Title of Invention	"A MODIFIED POULTRY FEED FOR PRODUCTION OF EGGS"
Abstract	This invention relates to the use of minerals and vitamins in poultry eggs, through utilization the composition of minerals and vitamins mixtures in poultry feed, to ameliorate their deficiencies in diabetics. The modified egg contains appropriate concentrations of vitamins and minerals which on consuming helps to restore the depleted minerals almost close to their normal levels in the tissues and reduces the severity of hypercholesterolemia, dyslipidemia, hyperinsulinemia, hyperglycemia and arterial hypertension of diabetic consumers.

Title of Invention

"A MODIFIED POULTRY FEED FOR PRODUCTION OF EGGS"

Abstract

This invention relates to the use of minerals and vitamins in poultry eggs, through utilization the composition of minerals and vitamins mixtures in poultry feed, to ameliorate their deficiencies in diabetics. The modified egg contains appropriate concentrations of vitamins and minerals which on consuming helps to restore the depleted minerals almost close to their normal levels in the tissues and reduces the severity of hypercholesterolemia, dyslipidemia, hyperinsulinemia, hyperglycemia and arterial hypertension of diabetic consumers.

Full Text	Filed of the invention The present invention relates to a poultry feed for use in the production of eggs with optimal concentration of minerals and vitamins More particularly, the present invention relates to an improved poultry feed for production of eggs with a capacity of restoration of mineral composition in mammals with diabetes, hypertension, obesity, hypercholesterolemia and hyperinsulinemia The present invention also relates to a method for the manufacture of such unproved poultry feed The present invention particularly relates to an improved egg with a capacity of restoration of mineral composition in mammals with diabetes, hypertension, obesity, hypercholesterolemia and hyperinsulinemia and a method for the production thereof The present invention also relates the use of minerals and vitamins in poultry eggs, through optimizing the composition of minerals and vitamins mixtures in poultry feed, to ameliorate their deficiencies in diabetics The modified egg contains appropriate concentrations of vitamins and minerals which on consuming helps to restore the depleted minerals almost close to their normal levels in the tissues and reduces the severity of hypercholesterolemia, dyslipidemia, hyperinsulinemia, hyperglycemia and arterial hypertension of diabetic consumers Background of the invention In diabetics, the body tissues tend to lose their minerals and the severity of the disease increases with further deterioration of mineral homeostatic mechanism notably of copper, zinc, magnesium, manganese and chromium etc with passage of time Some of these minerals are the constituent of either cytosohc or intramitochondrial superoxide dismutases enzyme (SOD), catalases glutathione-s-transferases and glutathione reductases which disarm the free reactive oxygen species (ROS) and protect the cell components from peroxidation Their altered levels in the diabetics lead to the deficiency of either one or more metals resulting in a reduction of antioxidative defense system As a consequence of this, ROS generated get accumulated in the cells leading to the severity of the disease A number of dietary supplement formulations of vitamins and minerals combinations in the form of tablets/capsules have been developed for human consumption to restore the altered minerals and vitamins status in the body so as to prevent the complications in the disease occurring due to their deficiencies/excess However, the results of the available dietary supplements are not as satisfactory as are expected particularly in diabetics since they are in synthetic forms whose absorption rate is less than those of natural forms or either their binding legends get depleted during the course of the disease or they are not as complete as they are thought of The poultry eggs can be conveniently used as a material for introduction of vitamins and minerals to restore their composition in the body since they serve as the natural source of vitamins and minerals which exist in bound forms They are easy to absorb and the concentrations of vitamins and minerals in eggs can be manipulated by incorporating appropriate changes in the birds feed The prior art methods had considerable disadvantages One of the drawbacks of the conventional eggs for their use as source of vitamins and minerals is that they contain high cholesterol content (>210 mg/50g egg) which is an accepted high risk factor of cardiovascular diseases, such as atherosclerosis, myocardial infarction and hypertension usually associated with NIDDM The eggs as such, therefore, cannot be used as a source of vitamins and minerals for such patients unless modified Appropriate modification in dietary feed has led to the development of the eggs which have lower cholesterol (106-160mg/50g egg), less saturated fats (0 8-1 2g) more of omega-3 unsaturated fatty acids (70-400mg), more of vitamin E (0 5-10mg), folate (10-150 mg) and iodine (1-120 meg) than the standard eggs reported in technical publication which include US Patent No, 4128640, 4187294, 4197293, 4197294, 4394376, 4410541, 4739853, 4764531, 501276, 5091195, 5246717, 5415879, 5520938, 5520398, 5665375, 6316041 and 5759567 incorporated herein by reference No attempts have been made to modify the minerals in the feed of the birds (except copper and iodine as cholesterol lowering agents in eggs) for the production of eggs, which on consuming may rehabilitate the minerals status in diabetics Objects of the invention Accordingly, it is an object of the present invention to provide a poultry feed for use in the production of eggs with optimal concentration of minerals and vitamins It is another object of the present invention to provide an improved poultry feed for production of eggs with a capacity of restoration of mineral composition in mammals with diabetes, hypertension, obesity, hypercholesterolemia and hypermsulmemia It is another object of the present invention to provide a method for the manufacture of such improved poultry feed It is another object of the present invention to provide a method for producing modified eggs conducive to the healthy consumers It is another object of the present invention to provide poultry eggs, which restore the depleted minerals composition in the consumers It is another object of the present invention to provide poultry eggs with minerals, which lower the blood cholesterol, LDL-c, VLDL-c and increase triglycerides and HDL-c in diabetic consumers It is another object of the present invention to provide modified eggs, which lower the blood pressure in diabetic consumers It is another object of the present invention to provide modified eggs, which lower elevated blood glucose in diabetic consumers It is another object of the present invention to provide modified eggs, which lower the elevated insulin level in blood of diabetic consumers It is another object of the present invention to provide modified eggs, which lower the elevated creatinine in blood and urine of diabetic consumers It is another object of the present invention to lower the elevated cortisole and TSH and increase the depressed levels of T3 and T4 in blood of diabetic consumers It is another object of the present invention to provide modified eggs to lower the elevated aldosterone concentration in the blood of the diabetic consumers It is another object of the present invention to provide modified eggs to restore the normal level of glycogen in liver and muscles of diabetic consumers It is another object of the present invention to provide modified eggs to restore the disordered pathology of the tissue of diabetic consumers Summary of the invention The above and other objects of the present invention are achieved by the modified egg of the present invention wherein the modified egg is a whole shell poultry egg having minerals and vitamins composition sufficient enough to interact with the tissue minerals and vitamins so as to rehabilitate their altered status in diabetics and decrease the seventy of the disease The minerals and vitamins are delivered through a novel poultry feed of the present invention These minerals predominately include zinc, copper, magnesium and manganese Accordingly, the present invention provides an improved poultry feed for production of eggs with a capacity of restoration of mineral composition in a patient said feed comprising a major amount of dietary material such as herein described, from up to 30 % by wt of minerals of the kind such as herein described and up to 30 % by wt of vitamins of the kind such as herein described In a preferred feature, the minerals are selected from one or more of Sodium derivatives, Ferrous derivatives, Magnesium derivatives, Zinc derivatives, Copper derivatives and Iodine derivatives In a preferred feature, said minerals comprise from 0 2 to 2 0 g of sodium derivatives, from 0 2 to 3 0 g of ferrous derivatives, from 0 05 to 500 g of magnesium derivatives, from 0 02 to 0 05 g of zinc derivatives, form 0 50 to 1 g of copper derivatives, from 0 002 to 0 005 g of iodine derivatives, based on 1 kg of said poultry feed In a preferred feature, the minerals are selected from sodium bicarbonate, ferrous sulphate, manganese sulphate, copper sulphate, zinc sulphate, magnesium sulphate and potassium iodide or mixtures thereof In a preferred embodiment, said vitamins are selected from ascorbic acid, calcium pentathenate, choline chloride, folic acid, ergocalciferol, nicotinic acid, cyanocobalamine, pyndoxine hydrochloride, thiamine hydrochloride, riboflavin, a-Tocopherol, carotenoid and mixtures thereof In a preferred feature, said vitamins comprise from 200 to 700 mg of ascorbic acid, from 320 to 500 mg of calcium pentathenate, from 200 to 500 mg of choline chloride, from 100 to 500 mg of Folic acid, from 0 05 to 0 200 mg of ergocalciferol, from 300 to 500 mg of nicotinic acid, from 0 01 to 100 mg of cyanocobalamine, from 50 to 200 mg of pyndoxine hydrochloride, from 200 to 500 mg of Thiamine hydrochloride, from 100 to 300 mg of riboflavin, from 200 to 500 mg of, a-tocopherol, from 15 to 40 mg of carotenoid, of each of said vitamins amount based on lKg of the said poultry feed The present invention also provides a modified egg wherein the egg is a whole shell poultry egg having a content of constituents comprising total protein,6 5-8g, carbohydrates 0 5-0 7g, total lipids, 5-6 5g, triglycerides, 1 5-2 5g, cholesterol, 95-125mg, phospholipids,! 5-2mg, Omega-3 fatty acids (Lenolenic acids), 70 - 80mg, vitamin E,10-12 5mg, vitamin D, 0 50-0 850 mcg, Bi,0 05- 0 10mg, B2,0 275-0 350mcg, Bi2,0 18 -0 25mcg, foliate, 0 03-0 1mg, B6,0 015-0 05mcg, carotenoids, 0 5-0 85mg, Ca++,30 - 40mg, Mg,0 8-2 5mg, Zn,0 5-1 5mg, Cu,2 0-3 5mg, Fe,l 75-2 5mg, iodine 0 75-2 5mcg per 50g liquid egg In a preferred embodiment, the poultry egg is selected from chicken, egg, duck egg and turkey egg The present invention also relates to a method for the production of a modified egg which comprises feeding poultry with a feed comprising of an improved poultry feed for production of eggs with a capacity of restoration of mineral composition in a patient said feed compnsmg a major amount of dietary material such as herein described, from up to 30 % by wt of minerals of the kind such as herein described and up to 30 % by wt of vitamins of the kind such as herein described, allowing said poultry to lay eggs and collecting said eggs Detailed description Genetically obese (ob/ob) mice, diabetic (db/db) mice and Zuker diabetic fatty rats (ZDF/Drt) and transgenic rodents serve as models of research to understand the etiology of obesity, diabetes, hypertension and cardiovascular diseases One of the important finding on these animals is that they absorb and retain greater amount of zinc in liver, adipose tissue and tnceps muscles than their lean control counter parts They have a greater potential of absorbing orally administered zinc from the lumen of gastrointestinal tract to blood plasma and can absorb 60% of more of glucose than those of their lean counter parts (Collip J A, Pediatrics Ann 13, 465-472, 1984, Begin - Heick, N, Daple - Scott J M, Rowe J and Heick M M Diabetes 34, 179-184,1985, Kennedy M L and Failla M L, J. Nutr 117, 886-893,1987, Lin W H, Chen M D and Lin P Y J. Formoson, Med. Assoc 91, 27-33, 1992] Supplementation of Zn promotes food intake, linear growth and body wt increase in children suffering from marginal to severe Zn-deficiency (Mc Gain C J, Kasarkis E J Jr Allen EJ, Prog. Food Nutr Sci. 9, 185-226, 1985) Investigatory studies on laboratory animals however provided evidences that anabolic effects of Zn result from elevated food consumption, higher absorption rate of nutrients l e amino acids (Moran J R and Lyerly A, Life Science 36 2551-2556 1985), fatty acids (Koo S I and Turk D E, J. Nutr 107 909-919, 1977) and glucose (Taneja S K and Arya P, Brit. J. Nutr 68 505-514, 1992) in addition to activation of protein and nucleic acids syntheses These anabolic effects of Zn are unlikely to dimmish after cessation of growth The continuous input of excess nutrients in tissues by excessive bioavailability of Zn after adolescence may create environment essential for obesity It has been reported that an increase of zinc to double the amount of recommended dietary allowance (RDA) increases the portion of the body fat during 25 weeks period of dietary treatment in healthy children (Prentice A, Nutr. Rev 51, 268-270, 1993) There exists a positive linear correlation between hair zinc and body wt in adult humans The obese of both the sexes possess higher zinc concentration in their hair than those with normal body wt/ for their height (Taneja SK, Mahajan M and Arya P Expenentia 52, 31-33,1996) A study on the relation of Zn absorption and deposition of fat in ob/ob mice and high fat diet induced ICR mice showed that the ICR mice fed a high fat diet containing Zn in greater amount resulted in significantly higher gain in body wt and more fat deposition identical to ob/ob mice than did their lean control counter parts fed on a diet containing high fat but low zinc [Chen M D , Lin P Y , Cheng L V and Lin W H Biol. Trace Elem. Res 52 125-132,1996] The proposed potential mechanism for development of obesity involving Zn based on the observations on the activation of ovine metallothionein - ovine growth hormone transgene (OMT - la - OG) by ZnSO4 is that it activates the genes of growth hormone during early period of growth and induces the relatively undifferentiated preadipacytes to be committed to become adipocytes This may increase their population The enhanced population of adipocytes responds by filling with tnacylglycerol due to increased absorption of nutrients caused by Zn and eventually leading to the observed state of obesity Alternately, the elevated growth hormone level may increase the insulin level which lead to increased glucose transport into adipocytes, thereby enabling them to increase hpogenesis Accordingly, the over supply of zinc during growth phase sets in the conditions for the onset of obesity [Pomp D, Oberbauer A M and Murray J D, Transgenic Res 5, 13-23,1996] However, the obese subjects possess either lower or normal plasma level of Zn compared to nonobese control (Chen M D , Lin P Y Sheu W W H, Biol. Trace Elem. Res. 60, 123-129, 1997) The lower plasma Zn level in obese individuals can return to normal following dietary therapy (Di-Martino G, Matera M G, Di-Martino B, Vacca C, Di-Martino S and Rossi F , J. Med. 24 177-183, 1993) In fact the plasma Zn status does not truly reflect the body Zn status as it tends to mamtain Zn homeostasis either by increasing Zn absorption or decreasmg Zn excretion depending upon Zn intake and metabolic requirements Therefore, no significant difference can exist between BMI and serum Zn in healthy subjects Tissue Zn status is thus more reliable to determine the relationship between the two It is evedent from a report that the hair Zn concentration increases with increase in BMI in Indian women (Taneja S K and Mahajan M, J. Punj. Acad. Sci., 1,211-216, 1999) Thus, there are strong reasons to believe that excessive bioavailability of Zn, either in food chain or due to genetic disorder, to the tissues results in obesity Obesity poses a formidable challenge to the growing population all over the world as it is etiologically linked to insulin resistance accompanying to non-insulin dependent diabetes melhtus, hypertension and coronary artery disease This classical clustering of disease is described as metabolic syndrome - X Insulin resistance seems to be an essential adaptation to prevent further weight gain in obese subjects (Eckel RH, Lancet 340, 1452-1453, 1992) and is developed through increased secretion of glucocorticoid from zona fasciculate of adrenal cortex Insulin resistance results in hyperglycemia, glucosuna, altered protein, fat and carbohydrate metabolisms and chronical complications resulting from macro and microvascular pathology Associated with it, altered metabolism of Cu, Zn, Mn and Mg are frequently seen in experimental animals and humans There have been numerous studies evaluating the mineral status of diabetic subjects These studies yield inconsistent results The most frequently reported features of diabetics in genetic diabetic animal model and humans are that they have higher Cu and Mg concentrations, low Zn concentration in their blood plasma and display high amount of Zn loss in urine The degree of increase in plasma Cu in diabetes melhtus is associated with its complications of hypertension or macrovascular disease The plasma magnesium level has been reported to be less when diabetes melhtus is associated with retinopathy (Walter R M Jr, Unu-Hare J Y, Olin K L, Oster M H, Anawalt B D , Cntch field J W and Keen C L, Diabetes Care 14 