US 2012025 1512A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2012/0251512 A1 Farmer et al. (43) Pub. Date: Oct. 4, 2012

(54) PROBOTC SPORTS NUTRITION Related U.S. Application Data COMPOSITIONS (60) Provisional application No. 61/469.924, filed on Mar. 31, 2011. (75) Inventors: Sean Farmer, Miami, FL (US); David Keller, University Heights, Publication Classification OH (US); Andrew R. Lefkowitz, (51) Int. Cl. Mayfield Heights, OH (US) A6II 35/74 (2006.01) A6IP 2L/00 (2006.01) (73) Assignee: Ganeden Biotech, Inc., Mayfield A2.3L L/48 (2006.01) Heights, OH (US) (52) U.S. Cl...... 424/93.46; 426/61 (21) Appl. No.: 13/434,401 (57) ABSTRACT The present application relates to nutritional compositions (22) Filed: Mar. 29, 2012 comprising lactic acid-producing bacteria. US 2012/025 1512 A1 Oct. 4, 2012

PROBOTC SPORTS NUTRITION benefit to individuals that are suffering from or at risk of COMPOSITIONS developing a muscle wasting condition, e.g., cachexia, as a result of disease such as cancer or infection. RELATED APPLICATIONS 0007. The invention provides a composition comprising a 0001. This application claims the benefit of priority under sports nutrition composition and an isolated Bacillus coagul 35 U.S.C. S119(e) to U.S. Provisional Application No. lans spore (e.g., in an amount of 1x10° to 1x10" colony 61/469.924, filed Mar. 31, 2011, which is incorporated herein forming units (CFU) of Bacillus coagulans per unit dose. For by reference in its entirety. example, the isolated Bacillus coagulans comprise between about 0.000001% to about 50% by weight of the composition, FIELD OF THE INVENTION e.g., about 1%, about 10%, about 20%, about 30%, about 40%, or about 50% by weight of the composition. For 0002 The present application relates to sports nutrition example, the isolated Bacillus coagulans comprise between compositions comprising lactic acid-producing bacteria. about 0.00001% and about 25% by weight of the composi tion, e.g., about 1%, about 2%, about 3%, about 4%, about BACKGROUND OF THE INVENTION 5%, about 6%, about 7%, about 8%, about 9%, about 10%, 0003. The gastrointestinal microflora plays a number of about 11%, about 12%, about 13%, about 14%, about 15%, Vital roles in maintaining gastrointestinal tract function and about 16%, about 17%, about 18%, about 19%, about 20%, overall physiological health. The growth and metabolism of about 21%, about 22%, about 23%, about 24%, or about 25% the many individual bacterial species inhabiting the gas by weight of the composition. In one example, one unit dose, trointestinal tract depend upon the substrates available to e.g., one serving, comprises one billion CFU Bacillus coagul them, most of which are derived from the diet. lans and 20-25 grams of protein (e.g., 23 grams of whey 0004 Probiotic organisms are non-pathogenic, non-toxi protein) in 30 grams of powder. The balance of the composi genic microorganisms that are beneficial to the host organ tion optionally comprises nutritionally inactive ingredients ism. Since probiotics do not generally permanently colonize Such as excipients, fillers, preservatives or nutritionally active the host, they are typically ingested regularly for health pro ingredients such as vitamins or minerals. moting properties to persist. 0008. The invention also provides bacterial species including Bacillus coagulans, e.g., Bacillus coagulans ham SUMMARY OF THE INVENTION mer, preferably Bacillus coagulans hammer Strain Accession 0005. The invention is based on the discovery that lactic No. ATCC 31284, or one or more strains derived from Bacil acid-producing bacteria, particularly Bacillus coagulans, lus coagulans hammer strain Accession No. ATCC 31284 improve the absorption of protein compositions as well as (e.g., ATCC Numbers: GBI-20, ATCC Designation Number micronutrient compositions. The microorganisms, e.g., PTA-6085; GBI-30 or BC, ATCC Designation Number spores remain viable, retain their beneficial probiotic proper PTA-6086; and GBI-40, ATCC Designation Number PTA ties, and increase metabolism and/or absorption of protein in 6087; see U.S. Pat. No. 6,849,256 to Farmer). sports nutrition compositions. Unlike other probiotic bacteria 0009 Optionally, the isolated Bacillus coagulans is in the that die in the stomach or Small intestine, the probiotic organ form of a spore. Alternatively, the isolated Bacillus coagulans isms described herein, e.g., Bacillus coagulans strain GBI-30 is in the form of a vegetative cell. In another aspect, the or BC, ATCC Designation Number PTA-6086, germinate isolated Bacillus coagulans is in the form of a mixture of in the stomach and/or Small intestine. Accordingly, the inven Vegetative cells and spores. The Bacillus coagulans is pre tion describes probiotic sports nutrition compositions. The dominantly in spore form, e.g., about 75%, about 80%, about invention provides an isolated Bacillus coagulans bacterium 85%, about 90%, about 95%, about 96%, about 97%, about (e.g., a spore) in sports nutrition compositions. In some cases, 98%, about 99%, or about 100% spores. Alternatively, the the composition comprises at least 15% protein. For example, Bacillus coagulans is predominantly in vegetative form, e.g., the composition comprises at least 15%, at least 20%, at least about 75%, about 80%, about 85%, about 90%, about 95%, 25%, at least 30%, at least 35%, at least 40%, at least 45%, at about 96%, about 97%, about 98%, about 99%, or about least 50%, at least 55%, at least 60%, at least 65%, at least 100% vegetative cells. 70%, at least 75%, at least 80%, at least 85%, at least 90%, at 0010 Provided is an isolated Bacillus coagulans and a least 95%, at least 98%, at least 99% or more of a purified or sports nutrition composition. In some cases, the compositions processed protein (e.g., protein extracted from a food-stuff) described herein comprise a large amount of calories per unit and a Bacillus coagulans spore. dose to assista Subject in gaining weight, e.g., muscle weight 0006. The compositions described herein are suitable for or fat weight. A unit dose of the compositions described consumption by a mammal, e.g., any mammal, e.g., a human, herein is the amount of composition administered to a con a primate, a mouse, a rat, a dog, a cat, a cow, a horse, or a pig. Sumer in a single dose, i.e., one serving. Unit-dose packaging In a preferred embodiment, the mammal is a human. For is the packaging of a single dose, e.g., in a non-reusable example, the invention includes a method of enhancing lean container. For example, a unit dose refers to a physically muscle development or increasinglean body mass by admin discrete unit Suitable as unitary dosages for an individual, istering to a Subject a composition comprising a Bacillus each unit containing a predetermined quantity of active mate coagulans spore in a composition comprising at least 85% rial calculated to produce the desired therapeutic effect, in purified, processed, and/or isolated protein. The Subject is a association with a suitable pharmaceutical carrier, diluent, or human being that desires to increase muscle development, excipient. Suitable packaging includes single or multiple unit strength, or lean body mass, or an animal, e.g., livestock or dosages. performance animal Such as a work animal or a racehorse, for 0011 Compositions that provide a large amount of calo which an increase in muscle development or strength is ries to assist a subject in gaining weight are referred to as desired. The compositions are also useful to confer clinical “weight gainers. In these cases, the composition comprises US 2012/025 1512 A1 Oct. 4, 2012

between about 100 and about 10,000 food Calories (kcal) per min A, Vitamin C, calcium, iron, vitamin D. Vitamin E, thia unit dose (i.e., serving), e.g., between 250 and 5,000 kcal, min, riboflavin, niacin, vitamin B6, folate, vitamin B12, between 500 and 3,000 kcal, between 750 and 2,500 kcal, or biotin, pantothenic acid, phosphorus, iodine, magnesium, between 1,000 and 2,000 kcal, e.g., about 1,000 kcal, about Zinc, Selenium, copper, manganese, chromium, and molyb 1,100 kcal, about 1,200 kcal, about 1,300 kcal, about 1,400 denum. kcal, about 1,500 kcal, about 1,600 kcal, about 1,700 kcal, 0015. An exemplary composition comprises glucose poly about 1,800 kcal, about 1,900 kcal, or about 2,000 kcal. One mer, a protein blend (i.e., whey protein concentrate, whey exemplary amount of calories is 1,230 kcal. Alternatively, the protein isolate, egg albumin, milk protein isolate, and par compositions do not comprise a large amount of calories. For tially hydrolyzed whey protein), rice protein concentrate, example, the compositions comprise between about 10 and brown rice concentrate, taurine, L-glutamine, non-dairy 500 kcal, e.g., between about 20 and 250 kcal, between about creamer (i.e., Sunflower oil, corn Syrup Solids, Sodium casein 50 and 200 kcal, or between about 100 and 150 kcal, e.g., ate, mono- and diglycerides, dipotassium phosphate, trical about 100 kcal, about 110 kcal, about 120 kcal, about 130 cium phosphate, Soy lecithin, and tocopherols), natural and kcal, about 140 kcal, or about 150 kcal. One exemplary artificial flavors, Xantham gum, calcium citrate, potassium amount of calories is about 150 kcal. citrate, dipotassium phosphate, cellulose gum, tricalcium 0012 Preferably, the composition comprises protein. For phosphate, magnesium aspartate, rice starch, carrageenan, example, the protein comprises about 1% to about 99% by Vitamin-mineral blend (i.e., ascorbic acid, niacinamide, d-Al weight of the composition, e.g., about 5%, about 10%, about pha tocopheryl Succinate, d-calcium pantothonate, Zinc cit 15%, about 20%, about 25%, about 30%, about 35%, about rate, pyridoxine hydrochloride, ferrous fumarate, thiamine 40%, about 45%, about 50%, about 55%, about 60%, about mononitrate, riboflavin, manganese amino acid chelate, beta 65%, about 70%, about 75%, about 80%, about 85%, about carotene, copper gluconate, folic acid, biotin, potassium 90%, or about 95% by weight of the composition. For iodide, chromium polynicotinate, molybdenum amino acid example, the composition comprises between 1 gram and 500 chelate, selenomethionine, cyanocobalamin, and cholecalcif grams of protein, e.g., about 10 grams, about 15 grams, about erol), Ganeden BC-30 (Bacillus coagulans GBI-30, ATCC 20 grams, about 25 grams, about 30 grams, about 35 grams, Designation Number PTA-6086), sucralose, acesulfame about 40 grams, about 45 grams, about 50 grams, about 55 potassium, and lactase. grams, about 60 grams, about 65 grams, about 70 grams, 0016. Another exemplary composition comprises the fol about 75 grams, about 80 grams, about 85 grams, about 90 lowing ingredients: whey protein concentrate, brown rice grams, about 95 grams, about 100 grams, about 150 grams, protein concentrate, whey protein isolate, egg albumin, milk about 200grams, about 250 grams, about 300 grams, about protein isolate, partially hydrolyzed whey protein, glucose 350 grams, about 400 grams, about 450 grams, or about 500 polymer, taurine, L-glutamine, nondairy creamer (i.e., Sun grams of protein. One exemplary composition comprises flower oil, corn syrup Solids, sodium caseinate, mono- and about 62% protein. For example, the composition