WO2004030632A2

WO2004030632A2 - Monascus derived poultry feed and by-products

Info

Publication number: WO2004030632A2
Application number: PCT/US2003/031510
Authority: WO
Inventors: Mark Peter Wach; Gary D. Walker; Ronald E. Sabourin
Original assignee: Sylvan Bioproducts, Inc.
Priority date: 2002-10-03
Filing date: 2003-10-03
Publication date: 2004-04-15
Also published as: AU2003282680A8; AU2003282680A1; WO2004030632A3

Abstract

The methods and compositions of the present invention produce cholesterol-lowering compounds including statins by colonizing Monascus species (red yeast) on a semi-solid substrate of the animal feed composition. The methods and compositions of the present invention further provide low cost production of Monascus species by inoculation and colonization of Monascus spp. on commercially-available poultry foodstuff. The methods and compositions provide for inoculation, incubation and colonization of Monascus spp. on a solid or semisolid substrate. The semisolid substrate is produced by combining poultry foodstuff which may be standard poultry foodstuff, soybean, cracked soybean, wheat, wheat bran and/or other appropriate components. The poultry foodstuff are aseptic to allow preferential colonization by Monascus spp. The Monascus colonization on the poultry foodstuff has been found to provide high levels of statins and other cholesterol-lowering components in the animal feed composition. When the animal feed composition is fed to an egg-laying animal, such as a chicken, duck, goose, turkey, or quail, levels of cholesterol in the eggs of the animal are significantly reduced.

Description

COMPOSITIONS AND METHODS FOR MONASCUS DERIVED ENHANCEMENT OF CHOLESTEROL-LOWERING COMPOUNDS

BACKGROUND OF THE INVENTION

[0001] Eggs are a staple food item in the human diet. They are relatively inexpensive and are an excellent source of dietary protein. Recently, however, the egg has come under increasing medical and dietary criticism due to the high level of cholesterol contained within the egg yolk It is desirable to reduce the cholesterol content because, for humans, reduction of dietary cholesterol and saturated fat is associated with a decreased risk of contracting certain diseases, including, for example, coronary artery disease, atherosclerosis, and stroke. In addition, reducing cholesterol and saturated/unsaturated fat ratios can significantly reduce the complications of diabetes.

[0002] Four major classes of human serum lipoproteins have been identified which are responsible for transport of cholesterol. Chylomicron (density less than 0.95 g/cm³) transports exogenous (dietary) triacylglycerol and cholesterol from the intestine to the liver and other tissues. Very low density lipoprotem (VLDL; density = 0.95 to 1.006 g/cm³) is released by the liver into the circulation. VLDL contains endogenous triacylglycerol and cholesterol. Low density lipoprotein (LDL; density = 1.006 to 1.063 g/cm³) is derived from VLDL by lipolysis and other processes. LDL transports cholesterol to the liver and other tissues and regulates de novo cholesterol synthesis. High density lipoprotein (HDL; density = 1.063 to 1.21 g/cm³) reverse transports endogenous cholesterol from peripheral tissues to liver for excretion. HDL provides a reservoir for apolipoproteins. Molecular Biology and Biotechnology, R.A. Meyers, Ed., Wiley-VCH, Inc., 1995. [0003] The relationship between elevated plasma cholesterol levels, especially plasma low density lipoprotein cholesterol, and an increased risk of contracting coronary heart disease has been well established. In light of this relationship, a dietary cholesterol intake of less than 300 mg/day has been recommended for all Americans. National Institutes of Health Consensus Development Panel, 1985. Foodstuffs containing lowered levels of cholesterol are therefore desired.

[0004] In general, poultry eggs contain between about 215 mg and 280 mg of cholesterol. Because the egg, and particularly the poultry egg, is an inexpensive source of dietary protein, considerable research has been carried out in an effort to develop an egg which has reduced quantities of cholesterol and saturated fats. This research has resulted in several methods for reducing the cholesterol and saturated fatty acid content of the egg. However, the methods for reducing the cholesterol and saturated fatty acid content currently in use have generally proven to be unsatisfactory. Efforts at reducing the level of cholesterol in intact chicken eggs have included genetic selection, use of low fat and high fiber diets, administration of pharmacological agents and various egg selection methods. Many of these methods are tedious and others have not led to large decreases in egg cholesterol.

[0005] Existing methods that reduce the level of cholesterol in intact chicken eggs by altering the diet of the laying hen are problematic. Existing methods are not cost-effective as a supplement to the chicken feed. One method involves the addition of HMG CoA reductase inhibitors, for example, statins such as atorvastatin, lovastatin or simvastatin, to the animal feed. HMG CoA reductase is a rate-limiting enzyme in cholesterol biosynthesis. Feed supplementation with low amounts of lovastatin or colestipol did not decrease the concentration of cholesterol in the egg yolk. Feed supplementation with higher amounts of lovastatin or colestipol did decrease the concentration of cholesterol in the egg yolk. Problems encountered with this animal feed composition make it prohibitive as a cost-effective feed supplement. These problems include the expense of producing the additives and the expense of processing the additives into the feed. Luhman et al., Poultry Science 69: 852-855 (1990); Mori, et al., J. Agric. Food. Chem. 47: 4731-4735 (1999); See U.S. Patent 6,177,121, incorporated herein by reference.

[0006] Monascus red yeast rice is a traditional Chinese medicine and food known for centuries to improve blood circulation. A known method for preparation of Monascus red yeast rice follows ancient methods by fermenting the fungal strain Monascus purpureus Went on moist and sterile rice. Analyses of the natural constituents of Monascus red yeast rice,, indicated the presence of a group of metabolites belonging to the monacolin family of polyketides, together with fatty acids, and trace elements. The presence of these compounds may explain in part the cholesterol-lowering ability associated with the traditional Chinese food. Ma, et al., J. Agric. Food. Chem. 48: 5220-5225, (2000).

[0007] A known method to reduce the level of cholesterol in intact chicken eggs involves costly dietary supplement of the animal feed with Monascus red yeast rice as a feed additive. Monascus red yeast rice preparation requires fermentation of Monascus species on sterile and moist rice. The Monascus red yeast rice is dried and added as a dried powder to commercial poultry feed, which included corn meal, soybean meal, limestone, canola meal, canola oil, dicalciu phosphate, salt, DL-methionine, vitamin mix, mineral mix. When fed to poultry, the Monascus red yeast rice-poultry feed mixture resulted in a reduced level of cholesterol in intact chicken eggs. The expense of adding Monascus red yeast rice to the animal feed composition make it prohibitive as a cost-effective feed supplement. See U.S. Patent 6,316,041, incorporated herein by reference.

[0008] Known methods for preparation of Monascus red yeast rice require fermentation of the fungal strain Monascus spp. on rice. The rice may be Oryza species, for example, Oryza sativa or Oryza sativa L. Ma, et al., J. Agric. Food. Chem. 48: 5220-5225, (2000).

[0009] Another known method to reduce the level of cholesterol in intact chicken eggs involves costly dietary supplement of the animal feed with a combination of phytosterol, Monascus red yeast rice, and a copper derivative as feed additives. Problems encountered with this animal feed composition make it prohibitive as a cost-effective feed supplement. These problems include the expense of producing the additives and the expense of processing the additives into the animal feed. See U.S. Patent 6,316,041, incorporated herein by reference.

[0010] A need exists in the art for animal feed compositions effective in reducing the amount of cholesterol in an egg when fed to an egg-laying animal and methods of producing compounds that will lower cholesterol levels in the blood serum of an animal. The animal feed composition must include a low cost commercial poultry feed composition that is more effective in reducing the amount of cholesterol in an egg, when compared to more complex and expensive poultry feed compositions. The method of producing the animal feed composition should minimize labor intensive steps and provide a less expensive production process. The present invention addresses these needs. SUMMARY OF THE INVENTION

[0011] The present invention fulfills the foregoing needs and satisfies additional objects and advantages by providing novel, effective animal feed compositions and methods of producing animal feed compositions effective in reducing the amount of cholesterol in an egg when the composition is fed to a poultry or other egg-laying animal. The methods and compositions of the present invention produce cholesterol-lowering compounds including statins by colonizing Monascus species (red yeast) on a semi-solid substrate of the animal feed composition. The methods and compositions of the present invention further provide low cost production of Monascus species by inoculation and colonization of Monascus spp. on commercially-available poultry foodstuff. The commercially-available poultry foodstuff are aseptic poultry foodstuff which allow preferential colonization by Monascus spp. The Monascus colonization on the aseptic poultry foodstuff has been found to provide high levels of cholesterol-lowering compounds including statins in the animal feed composition. When the animal feed composition is fed to an egg-laying animal, such as a chicken, duck, goose, turkey, or quail, levels of cholesterol in the eggs of the animal are significantly reduced.

