NOVEL SUPPLEMENTS FOR POULTRY AND LIVESTOCK

Field Of Invention

The subject matter of the present invention relates to novel poultry and livestock supplements that can replace the use of antibiotics, and at the same time provide nutritive value that can enhance the growth and productivity of poultry and livestock. Background Of The Invention

The present invention is directed to novel supplements for poultry and livestock. The supplement formulation comprises of synergistic combinations of probiotics, methylsulfonylmethane, saccharides, vitamins, carotenoids, xanthophylls, minerals and electrolytes. The supplements can effectively replace the use of antibiotics in poultry and livestock and at the same time serve as a source of pronutrients that enhance growth, weight gain and overall health of the same.

Antibiotics are widely used in poultry and livestock feeds and water. Apart from their direct medicinal value in treating and preventing infections, antibiotics have also been used as "pronutrients" to increase the efficiency of feed utilization, stimulate growth in intensive poultry and livestock farming and reduce bird and animal morbidity mortality.

It is important that poultry and livestock achieve rapid growth and body weight gain so that they may be shipped earliest for markets. Activities such as general handling, debeaking, tagging, medications, vaccinations, transportation, any kind of nutritional inadequacy, water shortage etc. result in a great deal of stress in birds and livestock. In addition, feeding and rearing of poultry and livestock are often carried out in intensive farming, often under such conditions of great stress for the birds and animals; a consequence of which is that the birds and animals may become greatly susceptible to contracting and transmitting infections. Also, rapid spread of infections amongst the birds and livestock in the feeding facility can be caused by feed contamination by fecal droppings. In addition, Clostridium perfringens, Escherichia coli and Salmonella species that colonize the gastrointestinal tract of chickens and livestocks are food borne pathogens that are not only responsible for major epidemics, but are also associated with processed poultry and animal products and may cause severe illness, and even death, hi human beings (Ref: Poultry: Feeds and Nutrition, 2 ^nd Ed, Authors Patrick, H and Schaible

P.J, AVI Publishing Company INC and Microbes Infect. 1999 Jul;l(8):639-44.). Thus antibiotics are routinely included in the feeds of poultry and livestock to inhibit the growth of microbial pathogens, as well as to prevent their spread in feeding facilities. Apart from their direct role in treating and preventing infections, antibiotics, often in subtherapeutic doses, also act as growth promoters and pronutrients in poultry and livestock (Ref: Poultry: Feeds and Nutrition, 2 ^nd Ed, Authors Patrick, H and Schaible PJ, AVI Publishing Company INC and Drugs. 1999 Oct;58(4):589-607.). Antibiotics increase the rate of growth in poultry and livestock and facilitate more efficient utilization of feed. They are also known to enhance the utilization of calcium, phosphorus and manganese, possibly because of increased acidity of the small intestine and ceca (Ref: and Drugs. 1999 Oct;58(4):589-607 and Poultry: Feeds and Nutrition, 2 ^nd Ed, Authors Patrick, H and Schaible PJ, AVI Publishing Company INC. ). Thus under average sanitary conditions, and with balanced diet, antibiotic supplementation can result in nearly 2 to 8% increase in growth, and 2 to 4% increase in feed conversion. (In fact, when antibiotics were first used in feeding, researchers obtained differences of as much as 18 to 25% increase in growth rate of chicks). Antibiotics have also been shown to counteract marginal deficiencies and imbalances in the diet. Though antibiotics are not nutrients by themselves, but chemicals produced by living organisms that destroy other organisms, they do not significantly change the total number of microorganism in the intestines. Rather, they may manifest their growth promoting, pronutrient effects by sparing the requirement for proteins, certain vitamins and minerals, making the intestinal pH more acidic, helping in higher absorption of nutrients, and stimulating growth (Ref: Animals on Drugs - antibiotic usage in livestock, The Environmental Magazine, Nov, 2000 by Kathleen O ¹ NeU and Poultry: Feeds and Nutrition, 2 ^nd Ed, Authors Patrick, H and Schaible PJ, AVI Publishing Company INC.). It is not known exactly how they react on the microorganisms in the intestines of normal birds and healthy livestock and improve nutrition, but it has been observed that in a "new" environment (such as a new facility where no poultry/animals had been raised earlier) the positive manifestation of antibiotic effects are not noticed and, in particular in case of chicks raised under germ-free conditions, are not benefited by feeding antibiotics (Ref: Poultry: Feeds and Nutrition, 2 ^nd Ed, Authors Patrick, H and Schaible PJ, AVI Publishing Company INC.). Thus it seems that the antibiotic effects could be related to destruction of harmful microorganisms that otherwise sequester or destroy nutrients, while at the same time either sparing the ones which have no such effects on nutrients, or actually stimulating those organisms that may synthesize some

growth promoting factors. The modes of pronutrient action may differ amongst various antibiotics, but in general they are believed to be one or more of the following:

