NZ786165A - A Transient Commensal Microorganism for Improving Gut Health - Google Patents

Abstract

The invention relates generally to administering compounds to promote mucosal healing in mammals in need thereof including, but not limited to humans. The compounds may include bifidobacteria and mammalian milk oligosaccharides.

Description

A Transient Cnrnrnensal hillernnrganisrn far lrnprnying Gut llealth WELD 0i?" THE lNVENTlON [Willi] 'l‘he embodiments described herein relate lly to healthcare, and mere particularly, to administrating compounds to te rnncosal g in mammals in need thereef including, but not limited tn hurnans BACKGROUND lll The intestinal tnicrebionie is the community of rnicroerganisrns that live Within the gastrointestinal tract, the rnajnrity of which is found in the large ine er colon, in a healthy individual, inest dietary nutrients that are consumed are absorbed by the body before they reach the colon, Many foods, however, contain indigestible carbohydrates (ie dietary fiber) that rernain intact and are not absorbed during transit through the gut to the colon. The c rnierobiorne comprises certain ial species that are able to partially consume these fibers and utilize the constituent sugars (free sugar ntnnorners or FSh/ls released by microbial digestion 0f the fibers) for energy and metabolism, as well as a larger number of hacterial species that simply thrive on the FEMS produced by these fiber degraders. Methods for measuring dietary fiber in various foods are well known to one of ordinary sltill in the art. [9993} in mammalian species, the nursing infant’s intestinal rnicrobiente is quite different front that of an adult niicrnbieine in that the adult gut inicrobionie generally contains a great ity of Organisms all present in a low percentage of the total populatien. The nursing hurnan infant’s niierebinine, on the ether hand, can be made up almost exclusively {up to 80%) of a single species. Diet drives the nce, complexity and diversity of rnicrnbial species in the inicrobionie. The abundance 0f different species in the celonic rnicrobionre is the result of the diversity Of the fiber in the diet of a typical adult whereas the nursing infant has only a single snuree of dietary fiber-------niarnnialian fililli accharides -------and hence the resultant infant niicrnbieine can be ted by ergani,srn(s) that preferentially utilize that type (if dietary fiber.

[Gilli-4} The transition front the sirnple, non—diverse thierobionie of the nursing infant tn a complex, diverse nticrobiorne of an adult reflects the rnarnntal’s transition from a single nutrient source of a x fiber (cg, mammalian milk oligosaccharides-—-lv’llvl0s) to more diverse dietary fiber sources. The post—weaning through adult mammalian microbiome contains a more diverse number of microbial species able to compete in the variable food niches that are generated by the diversity of the fibers in the complex diet of an adult relative to that of the infant.

EitltltlSl Pre—weaned mammalian infants have only one sou fee of nutrition: mammalian milk. Components in mammalian milk, namely mammalian milk oligosaccharides, have, over the course of evolution, selected for a small number of sms that are particularly suited to the inal environment, grow selectively on MMU, and confer ts to the host. ln the case of the breastfed infant, the indigestible portion of the millr is effectively broken down and ed by these ed organisms. As a result, these organisms are able to outcornpete and increase their abundance compared to ether environmental species, and this has the overall effect of Teducing complexity of the microbiome. [9996} For example, the HMOs represent about l5% of total dry weight (and energy) and are the third most abundant family of nutrients in human milk. These oligosaccharides se sugar residues in a complex and branched form that is not usable directly as an energy source for the baby or an adult, or for most of the microorganisms in the gut of that baby or adult. A distinct few microorganisms, such as Bifidubacierium longum subsp. infhntis (B. infer/iris), and Bifodobacterittm breve, have the unique capability to consume ic MMOs, such as those found in human or bovine milk (see, (2.3,, US Patent No, 8,l98,872 and US Patent ation Nos. lit/809,556 and 62/307,425, the disclosures of which are incorporated herein by reference in their entireties). When Bu infantry comes in contact with c n‘tain mammalian mill; oligosaccharides, a number of genes are specifically induced which are responsible for the uptake and internal truction of those mammalian milk oligosaccharides, and the individual sugar components are then catabolized to provide energy for the growth and reproduction of that organism (Sela et al, 2008). [lltlll'l’l if the appropriate bacteria are not present in the body of the mammal, the indigestible carbohydrates of, for example, mammalian milk become susceptible to non-specific hydrolysis, releasing FSMs capable of promoting the growth of opportunistic or highly ctive ens that would not have flourished otherwise, or are otherwise excreted from the body in the feces, The consequence of a tie microbionre is that is skewed towards int‘eetion, inflammation, intestinal damage, and pathogenesis. [9908} Conventional teaching with regards to the mammalian microbiome is that complexity provides stability, To be able to effectively consume the complex non—infant diet, maintaining a diversity of microorganisms in the microbiome is thought to he the key to promoting gut health. l.ozupone, Nature, Vol, 489, pp. 220—230 (20M). The inventors have ered that this is not necessarily the case and that the simplest microbiome may be of great benefit to the stabilization and recovery of the gut damaged by mation and/or agitation. [9999} The inventors have discovered that mucosal healing in the dysbiotie gut can be promoted by tic microorganisms driving the intestinal microbioine towards an intant~lilte state, which is simple, less diverse, and less pro-inflammatory than an adult gut microbiome particularly when inllamed by disease or dysbiosis. The inventors have also discovered that use of prohiotic microorganisms for which MMGS serve as a selective energy source (carbon source), is ularly beneficial for mucosal healing. Dysbiotic mammals in need of l healing would include humans that exhibit conditions such as, but not limited to, irritable Bowel Syndrome (lBS), s Disease (CD), Ulcerative Colitis (UC) (collectively irritable Bowel Disease or liBD), Short Gut Syndrome, colic, General diarrhea, overgrowth of certain pathogenic bacteria such as C’Zostridutm dificiic, or any other condition (such as extensive antibiotic use) where the gut is made susceptible to intection by pathogens, such as, but not limited to Escherichia, Ciastridium, la, Campylobacter, and Salmonella. [lltl‘llll This invention es methods of treating gastrointestinal dyshiosis by providing a patient with a ition comprising (i) a complex carbohydrate from a mammalian mill: source and (ii) bacteria which internalize the MMO prior to its hydrolysis; lly the composition is administered for a period of time tog, for at least 5 days). While the rganisms and oligosaceharides are normal components of infant nutrition, the methods of this invention are targeted at subjects other than int‘ants (ie. beyond 6 months of life); therefore, the compositions of this invention are formulated for more mature (and typically larger) subjects and for ibility with more complex, diets. The dysbiosis may be the result ot" irritable Bowel Syndrome, Crohnls Disease, Ulcerative colitis, Necrotizing Enterocolitis, bacterial overgrowth, bacterial induced diarrhea, antibiotic treatment, eating ers, obesity, or low diversity in dietary intake. The Bifidobacrerium is preferably selected from B. longttm, B. pseudocaz‘anclarion, B. adolescemis, B. animalis (eg, 3. animalis subsp. arziimziis, B, animalis subsp. Zach's), B. Zongum subsp, iongnm, B. pseudolongum, and B. brave, and more prelei‘ably, the B. {angina is B. iongzmt subs. injhntisi The mammalian milk sources may be human, caprine, porcine, ovine, equine, or bovine, and preferably, the bovine source is troin bovine colostrum. The Mth may be lrom whey, whey mother liquor, whey powder, whey tes, and/or the mammalian milk oligosaccharide may be fucosylated, sialylated or be tives thereof. in particular, the MMO may se Z’Ufucosyllactose, 3’—fucosyllactosc, difucosyllactose, lacto-N—fucosylpentose l, lacto~N—fucosylpentose ll, lacto—N-i'ucosylpentose ill, lacto—Nvfucosylpentose Vt 3’—sialyllactose, 6’—sialyllactose., 3‘—sialyl~3"fucosyllactose, sialyllacto~ N-tetraose, 3’~sialyllactoseamin, and b’-sialyllactosamine, produced synthetically and purified or isolated from natural sources or recombinant microorganisms. The Bifidobacrerium is typically provided in a daily dose of broth l0 million to l trillion chi, pre 'erably ll) million to ltlt') billion cfu, and more preferably from 4 n to 50 billion cfu. The lvll‘le is provided in a daily dose of front l to 60 g, ably from 2 to Zll) g.