[0012] The methods and compositions of the present invention provide for enhanced production of an animal feed composition effective in reducing the amount of cholesterol in an egg when fed to an egg-laying animal. The present invention provides methods and compositions for inoculation, incubation and colonization of Monascus spp. on a solid or semisolid substrate. The semisolid substrate is produced by combining, preferably, poultry foodstuff which may be standard poultry foodstuff, soybean, cracked soybean, wheat, wheat bran and/or other appropriate components. The poultry foodstuff are aseptic to allow preferential colonization by Monascus spp. The water content of the aseptic poultry foodstuff is preferably adjusted to obtain a final moisture content between about 25% to about 65% by weight of the aseptic poultry foodstuff to form a semi-solid substrate. Monascus spp. is inoculated and incubated with the aseptic poultry foodstuff, for a time sufficient to allow Monascus colonization. Monascus colonization results in production of statins and other cholesterol- lowering components in the poultry foodstuff. Monascus colonization results in production of statins, monacolin family of polyketides, fatty acids, trace elements, and other cholesterol- lowering components in the poultry foodstuff. In one aspect of the invention, the statins can include, but are not limited to, lovastatin, simvastatin, and/or storvastatin. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 depicts the production of statins by Monascus purpureus on components of poultry feed.

[0014] Figure 2 depicts the effect of Monascus purpureus-colomzed feed on the cholesterol content of chicken eggs.

[0015] Figure 3 shows HPLC chromatograms of statins produced by Monascus purpureus on soy and standard poultry feed.

[0016] Figure 4 depicts the effect of Monascus purpureus-colomzed feed on the cholesterol content of chicken eggs.

[0017] Figure 5 depicts the effect of Monascus purpureus-colomzed feed on the cholesterol content of chicken eggs.

DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

[0018] In one embodiment of the present invention, an animal feed for poultry comprises a poultry foodstuff and Monascus species colonized upon the foodstuff. The Monascus-colonized foodstuff is effective in reducing the amount of cholesterol in eggs of poultry fed the animal feed as compared to the amount of cholesterol in such eggs when the uncolonized foodstuff is fed to the poultry.

[0019] The Monascus-colonized foodstuff is an integral mixture of Monascus and poultry foodstuff. Monascus spp. is inoculated, incubated and colonized on the poultry foodstuff to produce an integral mixture that produces effective levels of Monascus spp. and Monascus products (for example, statins) effective in reducing the amount of cholesterol in eggs of poultry fed the animal feed as compared to the amount of cholesterol in such eggs when the uncolonized foodstuff is fed to the poultry.

[0020] As used herein, "Monascus-colonized poultry foodstuff or "Monascus- colonized foodstuff refers to the mixture comprising poultry foodstuff and Monascus colonized upon the poultry foodstuff. Monascus inoculation is not an additive to the poultry foodstuff, but rather a seed inoculation for growth of Monascus spp. yeast upon the poultry foodstuff. Monascus spp. inoculation, incubation, and colonization on the poultry foodstuff may occur at specified conditions of time, temperature, and moisture content. [0021] In a preferred aspect of the invention, the Monascus spp. colonizes the foodstuff in an amount effective in reducing the amount of cholesterol in eggs of poultry fed the animal feed of the present invention as compared to the amount of cholesterol in such eggs when uncolonized foodstuff is fed to poultry. In a further aspect, the effective amount of Monascus colonization by weight is between about 1% and about 20% by weight of the poultry foodstuff. In a further aspect, the effective amount of Monascus colonization by weight is between about 4% and about 17% by weight of the poultry foodstuff. In a further aspect, the effective amount of Monascus colonization by weight is between about 8% and about 10% by weight of the poultry foodstuff.

[0022] In a preferred aspect of the invention, the Monascus-colonized foodstuff reduces the amount of cholesterol in eggs of poultry fed the animal feed of the present invention as compared to the amount of cholesterol in such eggs when uncolonized foodstuff is fed to poultry. In one aspect, the amount of cholesterol in eggs of poultry fed the animal feed of the present invention is reduced at least about 5% per gram of yolk, when compared to yolks obtained from poultry fed uncolonized foodstuff. In a further aspect, the amount of cholesterol in eggs of poultry fed the animal feed is reduced at least about 15% per gram of yolk, when compared to yolks obtained from poultry fed uncolonized foodstuff. In a further aspect, the amount of cholesterol in eggs of poultry fed the animal feed is reduced at least about 25% per gram of yolk, when compared to yolks obtained from poultry fed uncolonized foodstuff.

[0023] In another aspect of the invention, Monascus colonization upon the foodstuff occurs upon a semi-solid or solid substrate of the foodstuff. In a preferred aspect of the invention, the Monascus-colonized foodstuff comprises water between about 25% and about 65% by weight of the poultry foodstuff.

[0024] In a preferred aspect of the invention, Monascus spp. is incubated with the foodstuff for a time and at a temperature for optimal Monascus colonization of the foodstuff. In one aspect, an incubation time between approximately 2 and 25 days achieves Monascus colonization of the food stuff. In another aspect, an incubation time between approximately 5 and 20 days achieves Monascus colonization of the foodstuff. In another aspect, an incubation time between approximately 13 and 17 days achieves Monascus colonization of the foodstuff.

[0025] In a further preferred aspect, an incubation temperature between approximately 15°C and 35°C achieves Monascus spp. colonization of the foodstuff. In another aspect, an incubation temperature between approximately 20°C and 30°C achieves Monascus spp. colonization of the foodstuff.

[0026] The poultry foodstuff on which Monascus spp. colonization occurs comprises soybean, wheat or combinations thereof. The poultry foodstuff further comprises cracked soybean, wheat bran, or combinations thereof. The poultry foodstuff further comprises corn, soybean, soybean meal, limestone, dicalcium phosphate, vitamin mix, trace mineral mix, biotin, sodium chloride, canola oil, methionine, or combinations thereof. In a further aspect of the invention the poultry foodstuff are aseptic poultry food stuff. The aseptic preparations preferably comprise poultry foodstuff, soybean, wheat, or a combination thereof. The aseptic preparations may also preferably comprise poultry foodstuff, cracked soybean, wheat bran, flax, corn (cracked, ground, whole, meal), oats, rice, barley, rapeseed, sorgham, millet, rye and combinations thereof. Additional materials known for use in the preparation of mushroom growth media may also be used such as rice, corn, rice, and other grains. Monascus spp. is inoculated, incubated and colonized upon the aseptic poultry foodstuff.

[0027] In another aspect of the invention, the Monascus spp. colonizing the foodstuff include, but are not limited to, Monascus purpureus, Monascus ruber, Monascus anka, Monascus vitreus, or Monascus paxii. m a further aspect, the poultry include, but are not limited to, chicken, duck, goose, turkey, quail, pigeon, guinea fowl, peafowl, pheasant, chukar, partridge, grouse, emu, rhea or ostrich.

