GB2079149A - A method of producing a treatment preparation for improving the fertility of mammals - Google Patents

Abstract

The invention relates to a new method of producing a treatment preparation for improving the fertility of mammals. The method involves using balancing agents containing substance(s), particularly enzyme activator(s) and/or substrate(s), present in the genital discharge in the period of oestrus of fertile female animals but partially or totally missing from the genital discharge in the period of oestrus of female animals showing signs of sterility and husbanded in the same geographical region, in amounts required to compensate the deficiency and/or containing substance(s) in amounts sufficient to inactivate (bind or neutralize) component(s), particularly calcium and/or magnesium ions, of the latter discharge present in excess as compared with the former one. The invention enables one to readjust the physiological conditions of the genital discharge of virtually sterile females, thereby providing favourable conditions for successful insemination.

Description

SPECIFICATION A method of producing a treatment preparation for improving the fertility of mammals This invention relates to a method of improving the fertility of mammals, by the production of a treatment preparation.

In the reproduction of mammals the females show well distinguishable signs of oestrus, which lasts for a shorter or longer period, depending on the species. Within this period females are ready to accept males, and this period is suitable for the insemination of the mature egg cells.

Although the females, with only a few exceptions, show the signs of oestrus at regular intervals, a considerable portion of females does not become pregnant in the life periods suitable for self reproduction, or the number of pregnancies is much lower than would in principle be possible.

The situation is more favourable with farm animals, particularly in large-scale farming, where planned and organised insemination of female animals frequently do not become pregnant. In this respect the practice of the last decades, replacing natural insemination almost exclusively by artificial insemination in breeding, has brought about considerable progress, leading to a significant percentage increase in the number of offsprings.

It still occurs, however, that some of the female animals do not become pregnant, although they show the normal signs of oestrus. This means that after the artificial insemination and the oestrus cycle period the animal starts rutting again. This phenomenon is termed in animal husbandry as repeat breeder.

These repeat breeder females were treated so far with various veterinary medicines and other compositions which influence pregnancy. The compositions applied so far can be classified essentially into three groups, on the basis of the observed or assumed pathologic conditions of the females to be treated.

The first group encompasses antiphlogistic and bactericidal preparations; the compositions belonging to the second group are to eliminate the harmful effects arising from the facilities of animal husbandry or foddering, whereas those of the third group are to normalize the abnormal ovarian functions.

When the conceptional impotence of the animals was attributed to a catarrh or an inflammatory disease of the urogenital tract, generally oxidizing agents or, more recently, antiseptics were applied. Of the known compositions belonging to this group e.g. the Mez6 solution, containing boric acid and potassium permanganate, furthermore various iodine preparations, combined most frequently with acidic agents (such as hydrochloric, boric or citric acid) to shift the pH value to the acidic region, are to be mentioned. Various combinations of antibiotics with sulfonamides are also known, which are utilized primarily in the treatment of infectious diseases.

In those instances where sterility, more particularly the so-called symptom-free sterility, is not bacterial in origin, but can be attributed to the technical conditions of animal husbandry (e.g. foddering or the conditions and milieu of large-scale farming), the animals are generally treated with substances which compensate for the deficiency in proteins and vitamins. It has been observed that the conditions of impregnation can be improved by applying uterine irrigating liquids which contain vitamins A, B, D and E and/or proteins (such as skimmed cow's milk, egg-yolk, etc.).

Intravenous hormone treatment is applied most frequently to normalize deficient ovarian functions. This method can be applied, however, only after a professional examination of the ovary and the uterine and a careful consideration of the clinical test results, requiring specially trained veterinarians for the treatment.

These methods of treatment proved to be, however, ineffective in a considerable number of the cases, and the required pregnancy could not be attained despite of the fact that the females had no catarrheal disease, their ovarian functions appeared to be normal, and the disturbances in physiological equilibrium due to the conditions of husbandry were counterbalanced by applying compensating substances.

The invention aims at the use of balancing agents in the insemination of mammals in order to improve their reproduction, which enable pregancy to occur principally on any race of mammals with a much higher probability than the natural one, and provoke a considerable increase of reproduction, particularly on animals bred under large-scale farming conditions.

It has been found, unexpectedly, that all the above advantages can be attained by the use of specific balancing agents.

The invention is based on the surprising recognition that the discharge of the genital tract of repeat breeder females is more or less different in chemical composition from the discharge of the genital tract of the well conceiving ones; thus the probability of the required pregnancy can be increased to a great extent by determining and then eliminating these differences. Our experiences show that on "repeat breeder" cows (i,e. on females with symptom-free sterility) the regulated equilibrium of the chemical components in the oestrus mucus becomes unbalanced, since both the nutrient and the energy content decreases, which affects the metabolic processes of both the sperms and the zygotes.

