FIELD OF THE INVENTION

The present invention relates to herbal compositions for providing complete fertility solution to solve the infertility problems of cattle by feeding either through the concentrate feed or feed supplement. More specifically, the present invention is related to an alternative remedy herbal based composition for improving natural fertility process in female cattle. The composition induces the fertility in cattle by enhancing the metabolic function of the entire body, resulting in an improvement in sexual function and an increase in reproductive function, thus effects as nutritional tonic and sexual function improvement. The composition comprises an effective amount of an extract and/or at least one bioactive fraction or powder from herbs such as Ruta graveolens, Moringa oleifera, Guizotia abyssinica, Tribuhis terrestris, Nigella sativa, Mucuna pruriens, Aloe vera etc.; one or more additive selected from dicalcium phosphate (DCP), chelated minerals, mineral mixture and bypass fat; and process for the preparation of such extracts and herbal composition.

BACKGROUND OF THE INVENTION

The term "fertility" refers to the quality or state of being fertile. The term further refers to a female cattle being capable of breeding or reproducing. Infertility is one of the major reasons of non-viable dairy farming. This may be attributed due to nutritional imbalance, hormonal imbalance, mineral deficiencies and faulty management.

Gamete generating processes in cattle are constantly subjected to stresses from physical, chemical, and biological sources that can result in problems whereby female may become infertile or sub-fertile due to poor egg quality. These problems may be compounded in the cases of under or mal nourished catties. Eggs are produced by a well-known process called oogenesis. The process results in the formation of a large cell having a variety of resources for the construction of the embryo. Accordingly, oogenesis requires adequate nutrition to provide the cellular building blocks to create the equipped cells. The basic cellular building blocks include, among other things, amino acids, carbohydrates, proteins, fats and saccharides. The present invention provides compositions having effective fertility promoting nutrients to enhance gametogenesis and the maintenance of egg cells, which ultimately promote fertility. Nutrients are required to play a role in the development and function of egg. Detailed knowledge of the role-played by various nutrients or the level required for optimal development and function of egg is not well understood. Therefore, correcting key nutrient deficiency through supplementation has been problematic. The present invention overcomes these difficulties by providing compositions containing the key nutrients needed to improve fertility in cattle.

The causes of female infertility are very various, and it is not easy to find these causes. Typical causes include the interruption of tubal patency, and ovulation disorders, such as amenorrhoea, anovulatory menstruation, and sporadic anovulatory disorders. The term infertile cattle with impaired fertility used herein to refer to cattle populations that have difficulties in getting pregnant.

Due to stress on egg cells proper nourishment is essential to facilitate fertility and promote adequate amounts of viable gametes. Accordingly what is needed is a composition, which alleviates, overcomes, or cures the problem of infertility and sub- fertility by ensuring that gamete-generating tissues are nourished and have access to the cellular building blocks necessary for oogenesis and gamete maintenance. The compositions of the present invention are applicable to female catties wherein the egg integrity may be affected by physical, chemical, or biological stressors to the egg or developing egg in the target organ.

A wealth of literature has been published on the treatment of infertility in man and animals. Reproductive medicine has revealed a great number of treatments that have good success rates for many different indications. Clomiphene citrate is a commonly prescribed fertility drug. It stimulates the brain to release luteinizing hormone, which causes egg maturation and release from the ovary. The drug is taken for just a few days and then stopped. A brief fertile period may follow. A drawback is that more than one egg may be released, resulting in a multiple pregnancy. In addition, nearly half of all pregnancies achieved in this way result in spontaneous abortion. Perhaps the miscarriage rate is so high because such medications only force ovulation without correcting underlying deficiencies or hormonal abnormalities. Conventional treatments for various mammalian infertility conditions include the administration of human chorionic gonadotrophins (hCG), human menopausal gonadotrophin (hMG)(consisting of equal amounts of follicle stimulating hormone, FSH, and luteinizing hormone LH), or luteinizing hormone releasing hormone (LHRH), also known as gonadotrophin releasing hormones (GnRH). Treatment with these hormones however, is generally expensive, and does not always yield satisfactory results.