[0028] In another embodiment of the present invention, a method of producing poultry feed comprises inoculating poultry foodstuff with Monascus species. The inoculated foodstuff is incubated for a time and under conditions effective to produce enhanced amounts of one or more cholesterol-lowering compounds including statins in the food materials. The incubation of the inoculated foodstuff gives rise to levels of Monascus spp. in the poultry feed effective in reducing the amount of cholesterol in eggs of poultry fed the poultry feed as compared to the amount of cholesterol found in such eggs when the uncolonized foodstuff is fed to the poultry. In a further aspect, the effective amount of Monascus colonization by weight is between about 1% and about 20% by weight of the poultry foodstuff. In a further aspect, the effective amount of Monascus colonization by weight is between about 4% and about 17% by weight of the poultry foodstuff. In a further aspect, the effective amount of Monascus colonization by weight is between about 8% and about 10% by weight of the poultry foodstuff. [0029] In one aspect of the method of the present invention, the poultry foodstuff comprises soybean, cracked soybean, wheat, wheat bran, poultry foodstuff, or combinations thereof. In a further aspect, the poultry foodstuff is sterilized to produce aseptic poultry foodstuff. The aseptic poultry foodstuff is inoculated with Monascus spp.

[0030] In a further aspect of the present invention, the method produces Monascus colonization resulting in production of statins and other cholesterol-lowering components in the poultry foodstuff. Monascus colonization results in production of statins, monacolin family of polyketides, fatty acids, trace elements, and other cholesterol-lowering components in the poultry foodstuff. In a further aspect of the present invention, the method produces statins which comprise lovastatin, simvastatin, storvastatin or a combination thereof.

[0031] In a preferred aspect of the invention, Monascus spp. is inoculated and incubated on poultry foodstuff for a time and under conditions effective to produce enhanced amounts of one or more cholesterol-lowering compounds including statins in the food materials. In one aspect, Monascus inoculation and incubation on poultry foodstuff occurs for sufficient time and under conditions effective to produce at least about 0.01% statins by weight of total foodstuff. In another aspect, Monascus inoculation and incubation on poultry foodstuff occurs for sufficient time and under conditions effective to produce at least about 0.1% statins by weight of total foodstuff. In another aspect, Monascus inoculation and incubation on poultry foodstuff occurs for sufficient time and under conditions effective to produce at least about 0.3% statins by weight of total foodstuff.

[0032] In another aspect of the method of the invention, the poultry foodstuff is inoculated and incubated with the Monascus spp. in discrete units. The discrete units are contained within sealable polypropylene containers.

[0033] In another embodiment, the present invention is a method of reducing the amount of cholesterol in eggs produced by poultry. The method comprises feeding to the poultry an animal feed which has been inoculated with Monascus species. The Monascus spp. is present in the feed in an amount effective to attain a reduction in the amount of cholesterol in eggs produced by poultry as compared to the amount of cholesterol in such eggs produced when the animal feed without the Monascus is fed to the poultry. In a preferred aspect of the method of the present invention, the Monascus spp. is present in the feed in an amount effective to attain a reduction in the amount of cholesterol in eggs produced by poultry as compared to the amount of cholesterol in such eggs produced when the animal feed without Monascus is fed to the poultry. In one aspect, the amount of cholesterol in eggs of poultry fed the animal feed is reduced at least about 5% per gram of yolk, when compared to yolks obtained from poultry fed uncolonized foodstuff. In a further aspect, the amount of cholesterol in eggs of poultry fed the animal feed is reduced at least about 15% per gram of yolk, when compared to yolks obtained from poultry fed uncolonized foodstuff. In a further aspect, the amount of cholesterol in eggs of poultry fed the animal feed is reduced at least about 25% per gram of yolk, when compared to yolks obtained from poultry fed uncolonized foodstuff.

[0034] In another embodiment of the invention, a poultry animal is fed on improved feed comprising a poultry foodstuff colonized by Monascus species. on&sc s-colonized foodstuff is in an amount effective to reduce the amount of cholesterol in eggs of the poultry, as compared to the amount of cholesterol in such eggs when unimproved uncolonized foodstuff is fed to a poultry.

[0035] In another embodiment of the invention, an egg is produced by a poultry animal wherein the egg is from the poultry animal fed a poultry foodstuff colonized by Monascus species. Monascus-colonized foodstuff is in an amount effective to reduce the amount of cholesterol in eggs of the poultry animal, as compared to the amount of cholesterol in such eggs when uncolonized foodstuff is fed to a poultry.

[0036] In another embodiment, the present invention is a poultry egg having reduced cholesterol from a poultry animal fed on improved feed when compared to an egg from a poultry fed on unimproved feed. The reduced cholesterol poultry egg is produced by feeding to the animal a poultry foodstuff and Monascus spp. colonized upon the foodstuff. The Monascus- colonized foodstuff is fed to the poultry animal in an amount effective to reduce the amount of cholesterol in eggs of the poultry animal, as compared to the amount of cholesterol in such eggs when the uncolonized foodstuff is fed to a poultry.

[0037] In another embodiment, the present invention is a comestible containing an egg or egg product having reduced cholesterol from a poultry animal fed on improved feed when compared to an egg from a animal fed on unimproved feed. The reduced cholesterol egg or egg product is produced by feeding to the animal a poultry foodstuff and Monascus spp. colonized upon the foodstuff. The Monascus-colonized foodstuff is fed to the poultry animal in an amount effective to reduce the amount of cholesterol in eggs of the poultry animal, as compared to the amount of cholesterol in such eggs when the uncolonized foodstuff is fed to a poultry. [0038] In one aspect of the invention, Monascus spp. is incubated with a poultry foodstuff comprising standard poultry feed, cracked soybean, wheat bran and/or other appropriate components. The foodstuff are aseptic to allow preferential colonization by Monascus spp. In one preferred aspect, Monascus spp. is incubated with aseptic foodstuff comprising standard poultry feed and cracked soybean. In a further preferred aspect, Monascus spp. is incubated with aseptic foodstuff comprising standard poultry feed and wheat bran.

[0039] Utilization of relatively low cost, commercial poultry foodstuff as a colonization medium for Monascus spp. results in a lower cost animal feed composition effective in reducing the amount of cholesterol in an egg, when fed to an egg-laying animal. The lower cost animal feed compositions of the present invention compare to more costly Monascus spp. feed compositions that contain cofactors, vitamins, and/or trace minerals.

[0040] The animal feed of the present invention comprises Monascus spp. colonized upon a poultry foodstuff. The standard poultry foodstuff of the present invention include, but are not limited to, corn, soybean meal, limestone, dicalcium phosphate, vitamin mix, trace mineral mix, biotin, sodium chloride, canola oil, and methionine. Jm certain preferred methods and compositions of the present invention, the animal feed compositions utilize standard poultry foodstuff in combination with soybean, cracked soybean, wheat, wheat bran, flax, corn (cracked, ground, whole, meal), oats, rice, barley, rye, rapeseed, sorgham, millet or other suitable grains, or combinations thereof.

[0041] The animal feed compositions of the present invention contrast with more costly animal feed compositions. The more costly feed compositions may include canola oil, copper citrate, chromium, iodine, vitamin E, and folate, among other additives. The more costly compositions may also include processed Monascus spp. grown on rice. The more costly feed compositions may also include phytosterol, a copper derivative, and processed Monascus spp. grown on rice. Colonization of the animal feed composition by Monascus spp. utilizing compositions and methods of the present invention, achieves highly effective statin levels in the absence of pre-colonization on rice. The animal feed compositions and methods of the present invention include soybean, cracked soybean, wheat, and/or wheat bran in combination with standard poultry feed in the aseptic foodstuff, and do not necessarily require rice as one of the aseptic foodstuff.

[0042] In one embodiment of the present invention, an animal feed composition is effective in reducing the amount of cholesterol in an egg when fed to an egg-laying animal. The animal feed composition comprises aseptic foodstuff including, but not limited to, standard poultry foodstuff, soybean, cracked soybean, wheat, and/or wheat bran. Water content of the animal feed composition is adjusted to comprise between about 25% and about 65% by weight of the aseptic foodstuff to form a semi-solid substrate. Monascus spp. is inoculated into the aseptic foodstuff and incubated for a time sufficient to allow a level of Monascus colonization. The level of Monascus colonization is sufficient to produce the animal feed composition effective in reducing the amount of cholesterol in an egg when fed to the egg-laying animal.

[0043] In one exemplary aspect, the animal feed composition and methods of the present invention combines poultry foodstuff and cracked soybean. In a further aspect, the animal feed composition and methods of the present invention combines poultry foodstuff and wheat bran. In a further embodiment, the poultry foodstuff, cracked soybean, and wheat bran are aseptic.

