WO1989002220A1

WO1989002220A1 - Method and composition for enhancing animal fertility

Info

Publication number: WO1989002220A1
Application number: PCT/US1988/003128
Authority: WO
Inventors: John Mcmichael
Original assignee: John Mcmichael
Priority date: 1987-09-10
Filing date: 1988-09-09
Publication date: 1989-03-23
Also published as: AU2485988A

Abstract

Fertility is enhanced in an animal undergoing artificial insemination in which an insemination medium is employed by identifying an insemination medium component to which the animal is hypersensitive, and the animal is subcutaneously injected with a unit dose of 2.8 x 10-5 mg egg white extract and 2.8 x 10-4 mg of milk extract, the dose being diluted to a volume of 0.5 cc with saline containing 0.4 % phenol on the day of service, and one dose for each of the two following days.

Description

METHOD AND COMPOSITION FOR ENHANCING ANIMAL FERTILITY

Background

The present invention relates in general to methods and compositions for enhancing animal fertility under conditions of artificial insemination. In particular, the present invention relates to the enhancement of bovine fertitity under conditions of artificial insemination by injection of low doses of components of the insemination medium. In the United States, the vast majority of dairy cattle, and some beef animals, are bred by artificial insemination (Al). When a heifer or cow comes into heat, a veterinarian or a technician introduces semen from a selected bull into the animal. The semen contains live sperm with which the recipient is fertilized. The procedure requires a farm call by the veterinarian or technician, purchase of the semen, and often a repeated attempt at breeding because of an initial failure. Successful fertilization at first service occurs in about 60-65% of treated animals as indicated by data obtained from the American Breeding Service (ABS) in Washington, D.C. Follow-up visits must thus occur for as high as 40% of cattle, and some cows will require four or more services before either "taking" (i.e., being successfuly inseminated) or being culled from the herd .

Although a regimen known as "provocative neutralization" therapy has been applied in human and animals in the treatment of allergies and other immunological disease states, it has not been suggested that problem of infertility in artifical insemination may be suspectible to such treatment. Miller, Annals of Allergy, 3_\, 185-191 (1977); Miller, "Food Allergy, Provocative Testing and Injection Therapy," Charles C. Thomas, Springfield, 111. (1972); Miller, Clinical Med. , 81, 16-19 (1974) and cMichael, U.S. Patent No. 4,521,405.

Summary of the Invention

The present invention provides a composition for enhancing fertility in an animal undergoing artificial insemination in which an insemination medium is employed. The composition includes a dose of a medium component, more specifically a milk extract or egg white extract or a combination thereof, which dose is lower than the lowest dose necessary to provoke a humoral immune response. A preferred composition according to the present invention includes an amount of egg white extract having a hypersensitivity neutralizing activity of about 2.8 x 10^-5 g egg white extract per dose, an amount of milk extract having the hypersensitivity neutralizing activity of about 2.8 x 10~⁴ mg of milk extract per dose, and an immunologically compatible diluent. Most preferred is a composition having a volume of about 0.5 cc wherein the diluent is physiological saline containing 0.4% phenol.

The present invention also provides a method for enhancing fertility in an animal . undergoing artificial insemination in which an insemination medium is employed. In the method, an insemination medium component is identified to which the animal is hypersensitive, and the animal is injected with the component in a dose lower than the lowest dose necessary to provoke a humoral immune response. Preferred is a ^' method wherein the injecting step includes the step of including in the dose 2.8 x 10^~5 mg egg white extract and 2.8 x 10^~4 mg of milk extract, the dose being diluted to a volume of 0.5 cc with a 0.9% aqueous saline solution containing 0.4% phenol. A most preferred method according to the present injection includes subcutaneous administration of one dose on the day of service, and one dose for each of the two following days.

Detailed Description

Bovine infertility may result, from infectiorf hormonal upsets, dietary disruption, failure to ovulate, unpredictable ovulation, or anatomical abnormalities. Although the above causes of infertility may be diagnosed and corrected, many infertile animals remain. It may be the case that these remaining animals are infertile due to a localized hypersensitivity to the semen itself or to the protein preservatives mixed with semen for .-stabilization. Egg white or milk or a mixture of egg white and milk are commonly used as protein preservatives.

In a method according to the present invention, a component of an insemination medium is administered to an animal undergoing artificial insemination in an amount which is believed to be less than the lowest amount necessary to provoke a humoral immune response (that is, to begin production of antibody) . In order to identify "a dose lower than that required to provoke a humoral immune response," a wheal produced upon subcutaneous injection of the therapeutic material is evaluated according to the procedure set forth in Moore, Clinical Medicine, 81, 16-19 (1974), wherein such evaluation is employed to identify a dosage of vaccine useful in the eradicatin of the symptoms of influenza. Upon subcutaneous injection, a wheal may be determined to be positive ten minutes after injection as blanched, hard, raised and discoid (regular, sharply demarcated edges, as though a disc has been cemented to the skin). A negative wheal, indicative of a dose below that necessary to provoke an immune response, is so absorbed at the end of ten minutes that it is softer and flatter than at injection, may have an irregular or ragged edge, and has grown less than an average of two millimeters in diameter.

The present invention is more specifically illustrated in the following examples. In Example 1, the results of testing "problem breeder" cows for hypersensitivity to components of artificial insemination media are reported. In Example 2, a composition and method according to the present invention are described. In Example 3, results of application of the present invention to problem breeder"cows are reported.