1050-1056, 1991) However, normocupremia, hypocupremia, or hyperzincemia, normozmcemia and hypozincemia have been reported in some studies The concentrations of metals in blood plasma at a given time represent a total of bound, exchangeable and catabolic components of metals They therefore, can lead to conflicting results and predictions, more so in case of chronic diseases where urinary losses are high The metals concentrations in tissues are more dependable indicators for assessing metal deficiencies since their concentrations are maintained through dynamic equilibrium between tissue metals and exchangeable metal components of the blood plasma Some investigators therefore, preferred other tissues such as liver, muscles, hair, nails etc The levels of zinc and copper in hepatic and renal tissues have frequently been seen either higher or comparable to normal in diabetic melhtus complicated with hypertension The Cu in hair and nails in diabetes melhtus, hypertension and coronary artery disease patients have been found to be low (Sukumar A and Subramamam R, Biol. Trace Elem. Res., 34 89-98, 1992, Taneja S K, Mahajan M, Gupta S and Singh K P , Biol. Trace Elem. Res., 62, 255-264,1998) Cu deficiency has been also reported in muscle biopsies of diabetics (Sjogren A, Edvinsson L , Floren C H and Abdulla M, Nutr. Res., 6, 147-154, 1986) It has been reported that the NIDDM and myocardial infarction susceptible individuals are more prone to obesity as they are genetically predisposed to excess Zn absorption at a younger age, which reduces Cu bioavailability to the tissues over a penod of time The manifestation of clinical symptoms depends upon the degree of ionic imbalance resulting from the degree of stress and the type of food the individual consumes during intervening period Higher stress and zinc rich food aggravate the condition and favour their early onset compared with low stress or low zinc intake in food Because fibers and phytates are known to reduce zinc bioavailability their liberal use delay the onset of obesity and obesity related diseases (Taneja S K, Mahajan M Gupta S and Singh K P Biol Trace Elem. Res 62, 255-264, 1998, Taneja SK Girhotra S and Singh KP, Biol Trace Elem. Res 75, 177-186,2000) The reported variations may represent the trace elements concentration in diet, the pattern of progression of disease and degree of depletion of respective mineral binding legends due to excessive loss of albumin protein, increased rate of proteolysis and lipolysis The mechanistic functional consequences of mineral deficiencies have been discussed differently by different investigators to explain the etiology of diabetes mellitus Since Zn deficiency adversely affects the glucose disposal as it provides stimulating effect of glucose transfer in muscles and adipocytes, the therapeutics benefit of Zn supplementation has been reported in subjects with diabetes mellitus and obesity which both have insulin resistance (Brun J, Guintrand - Hugret R, Fons C , Carvajal J , Fedou C, Fusselher M, Bradet L and Orsetti A, Biol. Trace Elem. Res., 47 385-391, 1995) However, the reality of zinc deficiency in diabetes mellitus has been criticized, as its supplementation further mcreases serum Hb Aic values in diabetes mellitus (Cunningham J J, Fu A, Mearkle P L and Brown R G, Metabolism, 43 1558-1562, 1994) The tissue copper deficiency aggravates hyperglycemia, hypercholesterolemia, hypertension and dyslipidemia and reduction in the activity of cytosolic superoxide dismutase resulting in alteration of antioxidant defence system The reduction in sehnogleutathione peroxidase (Se-GSHPX) activity and increased synthesis of cholesterol promotes lipoprotein oxidation under copper deficiency in particular These effects are important given the etiological role of oxidised low density lipoprotein in initiation of atherosclerosis [Kathenne L O Robert M W and Carl L K Am. J. Clin. Nutr 59, 654-658, 1997 Singal P K, Kharpar N Palace V and Kumar D , Cardiovascular Res 40, 426-432, 1998] Copper deficiency during pregnancy affects the cross linking of connective tissue protein causing abnormal growth and maturation of the foetus [Barone A Ebesh, O Harper R G and Wapmr RA J. Nutr 128 1037-1041, 1998] The reduction of Mg (Mg ATP 2") occurs due to the presence of Mg binding defect (Mg BD) and accounts for all aspects of type - II diabetes It therefore, has a conservative role in the development of the disease (Ibert C W , Devendra K and J A, Biol. Trace. Elem. Res 98, 97-107, 2004] And most of the complications of type-II diabetes such as arrhythmias, hypertension and sudden cardiac death may be prevented by prophylactic magnesium supplementation (Balon T W, Gu J J, Tokuyama Y , Jasman A P and Nadler J J Am. J. Physiol. 269, 745-752, 1995) Manganese is a constituent of mitochondrial Mn-superoxide dismutase Its deficiency results in the generation of free radical of oxygen (ROS) leadmg to the oxidative stress and supplements to adverse effect of the zinc and copper deficiency leading to the depletion of cellular antioxidants and increased production of peroxidation products which accelerate the complications in diabetics (Cederberg J Galh J Luthman H and Enkson U J Diabetes 49, 101-107, 2000, Cedarberg J, Martin S and Eriksson UJ, Pediat. Res., 49, 755-762, 2001) These finds have led to the development of dietary supplements containing minerals and antioxidative vitamins in the form of capsules/tablets to relieve the diabetics from the oxidative stress by ameliorating the relevant minerals deficiency and increasing the cellular antioxidants such as vitamins E, A and C and other vitamins related to general metabolism including Bl, B2, B6, B12, etc However, the results of the available dietary supplements are not as satisfactory as expected particularly in diabetics, since they are in synthetic forms whose absorption rate is less than those of natural forms or their binding legends get depleted during the course of the disease or they are not as complete as they are thought of The poultry eggs can be conveniently used as a material for introduction of vitamins and minerals to restore their composition in the body since they serve as the natural source of vitamins and minerals which exist in bound forms They are easy to absorb and the concentrations of vitamins and minerals in eggs can be manipulated by incorporating appropriate changes in the birds feed The draw back of the conventional eggs for their use as source of vitamins and minerals is that they contain high cholesterol content (>210 mg/50g egg) which is an accepted high risk factor of cardiovascular diseases, such as atherosclerosis, myocardial infarction and hypertension The eggs as such, therefore, cannot be used as a source of vitamins and minerals for such patients unless modified The modified egg accomplishes by the provision of having low cholesterol, appropriate minerals and vitamins composition to act as their natural source which exist in bound form They are easy to absorb and have the minerals and vitamins composition sufficient enough capable to rehabilitate their status in diabetics and decrease the severity of the disease A modified egg of the present invention typically has between about 2 5-4mg of copper, 0 5-2 5 mg of zinc, 0 8-4 5 mg of magnesium 50-100 mg of iodine per 50g of liquid egg A present invention modified egg has about 10-12mg of vitamin E and about 0 3-0 75 mg of edible carotenoids as antioxidants The term poultry egg as employed herein refers to an egg laid by chicken hen and in general to avian species which include turkey, duck, goose, pigeon, guinea fowl, quail, pheasant, chukar, patndge, grouse, emu, rhea and ostrich The term liquid/edible egg is employed here m refers to liquid/edible content of whole poultry egg which is deshelled The commercial chicken shelled eggs consist of 8-11% shell, 56-61% albumen and 27-30% yolk The whole edible egg is constituted by 36% yolk and albumen 64% The detail of the composition of the whole edible egg is described in chapter 6 of egg "Science and Technology" Second edition Avi p Publishing company 1977 A modified egg of the present invention provides health and nutritive benefits in general and specifically to diabetics in comparison with commercial eggs such as chicken eggs A modified egg of the present invention typically has a contents per 50g of edible eggs of between about Yolk weight 15 -17g Albumen weight 32 - 34g Total protein 6-8 g Carbohydrates 0 5 - 0 7g Triglycerides 1 5-2 5 g Total cholesterol 95-125 mg Omega - 3 fatty acids 70-80 mg (Lenolenic acid) Phospholipids 1 5 - 2g In another embodiment, this mvention provides a poultry feed for production of eggs for the consumption of diabetic consumers The important aspect of the present invention is the removal of the free fat and oil rich ingredients and animal products from the conventional poultry feed so as to reduce fat contents in it Only the basic ingredients of oil seeds such as flax seeds for Omega-3-fatty acids as a main fat providing material is used as whole seed while soybean, mustard, sunflower and groundnut seeds are used as their cakes left after the extraction of oil for increasing the protein contents of the feed The present invention further contemplates the modifications in the mineral mixture in quantities between about zinc, 0 025 - 0 05mg, while the magnesium, 10-500 mg, copper, 500-1000 mg, manganese, 250-400 mg, iron, 250-500 mg, and iodine 0 01 - 100 mg derivatives per kilogram of the basal diet Zinc herein refers to zinc oxide, zinc sulphate, zinc chloride, zinc stearate, zinc acetate, zmc bromide, zinc nitrate or any other derivative of zinc Magnesium herein refers to magnesium acetate, magnesium - L - aspartate, magnesium carbonate, magnesium hydroxide, magnesium chloride, magnesium citrate, magnesium gluconate, magnesium lodate, magnesium iodide, magnesium oxide, magnesium phosphate, magnesium sulphate, magnesium nitrate or any other derivative of magnesium Copper herein refers to copper chloride, copper nitrate, copper oxide, copper sulphate, copper iodide or any other derivative of copper Manganese here refers to manganese acetate, manganese chloride, manganese oxide, manganese sulphate or any other derivative of manganese Iron herein refers to iron sulphate, iron chloride, iron nitrate, iron phosphate, iron oxide or any other derivative or iron Iodine herein refers to calcium iodide, calcium lodate potassium iodide, sodium iodide, cuperous iodide or other iodine bound organic compound or any other iodine derivative Vitamin mixture include Ascorbic acid, 200-700mg, calcium pentathenate, 300-500mg, ergocalciferol, 0 05-200mg, a-D-tocopherol, 200-1000mg, nicotinic acid, 300-500g, cyanacobalamine, 0 l-500mg, pyndoxine hydrochloride, 50-200mg, thiamine hydrochloride, 200-500mg, riboflavin, 100-300mg, choline chloride 200-500mg and folic acid 100-500mg As an additional feature a xanthophyll compound (20-50mg/kg) is added for providing the Roche colour scale value of the yolk in the absence of sufficient carotenoid source in the poultry feed to mcrease the acceptance of the egg by the consumer The Following examples illustrate the present mvention Example-I This example illustrates the production of modified eggs with lower content of cholesterol, modified minerals and vitamins composition in the egg The common practice of poultry breeding system is employed in the production of modified eggs Only the feed of the layers has been modified The composition of the experimental diet is listed in table-I (Table Removed) Mineral mixture include Sodium bicarbonate, 750 mg, Ferrous sulphate, 300 mg, Manganese sulphate, 500 mg, Copper sulphate, 750 mg, zinc sulphate, 50 mg, magnesium sulphate, 200 mg, potassium iodide, 0 005 mg in a kilogram of the basal diet Vitarmn mixture include Ascorbic acid, 500 mg, Calcium pentathenate, 400 mg, Choline chloride, 1000 mg, Folic acid, 10 mg, ergocalcrferol, 0 1 mg, pyndoxme hydrochloride, 80 mg, cx-D-tocopherol acetate, 500 mg, carotenoid, 20mg, of one kg of basal diet Example - II The example further illustrates the production of modified poultry eggs with a lowered content of cholesterol and more with vitamins and minerals in accordance with the present invention Following the example-I, 200 white layers, B V 300 Babcock strain, are fed adhbitum to the growers for 4 weeks prior to their ovulation and continued to feed on this diet throughout their ovulatory period After 2 weeks of ovulation and thereafter, the eggs were randomly collected for analysis The results of the analysis of these eggs are summarized in Table-II There are no differences in the, egg production or egg weight in comparison to the flocks of the birds feeding on conventional diet Table - II Values of the constituents of modified egg weighing 59 g (Table Removed) The following illustrations show the efficacy of the modified eggs herein albino rats (Wister strain) are used as test animal model fed on an established semi synthetic basal diet for feeding the animals which is given in table-Ill Table-in Semi synthetic basal diet (g/kg) of Wister rats. (Table Removed) Vitamins mixture (mg/kg) Ascorbic acid, 500, Biotm,4, calcium pentothenate,320, choline chlonde,2500, folic acid,10, Inositol,1000, Ergocalciferol 0 003, Nicotimc acid,300, Cyanocobalamin,0 40 Pyndoxme hydrochlonde,200, a-Tocoferol acetate,60, Retmol 0 30 *Minerals mixture (g/kg) Ca H2 P04, 25 3, MgSO4 H2O, 4 05, KCl, 3 43, K1,0 004, Na2 CO3 1 15, Fe SO4 H2O, 0 60, MnSO4H2O, 0 31, CuCl, 0 10, CoCl3 0 04, Zn SO4 7H2O 0 088, NaF, 0 008 This diet served as control (Diet -1) The non insulin dependant diabetes melhtus (NTDDM) is induced in rats by increasing ZnSo4 7H20 from 0 088mg/kg diet to 0 176mg/kg (Db-diet-I) and 0 352mg/kg (Db-diet-H) in the semi synthetic basal diet keeping all other ingredient constant This method is employed on the basis of the reports that excessive zinc in diet increases body fat in lean mice close to the genetically obese ob/ob mice (Chen M D, Lin PY Chen V and Lin W H Biol Trace Elem Res 52, 125-132, 1996) The rats when fed on three separated diets I e with 20mg Zn/kg diet (Control, Group -1) had a normal growth and blood profile pattern comparable to the rats fed on commercial standard rat pellet diet, while rats on 40mg Zn/kg diet (Group - II) and 80mg ZN/kg diet (Group - HI) showed mild and severe metabolic syndrome - X characterized by higher gain in body wt, hypertension, hyperglycemia, hypennsuhnemia and dyslipidemia comparable to those of human metabolic syndrome - X during 180 days of dietary treatment Zn being a part of human diet, this method of induction of NIDDM was preferred over streptozotocin or alloxan treated diabetic rats The following data illustrate the body wt, blood pressure, blood profile and Zn, Cu and Mg status in serum, liver, hair, abdominal muscle of male and female rats fed on basal diet containing 20mg Zn (Control, Group - I), 40mg Zn (Group -II, Db I) and 80mg Zn/kg diet (Group - III, Db II) for a period of 180 days The results of this experiment are summarized in tables IV, V, VI, VII and VIII Table IV Mean month wise body wt. and blood pressure in rats fed on basal diet (Group - I, control) and on Db diet -1 (Group - II) and Db diet - II (Group - ITI) for 120 days. (Table Removed) Values are mean + SEM of n = 10 Unne in rats on Db diet -1 (group-II) gives positive benedict test on day 90 and those on Db diet - II (group-Ill) on day 60 and there after until the termination of the experiment Table IV shows that the rats that are fed on Db diet- I (Group - II) and Db diet - II (Group - IE) gain more weight during the first 150 days of dietary treatment while it declined on 180 day Their systolic blood pressure shows an increments approximately 70% or more after 3 months of dietary treatment and onward than those of the rats fed on semi synthetic basal diet (Control) Table V Blood profile of male rats fed on basal diet (Control Group - I) and on Db diet - I (Group - II) and Db diet - II (Group - ID) after 180 days of dietary treatment. (Table Removed) Values are mean + SEM of n = 10 Table VI Blood profile of female rats fed on basal diet (Control Group - I) and on Db diet - I (Group - II) and on Db diet - II (Group - HI) after 180 days of dietary treatment. (Table Removed) Values are mean + SEM of n = 10 Data in the tables V and VI show that the blood profile of males and females rats on Db diet- I and Db diet - II show dyslipidaemia, hyperglycemia and high creatinine and hypennsulinemia, high cortisole, high aldosterone levels than those of control group of rats They therefore manifest typical noninsulin dependant diabetes mellitus complicated with hypennsulinemia and hypertension Table VII Mean Zinc (Zn), Copper (Cu) and Magnesium (Mg) concentrations in the serum, liver, hair, abdominal muscles and urine in male rats fed on basal diet (Control Group-I) and on Db diet-I (Group-II ) and on Db diet-II (Group-III) after 180 days of dietary treatment. (Table Removed) Values are mean ± SEM of n = 10 Table VIII Mean Zinc (Zn), Copper (Cu) and Magnesium (Mg) concentrations in the serum, liver, hair and abdominal muscles in female rats fed on basal diet (Control Group-I) and on Db diet-I (Group-II ) and on Db diet-H (Group-III) after 180 days of dietary treatment. (Table Removed) Values are mean + SEM of n = 10 Data in tables VII and VIII show that the mineral concentrations of Zn, Cu and Mg are significantly higher in serum of the group - II and III rats, whereas Zn concentration remains higher but those of Cu and Mg reduce drastically in other tissues such as liver, hair and abdominal muscles with their excessive loss in urine in spite of their adequate levels in their diets The altered mineral status in rats of groups - II and in is consistent with those of genetic diabetes mice/rats and human diabetic patients where an excessive loss of these minerals have been seen m urine leading to their deficiencies in the tissues Table IX Mean