WO2009109988A2

WO2009109988A2 - Animal feed composition

Info

Publication number: WO2009109988A2
Application number: PCT/IN2009/000045
Authority: WO
Inventors: Manget Ram Garg
Original assignee: National Dairy Developement Board (Nddb)
Priority date: 2008-01-16
Filing date: 2009-01-15
Publication date: 2009-09-11
Also published as: WO2009109988A3

Abstract

The present disclosure relates to an improved animal feed composition that enhances milk production and reproduction in cattle. The animal feed composition comprises of dicalcium phosphate, 50-70 % (w/w); calcium carbonate, 15-20 % (w/w); vitamin D3, 1-2 % (w/w) and lubricant, 3-10 % (w/w). The animal feed composition of the present disclosure has enhanced stability and can be stored for longer durations without the problem of rancidity.

Description

ANIMAL FEED COMPOSITION TECHNICAL FIELD

The present disclosure relates to an improved animal feed composition which enhances milk production and reproduction in dairy cattle. BACKGROUND OF ART

Under Indian conditions wherein the quality of forages and fodder available to animal is not good and most of the animals are fed on crop residues, supplementation of Calcium (Ca) assumes importance for productivity and fertility. Therefore, the need arises for additional supplementation of critical nutrients like calcium and phosphorous in order to address the deficiency of these nutrients and improve milk production. Requirement of calcium increases slowly during pregnancy and in high yielding cows; it increases further by 33 per cent or more with onset of lactation. Reduced blood calcium may delay uterine involution and increases the incidence of dystocia, retained placenta and prolapsed uterus. Calcium deficiency is directly not responsible to sub-fertility or fertility but disturbed ratio of calcium and phosphorous (P) has a blocking action on pituitary gland and consequently on ovarian action (Morrow, 1980). Higher ratio of Ca and P has been suggested to be associated with infertility in bovines whereas phosphorus deficiency in ruminants affects growth and reproduction (Kumar et al. 1992).

Non-availability of calcium in the diet results in calcium deficiency which is accentuated at the time of parturition results in clinical hypocalcaemia. Chronic deficiency however leads to a sub clinical syndrome characterized by production and fertility underperformances (Duffield et al., 2005). It is estimated that up to 60% of the herds in USA is suffering from subclinical hypocalcaemia (Horst et al., 1994). There appears to be unanimity in the criteria that if more than 50% of the animals in a herd show blood calcium level lower than 8 mg/dl, the herd can be classified as subclinical. Thus hypocalcaemia is a major field problem in dairy animals.

Moderate P deficiency leads to repeat breeding and low conception rates whereas severe deficiency may delay onset of puberty and post partum estrous because of inactive ovaries (Paugh et al, 1985). Phosphorus is directly related with reproduction and phosphorus supplementation has direct effect on release of gonadotrophins which ultimately induces cyclicity, oestrus and conception during post partum stage. Pasture- based diets are deficient in phosphorus whereas animal fed on grains have adequate P in their diet (Anonymous, 2006). Phosphorus deficient diet leads to phosphorus deficiency syndrome which is characterized by low blood P (less than 4 mg/dl), low conception rate, reduced feed intake, poor feed efficiency, low growth rate and reduced milk production and reproductive failures. The effects of P deficiency are mediated through deficiency of ATP (Samad, A., 1997).

The macro-nutrient deficiency problems are reported to be more common peri- parturient when the animal is in metabolic transition (Overtone and Waldron, 2004). It is estimated that more than 60% of the dairy cows experience ill effects of sub-clinical hypocalcaemia and this is linked to many reproductive disorders, retention of fetal membranes, delayed involution of uterus, ketosis, mastitis, etc (Duffield et ai, 2005). It is therefore important that dairy animals are supplemented with Ca and P especially during metabolic transition.

However, the composition and recommended dosage of most available calcium supplements does not stand the test of scientific evaluation. All the calcium preparations available presently are in liquid form and on critical analysis of the liquid preparation they show certain drawbacks. The first drawback of these liquid preparations is that the nutrients in the preparation including calcium and phosphorus do not remain in suspension for a long time and tend to precipitate. The farmers are forced to shake the container before use and in spite of this a lot of material remains at the bottom resulting in all dosage calculations going haywire. Another drawback is that the liquid preparations cannot hold more than specified quantities of minerals in suspension and hence manufacturers are forced to limit the quantities of these minerals in the preparation. This results in abysmally low levels of calcium and phosphorus. Yet another drawback is that the manufacturing of liquid preparations involves high investments on filling and sealing equipment which is invariably recovered from the farmer, without delivering any tangible benefit. Yet another drawback is that the administering liquid preparations to the animals are laborious, time consuming and often result in wastage.

Most of the issues concerning the liquids can be addressed by giving the product in a solid state. However, the powder form is also not perfect for animal feed and possess some drawbacks such as there is huge wastage of product, poor palatability, cause lots of dust and difficult to administer.

Tablet forms are also known in the art but they suffer from certain drawbacks like huge dosage requirement and high cost of manufacture. Pellet formulation is also known but due to the high content of minerals in the formulation some disadvantages like non formation and frequent breakage of the pellet were encountered along with unusually high die attrition in the pellet mill. The introduction of vegetable oil as a lubricant gives rise to the problems of rancidity. The use of antioxidants along with the vegetable oil is also not found effective in avoiding the problem of rancidity in animal feed compositions.