[0044] In another embodiment, the animal feed compositions of the present invention achieve a level of Monascus spp. that is between about 1% and about 20% by weight of the poultry foodstuff. The terms "Monascus colonization by weight is between about 1% and about 20% by weight of the poultry foodstuff or "Monascus spp. is between about 1% and about 20% by weight of the aseptic poultry foodstuff mean that production of Monascus spp. yeast and Monascus fermentation products by weight is between about 1% and about 20% by weight of the poultry foodstuff. Preferably, the animal feed composition of the present invention achieves a level of Monascus spp. that is between about 4% and about 17% by weight of the poultry foodstuff. In a further aspect, the animal feed composition of the present invention achieves a level of Monascus spp. that is between about 8% and about 10% by weight of the poultry foodstuff.

[0045] In accordance with preferred aspects of the invention, Monascus spp. is inoculated into the poultry foodstuff and incubated for a time and at a temperature sufficient to allow a level of Monascus colonization. In a further aspect, Monascus spp. is inoculated into aseptic poultry foodstuff. The level of Monascus colonization is sufficient to produce the animal feed of the present invention effective in reducing the amount of cholesterol in an egg when fed to the poultry or the egg-laying animal. The animal feed of the present invention is fed to an egg-laying animal and the amount of cholesterol in the egg is reduced by at least about 5% per gram of yolk, when compared to yolks obtained from an animal fed an animal feed in the absence of Monascus colonization. In another aspect, the animal feed of the present invention is fed to a poultry or an egg-laying animal and the amount of cholesterol in the egg is reduced by at least about 15% per gram of yolk, when compared to yolks obtained from an animal fed an animal feed in the absence of Monascus colonization. In a further aspect, the animal feed of the present invention is fed to an egg-laying animal and the amount of cholesterol in the egg is reduced by at least about 25% per gram of yolk, when compared to yolks obtained from an animal fed an animal feed in the absence of Monascus colonization.

[0046] The incubation time for Monascus spp. on the feed is preferably between about 5 and about 45 days, more preferably between about 10 and about 40 days, and even more preferably between about 20 and about 30 days. The incubation temperature for Monascus spp. is preferably between about 10°C and about 30°C, and more preferably between about 15°C and about 25°C.

[0047] In an aspect of the invention, Monascus species include, but are not limited to, Monascus purpureus, Monascus ruber, Monascus anka, Monascus vitreus, and Monascus paxii. All such Monascus spp. may be referred to as "Red Yeast". In another aspect of the invention, egg-laying animals include, but are not limited to, chickens, ducks, geese, turkeys, quail, pigeon, guinea fowl, peafowl, pheasant, chukar, partridge, grouse, emu, rhea and ostrich.

[0048] In another embodiment, the invention comprises a method of producing an poultry feed effective in reducing the amount of cholesterol in an egg when fed to an egg-laying animal. The method combines preferably poultry foodstuff including, but not limited to, standard poultry foodstuff, soybean, cracked soybean, wheat, wheat bran, or combinations thereof. The method further adjusts the water content of the poultry foodstuff to obtain a final moisture content preferably between about 25% and about 65% by weight of the poultry foodstuff. Monascus spp. is inoculated into the poultry foodstuff and incubated with the poultry foodstuff for a time sufficient to allow Monascus colonization. In one aspect Monascus spp. is inoculated into aseptic poultry foodstuff. Monascus colonization is sufficient to produce the poultry feed containing enhanced amounts of one or more cholesterol-lowering compounds including statins effective in reducing the amount of cholesterol in an egg when the animal feed is fed to the egg-laying animal.

[0049] Methods of producing poultry feed of the present invention can achieve levels of Monascus spp. that is between about 1% and about 20%> by weight of the poultry foodstuff. The terms "Monascus colonization by weight is between about 1% and about 20% by weight of the poultry foodstuff or "Monascus spp. is between about 1% and about 20% by weight of the aseptic poultry foodstuff mean that production of Monascus spp. yeast and Monascus fermentation products by weight comprises between about 1% and about 20% by weight of the poultry foodstuff. In another aspect, the methods of the present invention achieve a level of Monascus spp. that is between about 4% and about 17% by weight of the poultry foodstuff. In a preferred aspect, the methods of the present invention achieve a level of Monascus spp. that is between about 8% and about 10% by weight of the poultry foodstuff.

[0050] By methods of the present invention, Monascus spp. is inoculated into preferably aseptic poultry foodstuff and incubated for a time sufficient to allow a desired level of Monascus colonization. The level of Monascus colonization is sufficient to produce the animal feed effective in reducing the amount of cholesterol in an egg when fed to the poultry or egg- laying animal. Upon feeding the animal feed to the poultry or egg-laying animal, the amount of cholesterol in the egg is reduced by at least about 5% per gram of yolk, when compared to a yolk obtained from an animal fed the same feed but in the absence of Monascus colonization. In another aspect, the methods of the invention result in the reduction in the amount of cholesterol in the egg by at least about 15% per gram of yolk, when compared to a yolk obtained from an animal fed the same feed but in the absence of Monascus colonization. Reduction in the amount of cholesterol in the egg by at least about 25% per gram of yolk, when compared to a yolk obtained from an animal fed the same feed but in the absence of Monascus colonization is preferred.

[0051] In accordance with an aspect of the invention, methods of producing an animal feed compositions provide for inoculating and incubating Monascus spp. in discrete units of aseptic poultry foodstuff. Colonization of the aseptic poultry foodstuff by Monascus spp. occurs within the discrete units. Such units may be convenient containers such as plastic containers or bags, such as sealable polypropylene containers.

[0052] In another embodiment, the invention includes methods of reducing the amount of cholesterol in a poultry egg. The methods provide administering to an egg-laying animal an animal feed comprising a cholesterol-lowering Monascus colonized foodstuff effective in reducing the amount of cholesterol in the egg by at least about 5% or greater per gram of yolk, when compared to a yolk obtained from an animal fed the same feed but in the absence of Monascus colonization. The methods are effective in reducing the amount of cholesterol in the egg by at least about 15%, or by at least about 25%, when compared to a yolk obtained from an animal fed the same feed but in the absence of Monascus colonization. [0053] In another embodiment, the invention includes methods of producing cholesterol-lowering compounds including statins on a semi-solid substrate. The methods are effective in lowering levels of cholesterol in the blood serum of an animal. In addition, the methods are effective in a feed composition to reduce the amount of cholesterol in an egg when fed to an egg-laying animal. The methods provide production of Monascus-colonized feed. Monascus colonization is sufficient to produce statins, including but not limited to, lovastatin, simvastatin, and/or storvastatin.

[0054] Methods of producing cholesterol-lowering compounds including statins on a semi-solid substrate of the present invention can achieve levels of Monascus colonization by weight that is between about 1% and about 20% by weight of the poultry foodstuff. In another aspect, the methods of the present invention achieve levels of Monascus colonization by weight that is between about 4% and about 17% by weight of the poultry foodstuff. In another aspect, the methods of the present invention achieve a level of Monascus colonization by weight that is between about 8% and about 10% by weight of the poultry foodstuff. In a further aspect, the methods of the present invention provide Monascus colonization on aseptic poultry foodstuff.

[0055] The present invention provides improved methods and compositions effective in reducing the amount of cholesterol in an egg when fed to a poultry or an egg-laying animal. Examples of appropriate poultry or egg-laying animals for treatment according to the methods and compositions of the invention include, but are not restricted to, turkey, duck, goose, pigeon, guinea fowl, peafowl, quail, pheasant, chukar, partridge, grouse, emu, rhea and ostrich.

[0056] As described above, the present invention relates to an animal feed and a method of producing an animal feed or a poultry feed effective in reducing the amount of cholesterol in an egg when fed to a poultry. The compounds used in the composition or method are poultry foodstuff combining standard poultry foodstuff with soybean, cracked soybean, wheat, wheat bran, or combinations thereof; water comprising between about 25% and about 65% by weight of the poultry foodstuff; and Monascus spp. inoculated into the poultry foodstuff and incubated for a time sufficient to allow Monascus colonization. In one aspect, Monascus spp. is inoculated into aseptic poultry foodstuff. The Monascus colonization is sufficient to produce cholesterol-lowering compounds including statins in the animal feed.