While prescription or over-the-counter drugs may be efficacious in addressing the fertility problems, alternative medicine based on herbs and other natural ingredients appears to be more preferable from the overall health viewpoint. Botanical medicines on the other hand, nourish and support the female reproductive system to restore hormonal balance. Many dietary supplements have been described that function in improving the fertility of man and animals. In order to improve reproductive health of male and female animals it has been recommended to feed a good multivitamin preparation and include sufficient amounts of supplements in the diet, such as zinc, vitamin E and the amino acids arginine and taurine.

The following herbs are often recommended by naturopathic doctors and herbalists to help restore female fertility as given in WO20061 1 1546:

a) Chaste tree berry (Vitex agnitf-castus): This herb stimulates the release of luteinizing hormone (LH) from the pituitary gland in the brain. This, in turn, promotes ovulation. Chaste tree berry may restore normal periods in women with amenorrhea (lack of menstrual periods).

b) Dong quai {Angelica sinensis): Widely used for female complaints, including menstrual irregularities and infertility, dong quai can tone a weak uterus by promoting metabolism within the organ, 12 regulating hormonal control and improving the rhythm of the menstrual cycle.

c) Red clover blossoms (Trifolium pratense): These beautiful little flowers are categorized in many herbals as fertility promoters. Chemical analysis shows that the herb is rich in coumestans and isoflavones, estrogenlike compounds that may promote fertility, particularly in women who are deficient in estrogen. d) Licorice {Glycyrrhiza glabra): This plant contains hormonal Iy active compounds categorized as saponins. A Japanese study found licorice-based medicines improved menstruation in women with infrequent periods. The study also found that licorice helped women with elevated testosterone and low estrogen levels, as commonly occurs in polycystic ovary disease.

e) Siberian ginseng (Eleutherococcus senticosus): This and other tonic botanicals can improve fertility by enhancing overall health and vitality. Siberian ginseng also acts on the brain to promote regulation of reproductive hormones.

The patents of interest in which the natural agents used include US 20060068033, which discloses the alternative remedy based composition for enhancing fertility in which a plurality of herbs is combined to produce a composition that enhances and promotes fertility in females. The WO2005/041990 highlights the use of Shilajit or the extract for improvement of sexual function and an increase in reproductive function to treat infertility in animal. The invention in WO2006/1 1 1546 discloes the animal food combinations comprising beta-glucans and phytohormones like auxin or gibberellic acid; which improves the fertility of animals and increases their chances on a pregnancy. The US 6737084 discloses the compositions and methods for enhancement of sexual pleasure and disorders related to sexual pleasure especially in mammalian females. US 20060057234 disclose composition comprising pharmaceutical/nutraceutical agent and a bio-enhancer obtained from Glycyrrhiza glabra for fertility.

Despite all these products that may improve the fertility of animals, there is still room for alternative products that can play a role in treating the infertility of cattle without any undesired side effects. A need therefore exists for an alternative medicine-based, natural solution to the above difficulties experienced by female cattle. An alternative medicinal treatment to the drugs described here are low cost ^' natural ^' medicines such as various herbs, which are typically ingested over large periods of time to form part of one's diet.

Thus, the inventive composition will be useful as foods or drugs for the improvement of sexual function or the strengthening of reproductive function. Also, the inventive composition is derived from natural substances, available in India and other parts of Asia for a long time and thus, advantageous in that it has secured safety and no side effects. The present inventor aims to ^' develop the herbal compositions and specifically to those compositions with naturally occurring herb extracts or powder for promoting fertility in cattle without the undesired side effects. In particular, the herbal compositions which can be fed to cattle for treating infertility or sub-fertility. The present invention provides improved compositions that require little technical expertise to use, are rapid, and may be used either as a nutritional supplement, or as a medicinal treatment of infertile or sub-fertile female catties. The compositions are inexpensive and remarkably cost effective as compared to the existing compositions.

OBJECTS OF THE INVENTION

The primary object of the invention is to develop herbal compositions for providing complete fertility solution for infertility problems of cattle by feeding either through the concentrate feed or feed supplement.

It is another object of the invention composition to provide a complete fertility solution for fertility disorders like Anoestriis, Non-ovulation, repeat Breeding problems.

It is yet another object of the invention composition to provide solution for post parturient diseases like Prolapse of uterus.

It is yet another object of the invention to develop herbal composition which is helpful in prevention of pregnancy disorders like Mummification, maceration of Foetus, Distokia, Retention of placenta and Agalactia.

