Microbiome modulating formula for menopausal-aged women

Abstract

A method of improving health of menopausal aged women, including treatment of symptoms of menopause, the method including consumption by, or administration to, the subject of a probiotic composition. The composition includes four or more of the following components: (i) Lactobacillus rhamnosus, (ii) Bifidobacterium longum, (iii) Green tea or derivatives thereof, (iv) Cranberries, Cranberry extract or derivatives thereof, (v) Nichi glucan, glucans produced from Aureobasidium pullulans, polysaccharides produced from Aureobasidium pullulans, β-1,3-1,6-glucans, or derivatives of any of these, (vi) Bifidobacterium bifidum, and (vii) a composition comprising Lactobacillus acidophilus, Lactobacillus rhamnosus, Saccaromyces boulardii, Bifidobacterium breve, and alpha amylase. Bacteriophages that target E. coli may also be included.

Description

Attorney Docket No. BHMP107WO Microbiome Modulating Formula for Menopausal-Aged Women CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. provisional application 63 / 588,513, filed October 6, 2023, and titled “Microbiome Modulating Formula for Menopausal-Aged Women.” This application also claims priority to U.S. provisional application 63 / 664,505, filed June 26, 2024, and also titled Microbiome Modulating Formula for Menopausal-Aged Women.” Each of these prior applications are incorporated herein by reference. FIELD

[0002] This disclosure relates generally to a method and composition of improving or balancing a microbiome of a human gut. More particularly, the disclosure relates to a method of treating menopausal symptoms, and / or balancing the level of certain organisms in the gut of a subject in need thereof. BACKGROUND

[0003] Menopause is a natural part of women’s life, signaling the end of their reproductive phase. It occurs with the final menstrual period, which is known to occur after 12 months of amenorrhea (absence of menstrual period). The underlying cause of menopause is the cessation of ovarian follicular function and a decline in circulating estrogen levels in the bloodstream, thereby setting the stage for aging and accelerates the process of noncommunicable diseases. Most women worldwide experience menopause between the ages of 45-55 years, however it may also be triggered early as a consequence of surgical or medical procedures.

[0004] Some of the menopause-associated symptoms include hot flashes, digestive distress, weight gain, sleep disruption, and increased risk and occurrence of bacterial vaginosis and / or urinary tract infection. The majority of women do not need treatment for their menopause symptoms; however, hormone replacement therapy options are also available to help alleviate some of these symptoms.

[0005] The gut microbiome and its role in disease states and conditions, including menopause, have been explored. Hormonal changes can actually disrupt the balance of the gut microbiome, which in turn leads to some of the symptoms stated above. On the basis of this, probiotics, which are beneficial bacteria, have the potential to restore this microbial balance and alleviate symptoms. Probiotic supplements targeted for general women’s health are already available in the market, with claims to improve overall health. Typically, these products contain Lactobacillus and / or Bifidobacterium strains, which both have demonstrated the ability to restoreAttorney Docket No. BHMP107WO microbial balance in the gut and alleviate general digestive issues. However, evidence illuminating the microbiome differences in menopausal-aged women compared to reproductive- aged women has not been available and thus microbiome-related treatments have not been developed.

[0006] In addition, biofilms form in the gut and cause general health problems and are a hindrance to efforts to address microbiome imbalances. Biofilms are formed when unicellular microorganisms live together and form a community that is protected by an exopolysaccharide (EPS) matrix. This EPS matrix typically is a conglomeration of proteins, polysaccharides and extracellular DNA. Biofilm-associated microorganisms differ from their planktonic (freely suspended) counterparts. It is believed that biofilm-forming cells co-aggregate with each other to form coordinated groups attached to a biotic or abiotic surface, the cells are surrounded by a protective EPS matrix, communicate effectively through quorum sensing, and have low metabolic activity that limits the impact of conventional antimicrobials (both antifungal and anti- bacterial agents) acting against actively metabolizing cells or cells in stationary phase.

[0007] Microorganisms, including bacteria, fungi, and archaea form biofilms. For a given subject (e.g., a human) many microorganisms (termed microbiota) exist on or within various regions of said subject, e.g., the gut, the skin, the vagina, the respiratory tract, and the mouth. There is an inextricable link between a host subject’s gastrointestinal microbiota and digestion, immunity and metabolism. Gastrointestinal microbiota are important for vitamin and metabolites biosynthesis and digestion of complex macromolecules such as polysaccharides. Gastrointestinal microbiota, under normal conditions, aid in prevention of colonization of pathogenic microorganisms and in maintaining the integrity and function of the intestinal barrier, and supporting the immune system.

[0008] However, undesirable or unbalanced biofilms may be involved in a significant percentage of human microbial infections (Potera, C. (1999) SCIENCE 283:1837-8). It has been proposed that four criteria define biofilm etiology during an infection include whether the pathogenic bacteria or fungi are surface associated or adherent to a substratum; whether the bacteria are in clusters, encased in a matrix of bacterial, fungal or host constituents; whether the infection is localized; and whether the infection is resistant to antibiotic and anti-fungal therapies despite the antimicrobial sensitivity of the constituent planktonic organisms (Parsek, M. R. & Singh, P. K. (2003) ANN REV. MICROBIOL.57:677-701; Ghannoum, M., et al. (2015) MICROBIALBIOFILMS. Second Ed., Am. Soc. for Microbiology).Attorney Docket No. BHMP107WO

[0009] With respect to biofilms in the gut, it is now believed that bacteria and fungi can exist for example as biofilms on the colonic epithelium, within the mucus layer covering it, and on food particles in the lumen (MacFarlane, S. & MacFarlane, G. T. (2006) APPL. ENVIRON. MICROBIOL.72:6204-11; Probert, H. M. & Gibson, G. R. (2002) CURR. ISSUES INTEST. MICROBIOL.3:23-7). The biofilms in the GI tract populated at least with pathogenic bacteria form a dense filamentous film encased within an exopolysaccharide (EPS) matrix. Such biofilms are impenetrable to antimicrobials and host immune cells, and are difficult to remove or treat.

[0010] Various associations between microbiome gut flora and systemic or localized effects on other parts of the body have been theorized. However, little in the way of specific associations has been established with reasonable specificity and reproducibility. SUMMARY

[0011] A gut microbiome database was assembled from numerous subjects and test samples. A bioinformatics analysis of this gut microbiome database was performed focusing on the differences in the gut microbiome of young pre-menopausal women, and older women that would have undergone menopause. A differential abundance of several probiotic organisms of interest was observed that are associated with the young women cohorts when compared to the older women cohorts. Thus, it was discovered that there is need for improved compositions and methods for addressing such differences through the gut microbiome. It is theorized that through addressing these imbalances with the microbiome in older women that the symptoms of menopause can be improved.

[0012] As discussed herein, microbiome analysis was conducted on healthy subjects that were distributed into a menopausal-aged women cohort and a reproductive-aged women cohort. Differential abundance analysis using the ANCOMBC, ANCOMBC2, DESeq2, and ZicoDA methods were performed to determine the relative abundance of bacterial and fungal organisms between the cohorts.

[0013] The formulations disclosed herein were developed based on the aforementioned microbiome data and are designed to target specific microbiome organisms identified as differentially abundant between menopausal-aged and reproductive-aged women. This opens up new possibilities for personalized nutrition and microbiome modulation to address the specific needs of menopausal women. With these surprising discoveries, a composition comprising organisms designed to balance the microbiome to be closer to that of reproductive-aged women in menopausal-aged females for reduction of menopausal symptoms was developed.Attorney Docket No. BHMP107WO

[0014] In some aspects, the technology described herein relate to a method of improving health of menopausal aged women, including treatment of symptoms of menopause, the method including consumption by, or administration to, the subject of a probiotic composition including Lactobacillus rhamnosus, Bifidobacterium longum, Green tea, Cranberry extract, Nichi glucan, glucans produced from Aureobasidium pullulans, or polysaccharides produced from Aureobasidium pullulans, β-1,3-1,6-glucans, Bifidobacterium bifidum, and BIOHM FX (Lactobacillus acidophilus, Lactobacillus rhamnosus, Saccaromyces boulardii, Bifidobacterium breve, alpha amylase).

[0015] In some aspects, the technology described herein relate to a method of improving health of menopausal aged women, including treatment of symptoms of menopause, the composition including a live, freeze-dried probiotic composition including a combination of

[0016] Unless the context dictates otherwise, the terms composition, formulation, and dosage form can be used interchangeably herein. For example, it is contemplated that the composition or formulation consumed by, or administered to, a subject can be consumed or administered as a dosage form, for example, as a unit (for example, a spoonful) of the composition. Similarly, it is understood that a dosage form can, for example, comprise a composition or formulation described herein. For example, a dosage form can comprise, for example, a composition / formulation in the form of a powder described herein, or, for example, a capsule that contains such a composition / formulation.

[0017] In an embodiment, the composition disclosed herein further comprises one or more bacteriophages antagonistic to E. Coli and promoting growth of Bifidobacterium taxa.

[0018] Other features and advantages of the technology disclosed herein will be apparent from the following detailed description and claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] The figure is a schematic representation of an exemplary process for making a probiotic composition described herein. DETAILED DESCRIPTION

[0020] This disclosure pertains to a novel formulation for the modulation of specific targeted microbiome organisms, which includes both probiotic strains, dietary supplement and food ingredients. The formulation is the result of an innovative process that involves the analysis of microbiome data obtained from two distinct cohorts of women—menopausal-aged women andAttorney Docket No. BHMP107WO reproductive-aged women—allowing the inventors to identify organisms with differential abundance between these two groups.