comprises diglycerides, dopotassium phosphate, tricalcium phosphate, about 39 total grams, of which about 24 grams is protein. Soy lecithin, and tocopherols), dicalcium phosphate, natural Another exemplary composition comprises about 65% pro and artificial flavors, Xanthan gum, cellulose gum, carrag tein. For example, the composition comprises about 31 total eenan, lecithin, acesulfame potassium, Sucralose, lactase, and grams, of which about 20 grams is protein. Another exem Ganeden BC-30 (Bacillus coagulans GBI-30, ATCC Desig plary composition comprises about 15% protein. For nation Number PTA-6086). example, the composition comprises about 325 total grams, 0017. The compositions contain active ingredients, e.g., of which about 50 grams is protein. A typical protein compo protein and Ganeden BC-30 (Bacillus coagulans GBI-30, sition comprises about 50-100% protein such as whey pro ATCC Designation Number PTA-6086), and inactive ingre tein. For example, a unit dose comprises 1 billion CFU of dients, e.g., excipients, binders, or fillers. Fillers fill out the Bacillus coagulans and 23 grams of whey protein in a total of size of the compositions, making it practical to produce and 30 grams of powder. convenient for the consumer to use. By increasing the bulk 0013. In one aspect, the sports nutrition composition com volume, the fillers make it possible for the final product to prises purified or processed protein, such as soy protein, whey have the proper volume for consumer handling. Suitable fill protein, rice protein, hemp seed protein, casein protein or ers include Xantham gum, cellulose gum, lecithin, lactose, milk protein. Preferably, the protein comprises whey protein. Sucrose, glucose, mannitol, Sorbitol, calcium carbonate, and The protein comprises an amino acid selected from the group magnesium Stearate. consisting of isoleucine, alanine, leucine, arginine, lysine, 0018. In some cases, the compositions do not comprise aspartate, aspartic acid, methionine, cysteine, phenylalanine, certain ingredients. For example, the composition does not threonine, tryptophan, glycine, Valine, proline, histidine, include a Sugar (e.g., glucose, fructose, galactose, maltose or serine, tyrosine, asparagine, selenocysteine, pyrrolysine, lactose), gluten, aspartame, and/or artificial coloring. glutamate, glutamic acid, and glutamine. Optionally, the 0019 Exemplary forms for the compositions described sports nutrition composition comprises an isolated Bacillus herein include a protein powder, a ready to drink protein coagulans and protein, and further comprises creatine, cal shake, a proteinbar, a protein bite, and a protein gel. cium, Sodium caseinate, whey peptides, or lactoferrin. Alter 0020. The invention provides compositions comprising a natively, the invention provides an isolated Bacillus coagul dry mix for sports nutrition compositions comprising an iso lans and a sports nutrition composition selected from the lated Bacillus coagulans bacterium and a sports nutrition group consisting of creatine, calcium, sodium caseinate, composition. The dry mix is between 0.01% and 50% Bacil whey peptides, and lactoferrin. lus coagulans bacterium, e.g., about 1%, about 5%, about 0014. The compositions optionally further comprise addi 10%, about 15%, about 20%, about 25%, about 35%, about tional ingredients selected from the group consisting of 45%, or about 50% Bacillus coagulans bacterium. In some Sodium, potassium, Sugar, carbohydrates, dietary fiber, Vita cases, the dry mix is about 15% Bacillus coagulans bacte US 2012/025 1512 A1 Oct. 4, 2012

rium. For example, about 100 pounds of dry mix contains 0026. In other cases, the absorption of a chemical element about 15 pounds of Bacillus coagulans bacterium and about is increased in the subject after the administration of Bacillus 85 pounds of sports nutrition composition. Optionally, the coagulans in combination with protein as compared to the isolated Bacillus coagulans is in the form of a spore. Alter absorption of the chemical element after the administration of natively, the isolated Bacillus coagulans is in the form of a protein in the absence of Bacillus coagulans. Exemplary Vegetative cell. In another aspect, the isolated Bacillus coagul chemical elements include Sodium, potassium, calcium, iron, lans is in the form of a mixture of vegetative cells and spores. and zinc. Preferably, the isolated Bacillus coagulans is in the form of a 0027 Optionally, the Bacillus coagulans aids in the diges spore. More preferably, the bacterium is present as at least tion of the sports nutrition composition, e.g., protein. For 80% spores, e.g., at least 85%, at least 90%, at least 95%, at example, abdominal bloating, intestinal pain, passage of gas, least 98%, or at least 99% spores. loose bowel movements, and excessive abdominal gurgling 0021 Bacterial species suitable for use in the methods are reduced following consumption of protein and Bacillus describe herein include Bacillus coagulans, e.g., Bacillus coagulans compared to the consumption of protein without coagulans hammer, preferably Bacillus coagulans hammer Bacillus coagulans. However, Subjects ingesting the product strain Accession No. ATCC 31284, or one or more strains preferably have not been diagnosed with gastrointestinal dis derived from Bacillus coagulans hammer strain Accession ease or an inflammatory bowel condition. No. ATCC 31284 (e.g., ATCC Numbers: GBI-20, ATCC Des 0028. The compositions are also useful in the prophylactic ignation Number PTA-6085; GBI-30 or BC, ATCC Desig or therapeutic treatment of conditions associated with gas nation Number PTA-6086; and GBI-40, ATCC Designation trointestinal infection by various pathogens, thereby support Number PTA-6087; see U.S. Pat. No. 6,849.256 to Farmer). ing digestive health. The probiotic sports nutrition composi 0022. In some cases, the dry mix also includes a protein tions of the invention are also used in the methods described selected from the group consisting of soy protein, whey pro herein for boosting the immune system. In some cases, the tein, rice protein, hemp seed protein, and casein protein. In combination of Bacillus coagulans and protein works Syner other cases, the sports nutrition composition also includes gistically resulting in an enhanced inhibition of pathogens in creatine, calcium, Sodium caseinate, whey peptides, and the gastrointestinal tract of a Subject, as compared to the lactoferrin. administration of either Bacillus coagulans or protein alone. 0023 Methods of enhancing lean muscle development, In other cases, the combination of Bacillus coagulans and recovery, or repair are carried out by administering to a Sub protein works synergistically resulting in an enhanced boost ject desiring an enhancement of the lean muscle develop of the immune system, as compared to the administration of ment, recovery or repair the compositions described herein, either Bacillus coagulans or protein alone. e.g., a composition comprising a sports nutrition composition 0029 Optionally, the compositions of the invention addi and an isolated Bacillus coagulans spore, wherein the com tionally comprise L-alanine and/or inosine to promote the position comprises at least 80% protein. The composition is germination of the spores in the stomach and/or small intes optionally administered prior to or after an exercise period. tine. For example, the composition is administered within 60 min 0030 A probiotic lactic acid-producing bacteria suitable utes of an exercise period, e.g., within 15 minutes, within 30 for use in the methods and compositions of the invention minutes, or within 45 minutes. Alternatively, the composition produces acid and is non-pathogenic. Purified and/or isolated is administered within 2 hours, within 5 hours, or within 8 Bacillus coagulans is particularly useful as a probiotic in the hours of an exercise period. compositions described herein. By “purified’ or “substan 0024. The absorption of the protein is increased in the tially purified” is meant a Bacillus coagulans bacterium that Subject after the administration of Bacillus coagulans incom is Substantially free of contaminating microorganisms or bination with protein as compared to the absorption of the other macromolecules, e.g., polysaccharides, nucleic acids, protein after the administration of protein in the absence of or proteins. Bacillus coagulans. For example, the rate of absorption 0031 Purified defines a degree of sterility that is safe for (t) or overall absorption (from area under the curve administration to a human Subject, e.g., lacking infectious or (AUC)oat, and C) are increased after the administration toxic agents. Specifically, as used herein, an "isolated” or of Bacillus coagulans in combination with protein as com “purified Bacillus coagulans or protein is substantially free pared to after the administration of protein in the absence of of other cellular material, culture medium, chemical precur Bacillus coagulans, as measured by changes in amino acid sors, or other chemicals when chemically synthesized. Puri levels in, e.g., blood over a one hour test period, a two hour fied compounds are at least 60% by weight (dry weight) the test period, a four hour test period, an eight hour test period, compound of interest. Preferably, the preparation is at least a twelve hour test period, or a twenty-four hour test period. 75%, more preferably at least 90%, and most preferably at For example, protein absorption is increased by at least 10%, least 99%, by weight the compound of interest. Purity is 50%, 2-fold, 5-fold, 10-fold, or more following administra measured by any appropriate standard method, for example, tion of protein and Bacillus coagulans compared to protein by column chromatography, polyacrylamide gel electro without Bacillus coagulans. phoresis, or HPLC analysis. 