values of liver weight, total lipids, cholesterol, triglycerides and glycogen in rats fed on basal diet (Group I) and on Db diet-I and II (Group II and III) after 180 days of dietary treatment. (Table Removed) Values are Mean ± S E of n = 10 The data in table IX show reduction in total lipids, cholesterol, triglycerides and glycogen on Db diet -1 and Db diet - II rats which coincide well with their rise in the blood The data of body wt, blood pressure, blood serum profile and mineral status of rats in group-II and III clearly demonstrate that the diet containing higher Zn (even in pharmacological doses) when fed to rats for a longer duration induces diabetes in them, consistent with those of genetically db/db mice, Zukar diabetic fatty rats (ZDF/Drt) and in some of human diabetic patients The severity of the disease depends upon the amount of Zn m diet and degree of deficiencies of minerals in the tissues Even if the diet contains adequate levels of minerals and vitamins, they are not able to prevent the occurrence of these deficiencies in the tissues when Zn is excess in diet Example III This example illustrates that on feeding four fresh modified eggs mixed in diet to diabetic rats improve the metal status in tissues and reduce the severity of diabetes in them For this, two groups of rats are included in this study These groups are made out of the normal 21 days old (just after weaning period) males and females from a stock bred on the standard rat pellet diet They are fed on Db diet - I (group-II) and Db diet - II (group-Ill) respectively for a period of 120 days Their body wt blood pressure and urine glucose are monitored to check the onset of diabetes which are seen to manifest on 90th day in group-II and on 60 day in group-Hi fed diet groups similar to group II and III rats On the beginning 120th day, the animals of the group -II and III were shifted on the Db diet - I mixed with four fresh modified eggs and Db diet - II mixed with four fresh modified eggs/kg of semi synthetic diet respectively and continue to feed them on their respective diets for the period of 60 days prior to the termination of the experiment that lasted for 180 days Their body wt and blood pressure are monitored (Table X) Table X Mean month wise body weight and blood pressure in rats fed on basal diet (Control Group I) and on egg mixed Db diet-I & II (Group II* and III3) during the dietary treatment. (Table Removed) Values are Mean ±SE of n=10 Egg mixed Db diet -I and II are given for 60 days to rats (male and female) after 120 days of dietary treatment of Db diet-I and n without eggs The data of rats groups II* and III* show a lesser increase in their body wt and reduction in blood pressure compared to the group - II and group -III (refer Table IV) After the completion of the experiment, the blood serum and other tissues are bioassayed for the parameters investigated for group -1, II and III The result are summarized in table XI, XII, XIH, XIV and XV Table XI Blood profile of male rats fed on basal diet (Control Group -1) and on egg mixed Db diet -1 (Group - II)* and egg mixed Db diet - II (Group -III)* after the end of the dietary treatment. (Table Removed) Values are mean + SEM of n = 10 Egg mixed Db(EDb) diet I and II are given for next 60 days to male rats after 120 days of dietary treatment of Db diet-I and II Table XII Blood profile of female rats fed on basal diet (Group - I) and on egg mixed Db (EDb) diet - I (Group - II) and egg mixed Db (EDb) diet - II (Group -HI)* after the end of dietary treatment. (Table Removed) Egg mixed Db (EDb) diet-I and II are given for next 60 days to female rats after 120 days of dietary treatment of Db diet-I and II The data in table XI and XII show a significant reductions in total lipid, triglycerides, cholesterol, LDL-cholesterol, VLDL-cholesterol, glucose, insulin, cortisole, aldosterone , TSH, and rise in HDL-cholesterol occur in rats on egg mixed Db - I diet than those of (Table Removed) Db diet -1 group Their values are close to those of control group-I However, the recovery of these molecules are also significant in rats on egg mixed diet Db diet - II than those of Db diet - II group but their level approached close to the rats on Db diet -1 (group-II) and not close to the control group -1 both in male and female rats (Refer tables V and VI for comparison) Table XIII Mean Zinc (Zn), Copper (Cu) and Magnesium (Mg) concentrations in the serum, liver, hair, abdominal muscles and urine in male rats fed on basal diet (Control) and on egg mixed EDb diet I and II (Group II and III) after the end of dietary treatment. (Table Removed) Values are Mean ± S E M of n = 10 Egg mixed Db (EDb) diet I and II are given for next 60 days to male rats after the 120 days of dietary treatment of Db diet I and II Table XIV Mean Zinc (Zn), Copper (Cu) and Magnesium (Mg) concentrations in the serum, liver, hair, abdominal muscles and urine in female rats fed on basal diet (Control) and on egg mixed EDb diet I and II (Group n* and HI) after the end of dietary treatment. Values are Mean ±SEM of n =10 Egg mixed Db (EDb) diet-I and II are given for next 60 days to female rats after the 120 days of dietary treatment of Db diet I and II The data in tables XIII and XIV show that Zn concentration in serum and tissues significantly declines while the concentrations of Cu and Mg increase in the tissues and decline in the blood serum m rats on egg mixed Db diet - I than those of rats on Db diet -1 approaching close to the control values while their recovery is significant in rats on egg mixed Db diet - II rats approaching close to Db -1 group (Group-II) rats but not to the level of control group (Group-I) (For comparison refer tables no VII and VIII) Table XV Mean values of liver weight, total lipids, cholesterol, triglycerides and glycogen in rats fed on basal diet (Control) and on egg mixed Db diet-I and II (Group II* and III) after the end of dietary treatment. (Table Removed) Values are Mean ±SE ofn=10 Egg mixed Db (EDb) diet-I and II are given for 60 days to rats (male and female)' after 120 days of dietary treatment of Db diet-I and II The data in table XV show that the lipids, cholesterol, triglycerides and glycogen concentration the liver of both males and females show a remarkable recovery in rats on modified egg mixed Db diet -1 and egg mixed Db diet - II approaching close to the control values It is evident from the data per se that the inclusion of these modified eggs in diet of diabetics helps to restore the altered mineral composition of blood and tissues and reduce the severity of diabetes and protects them against diabetic induced complications if the individuals are feeding on diabetogenic diet Example IV This example illustrates that these modified eggs protect the embryos in womb, help to deliver normal neonates by preventing malformations in them due to diabetes complicated with hypertension and help towards the restoration of mineral composition in the body of the young ones For this study, three groups of rats are prepared from 21 days old pups just after their weaning period The males and females rats are separated from the main stock and divided into 3 groups Group-I is fed on the semi synthetic basal diet (Control) Group-II on Db diet -I and group-Ill on Db diet -II for a period of two months At this stage animals in group II and III are subdivided into two groups each One subgroup of group II is continued to feed on Db diet - I and the other on four fresh modified egg mixed Db diet - I/kg diet (Group - II) Like wise one subgroup of group - III is continued to feed on Db diet -II (Group -III) and the other on four fresh modified egg mixed Db diet - II/kg diet (Group - III) for a period of two months After a total of 4 months dietary treatment, the males and females in each group are mated within the group and continue to feed on their respective diet during their gestation and suckling periods The body wt and health of the neonates are recorded during their weaning period The result are summarized in tables XVI, XVII and XVIII Table XVI Mean Body Wt (g) of Pups of F1- Generation at birth and during the weaning period obtained after mating the Parents fed on basal diet (Group - I) and on Db diet - I (Group-II) and on Db diet-II (Group-III) (Table Removed) Values are mean + SEM Table XVII Mean Body Wt. (g) of pups of F1 - Generation at birth and during the weaning period obtained after mating the Parents fed on basal diet (Group-I) and on egg mixed Db diet -I (Group- II) and on egg mixed Db diet - II (Group - III) (Table Removed) Values are mean + SEM Table XVIII Mean Zinc (Zn), Copper (Cu) and Magnesium (Mg) concentrations in the liver, hair and abdominal muscles in pups of F1 generation just after the weaning period of pups (F-l Generation) (Table Removed) Values are mean + SEM of n = 10 Egg mixed Db diet-I and II was given for 60 days to male and female rat parents after 60 days of dietary treatment of Db diet-I and II respectively The Fl generation obtained after mating shows that the neonates of group -II and group-III have higher birth wt than those of their control, have malformed toes of the forehmb (curled toes), show restricted unbalanced body movements, gain lesser body wt during weaning period They are diabetic when their urine is tested on 10th and 19 day of weaning period by Benedicts test They possess extremely low concentration of Cu, Zn and Mg in their tissues and die after the completion of the weaning period (Table XVIII) The neonates of group-II and III* whose parents are fed on egg mixed Db diet- I and egg mixed Db diet -II also have higher body wt than those of control, gain more wt than group -II, and III during weaning period They remain healthy on completion of their weaning period But they are diabetic There is a partial rehabilitation of Cu, Zn and Mg concentrations in their tissues (Table XVIII) as their concentration are significantly higher than the groups II and III but still lower than the control group of rats The data of this set up experiment illustrate that the diabetes induced by this method in parents is genetically transmitted to the off spring of F1 generation and off springs die after weaning period due to malformations attributed by uncontrolled diabetes and hypertension But the inclusion of these modified eggs in diet of the dams protects the embryos in womb, allow them to deliver normal neonates and help towards restoration of minerals composition in their body Example V This example illustrates that the modified eggs provide protection against diabetes not only to those feeding on diabetogenic food but also against genetically transmitted diabetes For this study, the survived neonates of F1 generation of group -IE* are used After the weaning period of pups in group - IIP, they are subdivided into two subgroups One subgroup of group - IIP is fed on standard rat pellet diet (Group-EP) and other on egg mixed standard rat pellet diet (Group IE) for a period of 60 days Their body wt and systolic blood pressure are recorded on weekly basis through out the duration of the experiment The results of this study are summarized in table XIX Table XIX Mean body weight and blood pressure in Pups of F1 generation fed on pellet diet of control (Group - IF) on pellet diet of F-l generation of Db diet - III (Group - III) and on egg mixed pellet diet of F - 1 generation of Db diet - III* (Group - III*) after 60 days of dietary treatment. (Table Removed) Values are mean + SEM of n = 10 After 60 days of dietary treatment, they are sacrificed and bioassayed (Tables XX, XXI, XXII) Table- XX Blood profile of male rats of F1 generation fed on pellet diet of control (Group -1) on, pellet diet of F-1 generation of Db diet - III (Group - III) and on egg mixed pellet diet of F -1 generation of Db diet - III* (Group - III*) after 60 days of dietary treatment. (Table Removed) Values aremean+ S E M of n = 10 Table XXI Blood profile of female rats of F1 generation fed on pellet diet of control (Group -1) on pellet diet of F-1 generation of Db diet - III (Group - III) and on egg mixed pellet diet of F - 1 generation of Db diet - III* (Group - III*) after 60 days of dietary treatment. (Table Removed) Values are mean + SEM of n = 10 The data in table XX and XXI reveal that rats in group - III* on egg mixed pellet diet do not differ significantly m their serum glucose, total lipids, triglycerides, total cholesterol, HDL cholesterol, LDL cholesterol, VLDL cholesterol, creatinine, insulin, Cortisol, TSH, T3 and T4 than those of control rats However, the rats in group in display elevated concentrations of the blood parameters These rats are genetically predisposed to diabetes The normal diet can not rehabilitate the genetically altered physiology in them (Table XX and XXI) Table-XXII Mean Zinc (Zn), Copper (Cu) and Magnesium (Mg) concentrations in the serum, liver, hair and abdominal muscles in male rats of F1 generation fed on pellet diet of control (Group - I) on pellet diet of F-1 generation of Db diet - III (Group - III) and on egg mixed pellet diet of F - 1 generation of Db diet - III* (Group - III*) after 60 days of dietary treatment. (Table Removed) Values are mean + SEM of n = 10 Table XXIII Mean Zinc (Zn), Copper (Cu) and Magnesium (Mg) concentrations in the serum, liver, hair and abdominal muscles in female rats of Fi generation fed on pellet diet of control (Group - I) on pellet diet of F-1 generation of Db diet - III (Group - III) and on egg mixed pellet diet of F - 1 generation of Db diet - HI* (Group - III*) after 60 days of dietary treatment. (Table Removed) Values are mean + SEM of n = 10 There is a reduction in mineral concentrations of Zn, Cu and Mg in group III close to Db -I and Db-II rats in group -II and III compared to control rats (Refer to tables no VII and VIII) However, in group III*** the mineral composition in blood serum, abdommal muscles, liver and hair are close to the control rats This shows that the addition of these modified eggs rehabilitate the mineral composition in diabetics Example VI The following example illustrates that these eggs are safe to consume by healthy individuals For this purpose four liquid eggs were added in the pallet diet and healthy rats were fed on this egg mixed diet for a period of 60 days after their weaning period The data obtained is given in the following tables Table XXIV Mean body weight and blood pressure in rats fed on pellet diet and egg mixed pellet diet after two months of dietary treatment. (Table Removed) Values are mean + SEM of n = 10 Table XXV Blood profile of male rats fed on basal diet and egg mixed basal diet after two months of dietary treatment. (Table Removed) Values are mean+S EM of n = 10 Table XXVI Blood profile of female rats fed on basal diet and egg mixed basal diet after two months of dietary treatment. (Table Removed) Values are mean + SEM of n = 10 Table XXVII Mean Zinc (Zn), Copper (Cu) and Magnesium (Mg) concentrations in the serum, liver, hair and abdominal muscles in male rats fed on pellet diet and egg mixed pellet diet after two months of dietary treatment. (Table Removed) Values are mean + SEM of n = 10 Table XXVIII Mean Zinc (Zn), Copper (Cu) and Magnesium (Mg) concentrations in the serum, liver, hair and abdominal muscles in female rats fed on pellet diet and egg mixed pellet diet after two months of dietary treatment. (Table Removed) Values are Mean ± S E M of n=10 Table XXIX Mean values of liver weights, total lipids, cholesterol, triglycerides and glycogen in rats fed on pellet diet and egg mixed pellet diet after 2 months of dietary treatment. (Table Removed) Values are Mean ±SEM of n = 10 The data in tables XXIV, XXV, XXVI, XXVII, XXVIII, XXIX show that the inclusion of eggs m pallet diet of rats do not significantly alter the body wt, B P, blood profile and mineral composition in tissues They are therefore, safe for healthy consumers. We claim: 1) An improved poultry feed for the production of eggs with capacity of restoration of minerals composition in a patient, said feed comprising a dietary material of broken rice up to 35%, rye oat up to 15%, flax seed up to 2%, de-oiled rice bran up to 18%, de-oiled mustard cake up to 5%, de-oiled sunflower cake up to 2%, de- oiled groundnut cake up to 3%, di-hydrogen calcium phosphate up to 1.5%, marble powder up to 3%, marble grit up to 7%, mineral mixture up to 5%, vitamins mixture up to 5%, tyrosine powder up to 0.2% and multi-enzymes up to 0.05% by wt. in one kg of the feed such as here in described, wherein said minerals comprise from 0.2 to 2.0g of sodium derivatives, from 0.2 to 0.3g of ferrous derivatives, from 0.05 to 500g of magnesium derivatives, from 0.25 to 4g of manganese derivatives, from 0.02 to 0.10g of zinc derivatives, 0.05 to lg copper derivatives, from 0.002 to 0.005g of iodine derivatives and from 0.002 to 0.00lg of selenium derivatives and said vitamins comprise from 200 to 700mg of ascorbic acid, from 300 to 500mg of calcium pentathenate, from 200 to 500mg of choline chloride, from 100 to 500mg of folic acid, from 0.05 to 0.200mg of ergocalciferol, from 300 to 500mg of nicotinic acid, from 0.01 to 100mg of cyanocobalamine, from 50 to 200mg of pyridoxine hydrochloride, from 200 to 500mg of thiamine hydrochloride, from 100 to 300mg of riboflavin, from 200- 500mg of alfa-tocopherol, from 15 to 40mg of carotenoid per 1 kg of the poultry feed. 2) The poultry feed as claimed in claim 1, wherein the minerals are selected from one or more sodium derivatives, ferrous derivatives, magnesium derivatives, manganese derivatives, zinc derivatives, copper derivatives, iodine and selenium derivatives. 3) The poultry feed as claimed in claim 2, wherein the mineral mix is comprised of sodium bicarbonate, 0.75 to lg; ferrous sulphate, 0.3 to 0.5g; manganese sulphate, 0.5 to 0.7g, copper sulphate, 0.75 to lg,; zinc sulphate, 0.050 to 0.075g,; magnesium sulphate 0.2 to 0.5g; potassium iodide, 0.0005g-0.0075g; and sodium selenite, 0.0005-0.0008g.

