ES2217911B1 - A NEW COMPOSITION AND PROCEDURE TO INCREASE THE PRODUCTION OF USEFUL SPECIES FROM THE AGRICULTURAL VIEWPOINT. - Google Patents

Abstract

An economically effective process and a composition for improving the production of agriculturally useful animals, in particular mammals such as sheep and poultry such as chickens, by administering a new composition to increase fertility containing concentrations substantially low of an immunogen, which is a conjugated inhibin or a fragment thereof. These concentrations are preferably in the range of about 5 to 500 micrograms of immunogen per ml of composition and, more preferably, in the range of about 20 to about 300 micrograms of immunogen per thousand of composition. The increase in fertility is useful for mammals, in particular sheep, and for poultry, in particular chickens, and is effective in the conditions of commercial farms. The composition consists of inhibin or a fragment thereof, bound to a support protein and emulsified with an adjuvant. Preferably, the immunogen includes the N-terminal sequence (amino acids 1-29) of the bovine inhibin subunit conjugated to the keyhole hemocyanin (KLH) in a peptide ratio (kLH) of 1:40.

Description

A new composition and procedure for increase the production of useful species from the point of view agricultural.

Field and background of the invention

The present invention relates to a Improved procedure to increase species production agriculturally useful, particularly species of mammals such as sheep or avian species such as chickens, after administration of a composition comprising inhibin or a peptide derived therefrom, conjugated with a Protein support and emulsified with an adjuvant. The invention is also refers to an increase in both the fecundity of animals as the average litter size as a result of the treatment With the composition.

Due to the increasing demands from the  consumers of high quality products and safe for the environment environment, sheep are becoming increasingly important in the market. Sheep produce high quality milk; wool, a fiber with unique properties; and lamb meat, a meat of Low caloric content, nutritious and with an exquisite flavor. The sheep also offer a multitude of by-products used in the manufacture of many commercial products that vary from chalk to shampoos Therefore, an improved procedure to increase sheep production is important for the manufacture of products derived from these animals.

There is a general need for a procedure to increase ovulation and fertility in animal species economically and agriculturally useful, including mammals such as sheep and poultry species such as chickens, for example, by manipulating factors Hormonal in these animals to increase fertility. To the increase the level of certain of such factors and / or reduce the level of other factors, the overall fertility of the animal.

In mammals and birds, FSH (hormone follicle stimulant) plays an important role in the potentiation of follicular growth and development. Increasing of the ovulation ratio is a consequence of a level elevated FSH in the blood. However, it is known that the hormone inhibin acts as a feedback regulator inhibitor of FSH hormone (follicle stimulating hormone). By consequently, fertility can be increased by inducing a hyper-ovulation in mammalian and avian species through the immuno-neutralization of the endogenous inhibin (or its biologically active subunit).

Inhibin is secreted by granular cells  of the ovary in females and by the sertoli cells of the testicles in males. The inhibin consists of a subunit α (P m = 18 kDa) linked through a disulfide bond to one of two very homologous β subunits (of approximately 14 kDa) to form inhibin A (? -? A) or inhibin B (α-? B). The shape Fully processed molecule has a molecular weight of approximately 32 kDa. The inhibin selectively suppresses the Secretion of Follicle Stimulating Hormone (FSH) from the pituitary and also has local paracrine actions in the gonads The immuno-neutralization of inhibin endogenous by active or passive immunization produces an increase in Circulating FSH and may increase follicular development of the ovary and the number of oocytes.

Therefore, there is a need and it would be useful have a procedure to increase the fertility of agriculturally useful animals, including mammals such as sheep and poultry species such as chickens, through an immunization against inhibin that does not require large quantities or multiple injections of the immunogen and that it also promotes with high efficiency the hyperovulation of animal and, therefore, promote increased fertility.

Summary of the Invention

It is an object of the present invention provide an economically effective procedure to increase the production of agriculturally useful animals, in particular mammals such as sheep and poultry such as chickens, through the administration of a new Composition to increase fertility.