Furthermore the concentration of bivalent cations increases, whereupon the cells first swell and then agglutinate. The efficiency of enzymes indispensable for conception and implantation decreases, since the mucus contains only minimum amounts of the substrates and the respective activators.

Our tests have also shown that the biological equilibrium of the animal organism can be influenced by various trace elements. As known, these trace elements catalyze various enzymatic processes of vital importance.

Some of the trace elements, such as manganese, zinc and iodine, play an important role in the metabolic processes of reproduction biology itself. Other trace elements, such as iron, copper and cobalt, are, however, also indispensable for the living organism.

The chemical composition of the animal fodders (fodder plants), and thus also the extent of supplying the animal organism with microelements, is unequivocally determined by the soil composition characteristic for every region and varying geographically.

According to the invention the deficiency in trace elements, determined by chemical tests, is eliminated by the method claimed in claim 1 or claim 2 appended hereto.

Thus the object of the invention is, in one aspect thereof, to improve the reproduction of mammals by preparing a treatment solution, and in another aspect, using said solution, containing balancing agents, i.e. containing substance(s), particularly enzyme activator(s) and/or substrate(s), present in the genital discharge in the period of oestrus of well-conceiving (fertile) female animals but partially or totally missing from the genital discharge at the period of oestrus of female animals showing the signs of sterility and husbanded in the same geographical region, in amounts required to compensate the deficiency, and/or containing substance(s) in amounts sufficient to inactivate (bind or neutralize) component(s), particularly calcium and/or magnesium ions, of the latter discharge present in excess as compared with the former one.

According to the invention, first the chemical composition of the genital discharge of female animals which show signs of sterility and that of the genital discharge of fertile female animals husbanded in the same geographical region are examined, always in the period of oestrus, and on the basis of the discrepancies detected in chemical composition, the nature and amount of balancing agents required to eliminate the differences between the former and latter discharges, i.e. to compensate the deficiencies and/or to inactivate the excess, are determined.

In a preferred method, according to the invention, balancing agents which contain compounds composed of ions suitable for enzyme activation are applied.

It is also preferred to apply balancing agents which contain substances capable of supplying missing substrates, particularly lipids or fat-like substances, carbohydrates or proteins. As carbohydrates e.g. glucose and/or fructose can be applied. Further examples of substances capable of supplying missing substrates are amino acids, such as methionine (e.g. DL-methionine), arginine (e.g.

L-arginine), lysine, cysteine and 3,5-diiodotyrosine (e.g. 3,5-diiodo-L-tyrosine), furthermore mineral and organic phosphates, such as potassium dihydrophosphate, P-glycerol phosphates (e.g. sodium l3-glycerol phosphate) and lecithins, as well as mixtures thereof.

In a third preferred method, the balancing agents utilized according to the invention contain compounds which are indifferent to the organism and can bind the excess of calcium and/or magnesium ions present in the form of complex salts, particularly in chelated form.

Of course, any of the above methods of application can be combined with each other.

It is preferred to apply zinc, selenium, copper, iodine, iron and/or cobalt ions as ions capable of activating the enzymes. Zinc ions can be applied e.g. as zine sulfate (such as zinc sulfate heptahydrate) and/or as zinc acetate. Manganese ions can be introduced e.g. in the form of manganese(ll) sulfate, such as manganese(ll) sulfate heptahydrate. Iodine can be introduced e.g. as 3,5-diiodotyrosine.

It is preferred to apply polyaminocarboxylic acids and/or their salts as chelating agents capable of binding the excess of calcium and/or magnesium ions in the form of complex said. Of the chelate forming polyaminocarboxylic acids usable according to the invention e.g. ethylenediaminotetraacetic acid is to be mentioned. Of the salts of this compound the sodium salts, primarily the disodium salt, can be applied to advantage.

The balancing agents are utilized preferably in the form of solutions (hereafter: balancing solutions). It is preferred to apply water which is free of salts and micro-organisms, particularly twice distilled water, as solvent medium for the balancing solutions. The balancing agents can also be presented, however, in a form of two-phase systems, e.g. as powder compositions filled into ampoulles, to be mixed prior to use with a liquid also filled into ampoulles.