It is further object of the invention composition which regulates & initiates the process of ovulation by stimulation of the feedback mechanism and helps for proper formation of the uterine bed that harbors the zygote.

It is another object of the invention to develop an alternative remedy herbal based composition to maintain hormonal balance in reproductive system which brings animal in oestrus naturally and also to prevent early Embryonic death.

It is an object of the invention to develop herbal compositions with naturally occurring herb extracts or powdered from for promoting fertility in female cattle without undesired side effects. It is the further object of the invention herbal compositions to increase dietary intake of vitamins and minerals associated with the gametogenesis in cattle by improving the overall nutritional status of the cattle in a pre ovulation stage.

It is a further object of the present invention to provide herbal compositions having a long shelf life.

It is a further object of the present invention to develop herbal compositions, which are inexpensive and remarkably cost effective as compared to the existing compositions.

STATEMENT OF THE INVENTION

Herbal feed compositions for solving fertility problems in cattle comprising mixture of effective amount of an extract and/or bioactive fraction or powder of at least two medicinal herbs and one or more additives selected from dicalcium phosphate (DCP), chelated mineral mixture, mineral nutrients and bypass fat. The medicinal herbs are selected from the group of Ruta, Moringa, Guizotia, Tribulus, Nigella, Muc na, and Aloe. The medicinal herbs are essentially selected from the group of Ruta graveolens, Moringa oleifera, Guizotia abyssinica, Tribulus terrestris, Nigella sativa, Mucuna pniriens, and Aloe vera. The herbal mixture comprises at least four medicinal herbs preferably Ruta graveolens, Moringa oleifera or Guizotia abyssinica, Tribulus terrestris or Nigella sativa and Mucuna pruriens or Aloe vera. The effective amount of an extract or bioactive fraction or powder of selected herbs ranges 15 to 50% (w/w) Ruta graveolens, 15 to 50% (w/w) Moringa oleifera, 10 to 40% (w/w) Tribulus terrestris and 2.5 to 20% (w/w) Mucuna pruriens. The effective amount of an extract or bioactive fraction or powder of selected herbs preferably 30% (w/w) Ruta graveolens, 25% (w/w) Moringa oleifera, 20% (w/w) Tribulus terrestris and 10% (w/w) Mucuna pruriens. The herbal compositions are added with effective amount of chelated mineral mixture preferably ment omins chelated minerals to prepare the herbal feed compositions. The effective amount of chelated mineral mixture ranges from 10 to 25% (w/w) of total composition and preferably 15% (w/w) of total composition. The menthomins chelated minerals include zinc, manganese, copper, cobalt, selenium, chromium and iodine. DETAILED DESCRIPTION OF THE INVENTIO

The invention generally relates to herbal compositions for providing complete fertility solution to solve the most cases of infertility problems of cattle. More particularly, the present invention is related to an alternative remedy as herbal based composition for improving natural fertility process of female cattle and process for preparing herbal extracts or powdered form and compositions.

The herbal composition of the present invention upon feeding along with the concentrate feed induces the fertility in cattle by enhancing the metabolic function of the entire body, resulting in an improvement in sexual function and an increase in reproductive function, thus effects as nutritional tonic and sexual function improvement.

The invention is a synergistic formulation of an effective amount of an extract and/or at least one bioactive fraction or powder from herbs such as Ruta graveolens (Chalepenesis), Moringa oleifera, Guizotia abyssinica, Tribulus terrestris, Nigella sativa, Mucuna pruriem, Aloe vera etc. with one or more additive selected from dicalcium phosphate (DCP), chelated minerals mixture and mineral mixture which are used in functional foods. The chelated mineral mixture and DCP balance the diet of cattle and they nourish the cattle by improving the overall nutrition status of the cattle.

Surprisingly, it has now been found that the fertility of catties can be improved by feeding them with effective amount of extracts and/or at least one bioactive fraction or powder of naturally occurring herbs such as Ruta graveolens, Moringa oleifera or Guizotia abyssinica, Tribulus terrestris or Nigella sativa, Mucuna pruriens or Aloe vera with chelated mineral mixture, mineral nutrients and bypass fat or mineral mixture or dicalcium phosphate (DCP). Also, combinations of these herbs, in particular the combination with chelated minerals, were shown to have a synergistic effect in that they improved the fertility of the cattle, even more than these individual components on their own. The composition of invention proved particularly useful for getting cattle into heat earlier, so that they produced offspring earlier resulting in an increased number of offspring of the total population. It was also observed that the number of newborns per parent animal increased by using the composition of the invention. This is particularly useful in commercial stock farming where even small increases in the productivity of the stock may contribute significantly to the commercial profits.