[0057] The owαscws-colonized animal feed is an improved feed, effective in reducing the amount of cholesterol in an egg when fed to a poultry, or when the poultry is raised on the improved feed. When the poultry is raised on the improved feed of the present invention, the poultry produces improved eggs, poultry parts, egg components, especially yolks and other products which are reduced in the amount of cholesterol in the egg or poultry product, when compared to poultry raised on feed uncolonized by Monascus species.

[0058] In order to provide better understanding of the present invention, the following definitions are provided:

[0059] As used herein, "egg-laying animal" or "poultry" may refer to an egg-laying chicken hen, and generally to avian species which include, but are not limited to, turkey, duck, goose, pigeon, guinea fowl, peafowl, quail, pheasant, chukar, partridge, grouse, emu, rhea and ostrich.

[0060] As used herein, "egg" refers to liquid contents of a whole poultry egg which is de-shelled.

[0061] Commercial chicken shell eggs consist of 8-11% shell, 56-61% albumen, and 27-32% yolk. De-shelled whole egg is about 36% yolk and 64% albumen. The composition of albumen, yolk and whole egg is described in Chapter 6 of Egg Science & Technology, Second Edition, Stadelman and Cotterill, ed., Avi Publishing Company, 1977. Composition of the components of an egg are albumen (9.7-10.6% protein; 0.03% lipid); yolk (15.7-16.6% protein; 31.8-35.5%) lipid); whole egg (12.8-13.4% protein; 10.5-11.8% lipid).

[0062] The composition of yolk lipid is 65.5% triglyceride, 28.3% phospholipid and 5.2% cholesterol. The yolk phospholipid is composed of about 73% phosphatidylcholine, 15% phosphatidylethanolamine, 5.8% lysophosphatidylcholine, 2.5% sphinogomyelin, 2.1% lysophosphatidylethanolamine, 0.9% plasmalogen and 0.6% inositol phospholipid.

[0063] As used herein, "cholesterol" refers to a steroid compound found in animal cells. Cholesterol may be of dietary origin or may be synthesized de novo from acetate. The molecular structure is a 27-carbon structure based on four concerted rings, three of which contain six carbons and one contains five carbons. Cholesterol is a precursor to steroid hormone synthesis in the adrenal cortex and other organs of the animal. Cholesterol is a precursor to the synthesis of steroids including, but not limited to, aldosterone, corticosterone, cortisol, androstenedione, dehydroepiandrosterone, dehydroepiandrosterone sulphate. (Molecular Biology and Biotechnology, R.A. Meyers, Ed., Wiley- VCH, Inc., 1995).

[0064] As used herein, "a comestible containing an egg or egg product" refers to any food or edible product containing whole egg, egg yolk, egg white, or dehydrated whole egg, egg yolk, egg white, or dehydrated and reconstituted whole egg, egg yolk, egg white. A comestible may include a pre-packaged prepared food, egg in the shell, or a prepared food served in a restaurant, grocery store, or supermarket. Examples of prepared foods containing egg products are mayonnaise, bread, ice cream, cookie dough, custard, pudding, tapioca, or cooked eggs.

[0065] As used herein, "standard poultry feed" or "standard poultry foodstuff refer to a standard diet fed to poultry, for example, chickens, resulting in optimal growth and reproduction of the animals. The standard poultry foodstuff (percentage by weight) includes, but is not limited to, corn meal (approximately 55 to 60%), soybean meal (approximately 21 to 24%o), limestone (approximately 8 to 10%), canola oil (approximately 2 to 4%), dicalcium phosphate (approximately 1.7 to 1.8%), biotin supplement (approximately 1%), salt (approximately 0.3 to 0.6%), vitamin/trace mineral mix (approximately 0.15 to 0.25%), and methionine (approximately 0.15 to 0.2%).

[0066] As used herein, "poultry foodstuff or "aseptic poultry foodstuff or "poultry feed" or "animal feed" refer to all poultry foodstuff found in standard poultry feed. In addition, poultry foodstuff, aseptic poultry foodstuff, poultry feed or animal feed may include soybean, cracked soybean, wheat, wheat bran, or combinations thereof.

[0067] As used herein, "animal feed" or "poultry foodstuff also refers to Monascus- colonized poultry foodstuff. Poultry foodstuff refers to the mixture comprising poultry foodstuff and Monascus colonized upon the poultry foodstuff. Monascus inoculation is not an additive to the poultry foodstuff, but rather a seed inoculation for growth of Monascus spp. yeast upon the poultry foodstuff.

[0068] As used herein, "aseptic poultry foodstuff refers to sterile poultry foodstuff. Sterilization of poultry foodstuff may occur by any standard technique such as steam sterilization underpressure at 121°C. Alternatively, sterilization of poultry foodstuff may occur by any form of chemical sterilization or sterilization by UV irradiation that will leave the poultry foodstuff non-toxic when fed to poultry. o

[0069] As used herein, "water comprising between about 25% and about 65%> by weight of the poultry foodstuff refers to the final moisture content of the poultry foodstuff. Poultry foodstuff in varying degrees of hydration will have varying water content. Water may be added, if necessary, to achieve a final moisture content of 25% to about 65% by weight of the poultry foodstuff.

[0070] As used herein, "Monascus species" or "Monascus spp." refers to any number of species of the yeast genus Monascus. Monascus spp. include, but are not limited to, Monascus purpureus, Monascus ruber, Monascus anka, Monascus vitreus, and Monascus paxii. All such Monascus spp. may be referred to as "Red Yeast".

[0071] As used herein, "Monascus colonization" refers to the inoculation of Monascus spp., for example, Monascus purpureus or Monascus ruber, onto poultry foodstuff. Monascus spp. may be inoculated onto aseptic poultry foodstuff. Monascus spp. is incubated aerobically under controlled conditions of temperature and time. For example, Monascus spp. incubation may occur under controlled conditions for a time preferably between about 5 and about 45 days, more preferably between about 10 and about 40 days, and even more preferably between about 20 and about 30 days. The incubation temperature for Monascus spp. is preferably between about 10°C and about 30°C, and more preferably between about 15°C and about 25°C.

[0072] As used herein, "Monascus colonization by weight" or "Monascus-colonized poultry foodstuff further refers to Monascus colonization of the poultry foodstuff wherein production of Monascus spp. yeast and Monascus fermentation products by weight comprises between about 1% and about 20% by weight of the poultry foodstuff. Within further compositions and methods of the present invention, "Monascus colonization by weight" comprises production of Monascus spp. yeast and Monascus fermentation products by weight between about 4% and about 17%> by weight of the poultry foodstuff. Within further compositions and methods of the present invention, or "Monascus colonization by weight" comprises production of Monascus spp. yeast and Monascus fermentation products by weight between about 8% and about 10%> by weight of the poultry foodstuff.

[0073] In an exemplary aspect of the invention, Monascus colonization by weight comprises, for example, between about 8% and about 10%> by weight of the poultry foodstuff, and the Monascus-colonized animal feed is fed to a poultry. The amount of cholesterol in the egg of the poultry fed the animal feed of the present invention is reduced by at least about 5% per gram of yolk, when compared to a yolk obtained from a poultry fed an animal feed composition prepared in the absence of Monascus colonization. In another aspect, the amount of cholesterol in the egg of the poultry fed the composition of the present invention is reduced at least about 15% per gram of yolk, when compared to a yolk obtained from a poultry fed an animal feed prepared in the absence of Monascus colonization. In another aspect, amount of cholesterol in the egg of the poultry fed the composition of the present invention is reduced at least about 25% per gram of yolk, when compared to a yolk obtained from a poultry fed an animal feed prepared in the absence of Monascus colonization. [0074] The compositions and methods of the present invention provide a solid or semi- solid substrate of aseptic poultry foodstuff for Monascus colonization. The moisture content of the poultry foodstuff is between approximately 25% and 65% by weight of the poultry foodstuff. The moisture content provides an optimal solid or semi-solid substrate for Monascus colonization of the aseptic poultry foodstuff. Monascus colonization on the solid or semi-solid substrate of aseptic poultry foodstuff provides a low cost, efficient method for producing high levels of Monascus-colonized poultry foodstuff. The high levels of Monascus colonization are sufficient to produce the animal feed composition effective in reducing the amount of cholesterol in the egg when fed to an egg-laying animal.

