RU2062068C1 - Medium for diluting and preserving boar sperm - Google Patents

Abstract

FIELD: veterinary medicine. SUBSTANCE: medium has the following ingredients in 1000 ml of distilled water, in g: glucose - 53.0, trilon B - 3.5, sodium bicarbonate - 1.0, sodium citrate - 3.7, sodium salt of ampicyllin - 0.35-0.7, gentamycin sulfate - 0.35-0.2, polymixin sulfate - 0.025, levamysol hydrochloride or camysol - 0.06-0.15. EFFECT: prolonged storage time; improved biological properties. 2 tbl

Description

 The invention relates to biology, mainly to a zootechnical method of reproduction of pigs, and can be widely used in breeding enterprises, pig farms and other farms of pig production for storing germ cells of sperm outside the body in special synthetic media during artificial insemination of pigs.

A number of boar semen diluents are known: glucose-chelate-citrate (A.S. N 183901, 1964); glucose-chelate-citrate-sulfate (GOST -17637 72); "Environment for dilution and storage of boar semen" (a.s. 1466044.1986), which we developed and registered with the USSR State Committee for Inventions and Discoveries under the trademark "BIOKONSAN" (p. N 82839, 1987),
Inclusion for the first time by us in 1961-1963. the composition of Trilon B diluent (chelaton, selecton, complexon III, ethylenediaminetetraacetate -Na ₂ EDTA) has significantly improved the quality and effectiveness of the medium and, most importantly, stored (preserved) sperm at room temperature for 2-3 days without reducing the fertilizing ability of sperm (A.S. 183901, 1964).

 The situation in the world with diluents for storing boar semen is described in the recently published book by A. Gordon, "Control of the reproduction of farm animals." Translation from English. Moscow VO "Agropromizdat", 1988, p.290. In this book, it is noted that of the diluents used in modern production, the most widely known is a diluent based on ethylenediaminetetraacetate. This diluent is called "Kiev" or the diluent "Merka" and "Varoma", the drug is convenient for ease of preparation and the ability to use semen diluted by it for up to 3 days without reducing fertilization ability and future fertility. And further it is noted: "since 1970, the Kiev thinner replaced the Illinois one and began to use it mainly in Great Britain and other countries of Western Europe." The diluent developed by us back in the 60s of the MCC is called by the author of the book “Kiev”, “Merka” or “Varoma”, i.e. by the name of the companies producing this medium, for example, in Germany, the company "Merck".

 The aim of this invention is to extend the shelf life, increase the biological usefulness and fertilizing ability of spermatozoa, increase the fecundity of sows, as well as increase the protective ability of sows and born piglets with more equal and higher live weight.

 To achieve this goal in a known environment by ed. St. N 1466044 is additionally administered with natural antibiotics and immunomodulators, for example Levamisole or Camisole.

 It is known that the sanitizing effect (neutralization) depends on the dose of the antibiotic; the higher the dose, the higher the effect. However, increasing the dose leads not only to an increase in the antimicrobial effect of the drug, but also to an increase in its toxicity (3). Different animals have different species sensitivity of germ cells to drugs. For example, the toxicity of streptomycin and a combined mixture called spermosan -3 in doses of 75-100 thousand units is not visible on bull sperm. while for sperm of the boar and stallion, the harmless dose of a mixture of these preparations is only 30 thousand units / 100 ml. An increase in the dose of sanitizing drugs reduces sperm survival by 20%, and the fertility of animals decreases markedly (3). Therefore, the amount of penicillin and streptomycin (including spermosan) in media, according to the instructions and GOST, is reduced from 400-500 thousand units. up to 250-300 thousand units per 1000 ml of medium. However, at reduced doses of these drugs, along with a decrease in toxicity, the sanitizing effect also decreases.

 In this regard, we tested the combination of a semi-synthetic antibiotic (ampicillin sodium salt) with natural antibiotics (gentamicin sulfate, kanamycin sulfate, sigmamycin, polymyxin sulfate). Such combinations can reduce the amount of each of the antibiotics in the diluent without compromising the quality of the synthetic medium. With this combination of antibiotics, sperm storage time has increased.

Of the known media used for dilution and storage of boar semen, the composition of the ingredients closest to the claimed medium is the medium according to Auth. N 1466044 (1986), which we have chosen as a prototype:
Glucose, g 55.0 (55-60)
Trilon B, g 3.5 (2.45-3.5)
Sodium bicarbonate, g 1.0 (0.8-1.0)
Sodium citrate, g 3.7 (1.85-3.7)
Ampicillin sodium salt, g 0.7 (0.5-5.0)
Distilled water, ml 1000
The results of the influence of media (according to examples 1,2) with semisynthetic and natural antibiotics on sperm survival are shown in Table 1.