1) Inhibit the growth of nutrient sequestering or destroying microorganisms.

2) Facilitate the growth of beneficial microorganisms.

3) Destroy microorganisms that produce toxic products, such as excessive amounts of ammonia and other toxic nitrogenous waste products in the intestine; or suppression of production of toxic products by microorganisms - for instance, antibiotic effects appear to be correlated, not just to the elimination of Clostridium perfringens, but also to the suppression of its toxin production.

4) Improve availability of certain nutrients.

5) Provide good intestinal health for better absorption of nutrients.

6) Improve feed and/or water consumption.

7) Prevent clinical and sub-clinical diseases and conditions.

Thus for nearly 50 years antibiotics have been used extensively as poultry and livestock supplements, in therapeutic as well as sub-therapeutic doses, for its anti- pathogenic and pronutrient, progrowth effects. Commonly used antibiotics include penicillin, oxytetracycline, chlortetracycline, flavomycin, virginiamycin and bacitracin.

However continued use of antibiotics has been a cause for concern (Ref van den Bogaard et al, J Antibiotic Chemother. 2002 March; 49(3):497-505). Continued usage, even at sub-therapeutic levels, increases the possibility of antibiotic residue in poultry and livestock products, in the environment and, through the food-chain, in humans. This increases the possibility of development of drug-resistant bacteria, and a consequent reduction in the ability to cure infections in humans, birds and animals. In fact, the 50- year extensive use of antibiotics has seen a concurrent rapid rise in drug resistant microorganisms. Thus non-medical use of antibiotics in farming is being increasingly restricted. In the European Union for instance, precautionary legislation on the use of antibiotics as growth promoters is already in place. Further, consumer preferences for organic or "chemical free" products is also on the rise. The above factors make it imperative that efforts be made to identify alternatives to in-feed antibiotics that are safe, effective, as well as economical to user.

The present invention describes novel supplements that replace the use of antibiotics in poultry and livestock feeds. The supplement compositions are a source of nutrients, that can enhance the growth and productivity of poultry and livestock, as well as anti-microbial factors that effectively inhibit, if not completely eliminate, pathogenic

microorganisms. In particular the present supplement formulations can replace antibiotics in chick and livestock feeds when supplemented through drinking water.

The supplements according to the present invention comprise of combinations of probiotics and methylsulfonylmethane that effectively replace antibiotics in feed supplements. The supplements, according to the present invention, further comprise saccharides, vitamins, carotenoids, xanthophylls, minerals and electrolytes. The components of the supplements, according to the present invention, function synergistically to effectively replace antibiotics in poultry and livestock feeds.

The prior art describes feed supplements and components of feed supplements, as well as studies related to their effects on poultry and livestock.

US patent US5928686 and the PCT application WO96/39862 describes a process for enhancing the health, livability and cumulative weight gain of poultry by making available for consumption a high moisture solid containing 30% to 90% by weight water and 10 to 70 % dry matter, and wherein the dry matter contains at least 10 % carbohydrate, and between 15 to 50% by weight of an amino acid source.

British Poultry Science, 2004 October; 45(5): 684-94 describes effects of mushroom and herb polysaccharides on growth performance of broilers.

Berlin Munch Tierztl Wochenschr. 1999 Oct-Nov; 112 (10-11): 370-9 is an article on antibiotic growth promoter effects on animal nutrition.

British Poultry Science 1994, March; 35(1): 123-33, describes the effects of antibiotic growth promoters on monensin toxicity in broiler chicks.

Nutrition Journal 2004, 3: 19 reviews the anti-cancer effects of various antioxidants and probiotics.

US patent 6558718 describes nutrient clusters for food products.

US patent US4914135 describes the use of methylsulfonylmethane to treat parasitic infections.