[0021] The data-gathering process involves the collection of microbiome samples from the two cohorts of menopausal-aged and reproductive-aged women. For purposes of this study, the menopausal aged women cohort were defined as of the ages of 45-65, and the reproductive-aged women cohort were defined as of the age of 25-35.

[0022] Next generation sequencing and analysis of microbial composition data was performed on the microbiome samples to identify the organisms present in the microbiomes in the samples. Then data analysis of the microbiome organisms were identified that exhibited significant differential abundance between menopausal-aged and reproductive-aged women.

[0023] With this data, formulation development was performed to promote the growth and balance of the different target organisms in the menopausal-aged cohort. Formulation development included probiotic strains and dietary supplement ingredients that were known, found, or suspected to affect the target organisms.

[0024] The resulting formulation is intended to be used as a dietary supplement or functional food to support women's health during menopause by positively influencing their microbiome composition.

[0025] In addition, certain formulations disclosed herein are based, in part, upon the discovery that a composition or a dosage form comprising both non-pathogenic bacteria and fungi improves nutrient (for example, protein and / or vitamin) absorption by mammalian (for example, human) intestinal epithelial cells. Also provided herein is the discovery that it is possible to prevent or disrupt biofilms comprising pathogenic bacteria and / or fungi, and optionally archaea and / or protozoa, in a subject by administering to the subject a composition comprising both isolated and viable non-pathogenic bacteria and non-pathogenic fungi, which when administered promotes a more balanced microbiome and / or promotes the restoration of the natural microbiome in the subject.

[0026] Through the methods disclosed herein, several microorganisms were identified that were differentially abundant in the menopausal-aged women’s cohort. To address these changes and restore gut balance to promote overall health in women, the following ingredients and their combination were identified in order to positively modulate an aging menopausal aged woman’s microbiome back to the levels to that were observed in the reproductive-age cohort.Attorney Docket No. BHMP107WO

[0027] In an embodiment, the formulation comprises three or more or four or more of the following components: (i) Lactobacillus rhamnosus, (ii) Bifidobacterium longum, (iii) Green tea or derivatives thereof, (iv) Cranberries, Cranberry extract or derivatives thereof, (v) Nichi glucan, glucans produced from Aureobasidium pullulans, polysaccharides produced from Aureobasidium pullulans, β-1,3-1,6-glucans, or derivatives of any of these, (vi) Bifidobacterium bifidum, and (vii) a composition comprising Lactobacillus acidophilus, Lactobacillus rhamnosus, Saccaromyces boulardii, Bifidobacterium breve, and alpha amylase. Another component, e.g., grape seed extract or grape seed derivatives, may also be included.

[0028] This formulation is intended to improve overall health in women and provides a unique approach in achieving modulation of menopausal symptoms in menopausal aged women.

[0029] L. rhamnosus can be included in the composition in an amount of 0.1- 10 Billion CFUs (10-500 mg), or 0.01 to 15 billion CFUs, such as, 0.5 to 5 billion CFUS, or 1.5 to 3 billion CFUs. L. Rhamnosus is a lactic acid bacterium that is resistant to acid and bile and has growth characteristics that allow it to survive and persist within the human gastrointestinal tract. It also displays excellent adhesion to the intestinal epithelial layer to inhibit the growth and adherence of several known pathogens and has been reported to form biofilms.

[0030] B. longum, e.g., strain ES01, can be included in the composition in an amount of 0.1- 10 Billion CFUs (10-500 mg), or 0.01 to 15 billion CFUs, such as, 0.5 to 5 billion CFUs, or 1.5 to 3 billion CFUs. B. longum is a Gram-positive, catalase-negative rod-shaped bacterium. It is one of the 32 species that belong to the genus Bifidobacterium. It is a microaerotolerant anaerobe and is known to produce lactic acid.

[0031] Green tea can be included in the composition an amount of 100 to 750 mg, or 50 to 1000 mg, e.g., 75 to 800 mg, 150 to 650 mg. The green tea may be an extract or other derivation from Camellia sinensis leaves and buds. Green tea includes polyphenols such as epigallocatechin gallate (EGCG), epicatechin gallate, epicatechins and flavanols. Other components include three kinds of flavonoids, known as kaempferol, quercetin, and myricetin.

[0032] Cranberry extract can be included in the composition an amount of 100-1000 mg, or 50 to 1500 mg, e.g., 75 to 800 mg, 150 to 650 mg. Cranberries are fruit of the genus Vaccinium, and subgenuses include V. macrocarpon, V. erythrocarpum, V. japonicum, V. microcarpum, and V. oxycoccos. Cranberry extract includes phytochemical compounds, including one or more polyphenols – including proanthocyanidins, flavonols and quercetin.Attorney Docket No. BHMP107WO

[0033] Nichi glucan, glucans produced from Aureobasidium pullulans, polysaccharides produced from Aureobasidium pullulans, or β-1,3-1,6-glucans, can be included in the composition an amount of 10-100 mg, or 5 to 150 mg, e.g., 7 to 80 mg, 15 to 65 mg. Nichi glucan may be produced by one or more strains of Aureobasidium Pullulans (Black Yeast).

[0034] Grape seed derivatives or grape seed extract can also be included in the composition, for example, in an amount of 10 to 100 mg, or 5 to 150 mg, e.g., 7 to 80 mg, 15 to 65 mg. Grape seed extract includes procyanidins which are formed from proanthocyanidins in the extract. Proanthocyanidins are mainly composed of monomeric catechin and epicatechin, gallic acid and polymeric and oligomeric proanthocyanidins.

[0035] Bifidobacterium Bifidum (“B. bifidum”) can be used in amount of 10 to 100 mg, or 1 to 1000 mg, such as 5 to 300 mg, or 25 to 75 mg. B. bifidum is a gram-positive, anaerobic bacteria that is commonly found in the human gastrointestinal tract, particularly in infants and breast-fed babies. B. bifidum plays a role in the fermentation of carbohydrates and fibers to produce beneficial short-chain fatty acids which are associated with overall colon and gut health.

[0036] BIOHM FX is a commercial product comprising Lactobacillus acidophilus, Lactobacillus rhamnosus, Saccaromyces boulardii, Bifidobacterium breve, alpha amylase. BIOHM FX can be used in amount of 0.1 to 10 billion CFUs or 10-660 mg, such as, for example, or 0.01 to 15 billion CFUs, such as, 0.5 to 5 billion CFUs, or 1.5 to 3 billion CFUs.

[0037] The BIOHM FX component or sub-components of the BIOHM FX component may be included to aid in remediation of biofilms in the gut. Biofilms may exist in the GI tract and can block nutrient absorption and effectiveness of probiotics. They potentially may contribute to menopausal symptoms and could hinder efforts to remediate the decreased gut microorganisms found herein. In certain embodiments, compositions described herein can be used to disrupt a biofilm comprising, for example, pathogenic bacteria and fungi within the gastrointestinal (GI) tract (for example, esophagus, stomach, upper intestine, and / or the lower intestine) of a subject. In certain embodiments, the preselected region comprises the duodenum, the jejunum, and / or the ileum of the upper GI tract, whereas in other embodiments, the preselected region comprises the appendix, the proximal colon, and / or the rectum of the lower GI tract.

[0038] In alternative examples, derivatives, such as isolated extracts of one or more sub- components, of the listed components belong to may be used in place of or in addition to one or more of those disclosed above. In an embodiment, such substitutions will adhere to and colonize in the GI tract.Attorney Docket No. BHMP107WO

[0039] In an embodiment, the composition can also include non-pathogenic bacteria and fungi that improves nutrient (for example, protein, vitamin, mineral, and co-factor) absorption by mammalian (for example, human) intestinal epithelial cells. Some of these are listed below.

[0040] Live, freeze-dried powdered forms of the components disclosed herein of the composition can be selected for use in the formulation.

[0041] In an embodiment, the composition comprises four to seven components, or five to six components, selected from the group consisting of: (i) Lactobacillus rhamnosus, (ii) Bifidobacterium longum, (iii) Green tea or derivatives thereof, (iv) Cranberries, Cranberry extract or derivatives thereof, (v) Nichi glucan, glucans produced from Aureobasidium pullulans, polysaccharides produced from Aureobasidium pullulans, β-1,3-1,6-glucans, or derivatives of any of these, (vi) Bifidobacterium bifidum, and (vii) BIOHM FX, which is a composition comprising Lactobacillus acidophilus, Lactobacillus rhamnosus, Saccaromyces boulardii, Bifidobacterium breve, and alpha amylase.

[0042] In an embodiment, the composition comprises Lactobacillus rhamnosus, Bifidobacterium longum, Green tea, Cranberry extract, Bifidobacterium bifidum; and BIOHM FX. In an embodiment, the composition comprises Lactobacillus rhamnosus, Bifidobacterium longum, Green tea, Cranberry extract, and Bifidobacterium bifidum. In an embodiment, the composition comprises Lactobacillus rhamnosus, Bifidobacterium longum, Bifidobacterium bifidum; and BIOHM FX. In an embodiment, the composition includes any of the combinations above, except an amylase replaces the BIOHM FX component. In an embodiment, the composition includes any of the combinations above, except an amylase and Saccaromyces boulardii replaces the BIOHM FX component. In an embodiment, the composition includes any of the combinations above, except it includes Green tea, Cranberries, Cranberry extracts, or Nichi glucan, glucans produced from Aureobasidium pullulans, polysaccharides produced from Aureobasidium pullulans, β-1,3-1,6-glucans, or derivatives of any of these.