Full Text

Filed of the invention
The present invention relates to a poultry feed for use in the production of eggs with optimal concentration of minerals and vitamins More particularly, the present invention relates to an improved poultry feed for production of eggs with a capacity of restoration of mineral composition in mammals with diabetes, hypertension, obesity, hypercholesterolemia and hyperinsulinemia The present invention also relates to a method for the manufacture of such unproved poultry feed The present invention particularly relates to an improved egg with a capacity of restoration of mineral composition in mammals with diabetes, hypertension, obesity, hypercholesterolemia and hyperinsulinemia and a method for the production thereof The present invention also relates the use of minerals and vitamins in poultry eggs, through optimizing the composition of minerals and vitamins mixtures in poultry feed, to ameliorate their deficiencies in diabetics The modified egg contains appropriate concentrations of vitamins and minerals which on consuming helps to restore the depleted minerals almost close to their normal levels in the tissues and reduces the severity of hypercholesterolemia, dyslipidemia, hyperinsulinemia, hyperglycemia and arterial hypertension of diabetic consumers Background of the invention
In diabetics, the body tissues tend to lose their minerals and the severity of the disease increases with further deterioration of mineral homeostatic mechanism notably of copper, zinc, magnesium, manganese and chromium etc with passage of time Some of these minerals are the constituent of either cytosohc or intramitochondrial superoxide dismutases enzyme (SOD), catalases glutathione-s-transferases and glutathione reductases which disarm the free reactive oxygen species (ROS) and protect the cell components from peroxidation Their altered levels in the diabetics lead to the deficiency of either one or more metals resulting in a reduction of antioxidative defense system As a consequence of this, ROS generated get accumulated in the cells leading to the severity of the disease
A number of dietary supplement formulations of vitamins and minerals combinations in the form of tablets/capsules have been developed for human consumption to restore the altered minerals and vitamins status in the body so as to prevent the complications in the disease occurring due to their deficiencies/excess
However, the results of the available dietary supplements are not as satisfactory as are expected particularly in diabetics since they are in synthetic forms whose absorption rate is less than those of natural forms or either their binding legends get depleted during the course of the disease or they are not as complete as they are thought of
The poultry eggs can be conveniently used as a material for introduction of vitamins and minerals to restore their composition in the body since they serve as the natural source of vitamins and minerals which exist in bound forms They are easy to absorb and the concentrations of vitamins and minerals in eggs can be manipulated by incorporating appropriate changes in the birds feed
The prior art methods had considerable disadvantages One of the drawbacks of the conventional eggs for their use as source of vitamins and minerals is that they contain high cholesterol content (>210 mg/50g egg) which is an accepted high risk factor of cardiovascular diseases, such as atherosclerosis, myocardial infarction and hypertension usually associated with NIDDM The eggs as such, therefore, cannot be used as a source of vitamins and minerals for such patients unless modified
Appropriate modification in dietary feed has led to the development of the eggs which have lower cholesterol (106-160mg/50g egg), less saturated fats (0 8-1 2g) more of omega-3 unsaturated fatty acids (70-400mg), more of vitamin E (0 5-10mg), folate (10-150 mg) and iodine (1-120 meg) than the standard eggs reported in technical publication which include US Patent No, 4128640, 4187294, 4197293, 4197294, 4394376, 4410541, 4739853, 4764531, 501276, 5091195, 5246717, 5415879, 5520938, 5520398, 5665375, 6316041 and 5759567 incorporated herein by reference
No attempts have been made to modify the minerals in the feed of the birds (except copper and iodine as cholesterol lowering agents in eggs) for the production of eggs, which on consuming may rehabilitate the minerals status in diabetics Objects of the invention
Accordingly, it is an object of the present invention to provide a poultry feed for use in the production of eggs with optimal concentration of minerals and vitamins
It is another object of the present invention to provide an improved poultry feed for production of eggs with a capacity of restoration of mineral composition in mammals with diabetes, hypertension, obesity, hypercholesterolemia and hypermsulmemia
It is another object of the present invention to provide a method for the manufacture of such improved poultry feed
It is another object of the present invention to provide a method for producing modified eggs conducive to the healthy consumers
It is another object of the present invention to provide poultry eggs, which restore the depleted minerals composition in the consumers
It is another object of the present invention to provide poultry eggs with minerals, which lower the blood cholesterol, LDL-c, VLDL-c and increase triglycerides and HDL-c in diabetic consumers
It is another object of the present invention to provide modified eggs, which lower the blood pressure in diabetic consumers
It is another object of the present invention to provide modified eggs, which lower elevated blood glucose in diabetic consumers
It is another object of the present invention to provide modified eggs, which lower the elevated insulin level in blood of diabetic consumers
It is another object of the present invention to provide modified eggs, which lower the elevated creatinine in blood and urine of diabetic consumers
It is another object of the present invention to lower the elevated cortisole and TSH and increase the depressed levels of T3 and T4 in blood of diabetic consumers
It is another object of the present invention to provide modified eggs to lower the elevated aldosterone concentration in the blood of the diabetic consumers
It is another object of the present invention to provide modified eggs to restore the normal level of glycogen in liver and muscles of diabetic consumers
It is another object of the present invention to provide modified eggs to restore the disordered pathology of the tissue of diabetic consumers Summary of the invention
The above and other objects of the present invention are achieved by the modified egg of the present invention wherein the modified egg is a whole shell poultry egg having minerals and vitamins composition sufficient enough to interact with the tissue minerals and vitamins so as to rehabilitate their altered status in diabetics and decrease the seventy of the disease The minerals and vitamins are delivered through a novel poultry feed of the present invention These minerals predominately include zinc, copper, magnesium and manganese
Accordingly, the present invention provides an improved poultry feed for production of eggs with a capacity of restoration of mineral composition in a patient said feed comprising a major amount of dietary material such as herein described, from up to 30 % by wt of minerals of the kind such as herein described and up to 30 % by wt of vitamins of the kind such as herein described
In a preferred feature, the minerals are selected from one or more of Sodium derivatives, Ferrous derivatives, Magnesium derivatives, Zinc derivatives, Copper derivatives and Iodine derivatives
In a preferred feature, said minerals comprise from 0 2 to 2 0 g of sodium derivatives, from 0 2 to 3 0 g of ferrous derivatives, from 0 05 to 500 g of magnesium derivatives, from 0 02 to 0 05 g of zinc derivatives, form 0 50 to 1 g of copper derivatives, from 0 002 to 0 005 g of iodine derivatives, based on 1 kg of said poultry feed
In a preferred feature, the minerals are selected from sodium bicarbonate, ferrous sulphate, manganese sulphate, copper sulphate, zinc sulphate, magnesium sulphate and potassium iodide or mixtures thereof
In a preferred embodiment, said vitamins are selected from ascorbic acid, calcium pentathenate, choline chloride, folic acid, ergocalciferol, nicotinic acid, cyanocobalamine, pyndoxine hydrochloride, thiamine hydrochloride, riboflavin, a-Tocopherol, carotenoid and mixtures thereof
In a preferred feature, said vitamins comprise from 200 to 700 mg of ascorbic acid, from 320 to 500 mg of calcium pentathenate, from 200 to 500 mg of choline chloride, from 100 to 500 mg of Folic acid, from 0 05 to 0 200 mg of ergocalciferol, from 300 to 500 mg of nicotinic acid, from 0 01 to 100 mg of cyanocobalamine, from 50 to 200 mg of pyndoxine hydrochloride, from 200 to 500 mg of Thiamine hydrochloride, from 100 to 300 mg of riboflavin, from 200 to 500 mg of, a-tocopherol, from 15 to 40 mg of carotenoid, of each of said vitamins amount based on lKg of the said poultry feed
The present invention also provides a modified egg wherein the egg is a whole shell poultry egg having a content of constituents comprising total protein,6 5-8g, carbohydrates 0 5-0 7g, total lipids, 5-6 5g, triglycerides, 1 5-2 5g, cholesterol, 95-125mg, phospholipids,! 5-2mg, Omega-3 fatty acids (Lenolenic acids), 70 - 80mg, vitamin E,10-12 5mg, vitamin D, 0 50-0 850 mcg, Bi,0 05- 0 10mg, B2,0 275-0 350mcg, Bi2,0 18 -0 25mcg, foliate, 0 03-0 1mg, B6,0 015-0 05mcg, carotenoids, 0 5-0 85mg, Ca++,30 - 40mg, Mg,0 8-2 5mg, Zn,0 5-1 5mg, Cu,2 0-3 5mg, Fe,l 75-2 5mg, iodine 0 75-2 5mcg per 50g liquid egg
In a preferred embodiment, the poultry egg is selected from chicken, egg, duck egg and turkey egg
The present invention also relates to a method for the production of a modified egg which comprises feeding poultry with a feed comprising of an improved poultry feed for production of eggs with a capacity of restoration of mineral composition in a patient said feed compnsmg a major amount of dietary material such as herein described, from up to 30 % by wt of minerals of the kind such as herein described and up to 30 % by wt of vitamins of the kind such as herein described, allowing said poultry to lay eggs and collecting said eggs
Detailed description
Genetically obese (ob/ob) mice, diabetic (db/db) mice and Zuker diabetic fatty rats (ZDF/Drt) and transgenic rodents serve as models of research to understand the etiology of obesity, diabetes, hypertension and cardiovascular diseases One of the important finding on these animals is that they absorb and retain greater amount of zinc in liver, adipose tissue and tnceps muscles than their lean control counter parts They have a greater potential of absorbing orally administered zinc from the lumen of gastrointestinal tract to blood plasma and can absorb 60% of more of glucose than those of their lean counter parts (Collip J A, Pediatrics Ann 13, 465-472, 1984, Begin - Heick, N, Daple - Scott J M, Rowe J and Heick M M Diabetes 34, 179-184,1985, Kennedy M L and Failla M L, J. Nutr 117, 886-893,1987, Lin W H, Chen M D and Lin P Y J. Formoson, Med. Assoc 91, 27-33, 1992]
Supplementation of Zn promotes food intake, linear growth and body wt increase in children suffering from marginal to severe Zn-deficiency (Mc Gain C J, Kasarkis E J Jr Allen EJ, Prog. Food Nutr Sci. 9, 185-226, 1985) Investigatory studies on laboratory animals however provided evidences that anabolic effects of Zn result from elevated food consumption, higher absorption rate of nutrients l e amino acids (Moran J R and Lyerly A, Life Science 36 2551-2556 1985), fatty acids (Koo S I and Turk D E, J. Nutr 107 909-919, 1977) and glucose (Taneja S K and Arya P, Brit. J. Nutr 68 505-514, 1992) in addition to activation of protein and nucleic acids syntheses These anabolic effects of Zn are unlikely to dimmish after cessation of growth The continuous input of excess nutrients in tissues by excessive bioavailability of Zn after adolescence may create environment essential for obesity It has been reported that an increase of zinc to double the amount of recommended dietary allowance (RDA) increases the portion of the body fat during 25 weeks period of dietary treatment in healthy children (Prentice A, Nutr. Rev 51, 268-270, 1993) There exists a positive linear correlation between hair zinc and body wt in adult humans The obese of both the sexes possess higher zinc concentration in their hair than those with normal body wt/ for their height (Taneja SK, Mahajan M and Arya P Expenentia 52, 31-33,1996)
A study on the relation of Zn absorption and deposition of fat in ob/ob mice and high fat diet induced ICR mice showed that the ICR mice fed a high fat diet containing Zn in greater amount resulted in significantly higher gain in body wt and more fat deposition identical to ob/ob mice than did their lean control counter parts fed on a diet containing high fat but low zinc [Chen M D , Lin P Y , Cheng L V and Lin W H Biol. Trace Elem. Res 52 125-132,1996]
The proposed potential mechanism for development of obesity involving Zn based on the observations on the activation of ovine metallothionein - ovine growth hormone transgene (OMT - la - OG) by ZnSO4 is that it activates the genes of growth hormone during early period of growth and induces the relatively undifferentiated preadipacytes to be committed to become adipocytes This may increase their population The enhanced population of adipocytes responds by filling with tnacylglycerol due to increased absorption of nutrients caused by Zn and eventually leading to the observed state of obesity Alternately, the elevated growth hormone level may increase the insulin level which lead to increased glucose transport into adipocytes, thereby enabling them to increase hpogenesis Accordingly, the over supply of zinc during growth phase sets in the conditions for the onset of obesity [Pomp D, Oberbauer A M and Murray J D, Transgenic Res 5, 13-23,1996]