There is a need for a formulation which could overcome all the drawbacks of the liquid, powder, tablet and pellet forms known in the art as well as that which can bridge the gap between the requirement and availability of the critical minerals in dairy animals with vitamins for efficient absorption of the minerals.

SUMMARY

The present disclosure relates to an improved animal feed composition that enhances milk production and reproduction in cattle, said composition comprising; dicalcium phosphate in the range of 50- 70% (w/w); Calcium carbonate in the range of 15-20% (w/w);

Vit.D3 in the range of 1-2% (w/w); and lubricant in the range of 3-10% (w/w).

BRIEF DESCRIPTION OF ACCOMPANYING DRAWINGS

Fig.l shows total milk yield of buffaloes before start of study on early lactation group (15- 30 days calved)

Fig.2 shows average milk yield per day of buffaloes before start of the study on early lactation group (15-30 days calved).

Fig.3 shows total milk yield of buffaloes during study on early lactation group (30-45 days calved). Fig.4 shows average milk yield per day of buffaloes during study on early lactation group

(30-45 days calved).

Fig.5 shows total milk yield per week of buffaloes before start of study on mid lactation group (75-90 days calved).

Fig.6 shows average milk yield per day of buffaloes before start of study on mid lactation group (75-90 days calved).

Fig.7 shows total milk yield per week of buffaloes during study on mid lactation group (90-

105 days calved).

Fig.8 shows average milk yield per day of buffaloes during study on mid lactation group

(90-105 day calved). DETAILED DESCRIPTION

The present disclosure relates to an improved animal feed composition that enhances milk production and reproduction in cattle, said composition comprising dicalcium phosphate in the range of 50-70%(w/w); calcium carbonate in the range of 15-

20% (w/w); vit.D3 in the range of 1-2% (w/w); and lubricant in the range of 3-10% (w/w). In the present disclosure, the expression "services per conception" refers to the number of times that artificial insemination or natural service is required to be done before the female animal conceives.

In the present disclosure, the expression "days open" refers to the number of days between calving to the date of the next confirmed pregnancy.

In the present disclosure, the expression "per rectal examination" refers to a method of diagnosing disorders of the reproductive tract or confirming pregnancy used by medical veterinarians, wherein the veterinarians insert their hand through the rectum of the animal and feel the genitalia. In the present disclosure the expression "Vitamin D3"and "Vit.D3" is used interchangeably herein after.

An embodiment of the present disclosure relates to an animal feed composition in the form of pellet.

Another embodiment of the present disclosure relates to an animal feed composition wherein, the animal feed composition comprises a lubricant. The lubricant is selected from a group consisting of bypass fat and vegetable oils that include rice bran oil and rice polish.

Another embodiment of the present disclosure relates to an animal feed composition, wherein the animal feed composition comprises of bypass fat as a lubricant.

In an embodiment of the present disclosure relates to an animal feed composition is that the disadvantages of conventionally used pellets is overcome by introducing by pass fat as a lubricant in the animal feed composition. The bypass fat is inert, rancid free and contributes to the nutritive value of the pellet. It also enriches the milk fat and provides energy to the cattle.

Further, another embodiment of the present disclosure relates to an animal feed composition wherein Vitamin D3 is coated with isopropyl alcohol. The analysis of homoeostatic mechanisms in the body with respect to calcium indicates the influence of specific hormones like calcitonin and parathormone on calcium absorption. Vitamin D3 is also included in the formulation owing to its major role in the absorption of calcium from intestine. The vitamin D3 is susceptible to degradation and therefore to enhance the stability of VitaminD3 in the composition of the present disclosure, Vitamin D3 is coated with isopropyl alcohol.

Still another embodiment of the present disclosure relates to an animal feed composition, wherein dicalcium phosphate is in the range of 55-65 %( w/w). Yet another embodiment of the present disclosure relates to an animal feed composition wherein the dicalcium phosphate is 60 %( w/w).

Another embodiment of the present disclosure relates to an animal feed composition wherein the calcium carbonate in the range of 17-19 %( w/w). Another embodiment of the present disclosure relates to an animal feed composition wherein the calcium carbonate is 19 % (w/w).

Still another embodiment of the present disclosure relates to an animal feed composition wherein the vitamin D3 in the range of 1.2- 1.8 % (w/w).

Yet another embodiment of the present disclosure relates to an animal feed composition wherein the vitamin D3 is 1 %( w/w).

Another embodiment of the present disclosure relates to an animal feed composition wherein the lubricant in the range of 5-8 %( w/w).

Another embodiment of the present disclosure relates to an animal feed composition wherein the lubricant is in the range of 6-7 %( w/w). Further another embodiment of the present disclosure relates to an animal feed composition optionally comprises sweeteners in the range of 0.05-0.15 % (w/w).

Another embodiment of the present disclosure relates to an animal feed composition, wherein the sweetener includes saccharine.

Another embodiment of the present disclosure relates to an animal feed composition optionally comprises sweeteners in the range of 0.06-0.12 % (w/w).

Still another embodiment of the present disclosure relates to an animal feed composition optionally comprises starch in the range of 5-10% (w/w).

Another embodiment of the present disclosure relates to an animal feed composition optionally comprises starch in the range of 6-8 % (w/w). Yet another embodiment of the present disclosure relates to an animal feed composition optionally comprises of oil bran in the range of 6-10% (w/w).

Another embodiment of the present disclosure relates to an animal feed composition optionally comprises of oil bran in the range of 7-9 % (w/w).