[0043] Additional probiotic additives in contemplated embodiments include those selected from the following: Lactobacillus casei, Lactobacillus plantarum, Lactobacillus acidophilus, Bacillus subtillis, Lactobacillus rhamnosus, Lactobacillus helveticus, Bifidobacterium longum, Bifidobacterium lactis, Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium adolescentis, Bifidobacterium breve, Saccaromyces boulardii, Brevebacillus laterosporus, and combinations thereof. All such probiotic organisms could be included in the formulation at levels of 0.1-10 billion CFU, and / or 10-500 mg.Attorney Docket No. BHMP107WO

[0044] Further additional additive components in contemplated embodiments include those selected from the following: barley, xylan, acerola and or acerola extract, isoflavone(s), soy isoflavone, moringa oleifera, guava or guava extract, berberine, doenjang, fermented soybean, P95 scFOS, FOS (fructooligosaccharide), guarana seed powder, guarana seed extract, gurarana extract, Tianwang buxin granules, red clover, dried cranberry, inulin, lactulose, hesperidan, Novasoy, PREFORPRO, grape seed extract, black cohosh, magnesium, Winclove 607, and combinations thereof. All such additive components above may be included at levels of 5- 10,000 mg

[0045] In certain embodiments, instead of the full BIOHM FX component, the composition of comprises a non-pathogenic fungal species Saccharomyces boulardii and non-pathogenic bacterial species of one or more of Lactobacillus rhamnosus, Bifidobacterium breve, and Lactobacillus acidophilus. In certain embodiments, the composition comprises a non- pathogenic fungal species Saccharomyces boulardii and non-pathogenic bacterial species Lactobacillus rhamnosus and Bifidobacterium breve. In certain embodiments, the composition comprises a non-pathogenic fungal species Saccharomyces boulardii and non-pathogenic bacterial species Lactobacillus rhamnosus and Lactobacillus acidophilus. In certain embodiments, the composition comprises a non-pathogenic fungal species Saccharomyces boulardii and non-pathogenic bacterial species Bifidobacterium breve and Lactobacillus acidophilus. In certain embodiments, the composition comprises a non-pathogenic fungal species Saccharomyces boulardii and the non-pathogenic bacterial species Lactobacillus rhamnosus, Bifidobacterium breve, and Lactobacillus acidophilus.

[0046] In certain embodiments, instead of the full BIOHM FX component, the composition of the invention comprises a non-pathogenic fungal strain Saccharomyces boulardii SB 48 and non-pathogenic bacterial strains of one or more of Lactobacillus rhamnosus LB 20, Bifidobacterium breve S 46, and Lactobacillus acidophilus RP 32. In certain embodiments, the composition comprises a non-pathogenic fungal strain Saccharomyces boulardii SB 48 and non- pathogenic bacterial strains Lactobacillus rhamnosus LB 20 and Bifidobacterium breve S 46. In certain embodiments, the composition comprises a non-pathogenic fungal strain Saccharomyces boulardii SB 48 and non-pathogenic bacterial strains Lactobacillus rhamnosus LB 20 and Lactobacillus acidophilus RP 32. In certain embodiments, the composition comprises a non- pathogenic fungal strain Saccharomyces boulardii SB 48 and non-pathogenic bacterial strains Bifidobacterium breve S 46 and Lactobacillus acidophilus RP 32. In certain embodiments, the composition comprises a non-pathogenic fungal strain Saccharomyces boulardii SB 48 and theAttorney Docket No. BHMP107WO non-pathogenic bacterial strains Lactobacillus rhamnosus LB 20, Bifidobacterium breve S 46, and Lactobacillus acidophilus RP 32.

[0047] Including an enzyme in the composition (e.g., the amylase in BIOHM FX or another suitable enzyme) disrupts extracellular matrix polysaccharides (EPSs) present in the biofilm making the pathogenic organisms more open to challenge by the non-pathogenic bacterial and fungal strains in the dosage forms. In certain embodiments, upon administration of the composition, the non-pathogenic bacterial strains (i) displace the pathogenic bacteria in the biofilm, (ii) interfere with the attachment of the pathogenic bacteria or fungi to a substratum of the biofilm, (iii) displace the pathogenic bacteria / fungi from an extracellular polymeric matrix present in the biofilm, (iv) prevent filamentation of the pathogenic fungi in the biofilm, or (v) a combination of any of the foregoing. In certain embodiments, administration of the composition permits restoration of the natural microbiome of the region in the subject and reduces the growth and / or development of pathogenic bacteria and / or pathogenic fungi.

[0048] While present in the BIOHM FX component, in some embodiments, the probiotic composition may also comprise an additional enzyme or an enzyme may be included if the BIOHM FX component is not included. For example, the enzyme may be capable of disrupting a biofilm that may block or otherwise retard nutrient absorption. The inclusion of such an enzyme is particularly useful if subject has a biofilm of pathogenic bacteria and / or fungi. The microorganisms within the biofilm that are disrupted by the composition may comprise any type of pathogenic microorganism, e.g., bacteria (for example, Gram-positive bacteria and Gram- negative bacteria), fungi (for example, yeast and mold), archaea, and protozoa.

[0049] In certain embodiments, the probiotic composition further comprises an enzyme capable of disrupting a biofilm in a subject. In certain embodiments, the enzyme is selected from the group consisting of amylase, cellulase, hemicellulase, lysozyme, pectinase, DNase I, Serratia peptidase or Serratiopeptidase, hemicellulase / pectinase complex, β-1,3-glucanase, acid protease, alkaline protease, glucoamylase, endoglucanase, xylanase, lipase, lysozyme, protease / peptidase complex, dipeptidyl peptidase IV (DPP-IV), chitosanase, bromelain, papain, kiWi protease actinidi, a plant-derived protease, zymolase, nuclease.and phytase. In certain embodiments, the enzyme is an amylase selected from the group consisting of Bacillus stearothermophilus amylase, Bacillus amyloliquefaciens amylase, Bacillus subtilis amylase, Bacillus licheniformi amylase, Aspergillus niger amylase, and Aspergillus oryzae amylase.

[0050] In certain embodiments, the composition comprises about 500 SKB units of amylase, selected from α–Amylase, an endo-hydrolase that catalyzes the hydrolysis of internal α-1, 4-Attorney Docket No. BHMP107WO glycosidic linkages in starch to yield products like glucose and maltose, β-Amylase, an exo- hydrolase enzyme that hydrolyses α -1, 4-glucan linkages to yield successive maltose units, and γ-Amylase, which cleaves α -1, 6-glycosidic linkages, in addition to cleaving the last α -1, 4- glycosidic linkages to yield glucose, or a combination thereof.

[0051] In certain embodiments, the probiotic composition comprises 100 to 5,000 SKB units of amylase, 200 to 4,000 SKB units of amylase, from 300 to 2,000 SKB units of amylase, or 400 to 1,000 SKB units of amylase. In certain embodiments, the composition comprises 500 SKB units of amylase. An SKB or Sandstedt, Kneen, and Blish unit refers to the amount of amylase to catalyze 1 µmole substrate per minute. In certain embodiments, composition comprises a cellulose, for example, and comprises 100 to 300 CU (Cellulase unit) units per unit composition, for example, 200 CU. In certain embodiments, composition comprises a hemicellulose / pectinase complex, and comprises 60 to 100 HSU (Hemicellulose Specific Units) units per unit composition, for example, 80 HSU. In certain embodiments, the composition comprises a β- gluconase, and comprises 6 to 10 BGU (Beta Glucanase Unit), units per unit composition, for example, about 8 BGU. In certain embodiments, the composition comprises an acid protease, and comprises 15 to 25 SAP (Shrimp Alkaline Phosphatase) units per unit composition, for example, about 20 SAP units. In certain embodiments, the composition comprises alkaline protease, and comprises 15 to 25 HUT (hemoglobin unit Tyrosine base) units per unit composition, for example, about 20 HUT units.

[0052] The enzyme may be chosen depending upon the type of biofilm and the microorganisms disposed therein. For example, an amylase enzyme may be used to degrade or otherwise disrupt carbohydrate components of the biofilm, and a nuclease such a DNase I may be used for digest or otherwise disrupt DNA in the biofilm.

[0053] In certain embodiments, the composition comprises two or more (e.g., 2, 3, 4, 5, or more) different enzymes selected from amylase, cellulase, hemicellulase, lysozyme, pectinase, DNase I, Serratia peptidase, Serratiopeptidase, hemicellulase / pectinase complex, β-1,3- glucanase, acid protease, alkaline protease, glucoamylase, endoglucanase, xylanase, lipase, lysozyme, protease / peptidase complex, dipeptidyl peptidase IV (DPP-IV), chitosanase, bromelain, papain, kiWi protease actinidi, a plant-derived protease, phytase, zymolase and nuclease.

[0054] In certain embodiments, total amount of cellulase administered, for example, by the administration of one or more units of the composition, may be 1 to 10,000 CU, the total amount of hemicellulase / pectinase complex administered, for example, by the administration of one orAttorney Docket No. BHMP107WO more units of the composition, may be 1 to 8,000 HSU, the total amount of β-gluconase, for example, by the administration of one or more dosage units, may be 1 to 1000 BGU, the total amount of acid protease, for example, by the administration of one or more dosage units, may be 1 to 10,000 SAP, and the total amount of alkaline protease, for example, by the administration of one or more dosage units, may be 1 to 40,000 HUT.

[0055] In certain embodiments, a component of the composition described herein is used to disrupt a biofilm disposed within the GI tract of a subject with an elevated relative abundance of pathogenic bacterial species such as Solobacteria moorei, E. coli, Serratia marcescens, and Ruminococcus gnavus in the biofilm disposed within the GI tract of the subject when compared to levels typically found in healthy subjects. For example, the GI tract biofilm may comprise: Candida tropicalis and Escherichia coli; Candida tropicalis and Serratia marcescens; Candida tropicalis, Escherichia coli, and Serratia marcescens; other Candida yeasts, Klebsiella oxytoca, or Solobacteria moorei.