However, the obese subjects possess either lower or normal plasma level of Zn compared to nonobese control (Chen M D , Lin P Y Sheu W W H, Biol. Trace Elem. Res. 60, 123-129, 1997) The lower plasma Zn level in obese individuals can return to normal following dietary therapy (Di-Martino G, Matera M G, Di-Martino B, Vacca C, Di-Martino S and Rossi F , J. Med. 24 177-183, 1993)
In fact the plasma Zn status does not truly reflect the body Zn status as it tends to mamtain Zn homeostasis either by increasing Zn absorption or decreasmg Zn excretion depending upon Zn intake and metabolic requirements Therefore, no significant difference can exist between BMI and serum Zn in healthy subjects Tissue Zn status is thus more reliable to determine the relationship between the two It is evedent from a report that the hair Zn concentration increases with increase in BMI in Indian women (Taneja S K and Mahajan M, J. Punj. Acad. Sci., 1,211-216, 1999)
Thus, there are strong reasons to believe that excessive bioavailability of Zn, either in food chain or due to genetic disorder, to the tissues results in obesity Obesity poses a formidable challenge to the growing population all over the world as it is etiologically linked to insulin resistance accompanying to non-insulin dependent diabetes melhtus, hypertension and coronary artery disease This classical clustering of disease is described as metabolic syndrome - X
Insulin resistance seems to be an essential adaptation to prevent further weight gain in obese subjects (Eckel RH, Lancet 340, 1452-1453, 1992) and is developed through increased secretion of glucocorticoid from zona fasciculate of adrenal cortex Insulin resistance results in hyperglycemia, glucosuna, altered protein, fat and carbohydrate metabolisms and chronical complications resulting from macro and microvascular pathology Associated with it, altered metabolism of Cu, Zn, Mn and Mg are frequently seen in
experimental animals and humans There have been numerous studies evaluating the mineral status of diabetic subjects These studies yield inconsistent results
The most frequently reported features of diabetics in genetic diabetic animal model and humans are that they have higher Cu and Mg concentrations, low Zn concentration in their blood plasma and display high amount of Zn loss in urine The degree of increase in plasma Cu in diabetes melhtus is associated with its complications of hypertension or macrovascular disease The plasma magnesium level has been reported to be less when diabetes melhtus is associated with retinopathy (Walter R M Jr, Unu-Hare J Y, Olin K L, Oster M H, Anawalt B D , Cntch field J W and Keen C L, Diabetes Care 14 1050-1056, 1991)
However, normocupremia, hypocupremia, or hyperzincemia, normozmcemia and hypozincemia have been reported in some studies The concentrations of metals in blood plasma at a given time represent a total of bound, exchangeable and catabolic components of metals They therefore, can lead to conflicting results and predictions, more so in case of chronic diseases where urinary losses are high The metals concentrations in tissues are more dependable indicators for assessing metal deficiencies since their concentrations are maintained through dynamic equilibrium between tissue metals and exchangeable metal components of the blood plasma Some investigators therefore, preferred other tissues such as liver, muscles, hair, nails etc The levels of zinc and copper in hepatic and renal tissues have frequently been seen either higher or comparable to normal in diabetic melhtus complicated with hypertension The Cu in hair and nails in diabetes melhtus, hypertension and coronary artery disease patients have been found to be low (Sukumar A and Subramamam R, Biol. Trace Elem. Res., 34 89-98, 1992, Taneja S K, Mahajan M, Gupta S and Singh K P , Biol. Trace Elem. Res., 62, 255-264,1998) Cu deficiency has been also reported in muscle biopsies of diabetics (Sjogren A, Edvinsson L , Floren C H and Abdulla M, Nutr. Res., 6, 147-154, 1986)
It has been reported that the NIDDM and myocardial infarction susceptible individuals are more prone to obesity as they are genetically predisposed to excess Zn absorption at a younger age, which reduces Cu bioavailability to the tissues over a penod of time The manifestation of clinical symptoms depends upon the degree of ionic imbalance resulting from the degree of stress and the type of food the individual consumes during intervening period Higher stress and zinc rich food aggravate the condition and favour their early onset compared with low stress or low zinc intake in food Because fibers and phytates are known to reduce zinc bioavailability their liberal use delay the onset of obesity and obesity related diseases (Taneja S K, Mahajan M Gupta S and Singh K P Biol Trace
Elem. Res 62, 255-264, 1998, Taneja SK Girhotra S and Singh KP, Biol Trace Elem. Res 75, 177-186,2000)
The reported variations may represent the trace elements concentration in diet, the pattern of progression of disease and degree of depletion of respective mineral binding legends due to excessive loss of albumin protein, increased rate of proteolysis and lipolysis
The mechanistic functional consequences of mineral deficiencies have been discussed differently by different investigators to explain the etiology of diabetes mellitus
Since Zn deficiency adversely affects the glucose disposal as it provides stimulating effect of glucose transfer in muscles and adipocytes, the therapeutics benefit of Zn supplementation has been reported in subjects with diabetes mellitus and obesity which both have insulin resistance (Brun J, Guintrand - Hugret R, Fons C , Carvajal J , Fedou C, Fusselher M, Bradet L and Orsetti A, Biol. Trace Elem. Res., 47 385-391, 1995) However, the reality of zinc deficiency in diabetes mellitus has been criticized, as its supplementation further mcreases serum Hb Aic values in diabetes mellitus (Cunningham J J, Fu A, Mearkle P L and Brown R G, Metabolism, 43 1558-1562, 1994)
The tissue copper deficiency aggravates hyperglycemia, hypercholesterolemia, hypertension and dyslipidemia and reduction in the activity of cytosolic superoxide dismutase resulting in alteration of antioxidant defence system The reduction in sehnogleutathione peroxidase (Se-GSHPX) activity and increased synthesis of cholesterol promotes lipoprotein oxidation under copper deficiency in particular These effects are important given the etiological role of oxidised low density lipoprotein in initiation of atherosclerosis [Kathenne L O Robert M W and Carl L K Am. J. Clin. Nutr 59, 654-658, 1997 Singal P K, Kharpar N Palace V and Kumar D , Cardiovascular Res 40, 426-432, 1998]
Copper deficiency during pregnancy affects the cross linking of connective tissue protein causing abnormal growth and maturation of the foetus [Barone A Ebesh, O Harper R G and Wapmr RA J. Nutr 128 1037-1041, 1998]
The reduction of Mg (Mg ATP 2") occurs due to the presence of Mg binding defect (Mg BD) and accounts for all aspects of type - II diabetes It therefore, has a conservative role in the development of the disease (Ibert C W , Devendra K and J A, Biol. Trace. Elem. Res 98, 97-107, 2004] And most of the complications of type-II diabetes such as arrhythmias, hypertension and sudden cardiac death may be prevented by prophylactic magnesium supplementation (Balon T W, Gu J J, Tokuyama Y , Jasman A P and Nadler J J Am. J. Physiol. 269, 745-752, 1995)
Manganese is a constituent of mitochondrial Mn-superoxide dismutase Its deficiency results in the generation of free radical of oxygen (ROS) leadmg to the oxidative
stress and supplements to adverse effect of the zinc and copper deficiency leading to the depletion of cellular antioxidants and increased production of peroxidation products which accelerate the complications in diabetics (Cederberg J Galh J Luthman H and Enkson U J Diabetes 49, 101-107, 2000, Cedarberg J, Martin S and Eriksson UJ, Pediat. Res., 49, 755-762, 2001)
These finds have led to the development of dietary supplements containing minerals and antioxidative vitamins in the form of capsules/tablets to relieve the diabetics from the oxidative stress by ameliorating the relevant minerals deficiency and increasing the cellular antioxidants such as vitamins E, A and C and other vitamins related to general metabolism including Bl, B2, B6, B12, etc However, the results of the available dietary supplements are not as satisfactory as expected particularly in diabetics, since they are in synthetic forms whose absorption rate is less than those of natural forms or their binding legends get depleted during the course of the disease or they are not as complete as they are thought of
The poultry eggs can be conveniently used as a material for introduction of vitamins and minerals to restore their composition in the body since they serve as the natural source of vitamins and minerals which exist in bound forms They are easy to absorb and the concentrations of vitamins and minerals in eggs can be manipulated by incorporating appropriate changes in the birds feed
The draw back of the conventional eggs for their use as source of vitamins and minerals is that they contain high cholesterol content (>210 mg/50g egg) which is an accepted high risk factor of cardiovascular diseases, such as atherosclerosis, myocardial infarction and hypertension The eggs as such, therefore, cannot be used as a source of vitamins and minerals for such patients unless modified
The modified egg accomplishes by the provision of having low cholesterol, appropriate minerals and vitamins composition to act as their natural source which exist in bound form They are easy to absorb and have the minerals and vitamins composition sufficient enough capable to rehabilitate their status in diabetics and decrease the severity of the disease
A modified egg of the present invention typically has between about 2 5-4mg of copper, 0 5-2 5 mg of zinc, 0 8-4 5 mg of magnesium 50-100 mg of iodine per 50g of liquid
egg
A present invention modified egg has about 10-12mg of vitamin E and about 0 3-0 75 mg of edible carotenoids as antioxidants
The term poultry egg as employed herein refers to an egg laid by chicken hen and in general to avian species which include turkey, duck, goose, pigeon, guinea fowl, quail, pheasant, chukar, patndge, grouse, emu, rhea and ostrich
The term liquid/edible egg is employed here m refers to liquid/edible content of whole poultry egg which is deshelled
The commercial chicken shelled eggs consist of 8-11% shell, 56-61% albumen and 27-30% yolk
The whole edible egg is constituted by 36% yolk and albumen 64% The detail of the composition of the whole edible egg is described in chapter 6 of egg "Science and Technology" Second edition Avi p Publishing company 1977
A modified egg of the present invention provides health and nutritive benefits in general and specifically to diabetics in comparison with commercial eggs such as chicken eggs
A modified egg of the present invention typically has a contents per 50g of edible eggs of between about
Yolk weight 15 -17g
Albumen weight 32 - 34g
Total protein 6-8 g
Carbohydrates 0 5 - 0 7g
Triglycerides 1 5-2 5 g
Total cholesterol 95-125 mg
Omega - 3 fatty acids 70-80 mg (Lenolenic acid)
Phospholipids 1 5 - 2g
In another embodiment, this mvention provides a poultry feed for production of eggs for the consumption of diabetic consumers
The important aspect of the present invention is the removal of the free fat and oil rich ingredients and animal products from the conventional poultry feed so as to reduce fat contents in it Only the basic ingredients of oil seeds such as flax seeds for Omega-3-fatty acids as a main fat providing material is used as whole seed while soybean, mustard, sunflower and groundnut seeds are used as their cakes left after the extraction of oil for increasing the protein contents of the feed
The present invention further contemplates the modifications in the mineral mixture in quantities between about zinc, 0 025 - 0 05mg, while the magnesium, 10-500 mg, copper,
500-1000 mg, manganese, 250-400 mg, iron, 250-500 mg, and iodine 0 01 - 100 mg derivatives per kilogram of the basal diet
Zinc herein refers to zinc oxide, zinc sulphate, zinc chloride, zinc stearate, zinc acetate, zmc bromide, zinc nitrate or any other derivative of zinc
Magnesium herein refers to magnesium acetate, magnesium - L - aspartate, magnesium carbonate, magnesium hydroxide, magnesium chloride, magnesium citrate, magnesium gluconate, magnesium lodate, magnesium iodide, magnesium oxide, magnesium phosphate, magnesium sulphate, magnesium nitrate or any other derivative of magnesium
Copper herein refers to copper chloride, copper nitrate, copper oxide, copper sulphate, copper iodide or any other derivative of copper
Manganese here refers to manganese acetate, manganese chloride, manganese oxide, manganese sulphate or any other derivative of manganese
Iron herein refers to iron sulphate, iron chloride, iron nitrate, iron phosphate, iron oxide or any other derivative or iron
Iodine herein refers to calcium iodide, calcium lodate potassium iodide, sodium iodide, cuperous iodide or other iodine bound organic compound or any other iodine derivative
Vitamin mixture include Ascorbic acid, 200-700mg, calcium pentathenate, 300-500mg, ergocalciferol, 0 05-200mg, a-D-tocopherol, 200-1000mg, nicotinic acid, 300-500g, cyanacobalamine, 0 l-500mg, pyndoxine hydrochloride, 50-200mg, thiamine hydrochloride, 200-500mg, riboflavin, 100-300mg, choline chloride 200-500mg and folic acid 100-500mg
As an additional feature a xanthophyll compound (20-50mg/kg) is added for providing the Roche colour scale value of the yolk in the absence of sufficient carotenoid source in the poultry feed to mcrease the acceptance of the egg by the consumer
The Following examples illustrate the present mvention Example-I
This example illustrates the production of modified eggs with lower content of cholesterol, modified minerals and vitamins composition in the egg
The common practice of poultry breeding system is employed in the production of modified eggs Only the feed of the layers has been modified The composition of the experimental diet is listed in table-I
(Table Removed)