Patent application WO9736603A1 describes biocontrol agents for use in the treatment of opportunistic infection.

International Journal of Poultry Science 3 (9): 603-607, 2004 is a paper that evaluates the inhibition of pathogenic microbial growth on chicken feed by Lactobacilli.

MSM: The Definitive Guide, Authors Jacob, S. W and Appleton, J, Freedom Press is a review of the science of methylsulfonylmethane.

Poultry Science, 2004 M; 83(7): 1148-54 is an article that describes the effects of mannan oligosaccharide, bacitracin methylene disalicylate on the live performance and intestinal microbiology of turkeys.

J Nutrition 2004 Jun; 134 (6): 1487-92 is a paper that describes the role of dietary antibiotics in enhancing the bioavailability of alpha-tocopheryl acetate in broilers.

Infection 1999; 27 Suppl 2: S35-8 is a review antibiotic containing feed additives.

However most of the feed supplements, especially those of poultry and livestock include antibiotics as one of their components. The present invention describes novel supplements that replace the use of antibiotics in poultry and livestock feeds. Brief Description Of The Invention

The present invention relates to novel poultry and livestock supplements that comprise of probiotics and methylsulfonylmethane. The supplements further comprise saccharides, vitamins, carotenoids, xanthophylls, minerals and electrolytes. The components of the supplements described in the present invention function synergistically as nutrients sources, that enhance the growth and productivity of poultry and livestock, as well as anti-microbial factors that effectively inhibit, if not completely eliminate, pathogenic microorganisms. The supplements of the present invention can thus effectively replace antibiotics hi poultry and livestock feeds. Detailed Description Of The Invention

For nearly 50 years antibiotics have been widely used as poultry and livestock feed supplements, in therapeutic and sub-therapeutic doses. They have been used for their therapeutic value - for the treatment as well as prevention of infections, as well as for their "pronutrient" value in poultry and livestock. The pronutrient-growth promoting effects of antibiotics are manifested as increase in the rate of growth of poultry and livestock and greater utilization of feeds. They are also known to enhance the utilization of calcium, phosphorus and manganese, possibly because of favourable pH of the small intestine and ceca.

However over the years there has been increasing concern over the extensive use of antibiotics in poultry and livestock feed. Continued use of antibiotics increases the possibility of development of drug resistance by pathogenic microorganisms, thus posing a serious threat to the health of humans and animals. In fact, the 50-year extensive use of antibiotics has also seen a concurrent rapid rise in drug resistant microorganisms. Moreover there is increasing consumer preferences for organic or "chemical free" products, making it imperative that food products in general, and poultry and livestock

products in particular, are completely free of antibiotics (usage of antibiotics, even at subtherapeutic levels, increases the possibility of antibiotic residue in poultry and livestock products, in the environment and, through the food-chain, in humans). The above factors make it necessary that efforts be made to identify alternatives to feed-supplement antibiotics that are safe, effective, as well as economical to use.

The present invention describes novel supplements that replace the use of antibiotics in poultry and livestock feeds. The supplement compositions are a source of nutrients, that can enhance the growth and productivity of poultry and livestock as well as anti-microbial factors that effectively inhibit, if not completely eliminate, pathogenic microorganisms. The supplement compositions of the present invention comprise of probiotics and methylsulfonylmethane. Probiotics is the term used to describe microbes that have beneficial effects in the health of animals, including humans. They include some of normal flora of the intestines of animals and are known to be responsible for the healthy function and development of the gastro-intestinal tract. Methylsulfonylmethane is a compound that is used in animal nutrition. It has several beneficial biological effects such as anti-inflammatory effects, vasodilatory effects, inhibition of pain impulses, antimicrobial effects etc. So far however, probiotics and methylsulfonylmethane have not been used in combination, possibly the anti-microbial effects of methylsulfonylmethane was expected to reduce the beneficial effects of probiotics by inhibiting or killing the probiotic microorganisms. However, the supplement compositions of the present inventions comprise of combinations probiotics and methylsulfonylmethane. Rather than inhibiting the probiotic microorganisms, methylsulfonylmethane functions synergistically with the probiotics to improve the feed composition of poultry and livestock. The compositions further comprise saccharides, vitamins, carotenoids, xanthophylls, minerals and electrolytes. The components of the supplements described in the present invention function synergistically as nutrient/pronutrient sources, that enhance the growth and productivity of poultry and livestock; as well as anti-microbial factors that effectively inhibit, if not completely eliminate, pathogenic microorganisms. The supplement compositions of the present invention offer an excellent alternative to antibiotics in poultry and livestock feed.