Example 1

About two dozen problem breeder cows were tested via skin whealing to determine a dose lower than the lowest dose necessary to provoke a humoral immune response to their sensitivities to milk and egg white as well as to semen. Only one showed sensitivity to semen while all showed at least minimal sensitivity to the other two products. Example 2

Based upon the skin testing results of Example 1, a medicine was formulated which includes 2.8 x 10^~5 5 mg egg white extract per dose and 2.8 x 10^~4 mg of milk extract per dose.

Egg white extract and milk extract may be obtained from Allergy Laboratories, Inc., Oklahoma City, Oklahoma. Allergy supply houses commonly carry these

10. items. Milk, egg white or other extracts useful according to the present invention may also be prepared by using any of the extraction techniques commonly performed by allergy supply houses. Allergy Laboratories is believed to perform a 50% glycerin

15 extraction (i.e., an extraction solution contains 50% glycerin solution plus bicarbonate for buffering and sodium choloride for isotonicity) . Extraction is performed at a 1:20 ratio of weight (dry material to be extracted) to volume (extracting fluid) overnight. The

20 resulting material is filter-sterilized to remove large clumps of protein as well as to remove microbial contaminants.

Each dose of medicine, containing extracts of egg white and milk, is diluted to a volume of 0.5 cc

25 with saline (0.9% NaCl in H₂0) containing 0.4% phenol as a preservative. An animal is given one dose (0.5 cc) of medicine subcutaneously on the day of service, and one dose for each of the two following days, for a total of three doses.

30 Using this medicine, the rate of conception at first service has been increased to 80-85%.

No side effects have been observed. Furthermore, milk dumping is not required as it is for many other medicines used in lactating animals. 5 Example 3

In an application of the present invention, twelve cows were successfully treated with the composition according to Example 1 after skin tests as described therein which confirmed the appropriate dosage level. All animals received a series of three injections of 0.5 cc each, the first injection being on the day of service via Al, and then one injection on each of the next two days. The composition was injected subcutaneously, and no adverse side effects were observed. All twelve animals responded after one 3- injection series. The results appear in Table 1.

TABLE 1

Cow's Name Breed Location Number of Consecutive Unsuccessful Services Prior to Treatment

Delia Holstein Harrisburg, PA 7

Martha Holstein Harrisburg, PA 3

Quella Holstein Harrisburg, PA 3

Penny Holstein Harrisburg, PA 4

Lara Guernsey Spartansburg, PA 13

Sheri Guernsey Spartansburg,. PA 4

Top Hornet Guernsey Spartansburg, PA 6

Pandora Guernsey Spartansburg, PA 5

Dolly Holstein Centerville, PA 4

Cow #1 Holstein Woodcoc , PA 3

Betty Holstein Schoharie, NY 3

Bridgett Holstein Schoharie, N 4

Five other treated cows did not respond to the treatment, and a possible explanation for this lack of response is indicated in Table 2, where an explanation is known.

TABLE 2

Cow's Name Breed Location Reason for Treatment Failur

Emma Holstein Schoharie, New York Suspected chronic leptospir

RIN Holstein Harrisburg, Pennsylvania Unknown

Marcie Holstein Harrisburg, Pennsylvania Unknown

Linda Holstein Cambridge Springs, Steroid treatment following Pennsylvania fall in barn

Molly Guernsey Saegertown, Pennsylvania Unknown

Although the present invention has been described in terms of a particular embodiment, it is anticipated that variations and improvements will occur to those skilled in the art upon consideration of the present invention. For example, although a subcontaneous route of administration was employed herein, obvious appropriate modifications may be made in the composition or method of administration to permit intramuscular, sublingual and interperitoneal routes, including a variation involving injection with a polymer allowing for slow release ^"of the medium component. Likewise, any suitable physiological isotinic diluent may be used in place of the 0.9% saline used herein.

It is also contemplated that the present invention be applied to any animal undergoing artificial insemination, including horses and all other forms of livestock as well as laboratory and zoo animals.

Accordingly, it is intended that the present invention include all such variations and improvements which come within the scope of the invention as claimed.

Claims

1. A composition for enhancing fertility in an animal undergoing artificial insemination in which an insemination medium is employed, comprising a dose of a medium component which consists essentially of a dose lower than the lowest dose necessary to provoke a humoral immune response.

2. The composition as recited in claim 1 wherein said medium component comprises a milk extract.

3. The composition as recited in claim 1 wherein said medium component comprises egg white extract.

4. A composition forEnhancing fertility in an animal undergoing artificial insemination in which an insemination medium is employed consisting essentially of: an amount of egg white extract having the hypersensitivity neutralizing activity of 2.8 x 10^-5 mg egg white extract per dose; an amount of milk extract having the hypersensitivity neutralizing activity of 2.8 x 10~⁴ mg of milk extract per dose; and an immunologically compatible diluent.

5. The composition as recited in claim 4 wherein said composition has a volume of 0.5 cc, and wherein said diluent is aqueous 0.9% NaCl containing 0.4% phenol.

6. A method for enhancing fertility in an animal undergoing artificial insemination in which an insemination medium is employed comprising the steps of: identifying an insemination medium component to which the animal is hypersensitive; and injecting the animal with a dose of the component which dose is^' lower than the lowest dose necessary to provoke a humoral immune response.

7. The method as recited in claim 6 wherein said injecting step comprises the step of including in the dose 2.8 x 10~⁵ mg egg white extract and 2.8 x 10~⁴ mg of milk extract, the dose being diluted to a volume of 0.5 cc with saline containing 0.4% phenol.

8. The method as recited in claim 7 wherein said injecting' step comprises the step of subcutaneous administration of one dose on the day of service, and one dose for each of the two following days.