A further scope of the present invention will be refers to the use of substantially low concentrations of immunogen, which is a conjugated inhibin or a fragment of the same. These concentrations are preferably in the range from about 5 to about 500 micrograms of immunogen per ml of composition and, more preferably, in the range of about 20 to about 300 micrograms of immunogen per ml of composition.

It is another additional object of the present invention  provide a treatment to increase the fertility of mammals, particularly sheep, that is effective in conditions of commercial farms.

It is another object of the present invention provide a treatment to increase the fertility of poultry, particularly chickens, which is effective in the conditions of commercial poultry farms.

It is another object of the invention to provide a composition consisting of inhibin or a fragment thereof, associated with a support protein and emulsified with a adjuvant Preferably, the immunogen includes the sequence N-terminal (amino acids 1-29) of the bovine inhibin alpha subunit conjugated with hemocyanin of keyhole bark (KLH) in a peptide / KLH ratio of 1:40

In accordance with the present invention, provides a procedure to increase the production of a agriculturally useful species, comprising the procedure the stage of administering to the species a composition which contains an effective amount of inhibin or a fragment of the same.

In accordance with another embodiment of the present invention, a composition is provided to increase the production of an agriculturally useful species, comprising: (a) inhibin or a fragment thereof; (b) one support protein for conjugation with said inhibin; and (c) an adjuvant for the emulsion.

Hereinafter, the term "animal" will make reference to any species useful from the point of view agricultural, including, but not limited to, lower mammals such as cows, sheep, goats and pigs; and poultry such like chickens, ducks, geese and turkeys.

Description of preferred embodiments

The present invention relates to a economically effective procedure and to a composition, for improve the production of useful animals from the point of view agricultural, particularly mammals such as sheep and birds of pen such as chickens, by administering a new composition to increase the fertility it contains substantially low concentrations of an immunogen, which is a conjugated inhibin or a fragment thereof. These concentrations are preferably in the range of about 5 to about 500 micrograms of immunogen per ml of composition and, more preferably, in the range of about 20 to about 300 micrograms of immunogen per ml of composition.

The treatment to increase the fertility of the The present invention is useful for mammals, in particular sheep, and for poultry, in particular chickens, and is effective in conditions of commercial farms. The composition of the The present invention consists of inhibin or a fragment thereof, bound to a support protein and emulsified with an adjuvant. Preferably, the immunogen includes the sequence N-terminal (amino acids 1-29) of the bovine inhibin alpha subunit conjugated with hemocyanin of keyhole bark (KLH) in a peptide / KLH ratio of 1:40

The invention is illustrated by the following example which describes a preferred embodiment of the invention without imposing No limitation on its scope.

The procedure described below includes The following main stages. First, the immunogen The animal was subsequently immunized, such as, by example, a sheep, using the immunogen composition, which is tested to determine the amounts and programs of optimal dosage for administration. Then they were analyzed the results obtained to determine the specificity of the immunogen activated antibodies and the efficacy of the immunization.

1. Preparation of the immunogen (a) Preparation of the immunogen for injection

A synthetic peptide corresponding to  the N-terminal sequence 1-29 of the alpha subunit of bovine inhibin (usually synthesized by SYPEP Ltd. Dublin, CA, USA) (hereinafter referred to as "the immunogenic peptide ") with keyhole limpet hemocyanin (KLH, Sigma H-2133) with glutaraldehyde (Aldrich 34.085-5) at a peptide / KLH ratio of 1:40. He Conjugate was dialyzed overnight against water at 4 ° C. Water of dialysis was replaced and the conjugate was dialyzed for another 4 hours at 4 ° C.

To avoid aggregation of the material that is frequently produced during conjugation of KLH peptide with glutaraldehyde, the process of conjugation was performed in the presence of sodium dodecyl sulfate (SDS) at a final concentration of 0.1%. The presence of SDS in the conjugation process allowed to use relatively concentrations low immunogenic peptide in the composition. Without pretending limited by a single hypothesis, presumably, by preventing aggregate formation was made accessible more antigenic determinants peptide to the immune system of object animals, which in the experiments indicated below They were sheep. Therefore, a lower dose of the immunogenic peptide, with fewer booster injections, to induce A sufficient immune response.