[0075] In a further embodiment, inoculation and colonization by Monascus spp. occurs under controlled aerobic conditions of temperature and time within discrete units of the semi- solid substrate of aseptic poultry foodstuff. For example, the discrete units of the semi-solid substrate of aseptic poultry foodstuff are contained, e.g., within sealable polyethylene bags or sealable polypropylene containers.

[0076] As described above, the present invention relates to a composition and method of producing cholesterol-lowering compounds including statins on a solid or semi-solid substrate by Monascus colonization of the poultry foodstuff sufficient to produce statins effective in lowering levels of cholesterol in the blood serum of an animal. The compositions and methods are also sufficient to produce statins effective in animal feed to reduce the amount of cholesterol in an egg when fed to a poultry or egg-laying animal.

[0077] The compositions and methods of the present invention produce statins, for example, lovastatin, simvastatin, and/or storvastatin. Statins are inhibitors of HMG-CoA reductase, the rate-limiting enzyme in cholesterol biosynthesis. Statins produced by compositions and methods of the present invention are effective in an animal feed to reduce the amount of cholesterol in an egg when fed to a poultry or an egg-laying animal. Statins produced by compositions and methods of the present invention are also effective in lowering levels of cholesterol in the blood serum of an animal, for example, a human or a poultry.

[0078] The compositions and methods of the present invention include cholesterol- lowering compounds, in addition to statins, within the Monascus-colonized poultry foodstuff of the present invention that are responsible for increased effectiveness in lowering cholesterol in eggs when fed to an egg-laying animal. For example, Monascus-colonized poultry foodstuff of the present invention are more effective in lowering cholesterol in eggs when fed to an egg- laying animal than are purified statins added to standard poultry foodstuff. The comparison indicates that Monascus-colonized poultry foodstuff of the present invention are responsible for increased effectiveness in lowering cholesterol in eggs when fed to an egg-laying animal. Monascus colonization results in production of statins and other cholesterol-lowering components in the poultry foodstuff. Monascus colonization results in production of statins, monacolin family of polyketides, fatty acids, trace elements, and other cholesterol-lowering components in the poultry foodstuff. Monascus colonization results in production of monacolin family of polyketides, for example, monacolin K, monacolin K (hydroxy acid form), dihydromonacolin K, dehydromonacolin K, monacolin, and monacolin L. Monascus colonization results in production of fatty acids, for example, saturated fatty acids consisting mainly of palmitic, stearic, and arachidic acids, and unsaturated fatty acids, mostly represented by oleic liinoleic, and linolenic acids. Monascus colonization results in production of polyketide pigments, for example, monascidin A, ankaflavin, monascorubrine, and monascorubramine.

[0079] Utilizing compositions and methods of the present invention, poultry foodstuff that include cracked soybean and or wheat bran are particularly effective for Monascus colonization to produce statins and other components that lower cholesterol in eggs when fed to a poultry or an egg-laying animal. In an aspect of the invention, Monascus colonization on standard poultry foodstuff, soybean, cracked soybean, wheat, wheat bran, or combinations thereof, for a sufficient incubation period results in production of at least about 0.01% statins by weight of total poultry foodstuff. Jm another aspect, Monascus colonization on standard poultry foodstuff, soybean, cracked soybean, wheat, wheat bran, or combinations thereof, for a sufficient incubation period results in production of at least about 0.1% statins by weight of total poultry foodstuff. In another aspect, Monascus colonization on standard poultry foodstuff, soybean, cracked soybean, wheat, wheat bran, or combinations thereof, for a sufficient incubation period results in production of at least about 0.3% statins by weight of total poultry foodstuff.

Examples

[0080] The following examples are provided by way of illustration, not limitation.

EXAMPLE 1

Production of Statins by Monascus purpureus on Components of Poultry Foodstuff

[0081] In Figure 1 , Monascus colonization on cracked soybean and standard poultry foodstuff for a 15 day incubation period results in production of 0.21% statins by weight of total poultry foodstuff. As a further example, Monascus colonization on wheat bran and standard poultry foodstuff for a 15 day incubation period results in production of 0.13% statins by weight of total poultry foodstuff. By contrast, Monascus colonization on corn meal (or cracked corn) and standard poultry foodstuff for a 15 day incubation period results in production of only 0.04%_> statins by weight of total poultry foodstuff. The percentage by weight of cracked soybean or wheat bran in the total poultry foodstuff is between about 5% and about 20%.

EXAMPLE 2

Effect of Monascus purpureus on the Cholesterol Content of Chicken Eggs

[0082] In Figure 2, laying chicken hens were fed Monascus purpureus-colonized poultry foodstuff containing 2.1% Monascus-colonization, 4.2% Monαscus-coloniza on, 8.4% M-wαscws-colonization, or 17% MoMαscus-colonization by weight of total poultry foodstuff. The average cholesterol content of eggs was reduced to 142 mg of cholesterol per egg from hens fed 8.4% Monascus-colonized poultry foodstuff compared to control eggs containing 189 mg of cholesterol in uncolonized foodstuff. This was a reduction of 25% in the content of cholesterol in eggs from poultry fed Monascus-colonized foodstuff compared to controls, eggs from poultry fed uncolonized foodstuff. The average reduction in cholesterol in eggs from poultry fed all treatments of Monascus-colonized foodstuff (2.1%> to 17% Monascus-colonized poultry foodstuff) was 21.3%.

EXAMPLE 3

High-Performance Liquid Chromatogram (HPLC) of Statins produced by Monascus purpureus colonized on Soybean and Standard Poultry Foodstuff

[0083] High-performance liquid chromatography (HPLC) of Monascus purpureus colonizing cracked soybean and standard poultry foodstuff demonstrates production of statins and other compounds. The HPLC system consisted of an L-6200 pump and a Sig-Grad, RP-18 100 mm column. HPLC conditions used solvent A, H₂0; solvent B, 0.05% phosphoric acid; solvent C, acetonitrile. The sample consisted of a solvent extract of Monascus purpureus. HPLC chromatography was performed according to published protocols. Ma, et al., J. Agric. Food. Chem. 48: 5220-5225, (2000).

[0084] Figure 3 shows HPLC chromatography of statins and other compounds produced by Monascus purpureus colonizing cracked soybean and standard poultry foodstuff. Monascus colonization results in production of statins and other cholesterol-lowering components in the poultry foodstuff. Monascus colonization results in production of statins, monacolin family of polyketides, fatty acids, trace elements, and other cholesterol-lowering components in the poultry foodstuff. Monascus colonization results in production of statins, for example, lovastatin, simvastatin, and storvastatin. Monascus colonization results further in production of monacolin family of polyketides, for example, monacolin K, monacolin K (hydroxy acid form), dihydromonacolin K, dehydromonacolin K, monacolin, and monacolin L. Monascus colonization results further in production of fatty acids, for example, saturated fatty acids consisting mainly of palmitic, stearic, and arachidic acids, and unsaturated fatty acids, mostly represented by oleic liinoleic, and linolenic acids. Monascus colonization results further in production of polyketide pigments, for example, monascidin A, ankaflavin, monascorubrine, and monascorubramine.

[0085] Morcαscws-colonized poultry foodstuff of the present invention are more effective in lowering cholesterol in eggs when fed to an egg-laying animal than are purified statins added to standard poultry foodstuff. The comparison indicates that Monascus-colonized poultry foodstuff of the present invention are responsible for increased effectiveness in lowering cholesterol in eggs when fed to an egg-laying animal.