[0056] In certain embodiments, a composition described herein comprises non-pathogenic bacterial strains for disrupting or replacing a biofilm containing two pathogenic bacterial species. For example, the biofilm targeted for disruption or replacement may contain E. coli and a Bacteroides spp. (e.g., Bacteroides coprophilus, Bacteroides eggerthii, Bacteroides ovatus, Bacteroides fragilis, Bacteroides plebeius, or Bacteroides uniformis), or Solobacteria moorei.

[0057] In certain embodiments, the abundance of pathogenic fungus Candida tropicalis in a biofilm disposed within the GI tract of a subject is elevated when compared to levels typically found in healthy subjects. In certain embodiments, biofilms within the GI tract containing C. tropicalis or C. Albicans combined with E. coli and / or S. marcescens are enriched in fungal hyphae, a form of growth associated with pathogenic conditions. Fungal filamentation can sometimes be a virulence factor used by Candida to damage host tissues and to trigger a specific host immune response. In certain embodiments, the dosage form of the invention is used to disrupt a biofilm disposed within the GI tract of a subject with an elevated relative abundance of bacterial species such as species of the Firmicutes phylum. In certain embodiments, the dosage form of the invention is used to disrupt a biofilm disposed within the GI tract of a subject with an elevated relative abundance of fungal species such as species of the Ascomycota phylum and species of the Zygomycota phylum and species of the Basidiomycota phylum.

[0058] The bacteria within the biofilm exist in intimate contact with the fungus but may differ in their specific interactions with the fungus. In certain embodiments, the pathogenic bacteria (e.g., E. coli) may be fused to the fungal cells within the biofilm. Alternatively or in addition,Attorney Docket No. BHMP107WO the pathogenic bacteria and pathogenic fungi disposed within the biofilm may form a “digestive plaque,” where the bacteria and fungi are protected from antimicrobial drugs and host’s immune system. It is understood that digestive plaque can disrupt the normal or healthy microbiome of the GI tract, and cause or be otherwise associated with a GI disease or disorder.

[0059] While non-pathogenic bacterial strains are present in the BIOHM FX component, in some embodiments, the probiotic composition may also comprise or as a substitute for the BIOHM FX component comprise an additional at least two different non-pathogenic bacterial strains. In certain embodiments, the probiotic composition comprises three different non- pathogenic bacterial strains. In certain embodiments, the non-pathogenic bacterial strain is selected from: Lactobacillus rhamnosus, Bifidobacterium breve, Lactobacillus acidophilus, Bifidobacterium bifidum, and Lactobacillus reuteri. In certain embodiments, the non-pathogenic bacterial strain is selected from: Lactobacillus rhamnosus LB 20, Bifidobacterium breve S 46, and Lactobacillus acidophilus RP 32. In certain embodiments, the probiotic composition comprises Lactobacillus rhamnosus LB 20, Bifidobacterium breve S 46, and Lactobacillus acidophilus RP 32. In certain embodiments, the non-pathogenic fungal strain is selected from: Saccharomyces boulardii, Saccharomyces cerevisiae, Saccharomycetes sp HZ178, Saccharomyces bayanus, Pichia burtonii, Pichia jadinii, Pichia kudriavzevii, Pichia onychis, Pichia sp., and Picoa juniper. In certain embodiments, the non-pathogenic fungal strain is Saccharomyces boulardii SB 48.

[0060] In certain embodiments, the probiotic composition comprises 1 to 30, 3 to 25, 10 to 20, or 15 billion colony forming units of Bifidobacterium breve, about 10 billion colony forming units of Lactobacillus rhamnosus, 0.5 to 10, 1 to 8, 2 to 5 or 3.5 billion colony forming units of Saccharomyces boulardii, 0.5 to 5, 1 to 3.5, 1.25 to 2, or 1.5 billion colony forming units of Lactobacillus acidophilus, and 100 to 1000, 250 to 750, 450 to 550 or 500 SKP of Amylase. In certain embodiments, the probiotic composition comprises an admixture (blend) of one or more isolated and viable non-pathogenic bacterial strains and one or more isolated and viable non- pathogenic fungal strains is in the form of a powder. In certain embodiments, the probiotic composition comprises a powder of one or more isolated and viable non-pathogenic bacterial strains and one or more isolated and viable non-pathogenic fungal strains that is coated with functional coating (for example, coated with a controlled release coating) or a non-functional coating (for example, aesthetic coating). In certain embodiments, the probiotic composition is in an oral dosage form. In certain embodiments, the oral dosage form is a capsule, for example, a capsule that optionally is coated with a functional or non-functional coating.Attorney Docket No. BHMP107WO

[0061] In addition to the components listed above, the composition may comprise one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) different non-pathogenic bacterial strains.

[0062] In certain embodiments, exemplary bacterial strains to be included in the composition of the present invention may comprise bacterial strains of any one or more of the following bacterial species: Agrococcus jenensis, Alistipes indistinctus, Alistipes massiliensis, Alkalibacterium iburiense, Anoxybacillus kestanbolensis, Bacillus cereus, Bacillus clausii, Bacillus Coagulans, Bacteroides coprophilus, Bacteroides eggerthii, Bacteroides ovatus, Bacteroides fragilis, Bacteroides plebeius, Bacteroides uniformis, Bifidobacterium adolescentis, Bifidobacterium animalis, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium pseudolongum, Blautia obeum, Blautia product, Candidatus azobacteroides, Candidatus portiere, Candidatus Portiera, Clostridium celatum, Clostridium hiranonis, Clostridium neonatale, Clostridium perfringens, Clostridium tyrobutyricum, Collinsella aerofaciens, Collinsella stercoris, Coprococcus eutactus, Corynebacterium stationis, Desulfosporosinus meridiei, Desulfovibrio D168, Dorea formicigenerans, Eggerthella lenta, Erwinia oleae, Faecalibacterium prausnitzii, Lactobacillus agilis, Lactobacillus reuteri, Lactobacillus ruminis, Lactobacillus salivarius, Lactobacillus zeae, Listeria weihenstephanensis, Paenibacillus mucilaginosus, Parabacteroides distasonis, Pediococcus acidilactici, Peptostreptococcus anaerobius, Prevotella copri, Prevotella melaninogenica, Prevotella stercorea, Propionibacterium acnes, Pseudoramibacter eubacterium, Roseburia faecis, Rothia dentocariosa, Rothia mucilaginosa, Ruminococcus bromii, Ruminococcus callidus, Ruminococcus flavefaciens, Ruminococcus lavefaciens, Ruminococcus gnavus, Ruminococcus torques, Salinibacillus aidingensis, Staphylococcus sciuri, Streptococcus anginosus, Streptococcus sobrinus, Tissierella soehngenia, Veillonella dispar, and Veillonella parvula.

[0063] In certain embodiments, one or more of the bacterial strains listed in TABLE 1 below are included in the composition. TABLE 1 Bifidobacterium adolescentis Bacillus clausiiAttorney Docket No. BHMP107WO Bacillus cereus Candidatus Portiera Eggerthella lenta Coprococcus eutactus

[0064] E 2 below are inclue e co pos o . TABLE 2 Bifidobacterium adolescentis Eggerthella lenta

[0065] In certain embodiments, the non-pathogenic bacterial strains included in the composition of the present invention may comprise a combination of any two of Lactobacillus rhamnosus LB 20, Bifidobacterium breve S 46, and Lactobacillus acidophilus RP 32. In certain embodiments, the non-pathogenic bacterial strains included in the composition may comprise a combination of Lactobacillus rhamnosus LB 20 and Bifidobacterium breve S 46. In certain embodiments, the non-pathogenic bacterial strains included in the composition may comprise a combination of Lactobacillus rhamnosus LB 20 and Lactobacillus acidophilus RP 32. In certain embodiments, the non-pathogenic bacterial strains included in the composition may comprise aAttorney Docket No. BHMP107WO combination of Bifidobacterium breve S 46 and Lactobacillus acidophilus RP 32. In certain embodiments, the non-pathogenic bacterial strains included in the composition of the present invention may comprise a combination of Lactobacillus rhamnosus LB 20, Bifidobacterium breve S 46, and Lactobacillus acidophilus RP 32.

[0066] In certain embodiments, the composition comprises 10 billion to 40 billion, e.g., 15 billion to 40 billion, 20 billion to 40 billion, 10 billion to 30 billion, 15 billion to 30 billion, or 20 billion to 30 billion colony forming units of the non-pathogenic bacterial strain(s).