0025. The absorption of a vitamin is increased in the sub 0032. The Bacillus coagulans Hammer strains of the ject after the administration of Bacillus coagulans in combi invention are non-pathogenic and generally regarded as safe nation with protein as compared to the absorption of the for use in human nutrition (i.e., GRAS classification) by the vitamin after the administration of protein in the absence of U.S. Federal Drug Administration (FDA) and the U.S. Bacillus coagulans. Exemplary vitamins include vitamin A, Department of Agriculture (USDA), and by those skilled in Vitamin B, Vitamin B, Vitamin B vitamin Bs, vitamin B. the art. Furthermore, the Bacillus coagulans Hammer strains Vitamin B7, Vitamin Bo, Vitamin B, or vitamin C. vitamin D, of the invention germinate at or below human body tempera Vitamin E, and vitamin K. ture, rendering them useful as probiotics. Many Bacillus US 2012/025 1512 A1 Oct. 4, 2012 coagulans Strains outside the Hammer group have mostly 0037 Exemplary methods and compositions are industrial applications, little or no nutritional benefit, and described herein using Bacillus coagulans as a probiotic. environmental contaminants that have not been evaluated for Purified and/or isolated Bacillus coagulans is particularly safety. Moreover, many other non-Hammer strains of Bacil useful as a probiotic in sports nutrition compositions. Probi lus coagulans grow optimally at temperatures that exceed otic B. coagulans is non-pathogenic and is generally regarded human body temperature and, thus, do not germinate effi as safe (i.e., GRAS classification) by the U.S. Federal Drug ciently in the human body. Such strains are less or not suitable Administration (FDA) and the U.S. Department of Agricul as probiotics for human consumption. ture (USDA), and by those skilled in the art. 0033. The transitional term “comprising,” which is syn 0038 Bacillus coagulans is a non-pathogenic gram posi onymous with “including.” “containing,” or “characterized tive spore-forming bacteria that produces L(+) lactic acid by, is inclusive or open-ended and does not exclude addi (dextrorotatory) in fermentation conditions. It has been iso tional, unrecited elements or method steps. By contrast, the lated from natural sources, such as heat-treated soil samples transitional phrase “consisting of excludes any element, inoculated into nutrient medium (Bergey’s Manual off Sys step, or ingredient not specified in the claim. The transitional temic Bacteriology, Vol. 2, Sneath, P. H. A., et al., eds., Wil phrase “consisting essentially of limits the scope of a claim to liams & Wilkins, Baltimore, Md., 1986). Purified B. coagul the specified materials or steps "and those that do not mate lans strains have served as a source of enzymes including rially affect the basic and novel characteristic(s) of the endonucleases (e.g., U.S. Pat. No. 5,200,336), amylase (U.S. claimed invention. Pat. No. 4,980,180), lactase (U.S. Pat. No. 4.323,651), and 0034. Other features and advantages of the invention will cyclo-malto-dextrin glucano-transferase (U.S. Pat. No. be apparent from the following description of the preferred 5,102,800). B. coagulans has been used to produce lactic acid embodiments thereof, and from the claims. Unless otherwise (U.S. Pat. No. 5,079,164). A strain of B. coagulans (referred defined, all technical and scientific terms used herein have the to as L. Sporogenes; Sakaguti & Nakayama (ATCC 31284)) same meaning as commonly understood by one of ordinary has been combined with other lactic acid producing bacteria skill in the art to which this invention belongs. Although and B. natto to produce a fermented food product from methods and materials similar or equivalent to those steamed soybeans (U.S. Pat. No. 4,110.477). described herein can be used in the practice or testing of the 0039 Bacterial species include Bacillus coagulans, e.g., present invention, Suitable methods and materials are Bacillus coagulans hammer, preferably Bacillus coagulans described below. All publications, patent applications, pat hammer strain Accession No. ATCC 31284, or one or more ents, Genbank/NCBI accession numbers, and other refer strains derived from Bacillus coagulans hammer strain ences mentioned herein are incorporated by reference in their Accession No. ATCC 31284 (e.g., ATCC Numbers: GBI-20, entirety. In the case of conflict, the present specification, ATCC Designation Number PTA-6085; GBI-30 (BC), including definitions, will control. In addition, the materials, ATCC Designation Number PTA-6086; and OBI-40, ATCC methods, and examples are illustrative only and not intended Designation Number PTA-6087; see U.S. Pat. No. 6,849.256 to be limiting. to Farmer). 0040 Bacillus coagulans was previously mis-character DETAILED DESCRIPTION OF THE INVENTION ized as a Lactobacillus and labeled as Lactobacillus sporo genes. However, initial classification was incorrect because 0035. The probiotic organisms of the invention are non Bacillus coagulans produces spores and excretes L(+)-lactic pathogenic, non-toxigenic, retain viability during storage, acid through metabolism. Both of these characteristics pro and Survive passage through the stomach and Small intestine. vide key features to the utility of Bacillus coagulans. These Non-pathogenic lactic acid-producing bacteria (i.e., "lactic developmental and metabolic aspects required that the bac acid bacteria'). Such as the exemplary Bacillus coagulans, terium be classified as a lactic acid Bacillus. In addition, it is remain viable and retain their beneficial probiotic properties not generally appreciated that classic Lactobacillus species in sports nutrition compositions. Prior to the invention are unsuitable for colonization of the gut due to their insta described herein, probiotics were very susceptible to harsh bility in the harsh (i.e., acidic) pH environment of the bile, environment of the stomach and small intestine. Unlike other particularly human bile. By contrast, Bacillus coagulans probiotics that die in the stomach or Small intestine, the pro ATCC Designation Number PTA-6085; GBI-30 (BC') is biotic organisms described herein, e.g., Bacillus coagulans able to Survive and colonize the gastrointestinal tract in the strain GBI-30 or BC, ATCC Designation Number PTA bile environment and even grow in this low pH range. 6086, germinate in the stomach and/or small intestine. Spe Probiotic Activity of Bacillus coagulans cifically, the probiotic spores described herein can survive 0041. It is well-documented clinically that many species passage through the stomach and Small intestine. of bacterial, mycotic and yeast pathogens possess the ability to cause a variety of gastrointestinal disorders including, but Probiotic Lactic Acid-Producing Bacteria not limited to disruption of normal gastrointestinal biochemi 0036. The sports nutrition compositions include a lactic cal function, necrosis of gastrointestinal tissues, and disrup acid-producing bacterium, Such as a spore-forming Bacillus tion of the bioabsorption of nutrients, and like conditions. The species, e.g., B. coagulans. Preferably, the spore-forming probiotic microorganism-containing compositions described Bacillus species of the invention is B. coagulans Hammer. herein inhibit these pathogens. In the present case, Subjects There are many suitable bacteria identified as described are generally healthy, e.g., they have not been diagnosed with herein, although the invention is not limited to currently a gastrointestinal disease (e.g., irritable bowel syndrome or known bacterial species insofar as the purposes and objec Crohn's disease), an inflammatory bowel condition, or an tives of the bacteria is described. The property of acid pro autoimmune disease. The compositions and methods duction is important to the effectiveness of the probiotic lactic described herein are useful to increase absorption of protein acid-producing bacteria of this invention. and/or macro nutrients in a nutritional composition. US 2012/025 1512 A1 Oct. 4, 2012

0042. In one aspect, a Bacillus coagulans Strain is Clostridium species—excellent inhibition, where the Zone of included in the composition in the form of vegetative cells. In inhibition was consistently greater than 15 mm in diameter. another aspect, the Bacillus coagulans strain is included in Similarly, excellent inhibition was also seen for the opportu the composition in the form of spores. The invention also nistic pathogens Pseudormonas aeruginosa, and Staphyllo provides for including the Bacillus coagulans strain in the coccus aureus. Pathogenic enteric bacteria which were inhib composition in the form of a powder, a dried cell mass, a ited by Bacillus coagulans activity include, but are not limited stabilized paste, or a stabilized gel. to: Staphylococcus aureus, Staphylococcus epidermidis, 0043. Because Bacillus spores are heat and pressure-resis Streptococcus pyogenes, Pseudomonas aeruginosa, Escheri tant and can be stored as a dry powder, they are particularly chia coli (enterohemorragic species); numerous Clostridium useful for formulation into and manufacture of products Such species (e.g., Clostridium perfingens, Clostridium botulinum, as the various sports nutrition compositions described herein. Clostridium tributrycum, Clostridium sporogenes, and the Specifically, the probiotic organisms described herein, e.g., like); Gardnereia vaginails, Proponbacterium aenes, Aero Bacillus coagulans strain GBI-30 or BC, ATCC Designa monas hydrophia, Aspergillus species; Proteus species; and tion Number PTA-6086, survive passage through the stomach Klebsiella species. and small intestine. A Bacillus species is well suited for the present invention, particularly species having the ability to Micro-Encapsulation form spores which are relatively resistant to heat and other 0047. In one aspect, the lactic-acid producing bacteria are conditions, making them ideal for storage (shelf-life) in prod incorporated into a microcapsule coating prior to addition to uct formulations, e.g., sports nutrition compositions. Due to the sports nutrition composition, using any micro-encapsula the shelf-stable properties of the Bacillus coagulans strains tion process well-known in the art. The isolated Bacillus described herein, e.g., Bacillus coagulans strain GBI-30 or coagulans are packaged, or encapsulated, within another BC, ATCC Designation Number PTA-6086, the product material in order to protect the bacteria from the Surrounding formulations of the invention are not confined to a refrigerator environment. The capsules of the invention range in size from and may be stored at room temperature. The Bacillus coagul one-thousandth of a millimeter to seven millimeters. The lans of the invention survives storage (shelf-life) from about internal ingredients of the microcapsule are released from 12 days to about 2 years; from about 1 month to about 18 their shells in various ways, including mechanical rupture of months; from about 3 months to about 1 year; or from about the capsule wall, dissolution of the wall, melting of the wall 6 months to about 9 months. and diffusion through the wall. Thus, micro-encapsulation 0044. The probiotic organisms described herein, e.g., provides additional protection to the isolated Bacillus bacte Bacillus coagulans strain GBI-30 or BC, ATCC Designa rium during manufacturing and storage of the sports nutrition tion Number PTA-6086, promote digestive and oral health compositions of the invention. Physical methods of micro and support the immune system. The ability of Bacillus encapsulation include pan coating, air-Suspension coating, coagulans to inhibit various bacterial pathogens was quanti centrifugal extrusion, vibrational nozzle, and spray-drying. tatively ascertained by use of an in vitro assay. This assay is Chemical methods of micro-encapsulation include interfacial part of a standardized bacterial pathogen screen (developed polymerization, in-situ polymerization, and matrix polymer by the U.S. Food and Drug Administration (FDA)) and is ization. commercially available on solid support disks (DIFCOR) BACTROL(R) Antibiotic Disks). To perform the assay, potato 0048 Alternatively, the lactic-acid producing bacteria is dextrose plates (DIFCOR) were initially prepared using stan added to the sports nutrition composition without micro dard procedures. The plates were then individually inoculated encapsulation. with the bacteria (approximately 1.5x10°CFU) to be tested so as to form a confluent bacterial bed. Probiotic Sports Nutrition Compositions 0045. Inhibition of microorganisms (e.g. gastrointestinal 0049. The invention includes a method of increasing pro pathogens) by Bacillus