The immunogenic peptide / KLH conjugate is stored at -20 ° C until emulsion with an adjuvant comprising 85% (by volume) of mineral oil, 14.8% of Span (emulsifier) and 0.2% Tween-80 at concentration of 25 micrograms of peptide in 1 ml of adjuvant. The  Emulsified composition was maintained at 4 ° C.

(b) Evaluation of different concentrations of peptide immunogen to induce immunity in sheep

Titration of the antibody against the subunit Bovine inhibin alpha was quantified after administration of different concentrations of immunogenic peptide through injections. Five groups, each containing 5 Assaf dairy sheep, were injected twice via intramuscularly (i.m.) every two weeks. The injected compositions they contained 25, 50, 100 and 300 micrograms of immunogenic peptide. A control group received Phosphate Buffered Saline (PBS) Ten days after the second injection, blood was drawn Of all the animals. The serum was separated and kept at 20 ° C.

No substantial differences could be observed between the various doses of immunogenic peptide with the ELISA assay (described below). All animals, except animals of control, developed a high degree anti-inhibin compared to the control Hyper-immune serum positive shown in Figure 2.

(c) Quantification of antibodies anti-inhibin using the ELISA assay

Antibody titration specific anti-inhibin induced against various concentrations of injected immunogenic peptide will determined by an enzyme immunoassay (ELISA method) in which the wells were coated with the immunogenic peptide and the sheep antibodies were reacted with rabbit IgG anti-sheep labeled with peroxidase. The results They were read at 405 nm after substrate addition.

The positive control was a goat serum "hyper-immune" against the alpha subunit of the bovine inhibin (Biomakor Ltd, Israel) while controlling negative was the pre-immune serum pool from the sheep tested.

Figure 2 shows the test results ELISA for different concentrations of the immunogenic peptide. Even the relatively low dose of 25 micrograms of the peptide was able to induce a good immune response, as reflected for the high titration of the antibodies. All the results are compare with positive serum control hyper-immune against bovine inhibin and with the negative control serum obtained from the sheep before immunization

(d) Specificity of antibodies against aubunity inhibin alpha from bovine follicular fluid (BFF)

Antibody specificity was compared. induced by immunization of sheep against the fragment synthetic subunit alpha (1-29) of the bovine inhibin with the specificity obtained with inhibin natural from Bovine Follicular Fluid (BFF). How I know shown in Figure 1, the results demonstrate the binding specific for immunogen-induced antibodies in sheep with native inhibin. The experimental procedure was the next.

Ten sheep received two injections i.m separated by a period of two weeks. The injections contained synthetic 1-29 subunit of bovine inhibin conjugated with KLH and emulsified with an adjuvant. Ten days later from the second injection, blood was taken from the sheep and the serum It was separated and kept at -20 ° C.

(e) Western transfer in BFF

The BFF was separated on SDS polyacrylamide gel at 10% with a molecular weight marker and the fractions are transferred to a nitrocellulose sheet by electrotransfer Nitrocellulose was blocked with milk in skimmed and dehydrated powder in Tween buffer.

The blocked nitrocellulose was incubated with the inhibin alpha anti-subunit antibodies diluted with PBS. After washing in Tween buffer, stains transfer were incubated with rabbit IgG anti-sheep labeled with peroxidase. The spots of transfer were revealed using 4-chloro-1-naphthol as a substrate

As shown in the transfer spot of Figure 1, bands 1 and 2 contain BFF, while the band 3 contains the molecular weight marker (in thousands) and is indicated as 3. Two bands of bound antibodies are shown specifically, marked "P" and "I". The band "P" corresponds to the precursor of the inhibin subunit no processed (P.M. 56-58 kDa), while the band "I" is the alpha subunit of neutral inhibin (P.M. 32 kDa).