[0067] In certain embodiments, exemplary fungal strains to be included in the composition comprise any one or more of the following fungal organisms: Albatrellus syringae, Alternaria alli, Alternaria daturicola, Ambispora granatensis, Amyloathelia crassiuscula, Amylomyces rouxii, species of Ascomycota, Ascosphaera apis, Aspergillus flavus, Aspergillus fumigatus, Aspergillus oryzae, Aspergillus sp, Aspergillus terreus, Aspergillus versicolor, Bettsia alvei, Botryosphaeria mamane, Botryotinia fuckeliana, Candida arabinofermentans, Candida ernobii, Candida ethanolica, Candida glabrata, Candida humilis, Candida intermedia, Candida parapsilosis, Candida piceae, Candida quercitrusa, Candida tartarivorans, Candida temnochilae, Candida zemplinina, Candida zeylanoides, Chamonixia caespitosa, Cladonia polystomata, Cladosporium cladosporioides, Cladosporium halotolerans, Cladosporium sp JP67, Cladosporium sphaerospermum, Clavicorona taxophila, Clavispora lusitaniae, Craterellus sp, Dactylellina phymatopaga, Debaryomyces hansenii, Debaryomyces marasmus, Debaryomyces sp, Debaryomyces subglobosus, Dipodascus australiensis, Ectomycorrhizal, Emericella nidulans, Epulorhiza sp, Eremascus fertilis, Eremothecium gossypii, Eupenicillium cinnamopurpureum, Eurotium amstelodami, Eurotium cristatum, Exophiala dermatitidis, Filobasidiella neoformans, Fonsecaea monophora, Fusarium solani, Fusarium dimerum, Fusarium oxysporum, Fusarium sp, Galactomyces geotrichum, Galactomyces sp, Geomyces sp, Geotrichum cucujoidarum, Geotrichum sp, Glomus mosseae, Hanseniaspora sp, Helicobasidium longisporum, Helicostylum pulchrum, Hyphodontia flavipora, Hypochnicium cystidiatum, Inocybe sp, Kazachstania africana, Kazachstania unispora, Kluyveromyces lactis, Kluyveromyces yarrowii, Kodamaea ohmeri, Leptosphaeria biglobosa, Leptosphaerulina chartarum, Leucoagaricus sp, Lewia infectoria, Lichtheimia ramose, Macrophomina phaseolina, Monacrosporium coelobrochum, Mucor circinelloides, Mucor flavus, Mucor fuscus, Mucor sp, Myrothecium sp, Myxozyma melibiosi, Neocallimastix frontalis, Neocallimastix sp, Omphalotus nidiformis, Ophiocordyceps filiformis, Ophiocordyceps sinensis, Penicillium carneum, Penicillium chrysogenum, Penicillium concentricum, Penicillium crustosum, Penicillium digitatum, Penicillium granulatum, Penicillium griseofulvum, Penicillium griseoroseum,Attorney Docket No. BHMP107WO Penicillium polonicum, Penicillium psychrosexualis, Penicillium pulvillorum, Penicillium roqueforti, Penicillium sclerotigenum, Penicillium sp, Penicillium spinulosum, Penicillium verrucosum, Pezizomycetes sp genotype 323, Phaeophyscia exornatula, Phialocephala lagerbergii, Phlebia radiata, Phlebia sp, Phomopsis sp, Physcia stellaris, Physoderma maydis, Pichia burtonii, Pichia jadinii, Pichia kudriavzevii, Pichia onychis, Pichia sp, Picoa juniperi, Pilaira cesatii, Pilaira sp, Pirella circinans, Preussia sp, Reddellomyces donkii, Rhizomucor pusillus, Rhizopus oryzae, Rhodosporidium kratochvilovae, Rhodosporidium sp, Rhodotorula sp, Rinodina milvina, Saccharomyces bayanus, Saccharomyces bulderi, Saccharomyces cerevisiae, Saccharomyces mikatae, Saccharomyces sp, Saccharomycetes sp, Scleroderma sp, Sclerotinia sp, Scutellospora nodose, Sebacinales sp, Sporopachydermia sp, Stenocarpella maydis, Thamnidium elegans, Torulaspora delbrueckii, Trichosporon chiarellii, Tuber oligospermum, Umbelopsis isabellina, Wallemia sebi, Wallemia sp, and Zygoascus meyerae.

[0068] In certain embodiments, one or more of the fungal strains listed in TABLE 3 below may be included in the composition of the present invention. TABLE 3 Saccharomycetes sp HZ178 Pilaira sp Pi 3

[0069] In certain embodiments, the composition may comprise a non-pathogenic fungal strain selected from Saccharomyces boulardii, Saccharomyces cerevisiae, Saccharomycetes sp HZ178, Saccharomyces bayanus, strains belonging to the Pichia genus (for example, including but limited to Pichia burtonii, Pichia jadinii, Pichia kudriavzevii, Pichia onychis, and Pichia juniper).Attorney Docket No. BHMP107WO

[0070] In certain embodiments, the composition comprises a composition comprising a non- pathogenic fungal strain for disrupting a biofilm of a composition disclosed herein. In certain embodiments, the composition of the invention comprises one or more fungal strains listed in TABLE 3. In certain embodiments, the composition of the invention comprises one or more fungal strains listed in TABLE 3 and one or more of the bacterial strains listed in TABLE 1 or TABLE 2.

[0071] In certain embodiments, the composition comprises a composition comprising a non- pathogenic fungal strain Saccharomyces boulardii SB 48. In certain embodiments, the composition of the invention comprises a non-pathogenic fungal strain Saccharomyces boulardii SB 48 and one or more of the bacterial strains listed in TABLE 1 or TABLE 2. In certain embodiments, the composition comprises 1 billion to 10 billion, e.g., 2 billion to 8 billion, from 3 billion to 6 million colony forming units of a non-pathogenic fungal strain(s).

[0072] Recent developments have provided alternative effective means of selectively modifying gut microbiomes. Bacteriophages, provide an additional option to selectively reduce populations of undesirable microorganisms. Certain bacteriophages that can target E.coli were identified as being useful in the present composition to reduce E. coli in the guy and to promote growth of Bifidobacterium taxa, including B. longhum, B. bifidum, and B. breve, which as disclosed herein, were identified as deficient in the menopausal aged women cohort. Other benefits may also be realized from the reduction of E. coli in the microbiome, such as decrease in circulating pro-inflammatory cytokine interleukin 4 (Il-4), which has been associated with autoimmune and allergic responses in human population. Reference can be made to Febvre HP, Rao S, Gindin M, Goodwin NDM, Finer E, Vivanco JS, Lu S, Manter DK, Wallace TC, Weir TL. PHAGE Study: Effects of Supplemental Bacteriophage Intake on Inflammation and Gut Microbiota in Healthy Adults. Nutrients.2019; 11(3):666, which is incorporate herein by reference.

[0073] In an embodiment, the E. coli targeting phages are selected from myoviridae and / or siphoviridae and are effective to reduce E. coli in vitro or in the human gut. In an embodiment, the bacteriophage strains are selected from the group consisting of LH01-Myoviridae, LL5- Siphoviridae, T4D-Myoviridae, and LL12-Myoviridae. In an embodiment, the composition of PREFORPRO is included in the composition disclosed herein.

[0074] The bacteriophages may be present in a titer of, for example, 1x104to 1x108, such as 1x105to 1x107, or about 1x106phages per dose / capsule. In an embodiment, the bacteriophages may be present in the composition in an amount of 1 mg to 1000 mg, such as 5 mg to 750 mg, orAttorney Docket No. BHMP107WO 10 mg to 500 mg. The phages can be added to the composition as a freeze dried product, and can be processed along with the freeze-dried probiotics disclosed herein.

[0075] In addition to the above combinations, the composition may also include combinations of enzymes (bromelain, cellulase(es), lipase(es), papain, protease(es) amylase(es), galactosidase) in the 10-250 mg range, such as, for example, 25 to 200 mg, or 50 to 150 mg. Bacopa monnieri or Bacopa monnieri extracts can be present in a 10-500 mg range, such as 25 to 300 mg, or 100 to 200 mg. In addition, Lions Mane mushroom or Lion’s Mane mushroom extracts can be present in a 10-500 mg range, such as 25 to 300 mg, or 100 to 200 mg.

[0076] Other additives to any of the above combinations can be present in a total amount of 1 to 15000 mg, such as 100 mg to 10000, or 1500 to 5000 mg. Total fiber content can be present in a range of 500 mg to 5,000 mg), such as, for example, 1000 mg to 4000 mg, or 2000 mg to 3000 mg. Golden Kiwi Fiber can be present in an amount of 500 mg to 5,000 mg, such as, for example, 1000 mg to 4000 mg, or 2000 mg to 3000 mg.

[0077] In certain embodiments, the composition can be naturally sweetened, for example, with monk fruit, and preferably does not contain an allergen, artificial ingredient, or sweetener, and / or can be stored at room or ambient temperature, without refrigeration. The composition can be an engineered formulation that combines fruits, vegetables and herbal extracts with probiotics, prebiotics and enzymes to break down digestive plaque and achieve total gut balance. Depending upon the circumstances, the organic super green composition can further comprise organic plant extract (e.g., spirulina, barley grass, alfalfa leaf, wheat grass, chlorella, dulse, spinach leaf, broccoli (whole plant), parsley leaf, kale leaf, Echinacea angustifolia root, licorice root, milk thistle seed, Siberian Eleuthero root, beet root, rose hips, acai (fruit), green tea leaf, raspberry leaf, blueberry (fruit), goji berry, bilberry (fruit), ashwagandha root, rhodiola root, reishi mushroom, maca root, bee pollen, nettle leaf, ginko biloba (4:1 leaf extract), royal jelly (3x concentrate), grape seed (2:1 extract), ginger, plant-based protein (e.g., pea protein), and fiber (e.g., sunflower lecithin, apple (fruit), brown rice bran, inulin). A nutritional ingredient may be selected from one or more of: a prebiotic (e.g., inulin), collagen peptides, deglycyrrhizinated licorice (DGL), marshmallow root, vitamin A (e.g., retinyl form), vitamin C, and vitamin D3.

[0078] In certain embodiment, the composition comprises additives such as calcium carbonate, xylitol, cetyl alcohol, citric acid, natural flavor, monk fruit. In certain embodiments, the composition comprises additives such as cascara sagrada bark, psyllium husk, senna leaf, Flaxseed, aloe vera leaf, licorice root, medium chain triglyceride (MCT) oil. In certain embodiments, the composition comprises additives such as a dietary fiber, e.g., inulinAttorney Docket No. BHMP107WO (fructooligosaccharides FOS) and apple pectin. In certain embodiments, the composition comprises additives such as a blend of spirulina, barley grass, alfalfa leaf, wheat grass, chlorella, dulse, spinach leaf, broccoli, parsley leaf, kale leaf, echinacea angustifolia root, licorice root, milk thistle seed, Siberian eleuthero root, beet root, rose hips, acai (fruit), green tea leaf, raspberry leaf, blueberry (fruit), goji berry, bilberry (fruit), ashwagandha root, rhodiola root, reishi mushroom, maca root, bee pollen, nettle leaf, ginko biloba (leaf extract), royal jelly (3x concentrate), grape seed. In certain embodiments, the composition further comprises a vitamin, such as, vitamin C.