coagulans was Subsequently ascer tein absorption, enhancing lean muscle development, and/or tained by placing approximately 1.8x10 CFU of Bacillus increasing lean body mass by administering to a subject a coagulans in 10 ul of broth or buffer, directly in the center of composition comprising a Bacillus coagulans spore in a com the potato-dextrose plate with one test locus being approxi position comprising at least 50% purified or processed pro mately 8 mm in diameter per plate. A minimum of three test tein, e.g., a composition that contains at least 75% whey loci were used for each assay. The negative control consisted protein or other protein Source. The sports nutrition compo of a 10 ul volume of a sterile saline solution, whereas the sitions are suitable for human or animal consumption. A positive control consisted of a lul Volume of glutaraldehyde. typical Subject is a healthy adult, e.g., greater than 16 years The plates were then incubated for approximately about 18 hr old. Alternatively, the sports nutrition compositions may also at 30° C., at which time the Zones of inhibition were mea be administered to children under 18 years of age, e.g., under sured. As designated herein, “excellent inhibition' means the 15 years of age, under 10 years of age, or under 5 years of age. Zone was 10 mm or greater in diameter; and “good inhibition' Alternatively, the sports nutrition compositions are adminis means the Zone was greater than 2 mm in diameter but less tered to children and adults of allages. The Subject is a human than 10 mm in diameter. being that desires to increase muscle development, strength 0046. As expected, no “inhibition' was seen with the or lean body mass, or an animal, e.g., livestock or perfor negative, saline control, and excellent “inhibition' (approxi mance animal Such as a work animal or a racehorse, for which mately 16.2 mm diameter, average of three tests) was seen an increase in muscle development or strength is desired. The with the positive, glutaraldehyde control. For the enteric compositions are also useful to confer clinical benefit to indi microorganisms tested, the following inhibition by Bacillus viduals that are suffering from or at risk of developing a coagulans was found: (i) Clostridium species—excellent muscle wasting condition, e.g., cachexia, as a result of disease inhibition; (ii) Escherichia coli excellent inhibition; (iii) Such as cancer or infection. US 2012/025 1512 A1 Oct. 4, 2012

0050 Sports nutrition compositions are prepared by mix milk or juice (e.g., grapefruit juice, grape juice, orange juice, ing dry powder ingredients (e.g., protein and Bacillus coagul etc.), and often flavoring, resulting in a form known as a lans). Sports nutrition compositions include bodybuilding or “protein shake' (as in milkshake) or "pudding. Protein pow body maintenance foods, e.g., weight gainers, commonly der is generally consumed immediately before or after exer used by those involved in bodybuilding and athletics. They cising, or in place of a meal. The theory behind this regimen include protein (the most widely used such sports nutrition is that having a sufficient protein intake allows for efficient composition), amino acids, glutamine, essential fatty acids, growth and repair of muscle tissue. Protein powders also aid meal replacement products, prohormones, creatine, ther in fat loss, muscle building, and slow the aging process. mogenic products, testosterone boosters, and branched-chain 0054 Meal replacement products (MRPs) are either pre amino acids (BCAA). The three branched-chain amino acids packaged powdered drink mixes or edible bars designed to (BCA As) include leucine, isoleucine, and valine. Unlike replace prepared meals. MRPs are generally high in protein, otheramino acids, BCAAs are metabolized in the muscle and low in fat, have a low to moderate amount of carbohydrates, have an anabolicfanti-catabolic effects. BCAAs account for and contain a wide array of vitamins and minerals. The major 33% of muscle protein. Amino acids are the building blocks ity of MRPs use whey protein, casein (often listed as calcium of protein, which the body metabolizes in the stomach and caseinate or micellar casein), soy protein, and/or egg albumin intestines. Protein powder contains amino acids such as iso as protein sources. Carbohydrates are typically derived from leucine, alanine, leucine, arginine, lysine, aspartate, aspartic maltodextrin, oat fiber, brown rice, and/or wheat flour. Some acid, methionine, cysteine, phenylalanine, threonine, tryp MRPs also contain flaxseed oil powder as a source of essen tophan, glycine, Valine, proline, histidine, serine, tyrosine, tial fatty acids. asparagine, Selenocysteine, pyrrolysine, glutamate, glutamic 0055 Sports nutrition compositions may also contain acid, and glutamine. other additional ingredients that are beneficial for bodybuild 0051. Glutamine is the most abundantamino acid found in ing, including calcium, Sodium caseinate, whey peptide, human muscle; however, the body's natural glutamine stores glutamine peptides, L-glutamine, calcium alpha-ketoglut are depleted during anaerobic exercise. Thus, the sports nutri arate, additional amino acids, lactoferrin, conjugated linoleic tion compositions of the invention optionally include acid, medium chain triglycerides, and creatine (e.g., creatine glutamine. Serum glutamine is used by the body to counteract monohydrate). Creatine is a nitrogenous organic acid that is acidosis resulting from exercise. In order to replenish the loss found in the muscle tissue of vertebrates mainly in the form of of glutamine from the bloodstream, the body catabolizes phosphocreatine replenishment of ATP, and Supplies energy glutamine from the muscle. Thus, ingestion of glutamine for muscle contraction. Creatine improves strength, energy, combats muscle tissue wasting. muscle mass, recovery times, brain function, and reduces 0052 Suitable types of protein include soy protein, whey mental fatigue. Creatine is available in a variety of forms, protein, rice protein, hemp protein, casein protein, and egg including creatine monohydrate and creatine ethyl ester. white protein (e.g., egg albumin). Soy protein isolated from 0056 Although it is known that athletes and bodybuilders Soybeans contains isoflavones, a type of phytoestrogen. Whey may need an increased intake of protein, the exact amount is protein contains high levels of all the essential amino acids highly individualized and dependent on the type and duration and branched-chain amino acids. For example, the amino of the exercise as well as the physiological make up of the acids regarded as essential for humans are phenylalanine, individual. Research by Tarnopolsky etal. (1988) showed that Valine, threonine, tryptophan, isoleucine, methionine, leu for bodybuilding individuals, 1.97g of protein per kg of body cine, lysine, and histidine. Whey protein also has the highest weight per day is recommended, whereas endurance athletes content of the amino acid cysteine, which aids in the biosyn require 1.37 g/kg/d of protein. Studies Suggest that there are thesis of glutathione. For this reason, whey protein provides different protein requirements for anaerobic and aerobic exer amino acids to aid in muscle recovery. Whey protein is cise. Endurance athletes in aerobic activity may have derived from the process of making cheese from milk. There increased daily protein intake at 1.2-1.4g per kg body weight are two types of whey protein: whey concentrate (29%-89% per day, whereas strength training athletes performing protein by weight) and whey isolate (90%+ protein by anaerobic activity may have increased daily protein intake weight). Another type of protein includes rice protein, which needs at 1.4-1.8g per kg body weight so as to enhance muscle is made from whole grain, a complete protein source that is protein synthesis or to make up for the loss of amino acid highly digestible and allergen free. Hemp protein from hemp oxidation during exercise. seed contains complete and highly-digestible protein. Hemp 0057 Preferably, the sports nutrition composition ingre oil is high in essential fatty acids, i.e., alpha-linolenic acid (an dients are blended together as dry ingredients. Exemplary omega-3 fatty acid) and linoleic acid (an omega-6 fatty acid). ingredients of the sports nutrition compositions described Casein protein (or milk protein) has glutamine, and casomor herein include protein and Bacillus coagulans. An exemplary phin. Finally, egg-white protein is a lactose- and dairy-free strain of Bacillus coagulans Suitable in the sports nutrition protein. compositions described herein includes GBI-30 or BC'. 0053 Proteins come in various forms, including protein ATCC Designation Number PTA-6086. powder, ready to drink protein shakes, bars, bites, oats, and 0058. The dry mix for sports nutrition compositions com gels. Protein powders are available in a wide variety of flavors prises an isolated Bacillus coagulans bacterium. The dry mix including pineapple, orange, fruit punch, mixed berry, is approximately 1 billion CFU of Bacillus coagulans bacte mango, cookies and cream, Strawberry, Strawberry banana, rium per unit dose. Optionally, the dry mix is about 15% French Vanilla, Vanilla, Vanilla ice cream, Vanilla milkshake, Bacillus coagulans bacterium and 85% protein. For example, banana, banana cream, Dutch chocolate, mocha cappuccino, about 100 pounds of dry mix contains about 15 pounds of double rich chocolate, chocolate caramel, chocolate milk Bacillus coagulans bacterium and about 85 pounds of pro shake, extreme milk chocolate, chocolate mint, chocolate tein. The dry mix is between about 1% and about 50% by chip, and chocolate. The protein powder is mixed with water, weight of the sports nutrition composition, e.g., about 1% to US 2012/025 1512 A1 Oct. 4, 2012