Description of the essential components of the supplement follows: Probiotics

Probiotics are live microbial feed supplements that beneficially affect the host animal by improving its intestinal microbial balance.

The rumen in ruminant animals and the colon in monogastric animals or birds can be perceived as a metabolic chamber rich in microorganisms. Unlike the gut and the small intestine, the fermentative conditions in the rumen and colon are optimal for the development of a rich microbial flora. The rumen and colon are strictly anaerobic environments. Its bacterial ecosystem is composed of about 400 to 500 different types of bacteria most of which are "commensals" that participate in the fermentation of non- digestible food compounds, hi addition, the colonic flora also contains pathogenic bacteria such as Clostridium perfringens, certain types of Escherichia coli, Salmonella etc. Li imbalanced intestinal conditions these pathogens can develop and produce toxins, which can cause acute or chronic illness that could lead to severe gastro-intestinal disorders in the host. However, also present in the colon are groups of microorganisms, such as Bifidobacteria, Lactobacillus and yeasts, whose presence is associated with a well-functioning intestinal ecosystem of a healthy host, hi sufficient numbers they constitute a colonization barrier against pathogens and toxins and reduce their numbers and amounts to harmless levels. For instance Bifidobacteria has been suggested to protect against gastroenteritis associated with enteropathogenic bacteria (in particular the toxins produced by the same), as well as inhibit the growth of Salmonella, hi addition such "probiotic microbes" or "friendly microbes" help in the absorption and metabolism of carbohydrates, lipids, proteins and amino acids, as well as salvage energy and vitamins from such components that have escaped digestion and absorption in' the stomach and small intestine. They may also carry out detoxification and subsequent excretion of toxic factors that are either produced and/or released into the gastro-intestinal tract, provide additional uncharacterized factors, such as anti-microbial factors, and have anti- tumor/anti-carcinogenic effects. Thus proper maintenance of the gut flora is essential for the normal growth and development of an animal. However in newly hatched chicks and new born animals, the gut flora may be severely depleted of its probiotic microbes. In addition birds and livestock that have been subjected to stress conditions such as debeaking, tagging, transportation, any nutritional inadequacy and water shortage can result in greater susceptibility to infections. Also, Clostridium perfringens, Escherichia coli and Salmonella that often colonize the gastrointestinal tract of chickens, and livestock such as pigs, and are responsible for major poultry and porcine epidemics of devastating nature, are also major food borne pathogens that are associated with processed poultry and livestock products and may cause severe illness in humans as well. Thus in poultry and livestock, especially in newly hatched/born ones, depletion of gut

flora, could lead to serious conditions including reduced absorption of essential dietary components, and increased susceptibility to infections by pathogenic microorganisms. Moreover, the birds and animals may be highly susceptible to contracting and transmitting infection from the hatcheries and farms. This could seriously compromise the healthy development of the birds and livestock. Hence over the last few decades antibiotics have been used, in therapeutic and sub-therapeutic doses, in poultry and livestock feed and water. Antibiotics have been effective in combating pathogenic microorganisms, as well as having pronutrient, growth promoter effects. However, because of the ever-increasing incidence of antibiotic resistance by microorganisms, it has become increasingly necessary to reduce, or completely eliminate, the use of antibiotics in poultry and animal feeds. The present invention describes compositions that comprise probiotics as one of the components of poultry and livestock supplements. These probiotics eliminate the need for antibiotics. Also since newborn chicks and newborn animals are germ-free, feed supplements comprising probiotics competitively inhibit colonization by pathogenic microorganisms, and help establish a healthy gut microflora. Examples of probiotics used in the formulation include Lactobacillus acidophilus and Saccharomyces bouldardii. Lactobacillus acidophilus is used to inhibit the growth of enteropathogens like E.coli, Salmonella and Clostridium perfringens. Saccharomyces bouldardii is included as a source of cell wall mannanooligosaccharides. Methylsulfonylmethane