Yet another embodiment of the present disclosure relates to an animal feed composition, wherein the preservative premix is selected from the group consisting of methyl paraben and propyl paraben.

Another embodiment of the present disclosure relates to an animal feed composition optionally comprises of preservative premix in the range of 0.5-2 %( w/w) Still another embodiment of the present disclosure relates to an animal feed composition optionally comprises of preservative premix in the range of 1-1.5 %( w/w).

Further another embodiment of the present disclosure relates to an animal feed composition, wherein the pellet has diameter ranging from 3-8 mm. Another embodiment of the present disclosure relates to an animal feed composition, wherein the pellet has 3 mm diameter.

Yet another embodiment of the present disclosure relates to an animal feed composition, wherein the pellet has 6 mm diameter.

Another embodiment of the present disclosure relates to an animal feed composition, wherein the pellet has 8 mm diameter.

In another embodiment, the present disclosure provides an animal feed composition that has extended shelf life. The composition of the present disclosure can be stored for long duration which can be over 12 months.

In an embodiment, the present disclosure provides an animal feed composition having surprisingly improved conceptions in cattle.

In an embodiment, the present disclosure provides an animal feed composition, wherein of interactions studies of the animal feed composition on the various minerals in the digestive tract is carried out {Handbook of animal nutrition. G.C.Banerjee). The optimal ratio of the various minerals which includes phosphorous and calcium required by cattle is analyzed and used for formulation of the composition of the present disclosure. The optimal ratio of calcium and phosphorus analysed and used for formulation of the composition of the present disclosure ranges from 1 : 1 to 2: 1.

In an embodiment, the present disclosure provides an animal feed composition provides finalization of the source of the minerals in the animal feed composition. A host of materials like tricalcium phosphate, dicalcium phosphate, monocalcium phosphate, phosphoric acid and calcium carbonate were used to evaluate the bioavailability data, cost data and mode of their action to formulate the composition of the present disclosure.

In an embodiment, the present disclosure provides an animal feed composition, wherein the phosphate source used is selected from a group ranging from dicalcium phosphate, tricalcium phosphate, monocalcium phosphate and phosphoric acid.

In an embodiment, the present disclosure provides an animal feed composition, wherein the calcium source used is selected from a group ranging from calcium carbonate, dicalcium phosphate, tricalcium phosphate and monocalcium phosphate. In an embodiment, the present disclosure provides an animal feed composition, wherein the dicalcium phosphate provides high phosphate and calcium bioavailability.

In an embodiment, the present disclosure provides an animal feed composition, wherein the phosphoric acid provides high calcium bioavailability. Further, another embodiment of the present disclosure provides that the pellet of present disclosure resulted in better milk composition when compared to commercial liquid supplement. Serum profile data as discloses in example 5, shows significantly higher serum calcium in cows given pellets of present disclosure than the cows given commercial liquid supplement. Still another embodiment of the present disclosure provides that the pellet of present disclosure is more effective for increase in milk production. The increase is 19.04 per cent in cows and 31.76 per cent in buffaloes and there is significant difference in milk production found in both cows and buffaloes after feeding of the pellets of present disclosure as compared with that of commercial liquid supplement. The details are provided in example 6.

Another embodiment of the present disclosure is that the pellet of present disclosure is better in high yielding buffaloes because of its positive impact in milk yield and consistency in comparison with the commercial liquid supplement. The pellet of present disclosure shows significant (p<0.05) high milk yield in early lactation buffaloes and mid lactation buffaloes as is provided in example 7.

Yet another embodiment of the present disclosure is that the cows and buffaloes given pellet of present disclosure had better fertility indices in terms of calving to conception and services per conception as is provided in example 8.The number of animals open in group feeding with commercial liquid supplement was also higher than the group feeding with the pellet of present disclosure.

Another embodiment of the present disclosure is that the post partum ovarian rebound and conceptions were found to be higher in cows and buffaloes on supplementation of the pellets of present disclosure as compared to commercial liquid supplement as provided in example 9. From this example it can be concluded that supplementation of proper Ca: P ratio is useful in lactating animal where Ca helps to increase milk production and P improves reproductive efficiency in post partum animals.

The following examples are given by way of illustration of the present invention and should not be construed to limit the scope of present disclosure. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are intended to provide further explanation of the claimed subject matter.

EXAMPLES Example 1 Preparation of animal feed composition

The components of the formulation were weighed with the help of electronic scale. Dicalcium phosphate was used as the phosphorous and calcium source. Calcium carbonate was further used as the source of calcium. The Vitamin D3 used in preparation of the animal feed composition was coated with isopropyl alcohol. The components used for preparing the composition was in the ratio, dicalcium phosphate 60% (w/w), calcium carbonate 19% (w/w), starch 5% (w/w), preservative premix 0.6-0.8% (w/w), Vit.D3 1 % (w/w), sweetener 0.08-0.1% (w/w), lubricant 6-7% (w/w), Oil bran 8% (w/w). The lubricant used was bypass fat. The components of the animal feed composition were dumped into the dumping pit of the feed mill followed by transportation of the components to the mixer via the bucket elevator. The resultant mixture was blended for 10-15 minutes. After blending the mixture was transferred to the conditioner through another bucket elevator. In the conditioner steam was injected at 80-100⁰C and cooking was done for 1 minute. After the conditioning the mixture was passed through the pellet mill and forced out through an 8mm die. This resulted in the formation of 8mm pellets. These pellets were then passed through a cooler and kept for 1 hour. After discharging from the cooler, the pellets were ready for bagging and dispatch. Example 2 Preparation of animal feed composition