about 20%, about 5% to about 15%; about 6%, about 7%, package, an 18-ounce package, a 24-ounce package, a about 8%, about 9%, or about 10% by weight of the sports 48-ounce package, 80-ounce package, or 100-ounce package. nutrition composition. For example, a 3 gram sports nutrition In another example, the dried powder is packaged in unit dose composition contains about 7% dry mix by weight of the quantities, e.g., 5 grams, 10 grams, 20 grams, 30 grams, 40 sports nutrition composition. A 3.8 to 4 gram sports nutrition grams, 50 grams, 60 grams, 70 grams, 80 grams, 90 grams, or composition contains about 8-9% dry mix by weight of the 100 gram packets. Alternatively, the dried powder is pack sports nutrition composition. aged in bulk, e.g., about 500 grams, about 600 grams, about 0059. As the recommended dietary allowances (RDA or 700 grams, about 800 grams, about 900 grams, about 1,000 recommended daily intake; RDI) is about 1x10 bacterium grams, about 1,250 grams, about 1,500 grams, about 1,750 (according to EU guidelines), preferably, the sports nutrition grams, about 2,000 grams, about 2,250 grams, about 2,500 composition comprises at least about 1x10 viable bacteria. gram, or about 3,000 gram containers. In one aspect, the In another aspect, the sports nutrition composition comprises invention provides for storing the sports nutrition composi at least about 1x10° to 1x107; at least about 1x107 to 1x10: tion in a sterile package at room temperature prior to con or at least about 1x10 to 1x10 viable bacteria. Sumption. Alternatively, the composition is consumed imme 0060. The Bacillus and/or Bacillus coagulans in the form of a dried spore composition is mixed with dry whey powder diately. or is applied using any of a variety of known methods includ 0064. The active ingredients (i.e., Bacillus coagulans and ing, for example, applying a powder, spray-drying the probi protein), comprise between about 0.01% to about 10%: otic onto the sports nutrition composition, or soaking the 0.01% to about 1%; or about 0.05% to about 0.1% by weight composition in a solution containing the probiotic. Any of a of the probiotic sports nutrition composition. Optionally, the variety of methods for placing the bacterial composition into isolated Bacillus coagulans comprise about 1 mg to about 10 a sports nutrition composition can be used. In one aspect, a g; about 10 mg to about 1 g, or about 25 mg to about 75 mg by “spray-dry' method is used, in which the compositions are weight of the probiotic composition. Most preferably, the exposed in a low humidity chamber to an atomized mix amount of Bacillus coagulans bacteria is about 5x107 colony containing a liquid composition, where the chamber is Sub forming units (CFU) of bacteria per gram of food matrix. sequently exposed to approximately 80-110° F. to dry the liquid, thereby impregnating the material of the sports nutri 0065. In some cases, the Bacillus coagulans spores are tion composition with the components. incorporated into any type of dry or lyophilized product 0061. In some cases, Bacillus coagulans bacteria in the which is dissolved or mixed with water, milk, or juice, so long form of a spray-dried powder is included in or on the surface as the temperature of the Bacillus coagulans spore-contain of the sports nutrition compositions described herein. Prefer ing mixture is raised to the required heat-shock temperature ably, the isolated Bacillus coagulans is in the form of a spore, (i.e., 80° C. for 5 minutes) necessary for germination of the as the Bacillus coagulans spores of the invention are pro spores. The Bacillus coagulans spores may either be incor tected by a hardened coating that can withstand gastric acid porated into the dry or lyophilized product by the manufac and bile salts for delivery to the small and large intestines. The turer of the product or by the consumer during preparation. isolated Bacillus coagulans are at least 85%, at least 90%, at 0066. The Bacillus coagulans spores survive storage least 95%, or at least 99% pure spores. Alternatively, the (shelf-life), i.e., retain viability or the ability to germinate at isolated Bacillus coagulans is in the form of a vegetative cell. physiological conditions (e.g., ingestion), from about 12 days In one aspect, the isolated Bacillus coagulans are at least to about 2 years; from about 1 month to about 18 months; 85%, at least 90%, or at least 95% pure vegetative cells. In from about 3 months to about 1 year; or from about 6 months another aspect, the isolated Bacillus coagulans is in the form to about 9 months. of a mixture of vegetative cells and spores. The Bacillus coagulans mixture is 90% spores, 10% vegetative cells: 75% spores, 25% vegetative cells: 60% spores, 40% vegetative Example 1 cells; 50% spores, 50% vegetative cells; 60% vegetative cells, 40% spores; 75% vegetative cells; 25% spores; or 90% veg Preparation of Bacillus coagulans Cultures etative cells, 10% spores. 0062. In one aspect, the amount of bacteria is about 10 to 0067 Bacillus coagulans Hammer bacteria (ATCC 10' colony forming units (CFU) of bacteria per gram of Accession No. 31284) was inoculated and grown to a cell probiotic composition (i.e., vegetative cells and/or bacterial density of about 10 to 10 cells/ml in nutrient broth contain spores), preferably 10 to 10' CFU/g of sports nutrition com ing 5 g Peptone, 3 g Meat extract, 10-30 mg MnSO, and position. Alternatively, the concentrations are 10 to 10' 1,000 ml distilled water, adjusted to pH 7.0, using a standard CFU/g: 10 to 10 CFU/g; or 10' to 10' CFU/g of sports airlift fermentation vessel at 30° C. The range of MnSO nutrition composition. In one aspect, the amount of bacteria is acceptable for sporulation is 1 mg/l to 1 g/l. The vegetative about 1x10 CFU per gram of sports nutrition composition. cells can actively reproduce up to 45° C., and the spores are The actual amount in a sports nutrition composition will vary. stable up to 90° C. After fermentation, the B. coagulans A typical concentration is from approximately 1x107 to bacterial cells or spores are collected using standard methods 1x10' CFU; 1x10 to 1x10' CFU; or 1x10 to 1x10'CFU (e.g., filtration, centrifugation) and the collected cells and of viable bacterium or spores/g of sports nutrition composi spores can be lyophilized, spray-dried, air-dried or frozen. tion. 0068 A typical yield from the above culture is in the range 0063. Following drying, the sports nutrition composition of about 10 to 10" viable spores and more typically about is ready for immediate use or for storage in a sterile package, 100 to 150 billion cells/spores per gram before drying. Spores e.g., a 3-ounce package (e.g., a bag or a bottle), a 6-ounce maintain at least 90% viability after drying when stored at package, a 9-ounce package, a 12-ounce package, a 15-ounce room temperature for up to ten years, and thus the effective US 2012/025 1512 A1 Oct. 4, 2012 shelf life of a composition containing B. coagulans Hammer spores at room temperature is about 10 years. TABLE 1 Example 2 20 Minutes 120 Minutes Incubation Incubation Spore Count, Spore Count, Preparation of Bacillus coagulans Spores Sample CFU gram CFU gram 0069. A culture of dried B. coagulans spores was prepared Normal Saline - A 1.90E-10 188E-10 as follows. Ten million spores were inoculated into a one liter Normal Saline - B 2.12E-10 2.OOE-10 culture containing 24 g potato dextrose broth, 10 g of enzy Normal Saline - C 1.64E-10 2.06E-10 Average 189E-10 198E-10 mic-digest of poultry and fish tissue, 5 g of FOS and 10 g Saline pH 1.0 - D 2O8E--09 S.98E--O7 MnSO. The culture was maintained for 72 hours under a high Saline pH 1.0 - E 1.47E--09 O.OOE--OO oxygen environment at 37° C. to produce culture having Saline pH 1.0 - F 3.59E-09 O.OOE--OO about 150 billion cells per gram of culture. Thereafter, the Average 2.38E--09 1.99E-07 Saline pH 2.0 - G 3.63E--09 3.46E--09 culture was filtered to remove culture medium liquid, and the Saline pH 2.0 - H 4.47E--09 2.48E--09 bacterial pellet was resuspended in water and freeze-dried. Saline pH 2.0 - I 3.58E--09 2.82E--09 The freeze-dried powder is then ground to a fine powder using Average 3.89E-09 2.92E--09 standard good manufacturing practice (GMP). A dried com Saline pH 3.0 - J 1.65E-10 1.13E-10 Saline pH 3.0 - K 1.35E-10 1.11E+10 position that comprises at least about 75% (e.g., 80%, 85%, or Saline pH 3.0 - L. 180E-10 1.39E-10 90%) spores is then mixed with protein powder. Average 16OE-10 1.21E-10 Example 3 Bacillus coagulans Spores Survive in the Gastric Example 4 Environment The Effects of a Probiotic on the Absorption of Pro 0070 This study was performed in order to determine the tein and Micronutrients in Healthy Males Survivability rate of Bacillus coagulans spores as they pass through the stomach. Samples of Bacillus coagulans spores (0071. The effects of the probiotic GanedenBC on pro were subjected to a simulated gastric environment for varying tein and micronutrient absorption is evaluated in healthy lengths of time in order to attain their survivability rate. First, males following a 14-day period. The protocol outlined below a homogeneous sample of raw material Bacillus coagulans of determines the effect of a probiotic product (GanedenBC') at least 12 grams was prepared. Saline Solution at pH 1 was in combination with protein as compared to protein in the prepared using 3N HCl (150 mls each into six 250 ml media absence of Bacillus coagulans on protein absorption, both bottles) and sterilized. Additional saline solutions with pH 2 rate of absorption (t) and overall absorption (from area and 3 were prepared similarly, resulting in 6 sterile 250 ml under the curve (AUCo.) and C), as measured by bottles, each containing 150 ml pH adjusted saline. Six sterile changes in blood amino acid levels over a four hour test 250 ml media bottles each containing 150 ml normal saline period. The protocol outlined below is also designed to deter solution were prepared and sterilized. Phosphate buffer (~400 mine the effect of a probiotic product (GanedenBC') in ml) was prepared at pH 7.2. Test tubes (24) were prepared and combination with protein as compared to protein in the sterilized, each containing 9 ml of phosphate buffer pH 7.2. absence of Bacillus coagulans on vitamin and mineral Test tubes (120) were prepared, each containing 9 ml of absorption, both rate of absorption (t) and overall absorp normal saline. GYE (glucose-yeast extract) agar medium was tion (from AUCo., and C), as measured by changes in prepared and sterilized and cooled to 45° C. in a water bath. blood levels of select vitamins (vitamins B6, B12 and C) and Samples (24) of raw material were weighed, each ~500 mil minerals (calcium, iron and Zinc) over a four hour test period. ligrams (theoretically equivalent to 10 billion spores). The The study also determines the effect of a probiotic product samples were added to media bottles at 37° C. and incubated (GanedenBC') in combination with protein as compared to half for 20 minutes the other half for 120 minutes. After 20 protein in the absence of Bacillus coagulans on gastrointes and 120 minutes incubation, respectively, the samples were tinal (GI)-specific symptoms using a GI questionnaire after mixed to uniformity and pipet 1 ml into 9 ml of sterile phos 14 days of treatment. phate buffer pH 7.2. After all 12 samples from each time point 0072 The period of evaluation lasts approximately four were placed into test tubes containing sterile phosphate weeks with Subjects attending a screening/randomization buffer, serial dilutions were made until 6 tubes had been used visit and two follow-up test visits. Eligible subjects take for each sample. The final dilution for the final two test tubes GanedenBC' plus protein and protein alone, each for 14 were 3x107 and 3x10, which gave a count of roughly 300 and days, in random order. After each 14-day period, there is a test 30 CFU, respectively. The final 2 test tubes from each sample visit at which subjects take the test product (probiotic plus were placed into 70° C. water bath for 30 minutes. After 30 protein or protein alone) and a multiple vitamin and mineral minutes, they were cooled immediately to 45°C. Three sterile supplement. Blood is collected at baseline (prior to the test petri plates per tube were set out. 1.0 ml from the heat-treated product) and at 1, 2, 3 and 4 hours following product admin tube was added into each petri plate, then 15 ml of sterile istration. Levels of various amino acids, vitamins and miner molten GYEAgar medium (at 45°C.) was poured into each of als are measured so that the effects of the probiotic on protein the petri plates and mixed thoroughly. When solidified, the and micronutrient absorption are determined. The probiotic plates were incubated in an inverted position for 48 hours at increases the absorption of the protein and micronutrients by 40° C. The individual colonies were counted. Results were increasing the efficiency of the gut by balancing the intestinal expressed as CFU per gram as shown in Table 1 below. flora or by increasing the acidity of the stomach for more 1.OE+10=1x10'. efficient breakdown of the nutrients. US 2012/025 1512 A1 Oct. 4, 2012