The results confirm that the antibodies obtained from sheep immunized against the alpha subunit synthetic bovine inhibin specifically bind to natural bovine inhibin fragments.

2. Immunization of sheep with peptide composition immunogen

The sheep used in this study were sheep of a commercial cross breed (German Marino x local Assaf) of a Moshav farm in central Israel. The sheep were kept in the shadow throughout the year, they fed a diet commercially acceptable and had free access to water. All study sheep were tested at least 45 days after the last  Birth.

For the experimental study, they were divided Randomly 122 sheep in two groups of 61 each. The first group (treatment) was immunized with 1 ml of the composition emulsified immunogen peptide / KLH (25 micrograms peptide immunogen) through an intramuscular injection (i.m.) in the hip. The second group (control) was injected i.m with 1 ml of adjuvant alone. Two weeks later, he was injected via i.m. 1 ml of the emulsified composition of immunogenic peptide / KLH, which it also contained 25 micrograms of immunogenic peptide, to the group of treatment. The control group was given a second i.m. injection 1 ml adjuvant alone. On the same day, to all the sheep (control and treatment) were inserted intravaginal devices (EAZI BREED CIDR G ™ containing 300 mg  of progesterone, InterAg N.Z.) for 12 days. After withdrawing these devices, each sheep received 600 IU of gonadotropin from pregnant mare serum (PMSG) (Synchroject Vetimex, The Netherlands) by injection medium i.m .. Blood samples were taken from 10 sheep  from the treatment group and 3 sheep from the group of control. The blood was coagulated and the serum separated and stored at -20 ° C.

Two days after PMSG injection and 14 days after the reinforcement, rams were introduced to a relationship between rams and sheep of 1: 7. The rams remained with the Sheep for three weeks. Then the number of pregnancies Pregnancy rates were analyzed by one trial. Chi squared. The average litter size was analyzed by one trial Student's t

3. Results obtained after immunization of sheep with the composition containing immunogenic peptide (a) Anti-inhibin titers in serum

As shown in Figure 3, all sheep sampled from the treatment group who received injections from the immunogen had an anti-inhibin titer significant, while samples from the group of control had no such antibodies. The average value of ELISA results for the treatment group was 1.97 compared to 2.1 OD of the positive control, and for the control group it was of 0.22 versus 0.24 OD of the negative control.

(b) Proportions of pregnancy

The pregnancy rates of the groups are determined by actual deliveries between 140 and 175 days after the induction of the rams. The results were similar in The two groups. 68% (40/59) of the sheep of the group were pregnant of control and 66% (40/61) of the sheep in the treatment group, as shown in Table 1 presented below. In the end from the experimental period, two sheep were removed from the group of control due to illness. The calculation of the proportion of Pregnancy was based on actual births.

(c) Average litter size

The average litter size of the group of treatment was significantly greater than that of the group of control, 2.1 lambs per sheep versus 1.65, as shown in Table 1. This difference was reduced when only the live-born lambs, although the results of the group of treatment were still greater than those in the control group: 1.9 vs. 1.6 (Table 1).

Those figures reflect the number of sheep that they conceived in the first cycle or in the following cycle. The term three weeks during which the sheep were in contact with the rams he covered two cycles of the sheep, lasting each 17 day cycle

TABLE 1 Pregnancy ratios and average litter size of the two groups

Group Proportion of Average size of the pregnancy (%) Litter Control 68 (40/59) 1.65 a (66/40) * 1.6 to (65/40) Treatment 66 (40/61) 2.1 b (84/40) * 1.9 b (75/40) * Lambs born alive a, b P < 0.05

Eight of the nine lambs born dead from treatment group came from two sheep with a pregnancy quadruple and they aborted 5 days before the start of labor. Should indicate that the common practice for the treatment of sheep is to feed the pregnant sheep during the last month of pregnancy with nutritional levels corresponding to a pregnancy double (according to the NRC). The nutritional requirements for a Triple pregnancy outweigh those of a double pregnancy by 10%. How the treated and control sheep groups remained in the same nutritional conditions, the nine lambs of the group treated that they were born dead probably died because of some insufficient nutritional conditions.

