperm to make mixed semen, and this mixed semen is fertilized by contacting with an egg. In claim 1,
An artificial insemination method for animals, wherein the semen is frozen together with a freezing buffer containing ancient somatide and thawed at the time of insemination. In claim 1 or 2,
The ancient somatid has a fossil shellfish fossil shell fossil habitat 2000-25 million years ago with a particle size of 0.1-5 μm and is mixed with water at a rate of at least 5 cc per 1 g of the fine shell fossil. And holding the mixed liquid of fine particles and water for at least 4 hours and then containing ancient somatid-containing water consisting of a supernatant separated from a precipitate mainly composed of calcium carbonate, or powdered ancient somatid extracted therefrom An artificial insemination method for animals characterized by comprising:   In any one of Claims 1 thru | or 3, the weak acid water which melt | dissolved at least 1 type of the weak acid harmless to a human body containing acetic acid, dilute hydrochloric acid, a citric acid, malic acid is added to the said ancient somatid containing water, An artificial insemination method for animals, characterized in that ancient somatid water is weakly alkaline with a pH of about 7.5.   A fossil shellfish fossil that has been inhabited 2000 to 25 million years ago has a particle diameter of 0.1 to 5 μm, and is mixed with water at a ratio of at least 5 cc to 1 g of the fine shellfish fossil. Preserving sperm for free storage containing water containing ancient somatid consisting of a supernatant separated from a precipitate composed mainly of calcium carbonate after holding a water mixture for at least 4 hours Freezing buffer. In claim 5,
To the ancient somatid-containing water, weak acidic water containing at least one kind of weak acid harmless to human body, including acetic acid, dilute hydrochloric acid, citric acid and malic acid is added, and the ancient somatid water is weakened to about PH7.5. A refrigeration buffer for sperm freezing storage, characterized by alkali.   A fossil shellfish fossil that has been inhabited 2000 to 25 million years ago has a particle diameter of 0.1 to 5 μm, and is mixed with water at a ratio of at least 5 cc to 1 g of the fine shellfish fossil. After holding the water mixture for at least 4 hours, the ancient somatid-containing water consisting of the supernatant separated from the precipitate composed mainly of calcium carbonate, or the powdered ancient somatid extracted from this, is used to pistil the pollen. A method for artificial pollination of plants, characterized in that it is applied to any of the pistil roots and tips before, after or simultaneously.   A fossil shellfish fossil that has been inhabited 2000 to 25 million years ago has a particle diameter of 0.1 to 5 μm, and is mixed with water at a ratio of at least 5 cc to 1 g of the fine shellfish fossil. After holding the mixed liquid of water for at least 4 hours, the ancient somatid-containing water composed of the supernatant separated from the precipitate mainly composed of calcium carbonate, or the powdered ancient somatid containing pollen An artificial pollination method for plants, which is mixed with an artificial pollination solution and applied to a pistil.