Methylsulfonylmethane is a sulfur compound that is the primary oxidized metabolite of dimethyl sulfoxide. It belongs to a family of compounds present abundantly in the food chains of terrestrial and ocean life that provides bioavailable sulfur to as many as 85% of all living organisms. They are among the primary sources for sulfur. Biological effects of methylsulfonylmethane include its effects on connective tissue where, as a sulfur donor, it contributes to normal cross-linking or proteoglycans, a key component of connective tissue. Methylsulfonylmethane has also been shown to have antiinflammatory effects when administered orally, topically or intravenously. Other biological effects of methylsulfonylmethane include vasodilatory effects, inhibition of pain impulses and anti-microbial effects, muscle relaxation, antagonism of platelet aggregation, anti-oxidative effects, water retention effects etc. Methylsulfonylmethane has been shown to have several clinical uses including in the treatment of osteoarthritis, scleroderma, interstitial cystitis etc. It has also been shown to have beneficial effects in the treatment of poor hoof health in horses, sulfur deficiency, chronic obstructive

pulmonary disease in horses, canine arthritis, fibrous pleurisy in foals, as well as reduction in stress related deaths etc. Thus methylsulfonylmethane has general beneficial effects in the health of animals.

The compositions of the present invention include methylsulfonylmethane. It has good anti-stress action in poultry that increases its livability and productivity. It also acts as a critical methyl group donor, as well as a sulfur source, for which poultry and livestock have a very high requirement. In addition it acts as a water retention factor that prevents the loss of water from the body, thereby preventing weight loss in poultry and livestock. Despite its known anti-microbial effects, including bacteriostatic and bactericidal effects, most unexpectedly methylsulfonylmethane appears to have no effects on the viability of the probiotics used in the supplements of the present invention. Rather the combination of probiotics and methylsulfonyhnethane function synergistically as a source of growth-promoters, anti-stress agents and anti-pathogenic microbial factors, thus effectively replacing antibiotics in poultry and livestock supplements. The combination of probiotics and methylsulfonylmethane thus improves the overall health and performance of poultry and livestock. Saccharides

Saccharides are carbohydrates and include to mono, di, oligo and polysaccharides. Monosaccharides are molecules that have the general formula C _n H _2n O _n where n is an integer equal to 6 in case of dominant food monosaccharides. Examples of monosaccharides include glucose, fructose, galactose etc. Disaccharides are molecules that are composed of two monosaccharides units linked together by a "glycosidic" bond. Examples of disaccharides include sucrose, lactose, maltose etc. Oligosaccharides and Polysaccharides are saccharides that have more than three monosaccharide units. Examples of such oligo/polysaccharides include maltotrioses, dextrans, mannans, starch etc. Saccharides are the major source of energy for organisms. Di and oligosaccharide are first broken down to their individual monosaccharides in the gastro-intestinal tract by the action of specific enzymes. The monosaccharides are then metabolized in cells of tissues, and the energy released used to drive the metabolic processes required for the growth and maintenance of cells and tissues. Hence feed compositions require saccharides as a source of metabolic energy.

In additional to its nutritional function, saccharides, especially non-digestible ones such as fructo-oligosaccharides, also function as factors that encourage the development and multiplication of probiotics in the gut (hence called "prebiotics) and inhibit the

growth of pathogens such as Salmonella. In addition, saccrahrides adsorb pathogenic bacteria containing type 1 fimbriae thus inactivating them, promote mineral absorption, greatly improve lipid metabolism, improve gut and intestinal function (by increasing villi height, uniformity and integrity) and acts as immunomodulator that stimulates gut associated and systemic immunity by acting as a non-pathogenic microbial antigen giving an adjuvant like effect. The supplements of the present invention include saccharides, such as dextran-oligosaccharides. It is observed that in the presence of saccharides, the probiotic count in the supplements of the present invention is increased by a factor of two every ninety minutes, thus increasing the potency of the probiotic component of the supplement. Thus the overall performance of poultry and livestock is greatly improved. Vitamins

The supplement compositions of the present invention preferably include one or more vitamins, xanthophylls, carotenoids and minerals. The vitamins in the present invention include niacinamide, dl-α-tocopheryl acetate, dl-α-tocopherol (vitamin E), pyridoxine HCl, β-carotene (precursor of Vitamin A), riboflavin (vitamin B2), cyanocobalmin (vitamin B12), vitamin A, folic acid, thiamin HCl (vitamin Bl), menadione (vitamin K) and ergocalciferol (vitamin D2 and D3), Ascrobic acid (Vitamin C). Ascorbic acid in particular acts as an anti-oxidant and also enhances the absorption of methylsulfonylmethane. Carotenoids and Xanthophylls