(d) Litter size distribution

Litter size distribution between one, two, three or more offspring shows a clear tendency to litters of multiple lambs in the treatment group compared to the control group While only 5% of the sheep of control had a litter of three or more offspring, 27% of the sheep in the treatment group had three or more offspring. This difference is reduced when only born lambs are counted alive Table 2 and Figure 4 show the distribution of the litter sizes between groups.

TABLE 2 Distribution of litter sizes among groups

Group A baby Two offspring Three offspring > Three pups Control 47.5% 47.5% 2.5% 2.5% 47.5% * fifty%* Any* 2.5% Treatment 32.5% 40.5% fifteen% 12% 35% * 47.5% * 12.5% * 5%* * Lambs born alive

(e) Fertility

The overall fertility of each group, considered as lambs born from 100 sheep treated and calculated to from litter size, it was significantly larger in the Treatment group compared to the control. Table 3 It demonstrates the fertility results.

TABLE 3 Fertility of control groups and treatment

Group Fertility (Lambs born / 100 sheep) Control 112.2 108.8 * Treatment 138.6 125.4 * * Lambs born alive

The fertility calculation was performed with regarding the total number of sheep treated. The number calculation of lambs born from 100 sheep was performed as indicated to continuation:

100 x% Pregnancies x Average litter size = number of lambs born / 100 sheep

(f) Summary of the results

The results show that the procedure and the immunogen composition described herein increased the size of the litter of the sheep that were He had administered the immunogen. The total increase in the size of the litter was mainly due to the increase in the number of triplets and to the reduction of the number of births of a single offspring. In the normal conditions of a farm, this is a desired result, since sheep can easily withstand triple pregnancies and Lambs are born viable. The composition of the present invention it can also clearly induce a very antibody titer specific with very low doses (such as in the range of 20-300 micrograms per injection) of the peptide immunogen Field studies clearly demonstrate that the active immunization of sheep with the composition of the present invention led to an increase in litter size by triple.

4. Evaluation of the immunogenic peptide composition of the present invention to improve fertility in rams

Titration of antibody against quantified  the alpha subunit of bovine inhibin after administration of injections of a composition according to the present invention containing immunogenic peptide to rams Assaf The rams were a mixture of local cross breeds Awassi x Frezien. At the beginning of the test, samples were collected actively of the rams, by riding a sheep artificial, such that semen was collected with a vagina artificial. Every time semen was collected from a group of rams, all the rams in the group had also ridden the sheep artificial. At the beginning of the test the samples were examined for Check semen quality. The collected semen was diluted immediately with ram chemical diluent (RSD-1) and was checked on an SQA II B analyzer. The parameters examined included volume, concentration, Motility, TFSC (Total Functional Semen Concentration) and SMI (Semen Motility Index).

Six groups were examined, each containing 5  Assaf rams. The ram with the lowest score of each group it was treated by two intramuscular injections (i.m.) of 1 ml of the composition according to the present invention, separating injections for a period of ten days. In total they were treated Nine Rams The injected composition contained 50 micrograms of immunogenic peptide, while the other rams received Phosphate Buffered Saline (PBS). Fourteen days later from the second injection, semen was collected from all the rams and The quality was checked again with the same analyzer. Further, at that time, blood was taken from all treated animals and of an untreated ram. A ram during the trial period. The serum was separated and kept at 20 ° C.

The treatment of rams with the composition of the present invention clearly increased various factors related to fertility, as shown in Tables lA and 1B with a comparison of each ram before treatment (Table 1A) and after treatment (Table 1B ). Initial scores for the rams to be treated are provided in italics .