*Mineral mixture include Sodium bicarbonate, 750 mg, Ferrous sulphate, 300 mg, Manganese sulphate, 500 mg, Copper sulphate, 750 mg, zinc sulphate, 50 mg, magnesium sulphate, 200 mg, potassium iodide, 0 005 mg in a kilogram of the basal diet
**Vitarmn mixture include Ascorbic acid, 500 mg, Calcium pentathenate, 400 mg, Choline chloride, 1000 mg, Folic acid, 10 mg, ergocalcrferol, 0 1 mg, pyndoxme hydrochloride, 80 mg, cx-D-tocopherol acetate, 500 mg, carotenoid, 20mg, of one kg of basal diet Example - II
The example further illustrates the production of modified poultry eggs with a lowered content of cholesterol and more with vitamins and minerals in accordance with the present invention
Following the example-I, 200 white layers, B V 300 Babcock strain, are fed adhbitum to the growers for 4 weeks prior to their ovulation and continued to feed on this diet throughout their ovulatory period After 2 weeks of ovulation and thereafter, the eggs were randomly collected for analysis The results of the analysis of these eggs are summarized in Table-II There are no differences in the, egg production or egg weight in comparison to the flocks of the birds feeding on conventional diet
Table - II Values of the constituents of modified egg weighing 59 g
(Table Removed)
The following illustrations show the efficacy of the modified eggs herein albino rats (Wister strain) are used as test animal model fed on an established semi synthetic basal diet for feeding the animals which is given in table-Ill
Table-in Semi synthetic basal diet (g/kg) of Wister rats.