Carotenoids and Xanthophylls are also included in the compositions of the present invention. They are precursors of vitamins. Carothenoids are known to protect tissues from free radical damage. Both carotenoids and xanthophylls are free radical scavengers and have been shown to function as antioxidants in vitro, as well as reduce cancer cell formation. Marigold Oleoresin contains carotenoids and xanthophylls and can be used in the supplement as the source for the same. Minerals and Electrolytes

The supplement compositions of the present invention also comprises of Sodium, Potassium, Calcium, Chloride, Phosphorous and Sulfur. Calcium is an essential component of bones and is especially necessary for the proper growth and development of newly hatched chicks and newborn animals.

Thus compositions of the present invention comprises of synergistic components that not only serve as an excellent source of growth promoters to poultry and livestock,

but also possess effective anti-pathogenic microbial properties. In addition, the components of the present - methylsulfonylrnethane and probiotics - are surprisingly compatible with each other despite the anti-microbial effects of methylsulfonylmethane (which one would expect to effect the viability of the probiotics in the feed formulation). Moreover another unique aspect of this formulation is that when the supplement is added to water, since with the saccharide present as a diluent, the probiotics count increases very rapidly — it can double over a period of 90 minutes - thus greatly increasing the potency of the probiotic component of the supplement. The probiotics are also in an active growth phase to readily colonize the gut. To summarize, the formulations of the present invention:

1) safeguard the young chicks and livestock animals from infection and stress.

2) act as a strong performance enhancer.

3) are aqueous based formulation easily absorbed by the young birds and newborn livestock.

The supplements described in the present invention thus acts as an excellent alternative to antibiotics in poultry livestock feeds.

The following examples serve to illustrate the present invention. It is to be understood however, that the examples in no way constitute a limitation on the scope the present invention

Example 1

Ingredient Composition

(per kg)

Vitamin C 1.0 g

Pyridoxine (Vitamin B6) 552 mg

Cyanocobalamin (Vitamin B 12) 2.20 mg

Vitamin K3 1.104 g

Vitamin B2 2.864 g

Calcium D Pantothenate 4.408 g

Niacin 552 mg

Sodium Sulphate 88.08 g

Potassium Chloride 88.08 g

Vitamin A 4.0O g

Vitamin D3 10.00 g

Vitamin E 600 mg

Marigold Oleoresin 2.0O g

Oligo-dextran 2.0O g

Methylsulfonylmethane 10.00 g

Saccharomyces boulardii 10 x 10 ⁹ cells

Lactobacillus acidophilus 50 x l0 ⁹ cells

Carrier (sucrose) q.s

Example 2

Ingredient Composition

(per kg)

Vitamin C 5.0 g

Pyridoxine (Vitamin B 6) 5.0 g

Cyanocobalamin (Vitamin B 12) 22.0 mg

Vitamin K3 11.00 g

Vitamin B2 28.64 g

Calcium D Pantothenate 44.00 g

Niacin 5.00 g

Sodium Sulphate 100.00 g

Potassium Chloride 100.00 g

Vitamin A 20.00 g

Vitamin D3 15.0O g

Vitamin E 1.00 g

Marigold Oleoresin 5.00 g

Oligo-dextran 10.00 g

Methylsulfonylmethane 20.00 g

Saccharomyces boulardii 50 x l0 ⁹ cells

Lactobacillus acidophilus 10 x l0 ¹⁰ cells

Carrier (sucrose) q.s

Example 3

Ingredient Composition

(per kg)

Vitamin C 500 mg

Pyridoxine (Vitamin B6) 200 mg

Cyanocobalamin (Vitamin B 12) 1.10 mg

Vitamin K3 500 mg

Vitamin B2 1.35 g

Calcium D Pantothenate 2.0O g

Niacin 200 mg

Sodium Sulphate 5O g

Potassium Chloride 40 g

Vitamin A 2.0O g

Vitamin D3 5.00 g

Vitamin E 300 mg

Marigold Oleoresin 1.00 g

Oligo-dextran 1.00 g

Methylsulfonylmethane 5.00 g

Saccharomyces boulardii 5 x 10 ⁹ cells

Lactobacillus acidophilus 10 x l0 ⁹ cells

Carrier (sucrose) q.s

Example 4

Ingredient Composition

(per kg)