TABLE No. 1A Semen quality before treatment

Group Ram Volume Concent Motility SMI TFSC one 1458 one 151 70 372 774 one 1369 one 126 66 329 596 one 1396 0.6 140 68 351 684 one 121 one 137 68 346 664 Average 0.9 138.5 68 349.5 679.5 2 1929 one 104 61 288 448 2 1968 0.5 106 62 291 459 2 9934 0.6 125 66 327 588 2 Mines 0.6 138 68 348 672 2 1816 1.25 144 69 359 716 Average 0.79 123.4 65.2 322.6 576.6 3 Horns one 137 68 346 664 3 1163 one 154 71 378 801 3 212 1.1 110 63 300 490 3 OPEN 1.2 118 65 315 543 3 166 1.9 96 59 270 390

TABLE No. 1A (continued)

Group Ram Volume Concent Motility SMI TFSC Average 1.24 123 65.2 321.8 577.6 4 2417M 1.1 146 69 361 725 4 9888 1.1 128 66 331 604 4 Ear 1.9 166 74 412 954 4 419M 1.1 153 71 376 792 4 421M one 122 65 322 568 Average 1.24 143 69 360.4 728.6 5 7315 1.1 78 52 212 230 5 7279 one 132 67 339 636 5 Mines 1.9 121 65 320 560 5 123 0.7 137 68 346 664 5 6448 one 134 67 342 648 Average 1.14 120.4 63.8 311.8 547.6 6 5 0.7 96 59 270 390 6 FLUFF 1.1 100 60 279 417 6 one 1.2 128 66 331 608 6 108 0.8 123 5 324 576 6 24 2 125 66 327 588 Average 1.16 114.4 51.2 306.2 515.8

TABLE No. 1B Semen quality after treatment

Group Ram Volume Concent Motility SMI TFSC one 1458 one 183 79 462 999 one 1396 0.7 171 75 428 999 one 121 one 133 67 340 640 one 1369 1.5 133 61 340 999 Average 1.05 155 70.5 392.5 909.25 4 421M 2.7 146 69 361 725 4 Ear 1.5 173 76 432 999

TABLE No. 1B (continued)

Group Ram Volume Concent Motility SMI TFSC 4 419M 1.5 168 74 418 981 4 9888 2 190 81 482 999 Average 1,925 169.25 75 423.25 926 6 Fluffi 0.8 152 70 374 783 6 5 2 167 74 416 972 6 24 2 156 71 383 824 6 one 1.5 164 73 407 932 6 108 one 181 78 455 999 Average 1.46 164 73.2 407 902 5 Mines 2 204 85 522 999 5 7279 one 169 75 420 990 5 123 1.5 154 71 378 801 2 9934 0.8 194 82 493 999 5 6448 1.2 200 84 511 999 Average 1.3 184.2 79.4 464.8 957.6 3 166 2 163 73 405 923 3 OPEN 2 142 69 355 700 3 1163 one 94 58 266 378 3 212 1.5 162 73 399 896 Average 1,625 140.25 68.25 356.25 724.25 2 1816 0.75 86 55 242 306 2 Mines 1.2 85 55 240 300 2 1968 0.5 143 69 357 708 5 7315 one 120 65 318 553 2 1929 one 111 63 302 497 Average 0.89 109 61.4 291.8 472.8

Tables 2A and 2B demonstrate the relative quality  of the semen of each ram treated in the context of the group before of treatment (Table 2A) and after treatment (Table 2B). The relative quality was found as a ratio of the value of each parameter for the treated ram, divided by the average of these values for the group of rams to which the treated ram.

TABLE No. 2A Semen quality index before treatment

Ram Volume Concent Motility SMI TFSC 1369 1,111 0.91 0.97 0.94 0.87 1929 1.26 0.84 0.93 0.89 0.77 9934 0.75 1.01 1.01 1.01 1.02 166 1.5 0.78 0.9 0.84 0.67 9888 0.88 0.89 0.95 0.92 0.83 421M 0.8 0.85 0.94 0.89 0.78 7315 0.96 0.65 0.82 0.68 0.42 5 0.6 0.84 1.15 0.88 0.75 Fluffi 0.94 0.88 1.17 0.91 0.8

