JPS6236483B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to an agent for preventing infectious diseases in piglets. 45-70% of 2-3 week old suckling pigs in Japan
The pigs are affected by what is called white diarrhea, and a considerable proportion of the affected pigs die. Therefore, the economic loss is extremely large for the pig farming industry. This trend is similar in developed countries, and there are countless reports on prevention and treatment. These reports and the measures currently being taken include administering antibacterial agents such as various antibiotics and sulfur drugs, and some E. coli vaccines have also been developed, but their effectiveness varies and is not sufficient. I couldn't say yes. The present inventors conducted various studies with the aim of obtaining drugs for such diseases in piglets. As a result, it was thought that these diseases were mainly caused by the immunodeficiency of piglets immediately after birth, and this was prevented by first administering γ-globulin. (Special Publication No. 46-43899) By administering γ-globulin preparations, Escherichia coli,
It was found to be effective in preventing infectious diseases such as Japanese encephalitis and swine fever. However, it has not yet been sufficiently effective against white diarrhea, which is the most common disease. Therefore, the present inventors continued their research on enhancing the immunity of piglets, and as a result, when they administered a transfer factor obtained by separating white blood cells from the blood of adult pigs to piglets immediately after birth, it was found that Contrary to the common knowledge that transfer factors are ineffective, the present invention was completed by discovering that they surprisingly strengthen the immunity of piglets and non-specifically prevent various infectious diseases. The present invention is an agent for preventing infectious diseases of piglets, which contains as an active ingredient a transfer factor isolated and collected from white blood cells in the blood of adult pigs. The transfer factor, which is an active ingredient of the present invention, is obtained by collecting blood from an adult pig, preferably a large number of adult pigs, separating leukocytes from the blood, and subjecting the blood to a freeze-drying thawing method or a DNase treatment before or after freeze-drying and thawing. Separated and collected. White blood cells can be separated using commonly used methods. For example, pig blood is placed in a container containing an anticoagulant and saline, and white blood cells are separated by the dextran method, hemolysis method, or the like. The dextran method collects the upper layer of blood that contains many white blood cells, adds dextran saline solution to this, precipitates the red blood cells, collects the upper layer that contains many white blood cells, and centrifuges it to extract only the white blood cells. collect. In the hemolytic method, blood is collected at room temperature, red blood cells are precipitated, the plasma portion of the supernatant is collected, centrifuged, the supernatant is removed, and distilled water is poured over the precipitate, followed by a coagulant and saline solution while stirring. Add, centrifuge,
The precipitate is collected and this operation is repeated to collect pure white blood cells. Hemolysis is more suitable for obtaining the active ingredient of the present invention. The method for separating and collecting transfer factors from the leukocytes obtained as described above is the commonly used freeze-thaw method and the freeze-thaw method.
A method with ace processing is adopted. for example,
The leukocytes obtained as described above are frozen and thawed 6 to 12 times using methanol-added dry ice and a 37° C. constant temperature bath to completely destroy the leukocytes. This is dialyzed against distilled water, and the extra dialysate is freeze-dried to obtain a transfer factor. Alternatively, DN Ace and magnesium sulfate may be added to the frozen and thawed product and reacted at around 37°C for about 30 minutes, followed by subsequent dialysis and freeze-drying.
This DN Ace treatment may be performed before freezing and thawing operations. The transfer factor, which is the active ingredient of the present invention, obtained as described above is derived from 1 x 10 ⁹ white blood cells, and one unit of transfer factor is 1 unit of transfer factor, and the amount of polypeptide in one unit of transfer factor is 11.5 mg. , the amount of ribose is approximately 500μ
g, ion concentration is sodium 3.7mg, potassium
0.34mg, calcium 8μg, magnesium 1.1mg