(Table Removed)
*Vitamins mixture (mg/kg) Ascorbic acid, 500, Biotm,4, calcium pentothenate,320, choline chlonde,2500, folic acid,10, Inositol,1000, Ergocalciferol 0 003, Nicotimc acid,300, Cyanocobalamin,0 40 Pyndoxme hydrochlonde,200, a-Tocoferol acetate,60, Retmol 0 30
**Minerals mixture (g/kg) Ca H2 P04, 25 3, MgSO4 H2O, 4 05, KCl, 3 43, K1,0 004, Na2 CO3 1 15, Fe SO4 H2O, 0 60, MnSO4H2O, 0 31, CuCl, 0 10, CoCl3 0 04, Zn SO4 7H2O 0 088, NaF, 0 008 This diet served as control (Diet -1)
The non insulin dependant diabetes melhtus (NTDDM) is induced in rats by increasing ZnSo4 7H20 from 0 088mg/kg diet to 0 176mg/kg (Db-diet-I) and 0 352mg/kg (Db-diet-H) in the semi synthetic basal diet keeping all other ingredient constant This method is employed on the basis of the reports that excessive zinc in diet increases body fat in lean mice close to the genetically obese ob/ob mice (Chen M D, Lin PY Chen V and Lin W H Biol Trace Elem Res 52, 125-132, 1996) The rats when fed on three separated diets I e with 20mg Zn/kg diet (Control, Group -1) had a normal growth and blood profile pattern comparable to the rats fed on commercial standard rat pellet diet, while rats on 40mg Zn/kg diet (Group - II) and 80mg ZN/kg diet (Group - HI) showed mild and severe metabolic syndrome - X characterized by higher gain in body wt, hypertension, hyperglycemia, hypennsuhnemia and dyslipidemia comparable to those of human metabolic syndrome - X during 180 days of dietary treatment Zn being a part of human diet, this method of induction of NIDDM was preferred over streptozotocin or alloxan treated diabetic rats
The following data illustrate the body wt, blood pressure, blood profile and Zn, Cu
and Mg status in serum, liver, hair, abdominal muscle of male and female rats fed on basal
diet containing 20mg Zn (Control, Group - I), 40mg Zn (Group -II, Db I) and 80mg Zn/kg
diet (Group - III, Db II) for a period of 180 days The results of this experiment are
summarized in tables IV, V, VI, VII and VIII
Table IV
Mean month wise body wt. and blood pressure in rats fed on basal diet (Group - I, control) and on Db diet -1 (Group - II) and Db diet - II (Group - ITI) for 120 days.
(Table Removed)
Values are mean + SEM of n = 10
*Unne in rats on Db diet -1 (group-II) gives positive benedict test on day 90 and those on Db diet - II (group-Ill) on day 60 and there after until the termination of the experiment
Table IV shows that the rats that are fed on Db diet- I (Group - II) and Db diet - II (Group - IE) gain more weight during the first 150 days of dietary treatment while it declined on 180 day Their systolic blood pressure shows an increments approximately 70% or more after 3 months of dietary treatment and onward than those of the rats fed on semi synthetic basal diet (Control)
Table V
Blood profile of male rats fed on basal diet (Control Group - I) and on Db diet - I (Group - II) and Db diet - II (Group - ID) after 180 days of dietary treatment.

(Table Removed)
Values are mean + SEM of n = 10
Table VI
Blood profile of female rats fed on basal diet (Control Group - I) and on Db diet - I (Group - II) and on Db diet - II (Group - HI) after 180 days of dietary treatment.

(Table Removed)
Values are mean + SEM of n = 10
Data in the tables V and VI show that the blood profile of males and females rats on
Db diet- I and Db diet - II show dyslipidaemia, hyperglycemia and high creatinine and
hypennsulinemia, high cortisole, high aldosterone levels than those of control group of rats
They therefore manifest typical noninsulin dependant diabetes mellitus complicated with
hypennsulinemia and hypertension
Table VII
Mean Zinc (Zn), Copper (Cu) and Magnesium (Mg) concentrations in the serum, liver, hair, abdominal muscles and urine in male rats fed on basal diet (Control Group-I) and on Db diet-I (Group-II ) and on Db diet-II (Group-III) after 180 days of dietary treatment.

(Table Removed)
Values are mean ± SEM of n = 10
Table VIII
Mean Zinc (Zn), Copper (Cu) and Magnesium (Mg) concentrations in the serum, liver, hair and abdominal muscles in female rats fed on basal diet (Control Group-I) and on Db diet-I (Group-II ) and on Db diet-H (Group-III) after 180 days of dietary treatment.

(Table Removed)
Values are mean + SEM of n = 10
Data in tables VII and VIII show that the mineral concentrations of Zn, Cu and Mg
are significantly higher in serum of the group - II and III rats, whereas Zn concentration
remains higher but those of Cu and Mg reduce drastically in other tissues such as liver, hair
and abdominal muscles with their excessive loss in urine in spite of their adequate levels in
their diets The altered mineral status in rats of groups - II and in is consistent with those of
genetic diabetes mice/rats and human diabetic patients where an excessive loss of these
minerals have been seen m urine leading to their deficiencies in the tissues
Table IX
Mean values of liver weight, total lipids, cholesterol, triglycerides and glycogen in rats fed on basal diet (Group I) and on Db diet-I and II (Group II and III) after 180 days of dietary treatment.
(Table Removed)
Values are Mean ± S E of n = 10
The data in table IX show reduction in total lipids, cholesterol, triglycerides and glycogen on Db diet -1 and Db diet - II rats which coincide well with their rise in the blood
The data of body wt, blood pressure, blood serum profile and mineral status of rats in group-II and III clearly demonstrate that the diet containing higher Zn (even in pharmacological doses) when fed to rats for a longer duration induces diabetes in them, consistent with those of genetically db/db mice, Zukar diabetic fatty rats (ZDF/Drt) and in some of human diabetic patients The severity of the disease depends upon the amount of Zn m diet and degree of deficiencies of minerals in the tissues Even if the diet contains adequate levels of minerals and vitamins, they are not able to prevent the occurrence of these deficiencies in the tissues when Zn is excess in diet
Example III
This example illustrates that on feeding four fresh modified eggs mixed in diet to diabetic rats improve the metal status in tissues and reduce the severity of diabetes in them
For this, two groups of rats are included in this study These groups are made out of the normal 21 days old (just after weaning period) males and females from a stock bred on the standard rat pellet diet They are fed on Db diet - I (group-II) and Db diet - II (group-Ill) respectively for a period of 120 days Their body wt blood pressure and urine glucose are
monitored to check the onset of diabetes which are seen to manifest on 90th day in group-II
and on 60 day in group-Hi fed diet groups similar to group II and III rats
On the beginning 120th day, the animals of the group -II and III were shifted on the
Db diet - I mixed with four fresh modified eggs and Db diet - II mixed with four fresh
modified eggs/kg of semi synthetic diet respectively and continue to feed them on their
respective diets for the period of 60 days prior to the termination of the experiment that lasted
for 180 days Their body wt and blood pressure are monitored (Table X)
Table X
Mean month wise body weight and blood pressure in rats fed on basal diet (Control Group I) and on egg mixed Db diet-I & II (Group II* and III3*) during the dietary treatment.
(Table Removed)
Values are Mean ±SE of n=10
*Egg mixed Db diet -I and II are given for 60 days to rats (male and female) after 120 days of dietary treatment
of Db diet-I and n without eggs
The data of rats groups II* and III* show a lesser increase in their body wt and reduction in blood pressure compared to the group - II and group -III (refer Table IV)
After the completion of the experiment, the blood serum and other tissues are
bioassayed for the parameters investigated for group -1, II and III The result are summarized
in table XI, XII, XIH, XIV and XV
Table XI
Blood profile of male rats fed on basal diet (Control Group -1) and on egg mixed Db diet -1 (Group - II)* and egg mixed Db diet - II (Group -III)* after the end of the dietary treatment.

(Table Removed)
Values are mean + SEM of n = 10
*Egg mixed Db(EDb) diet I and II are given for next 60 days to male rats after 120 days of dietary treatment of
Db diet-I and II
Table XII
Blood profile of female rats fed on basal diet (Group - I) and on egg mixed Db (EDb) diet - I (Group - II)* and egg mixed Db (EDb) diet - II (Group -HI)* after the end of dietary treatment.
(Table Removed)

*Egg mixed Db (EDb) diet-I and II are given for next 60 days to female rats after 120 days of dietary treatment
of Db diet-I and II
The data in table XI and XII show a significant reductions in total lipid, triglycerides,
cholesterol, LDL-cholesterol, VLDL-cholesterol, glucose, insulin, cortisole, aldosterone ,
TSH, and rise in HDL-cholesterol occur in rats on egg mixed Db - I diet than those of
(Table Removed)

Db diet -1 group Their values are close to those of control group-I However, the recovery
of these molecules are also significant in rats on egg mixed diet Db diet - II than those of
Db diet - II group but their level approached close to the rats on Db diet -1 (group-II)
and not close to the control group -1 both in male and female rats (Refer tables V and VI
for comparison)
Table XIII
Mean Zinc (Zn), Copper (Cu) and Magnesium (Mg) concentrations in the serum, liver, hair, abdominal muscles and urine in male rats fed on basal diet (Control) and on egg mixed EDb diet I and II (Group II* and III*) after the end of dietary treatment.

(Table Removed)
Values are Mean ± S E M of n = 10
* Egg mixed Db (EDb) diet I and II are given for next 60 days to male rats after the 120 days of dietary
treatment of Db diet I and II
Table XIV
Mean Zinc (Zn), Copper (Cu) and Magnesium (Mg) concentrations in the serum, liver, hair, abdominal muscles and urine in female rats fed on basal diet (Control) and on egg mixed EDb diet I and II (Group n* and HI*) after the end of dietary treatment.

Values are Mean ±SEM of n =10
* Egg mixed Db (EDb) diet-I and II are given for next 60 days to female rats after the 120 days of dietary
treatment of Db diet I and II
The data in tables XIII and XIV show that Zn concentration in serum and tissues
significantly declines while the concentrations of Cu and Mg increase in the tissues and
decline in the blood serum m rats on egg mixed Db diet - I than those of rats on Db diet -1
approaching close to the control values while their recovery is significant in rats on egg
mixed Db diet - II rats approaching close to Db -1 group (Group-II) rats but not to the level
of control group (Group-I) (For comparison refer tables no VII and VIII)
Table XV
Mean values of liver weight, total lipids, cholesterol, triglycerides and glycogen in rats fed on basal diet (Control) and on egg mixed Db diet-I and II (Group II* and III*) after the end of dietary treatment.