TABLE No. 2B Semen quality index after treatment

Ram Volume Concent Motility SMI TFSC 1369 1.43 0.86 0.87 0.87 1.10 1929 1.12 1.02 1.03 1.03 1.05 9934 0.62 1.05 1.03 1.06 1.04 166 1.23 1.16 1.07 1.14 1.27 9888 1.04 1.12 1.08 1.14 1.08 421M 1.40 0.86 0.92 0.85 0.78 7315 1.12 1.10 1.06 1.09 1.17 5 1.37 1.02 1.00 1.02 1.08 Fluffi 0.55 0.93 0.96 0.92 0.87

Table 3 shows the semen quality index  for rams treated as a quality ratio before treatment (Table 2A) and quality after treatment (Table 2B). Clearly, semen quality increased after treatment for the parameters of Tables 2A and 2B. This particular relationship demonstrates the individual improvement of each ram treaty.

TABLE 3 Relationship between quality before treatment and quality after treatment for rams treated

Ram Volume Conc. Motility M: YES TFSC 1369 1.29 0.94 0.89 0.92 1.26 1929 0.89 1.21 1.10 1.16 1.37 9934 0.82 1.04 1.05 1.05 1.02 166 0.82 1.49 1.35 1.35 1.90 9888 1.18 1.26 1.24 1.24 1.30 421M 1.75 1.01 0.96 0.96 1.00 7315 1.17 1.69 1.60 1.60 2.78 5 2.28 1.21 1.16 1.16 1.44 Fluffi 0.58 1.05 1.01 1.01 1.09

Table 4 compares the average of rams treated and untreated before and after treatment. This form compares the average value of the group before and after treatment. The results were analyzed by a t-test to determine the meaning of the differences (a = P <0.005, n = not significant), demonstrating in this way that, before treatment, the rams to be treated had semen quality less than the control rams, the difference being statistically significant. After treatment, it was removed this difference. In some cases, after treatment, the treated rams had a slightly higher semen quality than the control rams, but this difference was not statistically significant.

Before treatment After treatment Category Rams Rams value Rams Rams from value from from p from treatment P control treatment control TFSC 660.9 470.1 0.0002 802.8 827.8 b 0.397 to SMI 344.2 292 0.0002 389.7 387.9 b 0.477 to Motility 64.5 61.5 0.2771 71.5 70.9 b 0.428 b Concent 135 108.3 0.0001 153.8 152.8 b 0.474 to Volume 1.09 1.06 0,3985 1.26 1.54 b 0.118 b

Tables 1-4 demonstrate clearly that treatment with the vaccine anti-inhibin of the present invention, as has been described previously, improved the semen quality of rams with a relatively low semen quality. This result is particularly significant, since improvement was demonstrated for a large number of rams that proved to have a quality of semen before treatment less than control rams, the difference being statistically significant, but that They had the same semen quality after treatment. This manner, composition and process of the present invention satisfactorily treat low semen quality in animals such as rams, which have great agricultural importance and commercial.

The importance of comparing each treated ram with control rams of the same group is that such a comparison solve any possible deviation from factors external, or to any internal variance within each group. The Table 3 shows the improvement of individual rams after treatment, while Table 4 shows the improvement within the groups of rams after treatment.

The parameters that were most influenced in the rams treated were motility, SMI and TFSC. Volume and the concentration were less affected, possibly because They were relatively good before treatment.

5. Evaluation of the immunogenic peptide composition of the present invention to induce immunity in hens layers

The anti-inhibin compositions and the process of the present invention have proven to be effective in mammalian species, such as sheep, as has been previously described. To examine the effectiveness of this invention in a non-mammalian species, specifically in poultry, the effect of the composition in hens was examined layers. A group of approximately 800 laying hens prematurely locked in a chicken coop and was divided into 3 groups of approximately 260-288 hens each. At the age of 18 weeks, before the start of laying eggs, Groups A and B were vaccinated with 0.1 ml (5 micrograms of peptide) of the anti-inhibin composition by means of intramuscular injections; Group B was vaccinated again a second time with this composition at the age of 20 weeks; and the Group C was left as an untreated control.