[0079] In certain embodiments, the composition comprises 100 mg to 2,000 mg of a prebiotic, such as inulin (e.g., 200 mg to 1500 mg, or 300 to 1000 mg). In certain embodiments, the composition comprises 1,000 mg to 20,000 mg of collagen peptides (e.g., 1,000 mg to 15,000 mg, or 2,000 to 10,000 mg). In certain embodiments, the composition comprises 100 mg to 1,000 mg of DGL or marshmallow root (e.g., 150 mg to 700 mg, or 200 to 500 mg).

[0080] In certain embodiments, the composition comprises 1,000 IU to 10,000 IU of vitamin A (e.g., 1,500 to 7,000, or 3,000 to 6,000 IU). In certain embodiments, the composition comprises 500 IU to 1,500 IU of vitamin D3 (e.g., 700 to 1,250 IU, or 850 to 1,100 IU). In certain embodiments, the composition comprises 100 mg to 1,000 mg of vitamin C (e.g., 200 mg to 800 mg, or 250 to 600 mg).

[0081] In certain embodiments, the composition comprises 1,000 mg to 20,000 mg of a plant- based protein, such as pea protein (e.g., 1,200 mg to 15,000 mg, or 2,000 mg to 5,000 mg). In certain embodiments, the composition comprises about 10 mg to about 200 mg of grape seed extract (e.g., 20 mg to 150 mg, 45 mg to 100 mg). In certain embodiments, the composition comprises 100 mg to 1,000 mg of ginger (e.g., 125 mg to 750 mg, or 250 mg to 550 mg).

[0082] In certain embodiments, a composition of the composition may further comprises one or more of the following ingredients, including spirulina, chlorella, dulse, barley grass, alfafa juice concentrate, wheat grass, spinach leaf, kale leaf, beet root, rose hips, acai fruit, raspberry fruit, blueberry fruit, goji berry, bilberry fruit extract, green tea leaf extract, ginkgo biloba, Echinacea purpurea root, licorice root extract, milk thistle extract, stinging nettle root, royal jelly, grape seed extract, bee pollen, maca root, reishi mushroom, rhodiola root, ashwagandha root, and / or fiber (e.g., sunflower lecithin, apple fiber, brown rice bran). In certain embodiments, a composition of the present invention can be flavored using stevia or monk fruit, or another natural flavor (e.g., natural blueberry flavor).Attorney Docket No. BHMP107WO

[0083] The compositions or dosage forms described herein, can be administered or consumed individually or in combination. For example, a composition that includes the Lactobacillus rhamnosus, Bifidobacterium longum, Green tea, Cranberry extract, Nichi glucan, glucans produced from Aureobasidium pullulans, polysaccharides produced from Aureobasidium pullulans, or β-1,3-1,6-glucans, and BIOHM FX may be administered or consumed individually as such, or may be co-administered or concurrently consumed with other nutrients or supplements.

[0084] In another aspect, the composition or method of treatment can comprise a nutrient in combination with the probiotic composition. The combination does not require that the nutrient and the probiotic composition be present in a single composition. In certain embodiments, the composition comprising a nutrient comprises both the nutrient and the probiotic composition.

[0085] In certain embodiments, the consumption by, or administration to the subject of the composition comprising a nutrient is contemporaneous with the consumption or administration of the probiotic composition. In certain embodiments, the consumption by, or administration to the subject of the composition comprising a nutrient is subsequent to or prior the consumption by, or administration of the probiotic composition.

[0086] In certain embodiments, a nutrient is included in the composition. In an embodiment, the nutrient is a protein. The protein may be selected from whey protein, soy protein, milk protein, egg protein, animal muscle protein, fish protein, brown rice protein, pea protein, hemp protein, cranberry protein, and artichoke protein. In certain embodiments, the protein is casein. In certain embodiments, the protein is a protein hydrolysate. In certain embodiments, the protein is comprised in a natural food source. In certain embodiments, the protein is an isolated and / or purified protein.

[0087] In another aspect, the nutrient may comprise a beverage product comprising a probiotic composition comprising the probiotic and enzyme components discussed above. The beverage can be a protein-rich beverage, e.g., comprising at least 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% of protein. The probiotic composition can include one or more of the compositions or dosage forms described herein.

[0088] Compositions or dosage forms are provided herein that can moderate or improve menopausal symptoms. In certain embodiments, the composition is formulated as a powdered blend of the components described above that optionally is coated with a functional coating (e.g., controlled release coating) or a non-functional coating (e.g., aesthetic coating). In certain embodiments, the powder blend of the composition is freeze-dried or spray-dried. In certainAttorney Docket No. BHMP107WO embodiments, the freeze-dried or spray-dried blend of the composition is further blended with an enzyme (for example, amylase). The freeze-dried or spray dried components may still be live. The components can be live micro-encapsulated, live spore probiotic, or for “postbiotic” organisms the organisms can be heat killed / heat inactivated.

[0089] Controlled release coatings can facilitate the continuous release, gradual release, prolonged release, and / or programmed release (e.g., pH-dependent release) of the microorganisms in the compositions or dosage forms disclosed herein. In certain embodiments, controlled release coating is hydroxypropyl methylcellulose (HPMC).

[0090] While typically the composition is designed for human subjects, primarily menopausal aged women, in certain situations, the subject may be a non-human mammal (e.g., human, a companion animal (e.g., dog, cat, or rabbit), or a livestock animal (for example, cow, sheep, pig, goat, horse, donkey, and mule, buffalo, oxen, or camel)). In certain embodiments, the subject is a healthy subject.

[0091] In certain embodiments, 30-50 of such capsules (e.g., 35-45 capsules) can be consumed over a period of 2-8 weeks (e.g., 3-6 weeks). In certain embodiments, the product can be consumed by a subject for up to one month (e.g., 30 days), or repeated cycles of one month. In each cycle, a single serving size (e.g., 1 scoop) may be consumed per day.

[0092] For example, one capsule can be consumed per day, with or without the simultaneous consumption of food. The probiotic composition preferably does not contain an allergen, artificial ingredient, or sweetener, and / or can be stored at room or ambient temperature, without refrigeration.

[0093] In certain embodiments, the composition may be consumed orally by the subject. In certain embodiments, the composition is consumed as a powder or product containing the powder. In other embodiments, the composition can be in the form of an oral dosage form, for example, where the composition in included, for example, within a capsule, cachet, pill, tablet, lozenge, powder, granule, or as a solution or a suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles, each containing the requisite number of colony forming units of the non-pathogenic bacteria and non- pathogenic fungus, and optionally the appropriate amount of enzyme.

[0094] In certain embodiments, the composition is disposed in a capsule or in a tablet. In certain embodiments, the composition is formulated as a tablet. In certain embodiments, the capsule is a vegetable cellulose capsule. In certain embodiments, the dosage form, for example, capsule or tablet, is coated with a coating, for example, a non-functional aesthetic coating or aAttorney Docket No. BHMP107WO functional coating, for example, a controlled release coating. The capsule or tablet may be formulated so as to provide slow or controlled release of the ingredients disposed therein.

[0095] The size of the dose delivered or consumed, will depend upon, among other things, the size and age of the subject, the indication or condition to be treated, and the mode of delivery of the composition. In certain embodiments, for example, oral compositions may be 1 mg to 10,000 mg or 50 mg to 10,000 mg of the active ingredients (for example, the non-pathogenic bacteria, the non-pathogenic fungus, and / or the enzyme). In certain embodiments, the composition may be 1 mg to 9,000 mg, 1 mg to 8,000 mg, 1 mg to 7,000 mg, 1 mg to 6,000 mg, 1 mg to 5,000 mg, 1 mg to 4,000 mg, 1 mg to 3,000 mg, 1 mg to 2,000 mg, 1 mg to 1,000 mg, 1 mg to 500 mg, 10 mg to 1000 mg, 20 mg to 1000 mg, 30 mg to 1000 mg, 40 mg to 1000 mg, 50 mg to 1000 mg, 100 mg to 1000 mg, 200 mg to 1000 mg, 300 mg to 1000 mg, 400 mg to 1000 mg, 500 mg to 1000 mg, 600 mg to 1000 mg, 700 mg to 1000 mg, 800 mg to 1000 mg, or 900 mg to 1000 mg of the active ingredients (for example, those disclosed above or in the prophetic Example 3, 4, or 5).

[0096] The compositions (either one or multiple units (for example 2, 3, 4, or 5 units, for example, capsules or tablets) may be administered once, twice or three times a day during the treatment period, for example, until the biofilm of interest has been disrupted, prevented from forming, and / or the subject’s normal microbiome has been restored which could take, for example, one week, two weeks, one month, two months, or three months and one year. In certain embodiments, one capsule, such as one enteric-coated capsule, that comprises about 30 billion colony forming units of non-pathogenic fungal strain(s) and non-pathogenic bacterial strain(s) and an enzyme, is administered once, twice, or three times a day during the treatment period, for example, until the biofilm of interest has been disrupted and / or the subject’s normal microbiome has been restored which could take, for example, one week, two weeks, one month, two months, or three months and one year.