The components of the formulation were weighed with the help of electronic scale. Dicalcium phosphate was used as the phosphorous and calcium source. Calcium carbonate was further used as the source of calcium. The Vitamin D3 used in preparation of the animal feed composition was coated with isopropyl alcohol. The components used for preparing the composition was in the ratio, dicalcium phosphate 55% (w/w), calcium carbonate 18% (w/w), starch 9% (w/w), preservative premix 1.7-1.9% (w/w), Vit.D3 2% (w/w), sweetener 0.13-0.15% (w/w), lubricant 5-6% (w/w), Oil bran 9% (w/w). The lubricant used was bypass fat. The components of the animal feed composition were dumped into the dumping pit of the feed mill followed by transportation of the components to the mixer via the bucket elevator. The resultant mixture was blended for 10-15 minutes. After blending the mixture was transferred to the conditioner through another bucket elevator. In the conditioner steam was injected at 80-100⁰C and cooking was done for 1 minute. After the conditioning the mixture was passed through the pellet mill and forced out through a 6mm die. This resulted in the formation of 6mm pellets. These pellets were then passed through a cooler and kept for 1 hour. After discharging from the cooler, the pellets were ready for bagging and dispatch. Example 3 Preparation of animal feed composition

The components of the formulation were weighed with the help of electronic scale. Dicalcium phosphate was used as the phosphorous and calcium source. Calcium carbonate was further used as the source of calcium. The Vitamin D3 used in preparation of the animal feed composition was coated with isopropyl alcohol. The components used for preparing the composition was in the ratio, dicalcium phosphate 65% (w/w), calcium carbonate 16% (w/w), starch 6% (w/w), preservative premix 1% (w/w), Vit.D3 1.5% (w/w), sweetener 0.1% (w/w), lubricant 4.4 % (w/w), Oil bran 6% (w/w). The lubricant used was bypass fat. The components of the animal feed composition were dumped into the dumping pit of the feed mill followed by transportation of the components to the mixer via the bucket elevator. The resultant mixture was blended for 10-15 minutes. After blending the mixture was transferred to the conditioner through another bucket elevator. In the conditioner steam was injected at 80-100⁰C and cooking was done for 1 minute. After the conditioning the mixture was passed through the pellet mill and forced out through a 3mm die. This resulted in the formation of 3mm pellets. These pellets were then passed through a cooler and kept for 1 hour. After discharging from the cooler, the pellets were ready for bagging and dispatch. Example 4 Preparation of animal feed composition

The components of the formulation were weighed with the help of electronic scale. Dicalcium phosphate was used as the phosphorous and calcium source. Calcium carbonate was further used as the source of calcium. The Vitamin D3 used in preparation of the animal feed composition was coated with isopropyl alcohol. The components used for preparing the composition was in the ratio, dicalcium phosphate 58% (w/w), calcium carbonate 17% (w/w), starch 8% (w/w), preservative premix 2% (w/w), Vit.D3 1.2% (w/w), sweetener 0.09-0.1% (w/w), lubricant 6-7 % (w/w), Oil bran 7% (w/w). The lubricant used was bypass fat. The components of the animal feed composition were dumped into the dumping pit of the feed mill followed by transportation of the components to the mixer via the bucket elevator. The resultant mixture was blended for 10-15 minutes. After blending the mixture was transferred to the conditioner through another bucket elevator. In the conditioner steam was injected at 80-100⁰C and cooking was done for 1 minute. After the conditioning the mixture was passed through the pellet mill and forced out through a 8mm die. This resulted in the formation of 8mm pellets. These pellets were then passed through a cooler and kept for 1 hour. After discharging from the cooler, the pellets were ready for bagging and dispatch. Example 5 Comparison of Milk production in dairy cows and buffaloes with the pellet of the present disclosure and commercial liquid supplement

The studies were conducted in two organized buffalo farms rearing 220 and 160 lactating buffaloes, respectively. The farms did not add mineral supplementation during last several years and were fed on grain-based concentrate and paddy straw. The farms had high incidence of mastitis and metabolic problems, such as, milk fever. For trials in cow three organized HF crossbred cow farms consisting of 80, 54 and 72 lactating buffaloes were selected. These farms were also detected with problem of sub-clinical calcium deficiency on routine metabolic profiling under a research project. In these farms mineral mixture supplementation was occasional. These farms encountered high incidence of mastitis and metabolic problems such as milk fever / ketosis. The studies were conducted on 60 lactating cows and buffaloes, each, (DIM less than 75 days). To analyze variations due to sampling, the animals were allocated to pellet of the present disclosure or commercial liquid supplement group on matching based on breed, DIM and lactation yield.

Trials period: The trials were conducted for 90 days with at least 10 days supplement acclimatization prior to start of the trails.

Recording of performance and sample analysis: Serum calcium and phosphorus was determined at the start of the trials and after the end of 90 days of supplementation period. The estimation was undertaken using the commercial kits for UV Spectrophotometer. Milk yield was recorded every 10 days (evening and next day morning record) and the pooled milk samples were also collected for fat, protein and total solids employing electronic milk analysis method. These farms were maintaining herd records on the software program 'Herdman', from where the data on heat, services and pregnancy was obtained. Data analysis: The data was analyzed for student't' test using the standard method (Snedecor and Cocheron). The Student's t-distribution (or also t-distribution) is a probability distribution that arises in the problem of estimating the mean of a normally distributed population when the sample size is small. It is the basis of the popular Student's t-tests for the statistical significance of the difference between two sample means, and for confidence intervals for the difference between two population means. The standard error of mean and the deviation in the study are measured by the parameters SEM and CD respectively.