At the age of 20 weeks, some chickens They started putting sporadically. Subsequently, they were collected the data related to the behavior of the 3 groups. The parameters examined included increasing the proportion of put; the increase in cumulative egg production; he increase in weight and mass of eggs per chicken; the reduction of the amount of food consumed per egg produced; and the persistence of the laying peak. With respect to the last point, it should be noted that the proportion of egg production varies cyclically during the life of the laying hen, such so that the peak set represents the maximum proportion of Egg production per day for that chicken. Typically such peak is reached at a daily egg production of 90%, or almost One egg a day.

The results of the study demonstrated a clear improvement of egg production compared to control, a significant reduction in egg mass per chicken locked up compared to the control, and a reduction significant amount of food consumed per egg produced, again compared to the control. So the Method and compositions of the present invention they also proved useful for increasing various parameters of Egg production from laying hens.

It will be appreciated that the above descriptions they only pretend to serve as examples, and that many are possible other realizations within the spirit and scope of the present invention

Claims (24)

1. A procedure to increase production of an agriculturally useful species, the procedure comprising the stage of administering to the species a composition that contains an effective amount of inhibin or a fragment thereof, said effective amount being in the range of about 20 to about 300 micrograms by dose. 2. The method of claim 1, in the that said inhibin is present at a concentration in the range of about 25 to about 50 micrograms by dose. 3. The procedure of claims 1 or 2, wherein said inhibin in a bovine inhibin. 4. The method of claim 3, in the that said inhibin is a synthetic peptide corresponding to the amino acids 1-29 of the sequence N-terminal of the alpha subunit of inhibin. 5. The procedure of any of the claims 1-4, wherein said composition Contains an emulsion adjuvant. 6. The procedure of any of the claims 1-5, wherein said inhibin is conjugates with a support protein. 7. The method of claim 6, in the that said support protein is keyhole hemocyanin (KLH). 8. The method of claim 7, in the that said inhibin is conjugated with said support protein with glutaraldehyde 9. The method of claim 8, in the that said conjugation is performed at an inhibin / KLH ratio of 1:40 10. The procedure of any of the claims 1-9, wherein the species is a mammal species 11. The procedure of any of the claims 1-10, wherein the species is the sheep. 12. The procedure of any of the claims 1-9, wherein the species is a Poultry species. 13. A composition to increase production of an agriculturally useful species, which understands:

(to)

inhibin or a fragment thereof, said inhibin being present in an amount of the range of about 20 to about 300 micrograms per dose;

(b)

a support protein for conjugation with said inhibin; Y

(C)

a emulsion adjuvant.

14. The composition of claim 13, in the that said inhibin is present at a concentration in the range of about 25 to about 50 micrograms by dose. 15. The composition of claims 13 or 14, wherein said inhibin is a bovine inhibin. 16. The composition of claim 15, in the that said inhibin is a synthetic peptide corresponding to the amino acids 1-29 of the sequence N-terminal of an alpha subunit of inhibin. 17. The composition of any of the claims 13-16, wherein said inhibin is conjugates with a support protein. 18. The composition of claim 17, in the that said support protein is keyhole hemocyanin (KLH). 19. The composition of claim 18, in the that said inhibin is conjugated with said support protein with glutaraldehyde 20. The composition of claim 19, in the that said conjugation is performed at an inhibin / KLH ratio of 1:40 21. The composition of any of the claims 13-20, wherein the species is a mammal species 22. The composition of any of the claims 13-21, wherein the species is a sheep. 23. A procedure to increase production of an agriculturally useful species, the procedure comprising the stage of administering to the species a composition that contains an effective amount of inhibin or a fragment thereof, said effective amount being in the range from about 0.31 micrograms to about 4.6 micrograms per kilogram of the species per dose. 24. A composition to increase production of an agriculturally useful species, which understands:

(to)

inhibin or a fragment thereof, said inhibin being present in an amount of the range of approximately 0.31 micrograms to approximately 4.6 micrograms per kilogram of the species per dose;

(b)

a support protein for conjugation with said inhibin; Y

(C)

a emulsion adjuvant.