The absorption ratio OD ₂₆₀ /OD ₂₈₀ is approximately 1.8. The agent for preventing infectious diseases for piglets of the present invention is prepared by dissolving the transfer factor in physiological saline.
This preparation is mainly administered to piglets of 3 weeks of age from immediately after birth, and the dosage is approximately 0.5 to 5 transfer factor units per injection subcutaneously or intramuscularly. Injecting the pigs several times a week will strengthen the piglets' immunity against various infectious diseases, improve their constitution, and produce healthy, well-developed pigs. Furthermore, when simultaneously administering the preparation of the present invention, in combination with gamma globulin preparations (preparations described in JP-A-46-43899 and JP-A-48-35021) in the serum collected from adult pigs in many pig pens, , it can strengthen immunity and completely prevent most infectious diseases in piglets shortly after birth. This prevents the conventional death of piglets immediately after birth, provides healthy and well-developed piglets, and is extremely useful in the pig farming industry. Next, an example of a method for collecting a pig transfer factor, which is the main component of the piglet infectious disease preventive agent of the present invention, will be described. Example Blood 25 collected from an adult pig was left at room temperature for about 1 to 2 hours, and when red blood cells were precipitated and white blood cells were in suspension, the plasma portion of the supernatant was collected and centrifuged at 2000 rpm for 7 minutes. After removing this supernatant, quickly pour distilled water into the precipitate, and while stirring, after 25 seconds, 0.5%
A 3.4% saline solution containing EDTA (ethylenediamine tartrate 2-sodium salt) was added, centrifuged at 1000 rpm for 5 minutes, and the precipitate was collected. The same operation was repeated 2 to 3 times to collect 77 x 10 ⁹ pure white blood cells. The leukocytes thus obtained were frozen and thawed for 12 hours using methanol-added dry ice and a 37°C thermostat.
Repeatedly, completely destroying white blood cells. Then
Add 0.2 mg of DNase per ¹⁰⁹ white blood cells and 0.2 ml of 10% magnesium sulfate solution, and let stand at 37°C for 30 minutes.
Dialyze the external solution with 50 times the volume of distilled water at 4°C with stirring for 48 hours. The external dialysis fluid is once freeze-dried.
637 mg of dry transfer factor is obtained. This was re-thawed in distilled water to give ¹⁰⁹ white blood cells/0.5ml, and dispensed into vials in 0.5ml portions while sterilizing with a 0.22μ Millipore filter.
Lyophilization is performed again to obtain a dry product of the transfer factor. One vial of the transfer factor obtained as described above is dissolved in an appropriate amount of physiological saline to obtain the drug of the present invention. Next, test results of the preventive effect on infectious diseases in piglets using the drug of the present invention will be shown. Test Example Eight piglets born from six sows were divided into four groups immediately after birth, with one group serving as a test group and the other group serving as a control group. Then, a preparation containing 1 unit of the transfer factor of the present invention was subcutaneously injected into the test group on the first day of birth, 1/2 unit on the third day of age, and 1/2 unit on the fifth day of age. The control group was kept under the same conditions without administering the transfer factor, and each animal was observed accurately by three viewers from birth to 28 days old. a) Loose stools lasting more than 3 days, (b)
Diarrhea was considered positive if watery fluid persisted for 2 days or more, and (c) shigellosis was observed for 1 day or more. Furthermore, no side effects were observed even when the drug of the present invention was administered. The test results were as shown in the table below.

【table】

[Table] In the fraction of the number of animals with diarrhea in this table, the denominator is the number of animals tested and the numerator is the number of animals with diarrhea. As is clear from the above test results, administration of the drug of the present invention significantly reduced the incidence of diarrhea in piglets.

Claims (1)

[Scope of Claims] 1. An agent for preventing infectious diseases of piglets, which contains as an active ingredient a transfer factor isolated and collected from white blood cells in the blood of adult pigs. 2. The agent for preventing infectious diseases in piglets according to claim 1, wherein the leukocytes are leukocytes in the blood of a large number of adult pigs.