Vitamin C 300 mg

Pyridoxine (Vitamin B6) 100 mg

Cyanocobalamin (Vitamin B 12) 2.20 mg

Vitamin K3 300 mg

Vitamin B2 500 mg

Calcium D Pantothenate 1.2O g

Niacin 150 mg

Sodium Sulphate 3O g

Potassium Chloride 30 g

Vitamin A 1.00 g

Vitamin D3 3.00 g

Vitamin E 200 mg

Marigold Oleoresin 500 mg

Oligo-dextran 500 mg

Methylsulfonyhnethane 10.00 g

Saccharomyces boulardii 2 x 10 ⁹ cells

Lactobacillus acidophilus 5 x 10 ⁹ cells

Carrier (sucrose) q.s

Example 5

Ingredient Composition

(per Kg)

Vitamin C 2.0 g

Pyridoxine (Vitamin B6) 800 mg

Cyanocobalamin (Vitamin B 12) 5 mg

Vitamin K3 3.00 g

Vitamin B2 5.00 g

Calcium D Pantothenate 4.408 g

Niacin 3 g

Sodium Sulphate 200 g

Potassium Chloride 175 g

Vitamin A 7 g

Vitamin D3 20.00 g

Vitamin E 1.00 g

Marigold Oleoresin 4.00 g

Oligo-dextran 4.00 g

Methylsulfonylmethane 20.00 g

Saccharomyces boulardii 50 x 10 ⁹ cells

Lactobacillus acidophilus 20 x 10 ¹⁰ cells

Carrier (sucrose) q.s

Example 6

Appropriate quantities of the components of the composition, other than Vitamin A,

Vitamin D3, Vitamin E and Marigold Oleoresin, were charged in a blender with an open air valve and inlet air pressure at 5 kg/cm ² or more. Vitamin A was dissolved in about

500 ml of warm water, Vitamin D3 and Vitamin E in about 10 ml and 200 ml respectively of warm polyethylene glycol 600 and Marigold Oleoresin in about 400 ml of

Tween 80. Dissolved Vitamin A, D3, E and Marigold Oleoresin were then added to the blender and blended for about 10 minutes. The material in the blender was then sieved with a 40 mesh sieve. The sieved material was then packed into liner bags and heat sealed.

Example 7

A pure culture of Lactobacillus acidophilus was grown in thioglycollate broth for 16 hours. 10 ⁸ cells were then inoculated in 10 mm diameter wells on nutrient agar plates that were spread with Escherichia coli, Salmonella typhi and Staphylococcus aureus, and incubated at 37 ⁰ C overnight. Zones of inhibition were observed around the Lactobacillus acidophilus spots. The data are presented in the table below.

From the above results it is clear that the probiotic Lactobacillus acidophilus has antimicrobial effects. Example 8

To study the antibacterial effects of the supplement compositions of the present invention, 1 g of the composition described in Example 1 was added to 50 ml of distilled water, of which 200 μl was inoculated in 10 mm diameter wells on nutrient agar plates that were spread with Escherichia coli, Salmonella typhi, Shigella flexneri and Staphylococcus aureus, and incubated at 37 ⁰ C for 24 hours. Zones of inhibition were observed around the supplement spots.

From the above results it is clear that the supplement compositions of the present have anti-microbial effects against common enteric pathogens. Example 9

To study the effects of the supplement compositions of the present invention, the feeds of 1100 birds, described as "Treated Group", were supplemented with the composition described in the Example 1. The effects on the "Treated Group" were compared with those of a "Control Group" of 1100, in which Methylsulfonylmethane and the probiotics Saccharomyces boulardii and Lactobacillus acidophilus were excluded from the supplement. The results are presented in the table below.

From the above results it clear that compositions of the present invention have greatly enhanced beneficial effects in birds supplemented with the same. Not only is the mortality levels in the "Treated Groups" much lesser then that of the "Control Group", both the average body weight, as well as the total body mass of the "Treated Group" are higher then that of the control group.