For the present trials the farms that were afflicted with problem of sub-clinical hypocalcaemia were selected in order to study comparative efficacy. It was hypothesized that when the animals in the herd have normal blood calcium levels, proving the biological availability is rather equivocal. For evaluating efficacy the criteria were: serum Ca and P levels at the end of supplementation, milk production in terms of yield and fat production, reproductive performance in terms of number of AI per conception and calving to conception period. Since the animal allocation to the two groups was based on 'matching' the comparison of the parameters at the herd level (as mean of the herd) was considered appropriate.

The milk production data in crossbred cow and buffalo suggest that the milk and fat yield in calcium pellet group was higher than the commercial liquid supplement group. The milk production, fat production and total solid yield in cow and buffalo herds supplemented with calcium pellet was found to be significantly higher than the commercial liquid supplement group (Table 1).

U Table 1: Effect of Pellet of present disclosure and commercial liquid supplement on Milk Production in Crossbred Cows and Buffaloes

*Data p > 0.05 when compared with paired't' test Group A and B is compared for total production in 90 days

There are a number of reports to suggest that supplementation of Ca in lactating cows or buffaloes has beneficial effects and the magnitude of these would depend on the net bioavailability of Ca, which largely depends on its composition (Keskar et. al, 2005). Since Ca is important for milk synthesis its supplementation would lead to better yield. It is rather difficult to compare the milk yield in these animals prior to supplementation since these were in varying lactation days. The previous lactation data of these animals was not available however based on the perception of the farmers it was deduced that after supplementation the variation in milk production and the yield was better.

Since the cow as well as buffalo farms selected for the trials were sub-clinically hypocalcemic, the serum calcium levels post-supplementation was a proof of comparative efficacy of the two supplements, namely the animal feed composition in the form of pellet (pellet of the present disclosure) and the commercial liquid supplement. Table 2 shows that the serum profile data, which suggests that the cows and buffaloes supplemented with the pellet of the present disclosure had significantly higher serum calcium level at day 100 when compared with that at the start of the trials. These cows and buffaloes also had significantly higher serum calcium than the commercial liquid supplement group at day 100. Serum calcium level in commercial liquid supplement group was higher when compared with at the start of the trial although it was not significant statistically. Similar results were obtained for serum phosphorous levels after supplementing the diet of the buffaloes of the present study with the animal feed composition. Animals of these farms were largely fed on grain-based concentrate diet and rice straw hence even though there was no supplementation of mineral mixture the blood P levels were normal even at the start of the supplementation. The data clearly indicate that pellet of the present disclosure had better bio-availability than commercial liquid supplement. The effect was attributed to better palatability of pellet of the present disclosure.

Table 2: Efficacy of Pellet of present disclosure and commercial liquid supplement on Replenishing Blood Ca and P in crossbred cows

* Data p > 0.05 when serum calcium at start of the supplement compared with 100^th day in the same treatment group.

# Date p> 0.05 when compared between the pellet of present disclosure and commercial liquid supplement

Example 6

Comparison of Milk production in dairy cows and buffaloes on administering commercial liquid supplement and pellet of the present disclosure

Studies were conducted to evaluate the efficacy of pellet of the present disclosure in fourty Holstein Fresian crossbred cows (HF cows/ HF)) and fourty Murrah buffaloes (MB) with mid-lactational stage from commercial dairy units located around college campus. Healthy and smoothly calved animals were selected for the present study only after carrying out general clinical examinations. All the animals selected under trial are being maintained on balanced and optimum diet as per the production performance. The trial was undertaken in animals with mid lactation stage only after confirming consistency of uniform milk production. Milk performance of the selected animals was assessed and recorded for a period of fifteen days. Twenty HF cows and twenty Murrah buffaloes were administered with 100 Gms special feed pellets (containing Ca-200000 mgs, P-100000 mgs Vit.D3- 40,000 IU per kg pellets carrying starch and sugar manufactured by Indian Immunologicals Limited, Hyderabad) daily for fifteen days which formed Group-HFl and MBl of the trial. Similarly, twenty HF cows and twenty Murrah buffaloes were administered with market preparation of 100 ml liquid mineral supplementation (containing Ca -1650 mg, P-85Omg, vit. D3-8000 IU, vit. B12-100 meg, Carbohydrate-40,000 mg per 100 ml liquid) daily for fifteen days which served as Group-HF2 and MB2 of the trial. Both the treatments were administered orally along with regular concentrates. AU the treated animals were followed daily and increment in milk performance was recorded for fifteen days. The recorded data was analyzed by't' test based on range (Snedecor and Cochran, 1967). The animals under trial were examined per rectally before initiation of treatment for assessment of ovarian activity. Animals were kept under keen observation to record oestrus during the trial period. Breedings were carried out in animals showing oestrus. Reproductive performance of the animals was also evaluated under the trial.

Lactational observations: The trial was emphasized on mid lactating animals with presumption that the period will not show any fluctuation in milk production due to intrinsic factors. The milk production performance prior to experimentation and after trial is detailed in the table 1 for cows and buffaloes. In case of cattle milk production was found to be increased by 19.04 per cent with special feed pellets while it was recorded as increased by only. 12.60 per cent with liquid mineral supplementation. The highest increase in the milk production i.e. 31.67 per cent was recorded in pellet of the present disclosure. The minimum increase in milk production of 4.90 per cent was recorded for known liquid supplementation group. The increase in milk production by administering pellet of the present disclosure group was found to be 50 per cent more than that for known liquid supplementation and statistically significant difference was recorded in the groups.