(Table Removed)
Values are Mean ±SE ofn=10
*Egg mixed Db (EDb) diet-I and II are given for 60 days to rats (male and female)' after 120 days of dietary
treatment of Db diet-I and II
The data in table XV show that the lipids, cholesterol, triglycerides and glycogen
concentration the liver of both males and females show a remarkable recovery in rats on modified egg mixed Db diet -1 and egg mixed Db diet - II approaching close to the control values
It is evident from the data per se that the inclusion of these modified eggs in diet of diabetics helps to restore the altered mineral composition of blood and tissues and reduce the severity of diabetes and protects them against diabetic induced complications if the individuals are feeding on diabetogenic diet Example IV
This example illustrates that these modified eggs protect the embryos in womb, help to deliver normal neonates by preventing malformations in them due to diabetes complicated with hypertension and help towards the restoration of mineral composition in the body of the young ones
For this study, three groups of rats are prepared from 21 days old pups just after their
weaning period The males and females rats are separated from the main stock and divided
into 3 groups Group-I is fed on the semi synthetic basal diet (Control) Group-II on Db diet
-I and group-Ill on Db diet -II for a period of two months At this stage animals in
group II and III are subdivided into two groups each One subgroup of group II is continued
to feed on Db diet - I and the other on four fresh modified egg mixed Db diet - I/kg diet
(Group - II*) Like wise one subgroup of group - III is continued to feed on Db diet -II
(Group -III) and the other on four fresh modified egg mixed Db diet - II/kg diet (Group -
III*) for a period of two months After a total of 4 months dietary treatment, the males
and females in each group are mated within the group and continue to feed on their
respective diet during their gestation and suckling periods The body wt and health of the
neonates are recorded during their weaning period The result are summarized in tables XVI,
XVII and XVIII
Table XVI
Mean Body Wt (g) of Pups of F1- Generation at birth and during the weaning period obtained after mating the Parents fed on basal diet (Group - I) and on Db diet - I (Group-II) and on Db diet-II (Group-III)

(Table Removed)
Values are mean + SEM
Table XVII
Mean Body Wt. (g) of pups of F1 - Generation at birth and during the weaning period obtained after mating the Parents fed on basal diet (Group-I) and on egg mixed Db diet -I (Group- II*) and on egg mixed Db diet - II (Group - III*)

(Table Removed)
Values are mean + SEM
Table XVIII
Mean Zinc (Zn), Copper (Cu) and Magnesium (Mg) concentrations in the liver, hair and abdominal muscles in pups of F1 generation just after the weaning period of pups (F-l Generation)

(Table Removed)
Values are mean + SEM of n = 10
*Egg mixed Db diet-I and II was given for 60 days to male and female rat parents after 60 days of dietary
treatment of Db diet-I and II respectively
The Fl generation obtained after mating shows that the neonates of group -II and group-III have higher birth wt than those of their control, have malformed toes of the forehmb (curled toes), show restricted unbalanced body movements, gain lesser body wt during weaning period They are diabetic when their urine is tested on 10th and 19 day of weaning period by Benedicts test They possess extremely low concentration of Cu, Zn and Mg in their tissues and die after the completion of the weaning period (Table XVIII)
The neonates of group-II* and III* whose parents are fed on egg mixed Db diet- I and egg mixed Db diet -II also have higher body wt than those of control, gain more wt than group -II, and III during weaning period They remain healthy on completion of their weaning period But they are diabetic There is a partial rehabilitation of Cu, Zn and Mg concentrations in their tissues (Table XVIII) as their concentration are significantly higher than the groups II and III but still lower than the control group of rats
The data of this set up experiment illustrate that the diabetes induced by this method in parents is genetically transmitted to the off spring of F1 generation and off springs die after weaning period due to malformations attributed by uncontrolled diabetes and hypertension But the inclusion of these modified eggs in diet of the dams protects the embryos in womb, allow them to deliver normal neonates and help towards restoration of minerals composition in their body Example V
This example illustrates that the modified eggs provide protection against diabetes not only to those feeding on diabetogenic food but also against genetically transmitted diabetes
For this study, the survived neonates of F1 generation of group -IE* are used After
the weaning period of pups in group - IIP, they are subdivided into two subgroups One
subgroup of group - IIP is fed on standard rat pellet diet (Group-EP*) and other on egg
mixed standard rat pellet diet (Group IE***) for a period of 60 days Their body wt and
systolic blood pressure are recorded on weekly basis through out the duration of the
experiment The results of this study are summarized in table XIX
Table XIX
Mean body weight and blood pressure in Pups of F1 generation fed on pellet diet of control (Group - IF) on pellet diet of F-l generation of Db diet - III** (Group - III**) and on egg mixed pellet diet of F - 1 generation of Db diet - III*** (Group - III***) after 60 days of dietary treatment.

(Table Removed)
Values are mean + SEM of n = 10
After 60 days of dietary treatment, they are sacrificed and bioassayed (Tables XX,
XXI, XXII)
Table- XX
Blood profile of male rats of F1 generation fed on pellet diet of control (Group -1) on, pellet diet of F-1 generation of Db diet - III** (Group - III**) and on egg mixed pellet diet of F -1 generation of Db diet - III*** (Group - III***) after 60 days of dietary treatment.

(Table Removed)
Values aremean+ S E M of n = 10
Table XXI
Blood profile of female rats of F1 generation fed on pellet diet of control (Group -1) on pellet diet of F-1 generation of Db diet - III** (Group - III**) and on egg mixed pellet diet of F - 1 generation of Db diet - III*** (Group - III***) after 60 days of dietary treatment.

(Table Removed)
Values are mean + SEM of n = 10
The data in table XX and XXI reveal that rats in group - III*** on egg mixed pellet
diet do not differ significantly m their serum glucose, total lipids, triglycerides, total
cholesterol, HDL cholesterol, LDL cholesterol, VLDL cholesterol, creatinine, insulin,
Cortisol, TSH, T3 and T4 than those of control rats However, the rats in group in** display
elevated concentrations of the blood parameters These rats are genetically predisposed to
diabetes The normal diet can not rehabilitate the genetically altered physiology in them
(Table XX and XXI)
Table-XXII
Mean Zinc (Zn), Copper (Cu) and Magnesium (Mg) concentrations in the serum, liver, hair and abdominal muscles in male rats of F1 generation fed on pellet diet of control (Group - I) on pellet diet of F-1 generation of Db diet - III** (Group - III**) and on egg mixed pellet diet of F - 1 generation of Db diet - III*** (Group - III***) after 60 days of dietary treatment.
(Table Removed)

Values are mean + SEM of n = 10
Table XXIII
Mean Zinc (Zn), Copper (Cu) and Magnesium (Mg) concentrations in the serum, liver, hair and abdominal muscles in female rats of Fi generation fed on pellet diet of control (Group - I) on pellet diet of F-1 generation of Db diet - III** (Group - III**) and on egg mixed pellet diet of F - 1 generation of Db diet - HI*** (Group - III***) after 60 days of dietary treatment.
(Table Removed)
Values are mean + SEM of n = 10
There is a reduction in mineral concentrations of Zn, Cu and Mg in group III** close to Db -I and Db-II rats in group -II and III compared to control rats (Refer to tables no VII and VIII) However, in group III*** the mineral composition in blood serum, abdommal muscles, liver and hair are close to the control rats This shows that the addition of these modified eggs rehabilitate the mineral composition in diabetics Example VI
The following example illustrates that these eggs are safe to consume by healthy individuals
For this purpose four liquid eggs were added in the pallet diet and healthy rats were
fed on this egg mixed diet for a period of 60 days after their weaning period The data
obtained is given in the following tables
Table XXIV
Mean body weight and blood pressure in rats fed on pellet diet and egg mixed pellet diet after two months of dietary treatment.

(Table Removed)
Values are mean + SEM of n = 10
Table XXV
Blood profile of male rats fed on basal diet and egg mixed basal diet after two months of dietary treatment.
(Table Removed)
Values are mean+S EM of n = 10
Table XXVI
Blood profile of female rats fed on basal diet and egg mixed basal diet after two months of dietary treatment.

(Table Removed)
Values are mean + SEM of n = 10
Table XXVII
Mean Zinc (Zn), Copper (Cu) and Magnesium (Mg) concentrations in the serum, liver, hair and abdominal muscles in male rats fed on pellet diet and egg mixed pellet diet after two months of dietary treatment.
(Table Removed)
Values are mean + SEM of n = 10
Table XXVIII
Mean Zinc (Zn), Copper (Cu) and Magnesium (Mg) concentrations in the serum, liver, hair and abdominal muscles in female rats fed on pellet diet and egg mixed pellet diet after two months of dietary treatment.

(Table Removed)
Values are Mean ± S E M of n=10
Table XXIX
Mean values of liver weights, total lipids, cholesterol, triglycerides and glycogen in rats fed on pellet diet and egg mixed pellet diet after 2 months of dietary treatment.

(Table Removed)
Values are Mean ±SEM of n = 10
The data in tables XXIV, XXV, XXVI, XXVII, XXVIII, XXIX show that the inclusion of eggs m pallet diet of rats do not significantly alter the body wt, B P, blood profile and mineral composition in tissues They are therefore, safe for healthy consumers.

We claim:
1) An improved poultry feed for the production of eggs with capacity of restoration
of minerals composition in a patient, said feed comprising a dietary material of
broken rice up to 35%, rye oat up to 15%, flax seed up to 2%, de-oiled rice bran
up to 18%, de-oiled mustard cake up to 5%, de-oiled sunflower cake up to 2%, de-
oiled groundnut cake up to 3%, di-hydrogen calcium phosphate up to 1.5%,
marble powder up to 3%, marble grit up to 7%, mineral mixture up to 5%,
vitamins mixture up to 5%, tyrosine powder up to 0.2% and multi-enzymes up to
0.05% by wt. in one kg of the feed such as here in described, wherein said
minerals comprise from 0.2 to 2.0g of sodium derivatives, from 0.2 to 0.3g of
ferrous derivatives, from 0.05 to 500g of magnesium derivatives, from 0.25 to 4g
of manganese derivatives, from 0.02 to 0.10g of zinc derivatives, 0.05 to lg
copper derivatives, from 0.002 to 0.005g of iodine derivatives and from 0.002 to
0.00lg of selenium derivatives and said vitamins comprise from 200 to 700mg of
ascorbic acid, from 300 to 500mg of calcium pentathenate, from 200 to 500mg of
choline chloride, from 100 to 500mg of folic acid, from 0.05 to 0.200mg of
ergocalciferol, from 300 to 500mg of nicotinic acid, from 0.01 to 100mg of
cyanocobalamine, from 50 to 200mg of pyridoxine hydrochloride, from 200 to
500mg of thiamine hydrochloride, from 100 to 300mg of riboflavin, from 200-
500mg of alfa-tocopherol, from 15 to 40mg of carotenoid per 1 kg of the poultry
feed.
2) The poultry feed as claimed in claim 1, wherein the minerals are selected from
one or more sodium derivatives, ferrous derivatives, magnesium derivatives,
manganese derivatives, zinc derivatives, copper derivatives, iodine and selenium
derivatives.
3) The poultry feed as claimed in claim 2, wherein the mineral mix is comprised of sodium bicarbonate, 0.75 to lg; ferrous sulphate, 0.3 to 0.5g; manganese sulphate, 0.5 to 0.7g, copper sulphate, 0.75 to lg,; zinc sulphate, 0.050 to 0.075g,; magnesium sulphate 0.2 to 0.5g; potassium iodide, 0.0005g-0.0075g; and sodium selenite, 0.0005-0.0008g.

Documents:

2264-del-2005-abstract.pdf

2264-DEL-2005-Claims-(03-08-2012).pdf

2264-DEL-2005-Claims-(15-07-2011).pdf

2264-DEL-2005-Claims-(16-08-2012).pdf

2264-del-2005-claims.pdf

2264-del-2005-Correspondence Others-(02-08-2011).pdf

2264-DEL-2005-Correspondence Others-(03-08-2012).pdf

2264-DEL-2005-Correspondence Others-(09-08-2011).pdf

2264-del-2005-Correspondence Others-(10-08-2011).pdf

2264-DEL-2005-Correspondence Others-(11-08-2011).pdf

2264-DEL-2005-Correspondence Others-(15-07-2011).pdf

2264-DEL-2005-Correspondence Others-(16-08-2012).pdf

2264-del-2005-Correspondence Others-(25-07-2011).pdf

2264-del-2005-correspondence-others.pdf

2264-del-2005-correspondence-po.pdf

2264-del-2005-description (complete).pdf

2264-del-2005-form-1.pdf

2264-del-2005-form-18.pdf

2264-del-2005-form-2.pdf

2264-del-2005-form-26.pdf

2264-del-2005-form-3.pdf

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Patent Number

253740

Indian Patent Application Number

2264/DEL/2005

PG Journal Number

34/2012

Publication Date

24-Aug-2012

Grant Date

22-Aug-2012

Date of Filing

24-May-2005

Name of Patentee

PANJAB UNIVERSITY

Applicant Address

PANJAB UNIVERSITY, CHANDIGARH 160014, INDIA.

Inventors:

#	Inventor's Name	Inventor's Address
1	TANEJA, SATISH KUMAR	DEPARTMENT OF ZOOLOGY, PANJAB UNIVERSITY, CHANDIGARH 160014, INDIA.

PCT International Classification Number

A61K 35/78

PCT International Application Number

N/A

PCT International Filing date

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1			NA