0073. Although the study population is comprised of gen as mean, standard deviation, count, median, and range (mini erally healthy adults that do not have GI disorders or symp mum to maximum value). Changes from the appropriate toms, there may be beneficial effects in GI health with the baseline are summarized and presented in the same way. administration of GanedenBC'. A six item questionnaire is Numerical variables and their changes from baseline are dis used to assess changes in GI health (e.g. abdominal pain, played graphically, as plots of mean value vs. time, with bloating and gas) with lower incidence indicating positive separate lines for the two products (probiotic VS. placebo). effects with regard to GI health. Categorical variables are presented as tabulations of counts Study Design and percentages of totals. I0085 For each continuous variable, the mean change from 0074 The study described herein is a prospective, ran baseline to each Subsequent time point are tested for nominal domized, double blind, placebo controlled, crossover clinical significance by the paired Student t test, or by the non-para trial. A total of 10 male subjects are enrolled, with each metric Wilcoxon test if non-normally distributed. The mean Subject receiving the two test products, in a randomly-as differences in the variable, or in the change in that variable signed sequence. Each subject serves as his own control. from baseline, between the different products are tested for Study Population: nominal significance by the paired Student t test or by the non-parametric Wilcoxon test if non-normally distributed. 0075. The study population includes 10 male subjects, For each categorical variable, difference in the distribution of aged 21 to 60 years, that do not have any gastrointestinal categories between the different product groups are tested for disease or inflammatory bowel condition, and who are not nominal significance by the Fisher Exact test if possible, or by taking on a regular basis any prescription or over-the counter the Chi-Square test if necessary. medications for any gastrointestinal problems. I0086 Adverse events (AEs) are listed, Medical Dictionary 0076 Test Products: for Regulatory Activities (MedDRA) encoded, grouped by Product 1 GanedenBC (Bacillus coagulans strain GBI-30 general type of event (gastrointestinal, neurologic, cardiac, or BC, ATCC Designation Number PTA-6086) plus Pro etc.), and cross-tabulated by event type and product. Differ tein. ences in AE patterns between the two products are tested by Product 2 Protein (placebo; no probiotic). the Fisher Exact test. Subjective remarks are categorized to the extent possible, and analyzed for pattern differences Duration of Study between the two products in the same way as AE’s. 0077. Excluding the screening visit, each subject complet 0087. The pharmacokinetic endpoints (C, t, and ing the study participates for approximately four weeks, AUCo.) after 14 days of product use are tested for signifi tthree days. cance between the two products by a repeated-measures analysis of variance (RM-ANOVA) for each endpoint, where Efficacy Assessments: the value of the variable at each point in time is the repeated 0078) Efficacy variables consist of protein absorption rate dependent variable, and the product identification of absorption (t) and overall absorption (AUCoa, and C) (GanedenBC'+Protein or Placebo) is the main factor. If the of blood amino acids, vitamin absorption rate of absorption efficacy variables are not normally distributed, and cannot be (t) and overall absorption (AUCo., and C) of vitamins normalized by logarithmic or other transformations, then B6, B12 and C. mineral absorption rate of absorption between-product testing are carried out using the Wilcoxon (W) and overall absorption (AUCo., and C) of min signed-ranks test. erals calcium, iron and Zinc, and GI questionnaire scores I0088 Assuming 15% attrition during the course of the (abdominal pain, abdominal bloating, gas, bowel movements, study, there are about 8 or 9 analyzable subjects remaining in gurgling noises and overall well-being). the per-protocol population. To have 80% power to obtain 0079 Efficacy endpoints generally consist of changes in significance (ps O.05) when testing a change over time (over efficacy variables from baseline to four hours post product 14 days of product use) within a product group, the mean administration following 14 days of product use. These change is about equal to about 15% larger than the standard changes (endpoints) are then be compared between the two deviation of the changes (a "1.15-sigma' effect size). To show products. that there was a significant difference between the two prod ucts, the mean difference is about 65% larger than the within Statistical Methods: group variability of the endpoint (a “1.65-sigma' effect size). I0089. Each efficacy endpoint is considered an indepen 0080 For each efficacy endpoint (amino acid, vitamin, dent question of interest, and is tested independently at the and mineral blood concentration), the following non-com 0.05 alpha level (ps0.05 required for a conclusion of statis partmental pharmacokinetic (PK) parameters are calculated: tical significance). Multiple testing is taken into account I0081. C. The maximum concentration observed dur when the results of the efficacy analyses are interpreted by the ing the four post-dose samples; statistician in the Final Statistical Report. I0082 to the time at which the maximum concentra tion was observed; Study Population I0083 AUC, the area under the concentration-vs time curve above the baseline (time 0) concentration, Inclusion Criteria integrated from time 0 to 4 hours post-dose, by trapezoi dal-rule quadrature. 0090 1. Male subjects, aged 21 to 60 years 0084 All safety and efficacy variables are summarized by 0091 2. Subject is able to understand and sign the time point and by product. Numerical variables are presented informed consent to participate in the study. US 2012/025 1512 A1 Oct. 4, 2012

0092. 3. Subject is willing and able to comply with the 0121 13. Subject has a history of drug or alcohol abuse protocol including: in the past 12 months. 0093 a. Attending three visits; 0.122 14. Subject has begun/stopped smoking sG 0094) b. Refraining from eating any yogurt or lacto months ago OR has plans to begin? quit Smoking. fermented beverages during the study: 0123 15. Subject has any condition or abnormality that, 0.095 c. Refraining from using any dietary supplements in the opinion of the investigator, would compromise the or nutritionals during the study; safety of the subject or the quality of the study data. 0096 d. Not taking any new vitamin and/or mineral 0.124 16. Subject is participating or has participated in Supplements until after study completion and refraining another research study within 30 days prior to the from taking any vitamin and/or minerals Supplements Screening visit. for 24 hours prior to the test visits. Concomitant Medications and Other Substances 0097 Exclusion Criteria Substances Permitted During the Study 0.098 1. Subject has any of the following medical con 0.125 Subjects may take the following substances during ditions: their participation in this study: 0099 a. active heart disease 0.126 Vitamins and/or minerals subject was taking prior 0100 b. uncontrolled high blood pressure (2 140/90 to starting the study (same frequency as prior to study mmHg) start). However, Subjects cannot take any vitamins and/ 0101 c. renal or hepatic impairment/disease or minerals for the 24 hours prior to the test visits. If the 0102 d. Type I or II diabetes Subject was not previously taking vitamins and/or min (0103 e. bipolar disorder erals, he is asked to not start taking any during the study. 0.104 f. Parkinson's disease 0127. Thyroid hormone replacement therapy, if drug 0105 g. unstable thyroid disease and dose are unchanged for at least 90 days before 0106 h. immune disorder (such as human immune Screening and throughout the study. deficiency virus (HIV)/acquired immune deficiency 0.128 Antihypertensive and antihyperlipidemic medi syndrome (AIDS)) cations, if drug and dose are unchanged for at least 90 0107 i. psychiatric disorders (hospitalized within the days before screening and throughout the study. past one year) Substances not Permitted During the Study 0.108 j. any medical condition deemed exclusionary I0129. Subjects may not take the following substances for by the Principal Investigator (PI) at least 60 days prior to the screening/randomization visit 0109 2. Subject has a history of cancer (except local (visit 1) or throughout the study. Any subject taking these ized skin cancer without metastases) within five years substances during the study is evaluated by the PI. prior to screening. 0.130 All oral antibiotics including: Amoxil R., Tri 0110. 3. Subject has a history of or currently has any mox(R) (amoxicillin), Cipro(R) (ciprofloxacin), Flagyl(R) gastrointestinal disease or disorder or any inflammatory (metronidazole), Doxy-100R) (doxycyline) and Bact bowel condition such as Crohn's disease, short bowel, rim R. Double Strength (trimethoprim-sulfamethox ulcerative colitis, or Irritable Bowel Syndrome (IBS). azole). 0111. 4. Subject is lactose intolerant (self-professed or 0131) Any medications that significantly interfere with diagnosed). bacterial flora Subjects may not take the following substances for at least 30 0112 5. Subject has had any stomach or intestinal sur days prior to the screening/randomization visit (visit 1) or gery (i.e. gastric bypass). throughout the study. Any subject taking these substances 0113. 6. Subject takes on a regular basis (defined as two during the study is evaluated by the PI. or more times per week) any prescription or over-the 0.132. Laxatives: psyllium husk (Metamucil), methyl counter medications for diarrhea, constipation, heart cellulose (Citrucel), polycarbophil, docusate (Colace, burn or any other gastrointestinal problems. Diocto), mineral oil, Sodium phosphate (and variants), 0114 7. Subject is currently taking laxatives or has magnesium citrate, magnesium hydroxide (Milk of taken laxatives within the 30 days prior to screening/ magnesia), magnesium Sulfate (which is Epsom salt), enrollment. glycerin Suppositories, Sorbitol, lactulose, and polyeth 0115 8. Subject is currently taking antibiotics (or any ylene glycol (PEG) drug that significantly interferes with bacterial flora) or 0.133 Digestive enzymes: CotaZym, Creon, Pancrease, has taken antibiotics within the 60 days prior to screen Digex, Dygase, GastrineX, Hi-Vegi-Lip, Ku-Zyme, ing/enrollment. Kutrase, Lactaid, Lapase, Lipram, Palcaps 10, Pan 0116 9. Subject is currently taking or has used in the 2400, Pancreatin, Pancrecarb, Pangestyme, Panocaps, Panokase, Sucraid, SureLac, Ultrase, Viokase, Zenpep past 30 days probiotics/prebiotics (including yogurt and 0.134 All dietary and herbal supplements including: lacto-fermented beverages) or any digestive enzymes 0.135 Probiotic products (including fortified foods): prescription or over-the-counter (OTC). Thirty-day Acidophilus, Bacid, Flora-Q, Florastor, Novaflor, washout allowed. RisaCuad, Superdophilus, Ganeden BC30, LAFTI 0117 10. Subject is on an unstable dose of medication B94, LAFTI L10, LAFTI L26, Bifiene, Align, (defined as fewer than 90 days at the same dose). Howaru Bifido, ProBactrix, Mutaflor, Actimel/Dan 0118 a. Anti-hypertensives and anti-hyperlipidemic Active, Cultura, Yakult, Lactobacillus fortis, Good medications ok if stable dose. Belly/ProViva/TuZen, LGG, Vi?it, Verum, DiarSafe, 0119 11. Subject is currently taking any medication Bion Flore Intime, Jarrow, Fem-Dophilus, Florajen3, deemed exclusionary by PI. Bio-K, CL 1285, A Biotica; I0120 12. Subject has an allergy to soy, milk, chocolate 0.136 Prebiotic products (including fortified foods): or any of the ingredients in the test product. lactulose, lactitol oligofructose, inulin, galacto-oli US 2012/025 1512 A1 Oct. 4, 2012 11