EXAMPLE 4

Effect of Monascus purpureus on the Cholesterol Content of Chicken Eggs

[0086] In Figure 4, laying chicken hens were fed Monascus purpureus-colonized poultry foodstuff containing 4.2% Monas cus-colonizaύon by weight of total poultry foodstuff. The average cholesterol content of eggs was reduced to 167 mg of cholesterol per egg from hens fed 4.2% Monαscws-colonized poultry foodstuff compared to control eggs containing 180 mg of cholesterol in uncolonized foodstuff. The average reduction in cholesterol in eggs from poultry fed the 4.2% Mowαscws-colonized foodstuff was 7.0%.

[0087] In Figure 5, laying chicken hens were fed Monascus purpureus-colonized poultry foodstuff containing 4.2%> Monascus-colo ization by weight of total poultry foodstuff. The average cholesterol content of eggs was reduced to 147 mg of cholesterol per egg from hens fed 4.2% Monascus-colonized poultry foodstuff compared to control eggs containing 167 mg of cholesterol in uncolonized foodstuff. The average reduction in cholesterol in eggs from poultry fed the 4.2% Monascus-colonized foodstuff was 12.1%).

[0088] Additional advantages and modifications of the invention disclosed herein will occur to those persons skilled in the art. Accordingly, the invention in its broader aspects is not limited to the specific details or illustrated examples described herein. Therefore, all departures made from the detail are deemed to be within the scope of the invention as defined by the appended claims.

Claims

What is Claimed:

1. An animal feed comprising: a poultry foodstuff; and colonized upon the foodstuff, Monascus spp. in an amount effective to reduce the amount of cholesterol in eggs of poultry fed the animal feed as compared to the amount of cholesterol in such eggs when the uncolonized foodstuff is fed to the poultry.

2. The feed of claim 1 wherein the poultry foodstuff comprises soybean, wheat, corn, flax, oats, rye, rice, barley, rapeseed, sorgham, millet, and combinations thereof.

3. The feed of claim 1 wherein Monascus spp. is selected from the group consisting of Monascus purpureus, Monascus ruber, Monascus anka, Monascus vitreus, and Monascus paxii.

4. The feed of claim 1 wherein the effective amount of Monascus colonization by weight is between about 1%> and about 20% by weight of the poultry foodstuff.

5. The feed of claim 4 wherein the effective amount of Monascus colonization by weight is between about 4% and about 17% by weight of the poultry foodstuff

6. The feed of claim 4 wherein the effective amount of Monascus colonization by weight is between about 8% and about 10% by weight of the poultry foodstuff.

7. The feed of claim 1 further comprising water between about 25% and about 65% by weight of the poultry foodstuff.

8. The feed of claim 2 wherein the soybean is cracked soybean.

9. The feed of claim 2 wherein the wheat is wheat bran.

10. The feed of claim 1 wherein the poultry foodstuff are aseptic poultry foodstuff.

11. The feed of claim 1 wherein the amount of cholesterol in eggs of poultry fed the animal feed is reduced at least about 5% per gram of yolk, when compared to yolks obtained from poultry fed uncolonized foodstuff.

12. The feed of claim 11 wherein the amount of cholesterol in eggs of poultry fed the animal feed is reduced at least about 15% per gram of yolk, when compared to yolks obtained from poultry fed uncolonized foodstuff.

13. The feed of claim 11 wherein the amount of cholesterol in eggs of poultry fed the animal feed is reduced at least about 25% per gram of yolk, when compared to yolks obtained from poultry fed uncolonized foodstuff.

14. The feed of claim 10 wherein the aseptic poultry foodstuff are poultry foodstuff and cracked soybean.

15. The feed of claim 10 wherein the aseptic poultry foodstuff are poultry foodstuff and wheat bran.

16. The feed of claim 2 wherein the poultry foodstuff further comprises corn, soybean meal, limestone, dicalcium phosphate, vitamin mix, trace mineral mix, biotin, sodium chloride, canola oil, and/or methionine.

17. The feed of claim 1 further comprising an incubation time to achieve Monascus spp. colonization between approximately 2 and 25 days.

18. The feed of claim 17 further comprising an incubation time to achieve Monascus spp. colonization between approximately 5 and 20 days.

19. The feed of claim 17 further comprising an incubation time to achieve Monascus spp. colonization between approximately 13 and 17 days.

20. The feed of claim 1 further comprising an incubation temperature to achieve Monascus spp. colonization between approximately 15°C and 35°C.

21. The feed of claim 20 further comprising an incubation temperature to achieve Monascus spp. colonization between approximately 20°C and 30°C.

22. The feed of claim 1 wherein the poultry is selected from the group consisting of chicken, duck, goose, turkey, quail, pigeon, guinea fowl, peafowl, pheasant, chukar, partridge, grouse, emu, rhea and ostrich.

23. A method of producing poultry feed comprising: inoculating poultry foodstuff with Monascus spp.; and incubating the inoculated foodstuff for a time and under conditions effective to produce enhanced amounts of one or more cholesterol lowering compounds in the food materials.

24. The method of claim 23 wherein the incubation gives rise to levels of Monascus spp. in the feed effective to reduce the amount of cholesterol in eggs of poultry fed the poultry feed as compared to the amount of cholesterol found in such eggs when the uncolonized foodstuff is fed to the poultry.

25. The method of claim 23 wherem the poultry foodstuff comprises soybean, wheat or combinations thereof.

26. The method of claim 23 wherein Monascus spp. is selected from the group consisting of Monascus purpureus, Monascus ruber, Monascus anka, Monascus vitreus, and Monascus paxii.

27. The method of claim 23 wherein the cholesterol-lowering compounds comprise statin, lovastatin, simvastatin, storvastatin or a combination thereof.

28. The method of claim 23 further comprising colonizing Monascus spp. by weight between about 1% and about 20% by weight of the poultry foodstuff.

29. The method of claim 28 further comprising colonizing Monascus spp. by weight between about 4%> and about 17%o by weight of the poultry foodstuff.

30. The method of claim 28 further comprising colonizing Monascus spp. by weight between about 8% and about 10% by weight of the poultry foodstuff.

31. The method of claim 23 further comprising adjusting the water content of the poultry foodstuff to obtain a final moisture content between about 25%> and about 65%> by weight of the poultry foodstuff.

32. The method of claim 25 wherein the soybean is cracked soybean.

33. The method of claim 25 wherein the wheat is wheat bran.

34. The method of claim 23 further comprising sterilizing the poultry foodstuff to produce aseptic poultry foodstuff.

35. The method of claim 24 wherem the amount of cholesterol in eggs of poultry fed the poultry feed is reduced at least about 5% per gram of yolk as compared to the amount of cholesterol found in such eggs when the uncolonized foodstuff is fed to the poultry.

36. The method of claim 35 wherein the amount of cholesterol in eggs of poultry fed the poultry feed is reduced at least about 15% per gram of yolk as compared to the amount of cholesterol found in such eggs when the uncolonized foodstuff is fed to the poultry.

37. The method of claim 35 wherein the amount of cholesterol in eggs of poultry fed the poultry feed is reduced at least about 25% per gram of yolk as compared to the amount of cholesterol found in such eggs when the uncolonized foodstuff is fed to the poultry.

38. The method of claim 34 wherein the aseptic poultry foodstuff comprises standard poultry foodstuff and cracked soybean.

39. The method of claim 34 wherein the aseptic poultry foodstuff comprises standard poultry foodstuff and wheat bran.

40. The method of claim 25 wherein the poultry foodstuff comprises corn, soybean meal, limestone, dicalcium phosphate, vitamin mix, trace mineral mix, biotin, sodium chloride, canola oil, and/or methionine.

41. The method of claim 23 further comprising inoculating and incubating the poultry foodstuff with Monascus spp. in discrete units.

42. The method of claim 41 wherein the discrete units are contained within sealable polypropylene containers.

43. The method of claim 23 wherein the incubation time for the inoculated foodstuff is between approximately 2 and 25 days.

44. The method of claim 43 wherein the incubation time for the inoculated foodstuff is between approximately 5 and 20 days.

45. The method of claim 43 wherein the incubation time for the inoculated foodstuff is between approximately 13 and 17 days.

46. The method of claim 23 further comprising an incubation temperature for the inoculated foodstuff between approximately 15°C and 35°C.