The compositions can be used in several forms: powdered feed form, concentrate form, blender form and base mix form.

The herbal feed composition is designed to provide a complete fertility solution for fertility disorders like Anoestrus, Non-ovulation, repeat Breeding problems. It incorporates both herbal ingredients and supplementation of chelated minerals which are required for normal reproductive processes and a deficiency of which can cause infertility. It regulates and initiates the process of ovulation by stimulation of the hormone feedback mechanism. The herbal feed composition helps for proper formation of the uterine bed that harbors the zygote. It also increases the blood circulation in the uterus thus reducing chances of Metritis and Pyrrometra. Herbal ingredients help to maintain hormonal balance in the reproductive system which brings animal in oestrus naturally and also helps to prevent early Embryonic death.

As used herein, the term "effective amount" means an amount necessary to achieve a desired result in solving fertility problems of cattle.

The herbal feed compositions for solving problems of fertility in cattle mainly comprises the mixture of effective amount of an extract or bioactive fraction or powder of following medicinal herbs:

1. Ruta graveolens : Family- Rutaceae

It is an ornamental, aromatic, culinary and medicinal plant, cultivated in gardens. It prefers rocky, well-drained soils and it resists dry weather. It is native to Europe, specially the Mediterranean region, but widely distributed into all the temperate and tropical regions. It is a very popular and attractive garden shrub in South America, where it is grown not only for ornamental and medicinal reasons but also because of the belief that it provides protection against evil.

2. Moringa oleifera: Family- Moringaceae

It is commonly referred to simply as Moringa is the most widely cultivated variety of the genus Moringa. It is an exceptionally nutritious vegetable tree with a variety of potential uses. The tree itself is rather slender with drooping branches that grows to approximately 10 m in height; however, it normally is cut back annually to one meter or less, and allowed to regrow, so that pods and leaves remain within arm's reach. Moringa is common in India, where its triangular, ribbed pods with winged seeds are used as a vegetable crop. It is particularly suitable for dry regions. The drumstick can be grown using rainwater without expensive irrigation techniques. The yield is good even if the water supply is not. The tree can be even grown on land covered with 10-90 cm of mud. Guizotia abyssinica Family: Asteraceae

It is an erect, stout, branched annual herb, grown for its edible oil and seed. Its cultivation originated in the Ethiopian highlands, and has spread to other parts of Ethiopia. Common names include: noog; niger, nyjer, or niger seed; ramtil or ramtilla; inga seed; and blackseed. The seed, technically a fruit called an achene, is often sold as bird seed as it is a favourite of finches, especially Goldfinch and Greenfinch. In the bird seed market, Nyjer is often sold or referred to as thistle seed. This is a misnomer due to early marketing of the seed as 'thistle' to take advantage of the finches' preference for thistle. Niger Seeds are also used in southern parts of India. In arnataka and Maharashtra, Niger seeds (called Uchellu and Karale in Kannada (local language of Karnataka) and Marathi respectively) are used to make a dry chutney which is used as an accompaniment with breads. They are also used as a spice in some curries. Tribulus terrest is : Family-Zygophyllaceae

It is a flowering plant and is native to warm temperate and tropical regions of the Old World in southern Europe, southern Asia, throughout Africa, and in northern Australia. It can thrive even in desert climates and poor soil. Like many weedy species, this plant has many common names. Puncture Vine, Caltrop, Yellow Vine, and Goathead are the most widely used. It is a taprooted herbaceous perennial plant that grows as a summer annual in colder climates. The stems radiate from the crown to a diameter of about 10 cm to over 1 m, often branching. They are usually prostrate, forming flat patches, though they may grow more upwards in shade or among taller plants. The leaves are pinnately compound with leaflets less than a quarter-inch long. The flowers are 4-10 mm wide, with five lemon-yellow petals. A week after each flower blooms, it is followed by a fruit that easily falls apart into four or five single-seeded nutlets. The nutlets or "seeds" are hard and bear two to three sharp spines, 10 mm long and 4-6 mm broad point- to-point. These nutlets strikingly resemble goats' or bulls' heads; the "horns" are sharp enough to puncture bicycle tires and to cause painful injury to bare feet. Nigella sativa: Family- Apiaceae