In case of buffaloes, milk production was found to be increased by 31.76 per cent in special feed pellets group while the same was found to be increased only by 17.34 per cent in known liquid mineral supplementation. The highest increase in the milk production i.e. 54.58 per cent was recorded in special feed pellets group whereas the minimum increase in milk production i.e. 5.83 per cent was recorded in known liquid mineral supplementation group. The increase in milk production in special feed pellets group was found to be double than the known liquid mineral supplementation group and the difference in both the groups was recorded as statistically significant.

Under field conditions, routinely animals are fed with market available different type of mineral supplements. It is common observation that any commercial mineral supplementation improves milk yield by 10 to 15 per cent. It was surprisingly found that administration of the pellet of the present disclosure improves milk yield by 31.76 per cent (Table 3-5).

Table 3: Comparative increase in milk production of lactating animals on feeding pellet of present disclosure and commercial liquid supplement.

Values against '+' shows standard error. Each value is mean of twenty replications. ** The values are significantly higher at 1% and 5% level of significance.

Table 4: Comparative increase in milk production due to pellet of present disclosure and commercial liquid supplement in cows.

** Value is significantly higher at both 1% and 5% level of significance. * Each value is an average of 20 replications.

Table 5: Comparative increase in milk production due to pellet of present disclosure and

commercial liquid supplement in Buffaloes.

** Value is significantly higher at both 1% and 5% level of significance.

* Each value is an average of 20 replications.

Example 7

Comparison of Milk production in dairy cows and buffaloes on administering pellet of the present disclosure and commercial liquid supplement

The present study was planned with the aim to see the comparative efficacy of pellet of the present disclosure versus commercial liquid supplement in high yielding buffaloes. For the present study 126 buffaloes were selected. Out of which, 63 were in early lactation or 30-45 days after calving (Group A) and 63 were in mid lactation or 90-105 days after calving (Group B). In each of the two groups A and B, 63 buffaloes were further subdivided into three groups, namely group fed with pellet of the present disclosure comprising 45 buffaloes, group fed with commercial liquid supplement comprising 6 buffaloes, group fed with typical concentrate ration comprising 12 buffaloes were taken.

All the buffaloes were subjected to same feeding regime consisting of the concentrate feed comprising of ground maize, wheat bran, pulse chuni, rice bran, mustard cake and common salt. The concentrate mixture consisted of ground maize 18%; wheat bran 22%; pulse chuni 20%, rice bran 15%; mustard cake 24%; common salt 1%. All the feed ingredients were soaked for 10-12 hours before being offered to buffaloes. Group A of buffaloes were offered 8 kg concentrate per day with 10 kg of wheat bhusa while group B of buffaloes were offered 10 kg concentrate with 10 kg wheat bhusa. No green fodder was available for the feeding of buffaloes, as the same was not available during the course of the present study. The buffalo fed with the pellet of the present disclosure were given a dose of 100 gm per day while the buffaloes fed with liquid supplement given a dose of 100 ml per day.

Prior to analyzing the efficacy of the pellet and commercial liquid supplement in the present study, milk yield of the buffaloes was recorded continuously for two weeks. After the initial recording, based on their milk production, buffaloes were randomly allotted to various groups by which average milk yield among different groups was more or less similar and statistically it did not differ significantly. After the initial milk recording of two weeks, the feeding commercial liquid supplement and the pellet, the milk yield of the buffaloes was recorded daily for four weeks (28 days). Data obtained was analyzed using Analysis of Variance and Duncan's New Multiple Range Test.

The milk production data recorded during the study is summarized and processed for statistical significance test. The result has been summarized for early lactation and mid lactation animals separately and has been presented as follows. Milk Yield in Early Lactation group

Period of study was 15 days before feeding pellet of the present disclosure and commercial liquid supplement. During first week of observation, the average weekly milk yield recorded for different groups ranged from 57.33 ± 0.88 to 55.00 ± 0.08 Its/ buffaloes / week. During the second week, milk yield ranged from 70.66 ± 0.66 to 67.66 ± 2.33 Its / buffaloes / week. It has been observed that after the calving of buffaloes, let down of milk gradually increases as a result of it in the second week there was increase in the milk production among buffaloes (Table 6).

Table 6: Milk yield of buffaloes before feeding pellet of the present disclosure and commercial liquid supplement (15-30 days calved)

Values bearing similar superscripts in the same row did not differ significantly (p<0.05).

During the first week after feeding pellet of the present disclosure and commercial liquid supplement, the milk yield ranged from 71.66 ± 0.44 to 73.66 ± 0.331ts / buffalo/ week. Significantly (p<0.05) higher milk yield was recorded in buffaloes fed with pellet of the present disclosure compared to buffaloes fed with commercial liquid supplement and control buffaloes (72.66 ± 0.33 vs. 71.66 ± 0.44 and 73.66 ± 0.33 lts/animal/week). Similarly, during the second week of administering pellet and commercial liquid supplement, the mean weekly milk yield of buffaloes fed with pellet of the present disclosure was significantly (p<0.05) higher than those fed with commercial liquid supplement and control group (78.00 ± 0.33 vs. 75.33 ± 0.33 and 73.00 ± 0.57 Its/ buffalo /week). The trend continued during the third and fourth week and comparatively higher milk yield was recorded in buffaloes fed with pellet of the present disclosure and those fed with commercial liquid supplement and control group (82.33 ± 0.88 vs. 78.66 ± 0.72, 73.66 ± 0.33 Its / buffalo/ week) and (84.00 ± 0.57 vs. 81.83 + 0.44, 73.00 + 0.33 Its / buffalo / week), respectively for third and fourth weeks.