47. The method of claim 46 further comprising an incubation temperature for the inoculated foodstuff between approximately 20°C and 30°C.

48. The method of claim 23 wherein the poultry is selected from the group consisting of chicken, duck, goose, turkey, quail, pigeon, guinea fowl, peafowl, pheasant, chukar, partridge, grouse, emu, rhea and ostrich.

49. A method of reducing the amount of cholesterol in eggs produced by poultry comprising: feeding to the poultry an animal feed which has been inoculated with Monascus spp., the Monascus being present in the feed in an amount effective to attain such reduction as compared to the amount of cholesterol in such eggs produced when the animal feed without the Monascus is fed to the poultry.

50. The method of claim 49 wherein the Monascus by weight is between about 1 and about 20% by weight of the animal feed.

51. The method of claim 50 wherein the Monascus by weight is between about 4 and about 17% by weight of the animal feed.

52. The method of claim 50 wherein the Monascus by weight is between about 8 and about 10% by weight of the animal feed.

53. The method of claim 49 wherein the Monascus is present in the feed in an amount effective in reducing the amount of cholesterol in the egg by at least about 5% per gram of yolk, as compared to the amount of cholesterol in such eggs produced when the animal feed without the Monascus is fed to the poultry.

54. The method of claim 53 wherem the Monascus is present in the feed in an amount effective in reducing the amount of cholesterol in the egg by at least about 15% per gram of yolk, as compared to the amount of cholesterol in such eggs produced when the animal feed without the Monascus is fed to the poultry.

55. The method of claim 53 wherein the Monascus is present in the feed in an amount effective in reducing the amount of cholesterol in the egg by at least about 25% per gram of yolk, as compared to the amount of cholesterol in such eggs produced when the animal feed without the Monascus is fed to the poultry.

56. The method of claim 49 further comprising sterilizing the animal feed to produce aseptic animal feed prior to inoculating with Monascus spp.

57. The method of claim 49 further comprising adjusting the water of the animal feed to obtain a final moisture content between about 25% and about 65% by weight of the animal feed.

58. The method of claim 49 wherein the animal feed comprises soybean, wheat or combinations thereof.

59. The method of claim 58 wherein the soybean is cracked soybean.

60. The method of claim 58 wherein the wheat is wheat bran.

61. The method of claim 49 wherein Monascus spp. is selected from the group consisting of Monascus purpureus, Monascus ruber, Monascus anka, Monascus vitreus, and Monascus paxii.

62. The method of claim 56 further comprising combining aseptic animal feed of standard poultry feed and cracked soybean.

63. The method of claim 56 further comprising combining aseptic animal feed of standard poultry feed and wheat bran.

64. The method of claim 49 wherem the poultry is selected from the group consisting of chicken, duck, goose, turkey, quail, pigeon, guinea fowl, peafowl, pheasant, chukar, partridge, grouse, emu, rhea and ostrich.

65. A poultry animal fed on improved feed comprising: the poultry animal fed a poultry foodstuff colonized by Monascus spp., in an amount effective to reduce the amount of cholesterol in eggs of the animal, as compared to the amount of cholesterol in such eggs when uncolonized foodstuff is fed to a poultry.

66. The poultry animal of claim 65 wherein the Monascus is present in the poultry foodstuff in an amount effective to reduce the amount of cholesterol in eggs of the animal by at least about 5% per gram of yolk, as compared to the amount of cholesterol in such eggs when uncolonized foodstuff is fed to a poultry.

67. The poultry animal of claim 66 wherein the Monascus is present in the poultry foodstuff in an amount effective to reduce the amount of cholesterol in eggs of the animal by at least about 15% per gram of yolk, as compared to the amount of cholesterol in such eggs when uncolonized foodstuff is fed to a poultry.

68. The poultry animal of claim 66 wherein the Monascus is present in the poultry foodstuff in an amount effective to reduce the amount of cholesterol in eggs of the animal by at least about 25% per gram of yolk, as compared to the amount of cholesterol in such eggs when uncolonized foodstuff is fed to a poultry.

69. An egg produced by a poultry animal comprising: an egg from the poultry animal fed a poultry foodstuff colonized by Monascus spp. in an amount effective to reduce the amount of cholesterol in eggs of the poultry animal, as compared to the amount of cholesterol in such eggs when uncolonized foodstuff is fed to a poultry.

70. The egg of claim 69 wherein the Monascus is present in the feed in an amount effective to reduce the amount of cholesterol in the egg by at least about 5% per gram of yolk, as compared to the amount of cholesterol in such eggs produced when uncolonized foodstuff is fed to the poultry.

71. The egg of claim 70 wherein the Monascus is present in the feed in an amount effective to reduce the amount of cholesterol in the egg by at least about 15% per gram of yolk, as compared to the amount of cholesterol in such eggs produced when uncolonized foodstuff is fed to the poultry.

72. The egg of claim 70 wherein the Monascus is present in the feed in an amount effective to reduce the amount of cholesterol in the egg by at least about 25% per gram of yolk, as compared to the amount of cholesterol in such eggs produced when uncolonized foodstuff is fed to the poultry.

73. The egg of claim 69 wherein Monascus colonization by weight is between about 1% and about 20%) by weight of the poultry foodstuff.

74. A poultry egg having reduced cholesterol from a poultry animal fed on improved feed when compared to an egg from a poultry fed on unimproved feed, produced by: feeding to the poultry animal the improved feed comprising a poultry foodstuff, and colonized upon the foodstuff, Monascus spp., the colonized foodstuff fed to the poultry animal in an amount effective to reduce the amount of cholesterol in eggs of the poultry animal, as compared to the amount of cholesterol in such eggs when unimproved uncolonized foodstuff is fed to a poultry.

75. The egg of claim 74 wherein the Monascus is present in the feed in an amount effective in reducing the amount of cholesterol in the egg by at least about 5% per gram of yolk, as compared to the amount of cholesterol in such eggs produced when unimproved uncolonized foodstuff is fed to the poultry.

76. The egg of claim 75 wherein the Monascus is present in the feed in an amount effective in reducing the amount of cholesterol in the egg by at least about 15%> per gram of yolk, as compared to the amount of cholesterol in such eggs produced when unimproved uncolonized foodstuff is fed to the poultry.

77. The egg of claim 75 wherein the Monascus is present in the feed in an amount effective in reducing the amount of cholesterol in the egg by at least about 25% per gram of yolk, as compared to the amount of cholesterol in such eggs produced when unimproved uncolonized foodstuff is fed to the poultry.

78. The egg of claim 74 wherein Monascus colonization by weight is between about 1% and about 20%) by weight of the poultry foodstuff.

79. A comestible containing an egg or egg product having reduced cholesterol from a poultry animal fed on improved feed when compared to an egg from a animal fed on unimproved feed, produced by: feeding to the poultry animal the improved feed comprising a poultry foodstuff, and colonized upon the foodstuff, Monascus spp., the colonized foodstuff fed to the poultry animal in an amount effective to reduce the amount of cholesterol in eggs of the poultry animal, as compared to the amount of cholesterol in such eggs when unimproved uncolonized foodstuff is fed to a poultry; adding the egg or egg product reduced in cholesterol to the comestible to produce a comestible reduced in cholesterol.

80. The comestible of claim 79 wherein the Monascus is present in the feed in an amount effective in reducing the amount of cholesterol in the egg by at least about 5% per gram of yolk, as compared to the amount of cholesterol in such eggs produced when unimproved uncolonized foodstuff is fed to the poultry.

81. The comestible of claim 80 wherein the Monascus is present in the feed in an amount effective in reducing the amount of cholesterol in the egg by at least about 15% per gram of yolk, as compared to the amount of cholesterol in such eggs produced when unimproved uncolonized foodstuff is fed to the poultry.

82. The comestible of claim 80 wherem the Monascus is present in the feed in an amount effective in reducing the amount of cholesterol in the egg by at least about 25% per gram of yolk, as compared to the amount of cholesterol in such eggs produced when unimproved uncolonized foodstuff is fed to the poultry.

83. The egg of claim 79 wherein Monascus colonization by weight is between about 1 % and about 20% by weight of the poultry foodstuff.