Example (recipe) 1. The medium is characterized by the following composition per 1000 ml of distilled or bidistilled water, g:
Glucose 55.0
Trilon B 3.5
Sodium bicarbonate 1.0
Sodium Citrate 3.7
Ampicillin sodium salt 0.35
Gentamicin Sulfate 0.35
Polymyxin sulfate 0.025 (250,000 units)
Example (recipe) 2. The medium is characterized by the following composition per 1000 ml of distilled or bidistilled water, g:
Glucose 53.0
Trilon B 3.5
Sodium bicarbonate 1.0
Sodium Citrate 3.7
Ampicillin sodium salt 0.7
Gentamicin Sulfate 0.2
Polymyxin Sulfate 0.025
In the second series of laboratory studies, we first studied the effect of immunomodulators (levamisole, camizole) included in the diluent on sperm survival with the aim of subsequent use of this medium for dilution and storage of sperm according to example (recipe) 3.

Example (recipe) 3. The medium is characterized by the following composition per 1000 ml of distilled or bidistilled water, g:
Glucose 55.0
Trilon B 3.5
Sodium bicarbonate 1.0
Sodium Citrate 3.7
Ampicillin sodium salt 0.35-0.7
Gentamicin sulfate 0.35-0.20
Polymyxin sulfate 0.025 (250,000 units)
Immunomodulator, e.g. levamisole or camizole, 0.06-0.15
As you know, levamisole (decaris, G. Richter, Hungary), tetramisole, camisole or other drugs of this series are usually recommended as an anthelmintic or in infectious and malignant diseases, as well as in immunodeficiency. These drugs are used both in medicine and veterinary medicine for therapeutic purposes, appointing them inside or intramuscularly. The use of drugs of this group in synthetic media intended for dilution and storage of sperm is unknown to us.

 In laboratory experiments, we set a goal to study the optimal doses of the immunomodulator in sperm storage media and its effect on the metabolic processes of germ cells, and in production experiments by introducing diluted sperm with immunomodulator additives into the uterine horns we were interested in the possibility of absorption of the latter through the mucous membrane of the genital tract into the body the animal and its effect on fertility, fertility and resistance of sows and offspring.

 The results of the second series of laboratory tests are also shown in table 1 (for example, recipe 3).

 From table 1 it is seen that the new environment (including from the immunomodulator) contributed to an increase in the survival time of sperm at the level of 60% or more by about two days.

 After establishing optimal doses of antibiotics and immunomodulators in the medium and obtaining positive results in laboratory experiments, scientific and production experiments were conducted. An immunomodulator, for example, levamisole or camisole (example 3), was added to the media (recipes 1 and 2). As a control, a known medium (prototype) was taken without the addition of an immunomodulator.

 Due to the fact that we planned, upon receiving positive results in this experiment, we did not disclose the recipe to the State Committee for Inventions on the subject of invention and the grant of a patent, i.e. no ingredients were announced in the media. Samples of media in plastic bags were prepared in the laboratory and delivered to the state farm plant. The well-known medium called "Bioconsan" was a control, and a new medium with the addition of an immunomodulator (levavisol or kamizol) was delivered to the farm under the conditional name "Bioconsan -2" (as in Example 3).

 1346 sows were inseminated with semen diluted with these media (673 animals in the control and experimental groups).

 As the data in Table 2 show, in the experimental group, sow fertility was 3.6% higher, and fertility by 1.1 pigs per farrow more than in the control group, in which sperm diluted with a known medium was used. It should be noted that the number of well-developed piglets in the litter with a live weight of at least 1000 g was 0.9 piglets more. TTT1 TTT2

Claims (1)

1. A medium for diluting and storing boar semen, including glucose, sodium bicarbonate, sodium citrate, antibiotics and distilled water, characterized in that it further comprises Trilon B and an immunomodulator, the antibiotic containing ampicillin sodium salt, gentamicin sulfate and polymyxin sulfate in the following ratio of components per 1000 ml of distilled water, g:
Glucose 55.0
Trilon B 3.5
Sodium bicarbonate 1.0
Sodium Citrate 3.7
Ampicillin sodium salt 0.35-0.70
Gentamicin sulfate 0.35-0.20
Polymyxin Sulfate 0.025
Immunomodulator 0.06-0.15
2. The medium according to claim 1, characterized in that as an immunomodulator contains levamisole or camizole.

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