The data in Table 7 provides performance of buffaloes fed with pellet of present disclosure to be superior in milk to those fed with commercial liquid supplement.

Table 7: Milk Yield of Buffaloes during 28 days of feeding pellet of present disclosure and commercial liquid supplement (30-45 Days Calved)

• Values bearing similar superscripts in the same row did not differ significantly

(p<0.05). The consistency of milk production in buffaloes in different groups has been presented graphically. For the period before the start of experiment, the amount milk production it is presented in figure 1 and 2 which disclose that with increase in time the milk yield of all the groups showed a similar rising trend and no significant difference was observed. Thus, in all the animals there was consistency in milk production. Similarly consistency of milk production in buffaloes in different groups after the start of experiment is depicted in figure 3 and 4.The graph (Fig 3 and 4) depicts that the trend observed in milk yield of buffaloes fed with the pellet of present disclosure shows an upward trend with the increase in lactation days. Similarly, somewhat upward trend was observed in buffaloes fed with commercial liquid supplement, but the milk yield in buffaloes fed with typical concentrate ration with no mineral supplement showed almost static milk yield trend without any upward growth. Therefore, there is significant difference in milk yield between animals given liquid supplement vis-a-vis animals fed with the pellets of the present disclosure. Milk Yield in mid lactation group

Period of study was 15 days before feeding pellet of the present disclosure and commercial liquid supplement. During first week of observation the average weekly milk yield recorded for different groups ranged from 85.33 ± 0.88 to 86.0 ± 0.33 Its/ buffalo/ week. During the second week, milk yield ranged from 86.33 ± 0.33 to 86.66 ± 0.33 Its / buffalo/ week. No significant difference was observed in the milk yield of various groups. Thus, the buffaloes were allotted to their respective groups. Table 8: Milk Yield of Buffaloes before feeding pellet of the present disclosure and commercial liquid supplement (75-90 Days Calved)

• Values bearing similar superscripts in the same row did not differ significantly (p<0.05).

During the first week of the study, the milk yield ranged from 86.66 ± 0.88 to 88.00 ± 0.33 Its / buffalo/ week as shown in Table 9. Significantly (p<0.05) higher milk yield was recorded in buffaloes fed with the pellet of present disclosure compared to buffaloes fed with the commercial liquid supplement and control animals (88.00 ± 0.33 vs. 87.83 ± 0.44 and 86.66 ± 0.88). Similarly, during the second week of the present experiment, the mean weekly milk yield of buffaloes fed with the pellet of present disclosure was significantly (p<0.05) higher then those fed with commercial liquid supplement and control group, 90.33 ± 0.33 vs. 88.50 ± 0.28 and 86.33 + 0.33 Its/ buffalo/week respectively. The trend continued during the third week and comparatively higher milk yield was recorded for buffaloes fed with the pellet of present disclosure that those fed with commercial liquid supplement and control group, 92.50 ± 0.33 vs. 88.66 ± 1.58, 86.66 ± 0.66 Its / buffalo/ week respectively. The trend continued during the fourth week and comparatively higher milk yield was recorded for buffaloes fed with the pellet of present disclosure that those fed with commercial liquid supplement and control group, 93.66 ± 0.33 vs. 89.50 ± 0.76, 86.66 ± 0.33 respectively for fourth week. Therefore, from the above it was found that that performance of buffaloes fed with the pellet of present disclosure was comparatively superior to those fed with commercial liquid supplement in mid lactation stage and similar trend was observed in early lactating stage. Table 9: Milk yield of buffaloes during trial (90-105 days calved)

The graph presented in figure 5 and figure 6 depicts that before start of trial the milk yield of buffaloes of different groups showed no significant increasing or decreasing trend and 5 were almost consistent.

In the mid lactation group almost consistent trend was observed in milk yield of buffaloes fed with the pellet of present disclosure over a period of four weeks. Similar trend was observed in control group without mineral supplement however milk yield in this group was significantly lower than those fed with the pellet of present disclosure. However,

10 somewhat upward trend was observed in group fed liquid supplement after second week but the mean milk yield was significantly lower than those fed Ca pellet. The graph presented above depicts that after the start of trial the milk yield of buffaloes of different groups increased but the increasing trend was highly significant in case of calcium pellet fed group as compared to either liquid supplement of without supplement group.

15 Example 8

Comparison of Reproduction or Fertility in dairy cows and buffaloes

The studies were conducted in two organized buffalo farms rearing 220 and 160 lactating buffaloes, respectively. These farms were detected with problem of sub-clinical calcium deficiency on routine metabolic profiling conducted to investigate fertility

20 problems. The farms did not add mineral supplementation during last several years and were fed on grain-based concentrate and paddy straw. For study in cow three organized HF crossbred cow farms consisting of 80, 54 and 72 lactating buffaloes were selected. In these farms mineral mixture supplementation was occasional. The studies were conducted on 60 lactating cows and buffaloes, each, (Days in milk, DIM less than 75 days). To analyze

25 variations due to sampling, the animals were allocated to pellet of the present disclosure or commercial liquid supplement group on matching based on breed, DIM and lactation yield. The studies were conducted for 90 days with at least 10 days supplement acclimatization prior to start of the trails.