[0097] In certain embodiments, the composition may be formulated for topical delivery, for example, as a liquid, emulsion, suspension, ointment, cream, gel, lotion, or powder. In certain embodiments, upon application to a skin of a subject, the composition may form a patch. Depending up the concentration of non-pathogenic organisms and the particular delivery device, the composition may be formulated to deliver from 1.0 mL / 5 cm2to 1.0 mL / 50 cm2, or from 1.0 mL / 5 cm2to 50 mL / 50 cm2, or from 1.0 mL / 5 cm2to 100 mL / 50 cm2.

[0098] The topical dosage form may also include a pharmaceutically acceptable carrier. Suitable carriers that may be useful in topical formulations include, but are not limited to,Attorney Docket No. BHMP107WO solubilizers such as C2-C8, straight and branched chain alcohols, diols and triols, moisturizers and humectants such as glycerin, amino acids and amino acid derivatives, polyaminoacids and derivatives, pyrrolidone carboxylic acids and its salts and derivatives, surfactants such as sodium lauryl sulfate, sorbitan monolaurate, emulsifiers such as cetyl alcohol, stearyl alcohol, thickeners such as methyl cellulose, ethyl cellulose, hydroxymethylcellulose, hydroxypropylcellulose, polyvinylpyrollidone, polyvinyl alcohol and acrylic polymers.

[0099] In certain embodiments, the composition may be formulated for nasal or oral administration in the form of a nasal spray and inhalation solution, and suspension, that is administered for example, by inhalation through the nose. The dose can be delivered to the nasal cavity via a spray pump or as nasal drops for local and / or systemic effects.

[0100] In certain embodiments, the composition may be an inhalation solution and / or suspension for delivery to the upper and lower respiratory tract (e.g. oropharynx, lungs) by oral inhalation for local and / or systemic effects and can be used with a specified nebulizer.

[0101] In certain embodiments, a composition of the present invention may be manufactured according to the process shown in Fig.1. The Lactobacillus rhamnosus, Bifidobacterium longum, Bifidobacterium bifidum; and optionally the bacterial and fungal components of BIOHM FX are sourced from various sources and are then each cultured separately in a small scale fermenter. For each strain, a culture sample is then used to inoculate a corresponding large scale production fermenter. The cultured fungal and bacterial strains are harvested via filtration and / or centrifugation, and can then be dried, for example, freeze dried. The dried organisms can then optionally be coated, for example, spray coated with a polymer such as hydroxylpropylmethyl cellulose (HPMC). The coated strains, for example, HPMC-coated strains, can then be subjected to further drying, such as, low temperature drying (e.g., room temperature). The resulting strains are then blended together at the appropriate concentrations for the specific formulation being manufactured. At this blending stage, if desired, other ingredients specific to the formulation (e.g., dried amylase, Nichi Glucan, green tea and cranberry extract, and optionally the dried BIOHM FX component) can be added. If desired, the blended composition can then be encapsulated, which can then be packaged into an appropriately labeled container. An exemplary manufacturing process is depicted in Fig.1.

[0102] The invention also provides a method of identifying a test subject suitable for treatment with a composition described herein. The method comprises: (a) quantifying the number or density and / or abundance of organisms of at least one non-pathogenic bacterial strain; and (b) comparing the number or density of organisms quantified in the sample against the number or density of corresponding organisms present in a corresponding sample of a corresponding regionAttorney Docket No. BHMP107WO of a healthy subject or a cohort of healthy female subjects of the age 25-35. When the number or level of organisms in the sample from the test subject is less than the number or level of organisms present in the corresponding region of the healthy female subjects of the age 25-35, e.g., at least 10% less than, at least 25% less than, or at least 50% less than the test subject is suitable for treatment with the dosage form. In certain embodiments, the non-pathogenic bacteria that are quantified may include those disclosed as decreased in the menopausal aged cohort in the Examples below.

[0103] Other compositions, components, concentrations, and dosages that can be combined with the teachings herein are disclosed in U.S.2019 / 0381119 and U.S.2021 / 0030818, incorporated herein by reference.

[0104] The following definitions are included for the purpose of understanding the present subject matter and for constructing the appended patent claims.

[0105] Throughout the description, where compositions are described as having, including, or comprising specific components, or where processes and methods are described as having, including, or comprising specific steps, it is contemplated that, additionally, there are compositions of the present invention that consist essentially of, or consist of, the recited components, and that there are processes and methods according to the present invention that consist essentially of, or consist of, the recited processing steps.

[0106] In the application, where an element or component is said to be included in and / or selected from a list of recited elements or components, it should be understood that the element or component can be any one of the recited elements or components, or the element or component can be selected from a group consisting of two or more of the recited elements or components.

[0107] Further, it should be understood that elements and / or features of a composition or a method described herein can be combined in a variety of ways without departing from the spirit and scope of the present disclosure, whether explicit or implicit herein. For example, where reference is made to a particular composition, that composition can be used in the various embodiments of such compositions of the present invention and / or in methods of the present invention, unless otherwise understood from the context. In other words, within this application, embodiments have been described and depicted in a way that enables a clear and concise application to be written and drawn, but it is intended and will be appreciated that embodiments may be variously combined or separated without parting from the present teachings andAttorney Docket No. BHMP107WO invention(s). For example, it will be appreciated that all features described and depicted herein can be applicable to all aspects of the invention(s) described and depicted herein.

[0108] It should be understood that the expression “at least one of” includes individually each of the recited objects after the expression and the various combinations of two or more of the recited objects unless otherwise understood from the context and use. The expression “and / or” in connection with three or more recited objects should be understood to have the same meaning unless otherwise understood from the context.

[0109] The use of the term “include,” “includes,” “including,” “have,” “has,” “having,” “contain,” “contains,” or “containing,” including grammatical equivalents thereof, should be understood generally as open-ended and non-limiting, for example, not excluding additional unrecited elements or steps, unless otherwise specifically stated or understood from the context.

[0110] Where the use of the term “about” is before a quantitative value, the present invention also includes the specific quantitative value itself, unless specifically stated otherwise. As used herein, the term “about” refers to a ±10% variation from the nominal value unless otherwise indicated or inferred.

[0111] Where a molecular weight is provided and not an absolute value, for example, of a polymer, then the molecular weight should be understood to be an average molecule weight, unless otherwise stated or understood from the context.

[0112] As a general matter, compositions specifying a percentage are by weight unless otherwise specified.

[0113] As used herein, the term “nutritional ingredient” refers to one or more ingredient(s) added to a composition of the present disclosure, which can be a natural ingredient having no known adverse physiological effect in a human, or known to be beneficial or necessary for optimal physiology and health of a human.

[0114] As used herein, the terms “treat,” “treating,” or “treatment,” and other grammatical equivalents as used herein, include alleviating, abating, ameliorating, or preventing a disease, condition or symptoms, preventing additional symptoms, ameliorating or preventing the underlying causes of symptoms, inhibiting the disease or condition, e.g., arresting the development of the disease or condition, relieving the disease or condition, causing regression of the disease or condition, relieving a condition caused by the disease or condition, or stopping the symptoms of the disease or condition, and are intended to include prophylaxis.Attorney Docket No. BHMP107WO

[0115] As used herein, the terms, “subject,” “patient,” “subject in need thereof,” and “patient in need thereof” are used interchangeably herein, and refer to a living organism, including animals and humans, suffering from or prone to a disease or condition that can be treated by the methods and compositions provided herein. The subject can be a human or non-human animal.

[0116] The foregoing examples are presented herein for illustrative purposes only, and should not be construed as limiting the invention in any way. EXAMPLES

[0117] The following Examples are merely illustrative and are not intended to limit the scope or content of the invention in any way.

[0118] EXAMPLE 1

[0119] A bioinformatics database was developed from patient fecal samples that were examined for various microorganisms. Data was developed with questionnaire data correlating to the various female patients based on their reported ages. Using data obtained from samples, a cohort of women in the 45-65 year age group were analyzed for gut microbiome diversity indices, differential abundance and statistical analyses. The same was done for a cohort of women with reported ages less than 45, i.e.,25-35.

[0120] EXAMPLE 2

[0121] The analysis revealed a differential abundance of several probiotic organisms of interest that are theorized to be associated with menopausal symptoms. By comparing results to healthy controls, women with ages less than 45, i.e., the reproductive-age cohort, several bacterial and fungal species were identified that were increased / decreased in the 45-65 cohort, i.e., the menopausal-age cohort.

[0122] Attempts were made to correlate the microorganisms present or absent in the microorganism data for the menopausal-age cohort. Microorganisms that were not in a normal range were identified and are shown in Table 4, where one or more species of the identified genuses were found to be different in menopausal-age women. TABLE 4 Increased in Menopausal-Age Women Decreased in Menopausal-Age WomenAttorney Docket No. BHMP107WO Psuedomonas Haemophilus Pantoea Prevotella

[0124] A composition was formulated to comprise multiple components that would address certain deficiencies in the menopausal-age cohort. This composition comprised: 1) Bifidobacterium longum—heat treated and post-biotic form 2) BIOHM FX 3) PREFORPRO

[0125] PROPHETIC EXAMPLE 1

[0126] A composition will be formulated to comprise multiple components that would address certain deficiencies in the menopausal-age cohort. This composition comprised: 4) Lactobacillus rhamnosus 5) Bifidobacterium longum 6) Green tea extract 7) Cranberry extract 8) Bifidobacterium bifidum 9) BIOHM FX

[0127] PROPHETIC EXAMPLE 2

[0128] A composition will be formulated to comprise multiple components that would address certain deficiencies in the menopausal-age cohort. This composition comprised: 1) Lactobacillus rhamnosus 2) Bifidobacterium longum 3) Green tea extractAttorney Docket No. BHMP107WO 4) Cranberry extract 5) Bifidobacterium bifidum 6) BIOHM FX 7) Nichi Glucan

[0129] PROPHETIC EXAMPLE 3

[0130] Testing and development formulation components will begin with in vitro assays. Growth promotion assays will be performed to identify an appropriate prebiotic fiber to optimize the growth of the composition of Example 3, or prophetic examples 1 or 2. Minimum inhibitory concentration (MIC) assays and or growth promotion assays will be completed to determine whether the select organisms are antagonistic to opportunistic pathogens.