For compensating the most frequent differences in chemical composition of the genital discharge of females which show the signs of sterility, i.e. to treat most of such animals, the individual constituents of the balancing agents, utilized according to the invention, to be administered per animal generally vary within the following limits:: Disodium salt of ethylenediamine- 15- 29 mg tetraacetic acid Sodium ,B-glycerol phosphate 4- 10 mg Lecithin 8- 11 mg Monosaccharides (such as glucose or 70-110 mg fructose), totally Manganese(ll) sulfate heptahydrate 250-350 rlg or an equivalent amount of another crystal form of manganese(ll) sulfate or of another manganese salt Zinc sulfate heptahydrate 200-300 ccg or an equivalent amount of another crystal form of zinc sulfate or of another zinc salt 3,5-Diiodo-L-tyrosine 100--1 50 lg DL-Methionine 90-120 Itg L-Arginine 10- 16,tt9 Of course, the amounts of the individual constituents of the balancing agents may be higher or lower than those mentioned above when treating virtually sterile females, if, in less frequent cases, their genital discharge differs in chemical composition less or more from the normal one.

From biological aspects, the major advantage of the method according to the invention is that no medicament is introduced into the organism of the animals, but only the deficiencies are supplemented or balanced which appear in the organism as a result of various exogenous and/or endogenous effects.

It is a further advantage that according to the invention an optimum physiological environment is provided for the sperm entering the organism of the females during insemination, on the one hand, and also for the zygotes and arising as a result of implantation, on the other hand, neither the individual constituents of the balancing agent nor the balancing agent as such represent the slightest risk of harm for the mother animal or for the zygotes.

It is an additional advantage that the method according to the invention can be performed by any person engaged with artificial insemination of animals, such as zootechnicians, husbandmen or other employees of the artificial insemination service.

The balancing agents utilized according to the invention, such as the balancing solutions, are introduced into the uterus at the time of insemination performed in a manner known per se. It is preferred to introduce the balancing agent as an infusion after insemination. The balancing agent is stored preferably at about-body temperature, or its temperature is adjusted to about body temperature directly prior to introducing it into the uterus.

The invention is elucidated in detail with the aid of the following non-limiting Examples, The tests described in the Examples were performed on cattles. It was assumed that the genital discharges of female animals differ in chemical composition from field to field, since both the nature and the amount of natural fodder plants grown on the specific area and consumed by the animals are different. The nutrient value of the natural green fodder depends on the composition of the soil. For this reason the tests described in the Examples, which verify that the balancing solutions are capable of readjusting the physiological conditions of the genital discharge, were performed in areas with different soil compositions. Within the individual tests, however, one area with one and the same characteristic soil composition was involved.The tests described in the Examples were performed in 3 state farms.

EXAMPLE 1 When examining the composition of the oestrus mucus of cows bred in state farm A it was established that it contains reducing monosaccharides, manganese and iodine in lower amounts than the required value. The analysis indicated that these components were present in the following amounts: monosaccharides: traces Mn2* ions: 1 0 g I- ions: traces Thus, a balancing solution, containing the missing amounts of glucose, fructose, iodine salt and manganese salt dissolved in water, was prepared and introduced into the uterus by infusion.The amounts of these balancing substances introduced per animal were as follows: glucose: 60 mg fructose: 45 mg manganese(ll) sulfate pentahydrate 270 ,ug [MnSO4 5H20] 3,5-diiodo-L-tyrosine: 100 At9 The three cows tested were so far inseminated 3, 5 and 7 times, respectively, without any positive result. After treating them with the above solution, all the three animals became pregnant. Based on the results of this test the solution was applied to treat a great number of virtually sterile animals, and 60% of the animals treated became pregnant.

EXAMPLE 2 When examining the composition of the oestrus mucus of cows bred in state farm B it was established that it contained inorganic phosphorous and zinc in lower amounts than required, whereas the amounts of calcium and magnesium where higher than necessary. The analysis indicated that these components were present in the following amounts: inorganic phosphorous: 110 zt9 zinc: 5 ffit9 calcium + magnesium: 10 mg In order to supply the missing components, zinc sulfate and mineral and organic phosphorous compounds (potassium dihydrophosphate and sodium glycernl phosphate) were dissolved in twice distilled water, and a chelating agent (disodium salt of ethylenediaminetetraacetic acid) was dissolved in the resulting solution in order to remove the excess of calcium and magnesium ions in complex form.