It is an annual flowering plant, native to southwest Asia. It grows to 20-30 cm tall, with finely divided, linear (but not thread-like) leaves. The flowers are delicate, and usually coloured pale blue and white, with 5-10 petals. The fruit is a large and inflated capsule composed of 3-7 united follicles, each containing numerous seeds. The seed is used as a spice. In English, Nigella sativa seed is variously called Black cumin, fennel flower, nutmeg flower, Roman coriander, blackseed, black caraway, or black onion seed. In English-speaking countries with large immigrant populations, it is also variously known as kalonji (Hindi kalaumji or kalomji). Mucuna pruriens: Family: Fabaceae

The plant is an annual, climbing shrub with long vines that can reach over 15 m in length. When the plant is young, it is almost completely covered with fuzzy hairs, but when older, it is almost completely free of hairs. The leaves are tripinnate, ovate, reverse ovate, rhombus shaped or widely ovate. The sides of the leaves are often heavily grooved and the tips are pointy. The stems of the leaflets are two to three millimeters long. Additional adjacent leaves are present and are about 5 mm long. The flower heads take the form of axially arrayed panicles. They are 15 to 32 cm long and have two to three, or many flowers. The accompanying leaves are about 12.5 mm long, the flower stand axes are from 2.5 to 5 mm. The bell is 7.5 to 9 mm long and silky. The sepals are longer or of the same length as the shuttles. The crown is purplish or white. The flag is 1.5 mm long. The wings are 2.5 to 3.8 cm long. In the fruit ripening stage, a 4 to 13 cm long, 1 to 2 cm wide, unwinged leguminous fruit develops. There is a ridge along the length of the fruit. The husk is very hairy and carries up to seven seeds. The seeds are flattened uniform ellipsoid, 1 to 1.9 cm long, 0.8 to 1.3 cm wide and 4 to 6.5 cm thick. 7. Aloe vera: Family- Liliaceae

It is also known as the Medicinal Aloe, is a species of succulent plant that probably originated in northern Africa. The species does not have any naturally occurring populations, although closely related Aloes do occur in northern Africa. The species is frequently cited as being used in herbal medicine since the beginning of the first century AD, because it is mentioned in the New Testament (John 19:39-40 And there came also Nicodemus, which at the first came to Jesus by night, and brought a mixture of myrrh and aloes... ). The natural range of A. vera is unclear, as the species has been widely cultivated throughout the world. It has been suggested that naturalised stands of the species occur through North Africa in Algeria, Morocco, and Tunisia, along with the Canary and Madeira Islands. The species was introduced to China, India, Pakistan and various parts of southern Europe in the 17th century.

Table 1: Details of the medicinal herbs used in herbal feed compositions for solving problems of fertility in cattle are as below:

Process for Preparation of Herbal feed compositions:

Method-I

The present invention herbal feed compositions are prepared by one type of method comprising the following steps:

a) Obtaining the part of medicinal herb from a group comprising leaves, bark, root and aerial parts;

b) drying the plant part of step (a);

c) powdering the dried plant material of step (b) to a coarse powder;

d) the dried and powdered plant material obtained in step (c) can be used directly to prepare the feed compositions by mixing the effective amount by weight of medicinal herb selected from the group of Ruta graveolens, Moringa oleifera, Guizotia abyssinic, Tribulus terrestris, Nigella sativa, Mucuna pruriens, and Aloe vera to obtain the herbal composition;

e) the above herbal composition is added with at least one of the ingredient selected from chelated mineral mixture or mineral nutrients or bypass fat or dicalcium phosphate to obtain the herbal feed composition.

Method- II

The present invention herbal feed compositions are prepared by another type of method comprising the steps as below:

a) Obtaining the part of medicinal herb from a group comprising leaves, bark, root and aerial parts;

b) drying the plant part of step (a);

c) powdering the dried plant material of step (b) to a coarse powder;

d) extracting the powdered dried plant material at a temperature in the range of 30 to 85° C;

e) extracting the plant material with water or alcohol or mixture of both for a period ranges from 6 hours to 6 days;

f) concentrating the obtained extract under reduced pressure at a temperature in the range of 40 to 85° C;

g) the concentrated extract is subjected to removal of solvent;

h) mixing the effective amount by weight of above concentrated extract of medicinal herb selected from the group of Ruta graveolens, Moringa oleifera, Guizotia abyssinica, Tribulus tcrrestris, Nigella sativa, Mucuna pruriens, and Aloe vera to obtain the herbal composition; i) the above herbal composition is added with at least one of the ingredient selected from chelated mineral mixture or mineral nutrients or bypass fat or dicalcium phosphate to obtain the herbal feed composition.