gosaccharides, tagatose, isomaltooligosaccharides, polydextrose, maltodextrin, fructo-oligosaccharides, -continued arabinogalactan, polyols-lactulose; Potassium 170 mg 59 0.137 Those supplements purported to improve GI Total Carbohydrate 3g 196 health including dandelion, devil's claw, feverfew, Dietary Fiber 1 g 4% ginger, goldenseal, lemon balm, peppermint, roman Sugars 1 g Protein 23 g chamomile, turmeric, Valerian, yarrow, mastic gum, Vitamin A dill, caraway, anise, cumin. Vitamin C Calcium Diet Iron 0.138. Subjects are required to fast (no food or beverage *Percent DailyValues are based on a 2,000 calorie diet. other than water, no caffeine) for eight hours prior to visits 2 and 3. Subjects are asked to refrain from alcohol for the 24 Ingredients: Whey Protein Concentrate (Whey Protein Con hours prior to visits 2 and 3. Subjects are not allowed to eat centrate, Soy Lecithin), Whey Protein Isolate, Cocoa Powder yogurt or to drink lacto-fermented beverages throughout the (Processed with Alkali), Natural & Artificial Flavors, study period. Examples of lacto-fermented beverages include Acesulfame Potassium and Sucralose. kefir (water and dairy), kambucha, kvass, ciderandgingerale. 0146. At the test visits, subjects are provided a multiple 0139 Subjects are not allowed to use protein powders (e.g. Vitamin with mineral Supplement. The Supplement that is whey, Soy, hemp, pea, rice) while participating in the study. used is the One A Day(R) Men's Health Formula. 0140. Subjects are not allowed to eat/drink foods and bev The ingredients in the One A Day Men's Health Formula erages high in Vitamin C for 24 hours prior to the test visits. 0147 Calcium Carbonate, Magnesium Oxide, Microcrys Examples of foods and beverages high in vitamin C include talline Cellulose, Ascorbic Acid, Croscarmellose Sodium, citrus fruits (such as oranges and grapefruit) and their juices, Gelatin, Maltodextrin; Less than 2% of Beta-Carotene, red and green pepper, kiwifruit, broccoli, strawberries, can Biotin, Cholecalciferol, Chromium Chloride, Crospovidone, taloupe, baked potatoes, and tomatoes. Cupric Oxide, Cyanocobalamin, D-Calcium Pantothenate, Other than this, subjects are allowed to continue their normal dl-Alpha-Tocopheryl Acetate. Folic Acid, Hydroxypropyl diet. Methylcellulose, Lycopene, Manganese Sulfate, Niacina mide, Phytonadione, Polyethylene Glycol, Pyridoxine Exercise Hydrochloride, Riboflavin, Silicon Dioxide, Sodium Sel 0141 Subjects are asked to refrain from exercise for the 24 enate, Soybean Oil, Starch, Stearic Acid, Thiamine Mononi hours prior to visits 2 and 3. Subjects are allowed to continue trate, Triacetin, Vitamin A Acetate and Zinc Oxide. their usual exercise routine. Product Dosing Product Ingredients 0148 Product administration instructions: Subjects are instructed to take one packet per day, at approximately the 0142. The following lists of ingredients are provided. same time of day. 0149 Preparation instructions: Mix one packet with six to Product 1: GanedenBC'+Protein eight ounces of cold water, milk or subject's preferred bever 0143. 1 billion CFU GandenBC30 and 23 g of whey pro age for 20 to 30 seconds in a shaker or blender. tein (whey protein isolate (WPI)>90) in 30 g of powder, Subjects are instructed to start taking the first study product chocolate flavor. the day after visit 1. Subjects are instructed to not take the study product prior to coming in for visits 2 and 3 (i.e. not take Product 2: Protein (Placebo) study product the morning of visits 2 and 3). Subjects are required to take the study product on site for visits 2 and 3. 0144) 23g of whey protein (WPI >90) in 30 g of powder, Subjects are instructed to start the second study product the chocolate flavor. day after visit 2. Subjects cannot take anything the evening of visit 2. Subjects are instructed to bring the remaining product Nutrition Facts for the Whey Protein Powder to all visits. (0145 Product Administration Instructions 0150 Take one packet per day, at approximately the same Serving Size: 1 Packet time of day; mix with 6 to 8 ounces of water, milk or favorite beverage and shake/blend for 20 to 30 seconds. Amount Per Serving 0151. The product is packaged so that the subject is pro Calories 130 vided only the quantity needed for the time between visits (14 Calories from fat 25 days) plus product to cover the window (additional three days of product). % Daily Value Total Fat 2.5g 4% Product Compliance Saturated Fat 1 g 59 Trans Fat Og 0152 Compliance is measured via the packet counting Cholesterol 65 mg 22% method. By documenting the number of calendar days Sodium 50 mg 296 between visits and the number of packets that should have been taken, compliance is calculated. The Subject's compli US 2012/025 1512 A1 Oct. 4, 2012 12 ance is recorded (as a % of prescribed amount) for each least five minutes prior to taking the measurements. Measure product and this compliance % is used to establish the sub ments are done with Subjects in a seated position and the ject's eligibility for inclusion in the populations (Safety, subject's left arm is used. Intent-to-Treat, and/or Per-Protocol) used for analysis of the Study Questionnaires study's results, in accordance with the criteria specified 0.161 Dichotomous Questionnaire the following is below. administered at visits 2 and 3. Each subject is asked: 0153. For visits 2 and 3, product administration are per (0162 Did you: formed on site. Documentation is done in each subject's 0163 Noteator drink any foods or beverages other Source confirming product administration. than water or have caffeine for the eight hours prior to the visit? 016.4 Eat or drink any foods or beverages high in Randomization vitamin C in the past 24 hours? 0.165 Drink any alcoholic beverages in the past 24 0154 The consulting statistician develops a randomiza hours? tion schedule and create a randomization log for use at the 0166 Take any vitamin and/or mineral supple investigative site. To minimize the effect of treatment ments in the past 24 hours? sequence (carry-over, training, fatigue, seasonality, etc.) on 0167 Exercise in the past 24 hours? safety and efficacy endpoints, the active product (A) and 0168 Use any new vitamin and/or mineral supple placebo (B) are administered to each subject in one of the two ments since the screening visit? possible sequences: A-B or B-A. Block-2 randomization is 0169. Use any protein powder other than the study used, so that the two possible product sequences are shuffled product since the screening visit? into random order for assignment for the first two subjects, 0170 Use any dietary supplements, any probiot then shuffled again into a different random permutation for ics, or prebiotics since 30 days prior to the screen assignment to the next two Subjects, and so on for the remain ing visit? ing Subjects to be enrolled (plus additional spare product). 0171 Eat or drink any yogurt or lacto-fermented beverages since 30 days prior to the screening visit? Study Procedures 0172 Did you take the study product today? 0173 If subject answers “yes” to any of the above Informed Consent questions, the Subject is discontinued as per PI discre tion and/or a protocol deviation is recorded. 0155. An informed consent form (ICF) is written in accor 0.174 GI Questionnaire—a six-part questionnaire dance with established criteria of the Institutional Review regarding general well-being and gastrointestinal symp Board (IRB) and the appropriate federal regulations (e.g., 21 toms is administered at visits 1, 2 and 3. Subjects are CFR Parts 50 and 56) to describe the study plan, procedures, asked, “how often in the past two weeks have you' and risks. The investigator and the IRB must all approve the experienced various GI symptoms with answer choices ICF and any ICF amendments or administrative changes of “all of the time, most of the time, sometimes, a little before they are used. and never'. Subjects are asked to rate how they feel 0156 Investigators ensure that each subject is clearly and overall with answer choices of “better, same and worse'. See, e.g., Worthley et al., 2009 Am J Clin Nutr, 90: fully informed of the purpose, potential risks, and require 578-586. For example, the following questions are ments of study participation. Written informed consent must asked: be obtained from each subject before performing any screen 0.175 1) How often in the past week have you had pain ing or other study procedures. in the abdomen? 0176) 2) How often in the past week have you experi Medical History enced abdominal bloating? (0177 3) How often in the past week have you been 0157. A Medical History is performed at the screening troubled by excessive passage of gas through the anus? visit to collect information on past and current medical con 0.178 4) How often in the past week have you been ditions, Surgical history, allergy information, and concomi troubled by frequent of loose bowel movements? tant or recently taken (in the past 90 days) medications includ (0179 5) How often in the past week have you been ing over the counter (OTC) non-prescription products, troubled by excessive gurgling noises from the abdo nutritional Supplements, herbals, and investigational prod men? uctS. 0180 6) In comparison to how you felt at the start of the study, over the past week, have you felt better, worse or Physical Examination the same? 0158. Subjects undergo a full physical examination at Protein and Micronutrient Absorption Test screening for determining eligibility. 0181. The following test is performed at visits 2 and 3 to assess the effects of the products on protein, vitamin and Vital Signs mineral absorption. The following is performed to standardize the testing condi 0159. At each visit, subjects have blood pressure and heart tions: rate measured. 0182. The test is performed after an eight hour fast; 0160 The following guidelines are used for the blood 0183 Subjects are not physically active during the test; pressure and heart rate: Subjects sit in a chair, resting, for at 0184. No Smoking is allowed during the test. US 2012/025 1512 A1 Oct. 4, 2012 13