The amounts of these balancing substances introduced per animal were as follows: sodium ,B-glycerol phosphate 4.5 mg (or an equivalent amount of lecithin) potassium dihydrophosphate (KH2PO4) 1.5 mg zinc sulfate heptahydrate (ZnSO4. 7H20) 200 89 disodium salt of ethylenediamine- 29 mg tetraacetic acid Four virtually sterile cows, so far inseminated 4, 5, 8 and 9 times, respectively, with no success, were treated with the above solution. The treatment was fruitless only on one cow, which was so far inseminated 4 times without any success, whereas the other three cows became pregnant.

Large-scale tests were also performed at the same state farm at a time when, because of the great drought, the biological value of the green fodder grown on the same area was very poor. Despite of this fact 30% of the treated cows regarded as sterile became pregnant.

In the above balancing solution disodium ethylenediaminetetraacetate can be replaced by an equivalent amount of an alkali carbonate to obtain similarly good results.

EXAMPLE 3 When examining the composition of the oestrus mucus of cows bred in state farm C it was established that it contained carbohydrates and proteins in concentrations lower than required. The analysis indicated that these components were present in the following amounts: monosaccharides: 8 mg methionine: traces arginine: traces A combination of glucose and fructose was applied to supply the carbohydrates, whereas the missing proteins were supplied in the form of the amino acids methionine and arginine. The amounts of these substances introduced per animal were as follows: glucose: 62 mg fructose: 30 mg DL-methionine: 95 xl9 L-arginine: 12 fug Two cows, which remained sterile even after several (8 and 10, respectively) inseminations, were selected for testing.After treating them with the balancing solution, both cows became pregnant.

Based on the results, all sterile cows of the state farm were treated for one year with the balancing solution, and then kept under observation. Within this period 57% of the sterile females became pregnant.

In the above test series a total amount of 1,295 sterile cows were treated with balancing solutions to readjust the physiological conditions of the oestrus mucus. Of the 1,295 sterile cows treated 628 (i.e.

48.5%) became pregnant. It should also be taken into account here that, as described in Example 2, several tests were performed under clearly unfavourable foddering conditions, which explains why the summarized result is not higher than the percentage given above.

Instead of, or along with, DL-methionine and L-arginine other amino acids, such as lysine and/or cysteine, can also be applied in an equivalent total amount to obtain similarly good results.

Claims (10)

1. A method of improving the reproduction of mammals by using balancing agents containing substance(s), particularly enzyme activator(s) and/or substrate(s), present in the genital discharge in the period of oestrus of well-conceiving female animals but partially or totally missing from the genital discharge in the period of oestrus of female animals showing the signs of sterility and husbanded in the same geographic region, in amounts required to compensate the deficiency, and/or containing substance(s) in amounts sufficient to inactivate (bind or neutralise) component(s), particularly calcium and/or magnesium ions, of the latter discharge present in excess as compared to the former one.

2. A method of producing a treatment preparation for use in improving the reproduction of mammals, comprising analysing or testing the respective genital secretions in the period of oestrus of fertile and partially or wholly sterile female animals husbanded in the same geographical region, comparing the results of the analyses of tests and preparing a treatment preparation made up of the substances in the amount required to balance or compensate the deficiency, and/or containing substance(s) in amounts sufficient to inactivate (bind of neutralize) component(s), particularly calcium and/or magnesium ions, of the latter discharge present in excess as compared to the former one.

3. A method as claimed in claim 1 or claim 2, wherein said balancing agent or preparation includes compounds composed of ions capable of activating enzymes.

4. A method as claimed in any preceding claim, wherein said balancing agent or said preparation contains substances capable of supplying missing substrates, preferably lipids or fat-like substances, carbohydrates or proteins.

5. A method as claimed in any preceding claim, wherein the said preparation or the said balancing agent contains compounds capable of binding the excess of calcium and/or magnesium ions in the form of complex salts, particularly a chelating agent.

6. A method as claimed in claim 3 or any claim appendant thereto, wherein zinc, selenium, copper, iodine, iron and/or cobalt ions is or are used as ion(s) capable of enzyme activation.

7. A method as claimed in claim 5 or claims 5 and 6, wherein a polyaminocarboxylic acid or a salt thereof is applied as a chelating compound capable of binding the excess of calcium and/or magnesium ions in the.form of complex salts.

8. A method as claimed in any preceding claim, wherein the said balancing agent or preparation is presented in the form of a solution (balancing solution).

9. A method as claimed in any preceding claim, wherein water free of salts and micro-organisms, preferably twice distilled water, is used as a solvent for forming a balancing solution from said agent or preparation.

10. A method as claimed in claim 1 or claim 2, substantially as herein described with particular reference to any one of Examples 1 to 3.