The cows and buffaloes were kept under observations for a period of around 100 days after start of the supplementation. None of the cow and buffalo when included in the studies was bred and the record of exhibition of estrus in these animals was not available. The data therefore was analyzed for the two groups after the end of the studies for calving to conception interval (C-CI) and number of services per conception. The percent cows and buffaloes remain open during the study period was also calculated. The data for crossbred cows and buffaloes is depicted in Table 10. The results indicate that crossbred cow group supplemented with the pellet of present disclosure had shorter C-CI (PO.05) when compared with commercial liquid supplement, whereas, in buffaloes the average C-CI was shorter in group supplemented with the pellet of present disclosure but the difference was not statistically significant. The parameter services per conception in groups supplemented with the pellet of present disclosure both in cow and buffalo herds were significantly lower suggesting better fertility in this group as shown in Table 10. When the number of cows and buffaloes open at the end of the present study was considered (open days >150 days). The proportion of such cows and buffaloes in group supplemented with the pellet of present disclosure was lower both in crossbred cows and buffaloes. The results also indicate that the average open days in buffalo group supplemented with the pellet of present disclosure and group supplemented with commercial liquid supplement were higher than crossbred cows.

Table 10: Effect of Pellet of present disclosure and commercial liquid supplement on Calving to Conception Interval and Services per Conception in Crossbred Cows and Buffaloes

• Data p > 0.05 when compared with paired 't' test Group A and B is compared for data at 100^th day post-supplementation Example 9 Comparison of Reproduction or Fertility in dairy cows and buffaloes Present studies were conducted to evaluate the reproductive efficacy of the pellet of the present disclosure in fourty Holstein Fresian crossbred cows and fourty Murrah buffaloes with mid-lactational stage from commercial dairy units. Healthy and smoothly calved animals were selected for the present study only after carrying out general clinical examinations. It was assured that all the animals selected under study were maintained on balanced and optimum diet as per the production performance. The cows and buffaloes under study were examined per rectally before initiation of treatment for assessment of ovarian activity. Cows and buffaloes were kept under keen observation to record oestrus during the study period. Breedings were carried out in cows and buffaloes showing oestrus. Observations on reproductive parameters are provided in table 11. The pellet of present disclosure resulted into reduction of anoestrus cases in cows and buffaloes from 65.00 per cent to 10.00 per cent and 80.00 per cent to 20.00 per cent respectively, whereas the anoestrus was reduced from 60.00 per cent to 15.00 per cent and 75.00 to 50.00 per cent respectively. Oestrus was found to be induced in 40.00 per cent cows and 35.00 per cent buffaloes after treatment with the pellet of present disclosure, whereas it was 25.00 per cent in cows and 10.00 per cent in buffaloes after commercial liquid supplementation. Present findings clearly indicate comparatively higher improvement in reproductive parameters after feeding with pellet of the present disclosure in cows and buffaloes.

The above results clearly indicate that for better production and fertility it is important that the animals are supplemented with calcium and phosphorus supplement that with right salt composition. From the present study it can deduced that Ca and P in the pellet of present disclosure has a better bio-availability in dairy cow and buffalo since its supplementation resulted in comparatively better production and fertility performance.

Table 1 1: Comparison of reproductive performance of cows and buffaloes fed with Pellet of present disclosure and commercial liquid supplement

Figures in parenthesis indicate percentages.

Claims

I/ We Claim:

1. An animal feed composition that enhances milk production and reproduction in cattle, said composition comprising;

Dicalcium phosphate in the range of 50-70 %( w/w); Calcium carbonate in the range of 15-20% (w/w);

Vitamin D3 in the range of 1-2% (w/w) and lubricant in the range of 3-10% (w/w).

2. The animal feed composition as claimed in claim 1, wherein said composition is in the form of pellet.

3. The animal feed composition as claimed in claim 1, wherein said lubricant is selected from a group consisting of bypass fat and vegetable oil.

4. The animal feed composition as claimed in claim 1, wherein said lubricant is bypass fat.

5. The animal feed composition as claimed in claim 1, wherein the Vitamin D3 is coated with isopropyl alcohol.

6. The animal feed composition as claimed in claim 1, wherein said dicalcium phosphate is in the range of 55 - 65 % (w/w).

7. The animal feed composition as claimed in claim 1, wherein said calcium carbonate is in the range of 17 - 19 % (w/w). 8. The animal feed composition as claimed in claim 1, wherein said Vitamin D3 is in the range of 1.2 - 1.

8 % (w/w).

9. The animal feed composition as claimed in claim 1, wherein said lubricant is in the range of 5 - 8 % (w/w).

10. The animal feed composition as claimed in claim 1 optionally comprising sweeteners in the range of 0.05 - 0.15 % (w/w).

1 1. The animal feed composition as claimed in claim 1 optionally comprising starch in the range of 5 - 10 % (w/w).

12. The animal feed composition as claimed in claim 1 optionally comprising oil bran in the range of 6 - 10 % (w/w).

13. The animal feed composition as claimed in claim 1 optionally comprising preservative premix in the range of 0.5 - 2 % (w/w).

14. The animal feed composition as claimed in claim 13, wherein said preservative premix is selected from the group consisting of methyl paraben and propyl paraben.