Preparation of Chelated Mineral Mixture:

The chelated minerals used in the present invention are as following:

The chelated minerals preferably menthomins chelated are mixed in the proportion of 8% (w/w) zinc, 2% (w/w) manganese, 1.2% (w/w) copper, 0.12% (w/w) cobalt, 0.05% (w/w) selenium, 0.09% (w/w) chromium and 0.04% (w/w) iodine. The chelated mineral mixture now added to the herbal composition in a proportion of 10 to 25% (w/w) of total composition and preferable 15% (w/w) of total composition.

Examples: The following specific examples presented to illustrate the herbal feed compositions for solving fertility problems in cattle prepared by above said method I but do not limit the scope of the invention and additional compositions are being prepared and tested.

TABLE-2: Specific combinations prepared for study are

A)

Experimental Details of the Trials:

The study is conducted on a sample of catties (buffaloes/cows) which are having the problem of fertility or anestrous from many days by one or the other reason. The above selected catties are fed by the herbal composition of present invention specifically composition no. I of Table-2A at a recommended dose of 20g/day/cattle for a period of 10 days and during the experiment and 10 days after the feeding, clinical observations like behavioral signs of estrous, pre-rectal examination for ovarian activity and other studies were conducted. The cattle exhibited heat within 6 to 8 days from the date of feeding and majority of cattle had confirmed their pregnancy.

Species and number of animals under the trials:

Cows: 39 Buffaloes: 109

Type of study:

Double blind Placebo Control Study: The Investigator as well as the AI workers using the product was not aware of the Drug or Placebo status of the product handed over to them. After finalization of the results, the sealed containing the information was opened before the Investigator, Coordinator and the Technical Coordinator of the study from the manufacturer side.

Study Design: Perspective Cohort Design:

The animals were first identified, evaluated and then included in the study. Both the Drug and Placebo cohorts were followed up during the period. All procedures done for Drug Group was also undertaken for Placebo Group.

Study Period: February 2009 to May 2009 SPECIFIC DETAILS OF THE TRIALS

1. Species and numbers of animals:

Buffaloes and Cows, irrespective of breeds.

2. Inclusion Criteria:

Two sets of studies were conducted. In the first set, 26 animals that had open period more than 120 days and were repeat breeders were included in the study.

In the second set studies animals with open period more than 45 days and not exhibited any estrus were included in the study .

3. Exclusion criteria:

Animals with any uterine pathology were excluded. Animals previously treated with any antibiotics, uterotropic drugs, etc., during the current parity were also excluded from the study.

Animals developing any major disease during the course of the trials were also excluded.

Animals with body score less than 3.0 were also not included in the study

4. Observation Period: Three months from the date of start of the treatment

5. Method of Administration: Orally daily once

6. Dose: 20 g daily

7. Frequency of Recording of Observation:

The data of the animals was maintained in Herdman -Server program, through which daily action and alerts were generated. The observation alerts were based on the list generated from the program.

8. Data Recording Procedure:

All the animals included in the study were already identified with a unique identification number and their registration was done in the computer program Herdman-server. The performance details of the animals were available through Herdman. The AI workers / para-veterinary staff were given daily action list / alerts which also included the animals due for heat / pregnancy detection. All events in the animals were recorded in its file. GENERAL METHODOLOGY OF THE TRIALS

Animal Allocation to Groups:

Initially through the custom designed report module the lists of animals with the inclusion criteria were generated. The animals belonged to the farmers spread across 23 different centers within the Chitale Dairy area. In order to avoid any confusion and mix up it was decided to include each of the centers either in Placebo group or Control Group. For example in Center X all eligible animals were included Drug Group whereas Y was included in Placebo group. This allocation was done on random basis to avoid any bias. Thus, the allocation to the Drug or Placebo group was based on the Center random allocation to the group. Since the inclusion and exclusion criteria were followed scrupulously, the animal allocation was found to be uniform. This was verified as comparison of average pen Period' of the 'Drug or the Placebo group was found to be similar (within two standard deviations of the mean).