Test Procedure: 0201 All p-values appearing in these Summarizations is 0185. Collect the baseline blood sample (0 hours) for considered descriptive, not inferential. No final statistical the evaluation of the baseline levels of various amino conclusions are drawn from them, but they are referred to in acids and select vitamins and minerals. the interpretation of the changes. 0186 Provide subject with the multiple vitamin with Statistical Analysis mineral Supplement (can be taken with water or the study product). Safety and Efficacy Variables and Endpoints 0187 Provide subject with the study product which is to 0202 Safety variables consist of adverse events and sub be consumed within 10 minutes. jective remarks. 0188 Timing of the test is from when the subject begins 0203 Safety endpoints consist of the changes in the safety to drink the study product. variables listed above with each product following 14 days of 0189 Collect blood samples at 1, 2, 3, and 4 hours for treatment. the evaluation of the absorption of the amino acids, 0204 There is no formal safety objective. Vitamins and minerals. 0205 Efficacy variables consist of protein absorption rate During the Time between Blood Draws: of absorption (t) and overall absorption (AUCo., and 0190. Subjects are primarily sitting. C) of blood amino acids, vitamin absorption rate of 0191 Subjects are allowed to walk around the site to use absorption (t) and overall absorption (AUCo., and C) the rest room and to stretch but are not allowed to do any of vitamins B6, B12 and C), mineral absorption rate of additional physical activity. absorption (t) and overall absorption (AUCo., and C) 0.192 Subjects are allowed to do restful activities such of minerals calcium, iron and Zinc, and GI questionnaire as read, listen to music with earphones, watch television, scores (abdominal pain, abdominal bloating, gas, bowel work on the computer or rest/sleep. movements, gurgling noises and overall well-being). 0193 Subjects are not allowed to eat or drink anything 0206 Efficacy endpoints generally consist of changes in other than water. efficacy variables from baseline to four hours post product administration following 14 days of product use. These Descriptive Summarization changes (endpoints) are then compared between the two 0194 For each efficacy endpoint (amino acid, vitamin, products. and mineral blood concentration), the following non-com partmental pharmacokinetic (PK) parameters is calculated: Other Embodiments (0195 C. The maximum concentration observed dur 0207. While the invention has been described in conjunc ing the four post-dose samples; tion with the detailed description thereof, the foregoing 0.196 t the time at which the maximum concentra description is intended to illustrate and not limit the scope of tion was observed; AUC, the area under the concen the invention, which is defined by the scope of the appended tration-vs-time curve above the baseline (time 0) con claims. Other aspects, advantages, and modifications are centration, integrated from time 0 to 4 hours post-dose, within the scope of the following claims. by trapezoidal-rule quadrature. 0208. The patent and scientific literature referred to herein 0.197 All safety and efficacy variables are summarized by establishes the knowledge that is available to those with skill time point and by product. Numerical variables are presented in the art. All United States patents and published or unpub as mean, standard deviation, count, median, and range (mini lished United States patent applications cited herein are mum to maximum value). Changes from the appropriate incorporated by reference. All published foreign patents and baseline are Summarized and presented in the same way. patent applications cited herein are hereby incorporated by Numerical variables and their changes from baseline are dis reference. Genbank and NCBI submissions indicated by played graphically, as plots of mean value vs. time, with accession number cited herein are hereby incorporated by separate lines for the two products (probiotic VS. placebo). reference. All other published references, documents, manu The graphs may contain Vertical error-bars around the mean scripts and scientific literature cited herein are hereby incor values, indicated Standard errors of the mean, if, in the opin porated by reference. ion of the statistician, this would make the graphs more infor 0209 While this invention has been particularly shown mative. Categorical variables are presented as tabulations of and described with references to preferred embodiments counts and percentages of totals. thereof, it will be understood by those skilled in the art that 0198 For each continuous variable, the mean change from various changes in form and details may be made therein baseline to each Subsequent time point are tested for nominal without departing from the scope of the invention encom significance by the paired Student t test, or by the non-para passed by the appended claims. metric Wilcoxon test if non-normally distributed. What is claimed is: 0199 For each continuous variable at each time point, the 1. A sports nutrition composition comprising a protein mean differences in the variable, or in the change in that Source and an isolated Bacillus coagulans spore, wherein said variable from baseline, between the different products are composition comprises at least 50% protein. tested for nominal significance by the paired Student t test or 2. The composition of claim 1, wherein said composition by the non-parametric Wilcoxon test if non-normally distrib comprises at least 75% protein. uted. 3. The composition of claim 1, wherein said isolated Bacil 0200 For each categorical variable, difference in the dis lus coagulans is Bacillus coagulans hammer StrainAccession tribution of categories between the different product groups No. ATCC 31284. are tested for nominal significance by the Fisher Exact test if 4. The composition of claim 1, wherein said isolated Bacil possible, or by the Chi-Square test if necessary. lus coagulans is selected from the group consisting of GBI-30 US 2012/025 1512 A1 Oct. 4, 2012

strain (ATCC Designation Number PTA-6086), GBI-20 20 strain (ATCC Designation Number PTA-6085), and GBI strain (ATCC Designation Number PTA-6085), and GBI-40 40 strain (ATCC Designation Number PTA-6087). strain (ATCC Designation Number PTA-6087). 22. The composition of claim 19, wherein said sports nutri 5. The composition of claim 1, wherein said composition tion composition comprises protein selected from the group comprises about 1x10° to 1x10" colony forming units (CFU) consisting of soy protein, whey protein, rice protein, hemp of Bacillus coagulans per unit dose. seed protein, or casein protein. 6. The composition of claim 1, wherein said composition 23. The composition of claim 22, wherein said protein comprises about 20grams of said protein to about 50 grams of comprises an amino acid selected from the group consisting said protein per unit dose. of isoleucine, alanine, leucine, arginine, lysine, aspartate, 7. The composition of claim 1, wherein said protein com aspartic acid, methionine, cysteine, phenylalanine, threonine, prises whey protein, brown rice protein, casein milk protein, tryptophan, glycine, Valine, proline, histidine, serine, egg albumin, and whey protein hydrolysate. tyrosine, asparagine, selenocysteine, pyrrolysine, glutamate, 8. The composition of claim 1, wherein said composition glutamic acid, and glutamine. comprises approximately 1 billion CFU Bacillus coagulans. 9. The composition of claim 8, wherein said protein is soy 24. The composition of claim 19, wherein said sports nutri protein, whey protein, rice protein, hemp seed protein, or tion composition is selected from the group consisting of casein protein. creatine, calcium, Sodium caseinate, whey peptides, and 10. The composition of claim 8, wherein said protein com lactoferrin. prises an amino acid selected from the group consisting of 25. A method of enhancing lean muscle development, isoleucine, alanine, leucine, arginine, lysine, aspartate, aspar recovery, or repair comprising administering to a subject tic acid, methionine, cysteine, phenylalanine, threonine, tryp desiring an enhancement of said lean muscle development, tophan, glycine, Valine, proline, histidine, serine, tyrosine, recovery, or repair a composition comprising a sports nutri asparagine, Selenocysteine, pyrrolysine, glutamate, glutamic tion composition and an isolated Bacillus coagulans spore, acid, and glutamine. wherein said composition comprises at least 50% protein. 11. The composition of claim 8, further comprising creat 26. The method of claim 25, wherein said composition ine, calcium, Sodium caseinate, whey peptides, or lactoferrin. comprises at least 75% protein. 12. The composition of claim 1, wherein said sports nutri 27. The method of claim 25, wherein the absorption of said tion composition is selected from the group consisting of protein is increased in said subject as compared to the absorp creatine, calcium, Sodium caseinate, whey peptides, and tion of said protein after administration of protein in the lactoferrin. absence of Bacillus coagulans. 13. The composition of claim 1, wherein said composition 28. The method of claim 27, wherein said absorption com does not comprise an Sugar. prises the rate of absorption or overall absorption. 14. The composition of claim 1, wherein said composition 29. The method of claim 25, wherein the absorption of a does not comprise gluten. Vitaminis increased in said Subject as compared to the absorp 15. The composition of claim 1, wherein said composition tion of said vitamin after administration of protein in the does not comprise aspartame. absence of Bacillus coagulans. 16. The composition of claim 1, wherein said composition does not comprise artificial coloring. 30. The method of claim 29, wherein said vitamin com 17. The composition of claim 1, wherein said composition prises vitaminA, Vitamin B, Vitamin B, Vitamin B vitamin is in the form of a protein powder, a ready to drink protein Bs, vitamin B vitamin B7, vitamin B. vitamin B, or shake, a proteinbar, a protein bite, or a protein gel. Vitamin C. vitamin D. Vitamin E, or vitamin K. 18. The composition of claim 1, wherein said composition 31. The method of claim 25, wherein the absorption of a comprises a weight gainer. chemical element is increased in said Subject as compared to 19. A composition comprising a dry mix for sports nutri the absorption of said chemical element after administration tion composition comprising a sports nutrition composition of protein in the absence of Bacillus coagulans. and an isolated Bacillus coagulans spore. 32. The method of claim 31, wherein said chemical ele 20. The composition of claim 1, wherein said protein com ment comprises sodium, potassium, calcium, iron, or zinc. prises whey protein. 33. The method of claim 25, further comprising adminis 21. The composition of claim 19, wherein said isolated tering a vitamin or mineral. Bacillus coagulans is selected from the group consisting of GBI-30 strain (ATCC Designation Number PTA-6086), GBI c c c c c