[0131] PROPHETIC EXAMPLE 4

[0132] Further in vivo assays in mice with the composition Example 3, or prophetic examples 1 or 2can be conducted to study the effect on menopausal symptoms.

[0133] PROPHETIC EXAMPLE 5

[0134] A human clinical trial will look at subjects in the two cohorts females 25-35 years old and females 45-65 years old. Subjects will have blood and stool samples taken at baseline for analysis. Blood samples will include (but not limited to) baseline safety work as well as markers for inflammation (Full list to be determined as part of protocol development). Stool will be analyzed for microbiome composition. Other stool and blood markers may be added as part of the study design. Questionnaires (e.g., GSRS for GI symptoms, Quality of Life score) including questions on menopausal type symptoms will be administered and reviewed. Subjects will then be randomized to either active or placebo arms for a period of time and the above tests repeated (based on design, may be repeated more than once). The active trial will involve taking a regular dosage of the composition of Example 3, or prophetic examples 1 or 2.

[0135] The entire disclosure of each of the patent and scientific documents referred to herein is incorporated by reference.

[0136] What has been described above includes examples of one or more embodiments. It is, of course, not possible to describe every conceivable modification and alteration of the above devices or methodologies for purposes of describing the aforementioned aspects, but one of ordinary skill in the art can recognize that many further modifications and permutations of various aspects are possible. Accordingly, the described aspects are intended to embrace allAttorney Docket No. BHMP107WO such alterations, modifications, and variations that fall within the spirit and scope of the appended claims. Furthermore, to the extent that the term “includes” is used in either the detailed description or the claims, such term is intended to be inclusive in a manner similar to the term “comprising” as “comprising” is interpreted when employed as a transitional word in a claim. The term “consisting essentially” as used herein means the specified materials or steps and those that do not materially affect the basic and novel characteristics of the material or method. Unless the context indicates otherwise, all percentages and averages are by weight. If not specified above, the properties mentioned herein may be determined by applicable ASTM standards, or if an ASTM standard does not exist for the property, the most commonly used standard known by those of skill in the art may be used. The articles “a,” “an,” and “the,” should be interpreted to mean “one or more” unless the context indicates the contrary.

Claims

Attorney Docket No. BHMP107WO WHAT IS CLAIMED IS:

1. A method of improving health of a female subject aged 45-65, the method comprising consumption by, or administration to, the subject of a probiotic composition comprising three or more of the following components (i)-(vii): (i) Lactobacillus rhamnosus, (ii) Bifidobacterium longum, (iii) Green tea or derivatives thereof, (iv) Cranberries, Cranberry extract or derivatives thereof, (v) Nichi glucan, glucans produced from Aureobasidium pullulans, polysaccharides produced from Aureobasidium pullulans, β-1,3-1,6-glucans, or derivatives of any of these, (vi) Bifidobacterium bifidum, and (vii) a composition comprising Lactobacillus acidophilus, Lactobacillus rhamnosus, Saccaromyces boulardii, Bifidobacterium breve, and alpha amylase.

2. The method of claim 1, wherein the probiotic composition comprises at least five of the components (i)-(vii).

3. The method of claim 1, wherein the probiotic composition comprises at least six of the components (i)-(vii).

4. The method of claim 1, wherein the probiotic composition comprises all seven components (i)-(vii).

5. The method of claim 1, wherein the probiotic composition comprises (i) Lactobacillus rhamnosus, (ii) Bifidobacterium longum, (iii) Green tea or derivatives thereof, (iv) Cranberries, Cranberry extract or derivatives thereof, and (vi) Bifidobacterium bifidum.

6. The method of claim 1, wherein the probiotic composition comprises an enzyme that disrupts extracellular matrix polysaccharides (EPSs) present in a gastrointestinal tract biofilm.

7. The method of claim 1, wherein the composition is consumed or administered in effective amounts to improve one or more symptoms of menopause selected from the group consisting of: hot flashes, digestive distress, weight gain, sleep disruption, or increased risk of occurrence of bacterial vaginosis, and increased risk of occurrence of urinary tract infection.

8. The method of claim 1, wherein the probiotic composition is freeze dried or spray dried.

9. A composition comprising a live, freeze-dried probiotic composition comprising four or more of the following components: (i) Lactobacillus rhamnosus, (ii) Bifidobacterium longum, (iii) Green tea or derivatives thereof, (iv) Cranberries, Cranberry extract orAttorney Docket No. BHMP107WO derivatives thereof, (v) Nichi glucan, glucans produced from Aureobasidium pullulans, polysaccharides produced from Aureobasidium pullulans, β-1,3-1,6-glucans, or derivatives of any of these, (vi) Bifidobacterium bifidum, and (vii) a composition comprising Lactobacillus acidophilus, Lactobacillus rhamnosus, Saccaromyces boulardii, Bifidobacterium breve, and alpha amylase.

10. The composition of claim 9, wherein the composition comprises at least five of the components.

11. The composition of claim 9, wherein the composition comprises at least six of the components.

12. The composition of claim 9, wherein the composition comprises all seven components.

13. The composition of claim 9, wherein the composition comprises (i) Lactobacillus rhamnosus, (ii) Bifidobacterium longum, (iii) Green tea or derivatives thereof, (iv) Cranberries, Cranberry extract or derivatives thereof, and (vi) Bifidobacterium bifidum.

14. The composition of claim 9, wherein the composition comprises an enzyme that disrupts extracellular matrix polysaccharides (EPSs) present in a gastrointestinal tract biofilm.

15. The composition of claim 9, wherein the components of the composition are present in effective amounts to improve one or more symptoms of menopause selected from the group consisting of: hot flashes, digestive distress, weight gain, sleep disruption, or increased risk of occurrence of bacterial vaginosis and increased risk of occurrence of urinary tract infection.

16. The composition of claim 9 further comprising one or more additional probiotic components selected from the group consisting of: Lactobacillus casei, Lactobacillus plantarum, Lactobacillus acidophilus, Bacillus subtillis, Lactobacillus rhamnosus, Lactobacillus helveticus, Bifidobacterium longum, Bifidobacterium lactis, Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium adolescentis, Bifidobacterium breve, Saccaromyces boulardii, and Brevebacillus laterosporus.

17. The composition of claim 9 further comprising one or more additional components selected from the group consisting of: barley, xylan, acerola and or acerola extract, isoflavone(s), soy isoflavone, moringa oleifera, guava or guava extract, berberine, doenjang, fermented soybean, P95 scFOS, FOS (fructooligosaccharide), guarana seed powder, guarana seed extract, gurarana extract, Tianwang buxin granules, red clover, dried cranberry, inulin,Attorney Docket No. BHMP107WO lactulose, hesperidan, Novasoy, PreforPro, grape seed extract, black cohosh, magnesium, and Winclove 607.

18. A composition in a dosage form comprising a live, freeze-dried probiotic composition comprising probiotic composition comprising four or more of the following components: (i) Lactobacillus rhamnosus in an amount of 0.01 to 15 billion CFUs, (ii) Bifidobacterium longum in an amount of 0.01 to 15 billion CFUs, (iii) Green tea or derivatives thereof in an amount of 50 to 1000 mg, (iv) Cranberries, Cranberry extract or derivatives thereof in an amount of 50-1500 mg, (v) Nichi glucan, glucans produced from Aureobasidium pullulans, polysaccharides produced from Aureobasidium pullulans, β-1,3-1,6-glucans, or derivatives of any of these, in an amount of 5 to 150 mg, (vi) Bifidobacterium bifidum in an amount of 1 to 1000 mg, and (vii) a composition comprising Lactobacillus acidophilus, Lactobacillus rhamnosus, Saccaromyces boulardii, Bifidobacterium breve, and alpha amylase in a total amount of 0.01 to 15 billion CFUs.

19. The method of claim 1, wherein the probiotic composition further comprises grape seed derivatives or grape seed extract.

20. The composition of claim 9, wherein the probiotic composition further comprises grape seed derivatives or grape seed extract.

21. The composition of claim 18, wherein the probiotic composition further comprises grape seed derivatives or grape seed extract.

22. The method of claim 1, wherein the composition further comprises one or more types of bacteriophages that target E. coli.

23. The method of claim 22, wherein the one or more types of bacteriophages are selected from the group consisting of: Myoviridae and Siphoviridae.

24. The composition of claim 9, wherein the composition further comprises one or more types of bacteriophages that target E. coli.

25. The method of claim 22, wherein the one or more types of bacteriophages are selected from the group consisting of: LH01-Myoviridae, LL5-Siphoviridae, T4D-Myoviridae, and LL12- Myoviridae.Attorney Docket No. BHMP107WO 26. The composition of claim 24, wherein the one or more types of bacteriophages are LH01- Myoviridae, LL5-Siphoviridae, T4D-Myoviridae, and LL12-Myoviridae, and the one or more types of bacteriophages are present in a dosage form in a titer of 1x104to 1x108.

27. The composition of claim 18, wherein the composition further comprises one or more types of bacteriophages that target E. coli.

28. The composition of claim 27, wherein the one or more types of bacteriophages are LH01- Myoviridae, LL5-Siphoviridae, T4D-Myoviridae, and LL12-Myoviridae, and the one or more types of bacteriophages are present in the dosage form in a titer of 1x104to 1x108.

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