Observation of Estrus:

In order to maintain uniformity and avoid variation due to variable estrus detection capability of farmers, the system of 'estrus prediction' by Herdman software was followed. On the due date Herdman generated action list which was communicated to the AI worker as well as the farmer (through the sms message). The AI worker visited the farmer and checked the estrus personally. Estrus detection was based on symptoms of estrus and per rectal examination. In case the animal was not found showing any symptom, the AI worker carried out per rectal examination to make the observation definite.

Per rectal Examination:

Per rectal examination of the reproductive tract was performed at each observation point, irrespective whether estrus was there or not. This was done following routine procedures. The AI workers have been trained in this procedure and were doing this kind of examination since at least last more than three years.

Efficacy Criteria:

In the first set study the efficacy criteria was based on both the estrus and pregnancy obtained after insemination. This study was done under farm conditions. In the second study however the efficacy criteria was solely based on anestrous animals exhibiting estrus and pregnancy was recorded later on but not considered as the efficacy criteria. This was done to avoid confounding effects due to insemination- and pregnancy-related related factors.

Statistical Tests:

Chi-square test was performed and value more than 3.84 was considered as significant RESULTS:

The summary results are given in Table 3 and 4, for cows and buffaloes whereas the combined results are depicted in Table 5.

Set 1:

Table 3: Efficacy of Herbal composition in Repeat Breeding Cows and Buffaloes

% Animals for pregnancy status

Pregnant Not pregnant

Drug Group (Herbal Composition) 62.5% 37.5 %

Placebo Group (Control) 40% 60 %

Set 2:

Table 4: Efficacy of Herbal composition in post-parturient anestrous cows and buffaloes

Cows

Number and % animals exhibiting estrus two 21 days cycle

Estrus No estrus Total

Drug Group (Herbal Composition) 14 (70%) 6 (30%) 20

Placebo Group (Control) 09 (50%) 09 (50%) 18

Total 23 15 38 Buffaloes

Number and % animals exhibiting estrus two 21 days cycle

Estrus No estrus Total Total

Drug Group (Herbal Composition) 22 (57.9%) 16 (42.1) 38

Placebo Group (Control) 14 (30.4 %) 32 (69.6%) 46

Total 36 48 84

Table 5: Combined Cows and Buffaloes

Number and % animals exhibiting estriis two 21 days cycle

Estrus No estrus Total

Drug Group (Herbal Composition) 36 (52%) 33 (45%) 69

Placebo Group (Control) 17 (32%) 36 (68%) 53

Total 53 69 122

CONCLUSION:

From the results it is evident that Herbal composition is an effective product in induction of estrus in post-parturient anoestrus syndrome in cows and buffaloes. The product is more effective in crossbred cows than buffaloes.

In the set 1 study the product Herbal composition resulted in 62.5% cases compared to 40% in Placebo group. From the Chi-square statistics these differences were found to be significant (P<0.05).

In the set 2 study which was done for anoestrus cows and buffaloes with an open period more than 120 days, the product was also found to be ^' effective. From Table 3 and 4 it is evident that estrus induction was found in 70% cows and 52% buffaloes compared to 50% and 30.4% in Placebo group. Herbal composition therefore can be recommended in cows and buffaloes with true anestrous syndrome, provided that these animals do not have any uterine pathology or gross energy malnutrition.

A synergistic effect is achieved when the above said herbs are included in a feed composition, preferably at least four herbs over those fed without the herbal feed composition. Thus, one embodiment of the invention provides a method of selecting herbs for compositions according to these principles..

It is an important aspect of the combination herbal preparation of the present invention that it contains a mixture of herbs or extracts of herbs rather than a single herb. An unexpected synergistic effect is exhibited by the various ingredients of the herbal preparation of the present invention. The strategic combination of herbs of the present invention exhibits beneficial effects when optimally combined as discussed above. The advantage of the polyherbal composition also minimizes the risk of development of drug resistance.

Acknowledgments: I sincerely express thanks to Dr. A. Samad, Dean, Bombay Veterinary College (BVC), Mumbai, India for conducting filed trials of the product. I greatly appreciate kind co-operation of Dr. Harish Ingle.