{Gilli} Some embodiments of the instant invention include compositions comprising a.

MMO and a microorganism wherein the MMO induces a change in the rganism such that the MMO then becomes an energy source for the organism, and when ingested by a mammal, the induced or ted microorganism provides a benefit to the gut of that mammal. Additional embodiments involve the maintenance of the induced rganism in the gut of the mammal by maintaining the y supply or" MMOS or other glycans that are ive for that microorganism. A further embodiment involves the subsequent clearance of the microorganism from the gut by the cessation of the supply or" the lleG to the mammal. [9912} in one mode of this invention, the MMO and the bifidobacteria are provided in a dry form which may also be enrobed in a material that would provide enteric protection. in another mode, the MMO and the bilidobactcria are ulated, and the e may further comprise an enterie coating such as a coating that is not ted by passage through the stomach. in another mode, the MMO is provided as a solution and the bifidobacteria is provided as an enteric—coated , tablet, or capsule. [9GB] in yet another mode, the method of this invention further comprises administering Lacrobacilius or Pediococcus conteinporaneously with the composition, The Lacrobacilius may be selected front L, plantarztm, L. easel", Lt amri, l. brat/is, L. ominis, L, tzmz, L. i, L. johnsonii, L. [Jamaal/is, L. sakci, L. .ralivarim, L. rhamnams (cg, LGG}, L. acidopltilztst L. cart/ants, L. mucosae, L, crispams, and/or Lt remcri, Preferably the anmbaciiins is L" reuteri. The occus may he selected from P5 aciditactici, P. stilesii, P. argcntinicns, P. damsel/iii and/or P. pentosacmst The Laclobacillus or Pediacoccus may he provided in a daily dose of trorn l0 million to l trillion chi, prelerahly the Lactobaciilus and Pediococcus is provided in a daily dose of from 5 billion to 50 billion cfu.

BRIEF DESCREP'EEON (ll? THE DRAWlNGS [Gill-4i Fig l: Chart showing the base l0 log change in B. iiy‘amis levels by day during niucosal healing diet ing BMO, G08, and B. infantis. The data are reported as CFU B. infantis per ug DNA divided. by CPU total bacteria per ug DNA. [titllSl tilt}. 2,: Chart showing the overall weight gain at 28 days for piglets receiving standard antibiotics at birth, no otics or the mucosal healing preparation bed in the application.

Development of the Neonatal Mierobiome {99%} Certain hifidohacteria, such as B. tor/23ml suhsp. infimris, possess n genes dually or in gene rs that are dedicated to the alization and deconstruction of lih/lUs (Sela and Mills, 20H), Trends in Microbint, l8:298~3ll7). When such bacteria interact with MMOs, like those found, in mammalian milk, these genes for transporting and catabolizing t'ucosylated and/or sialylated oligosaccharides, are upregulated (Kim, et al., 2M3, PLOS ONE, 8(2):e57535; Garrido, et al.., 20l5, Nature Scientific Reports), The inventors have recently discovered that certain hitfidobacteria ing, but not limited to B. intently can be "activated" by their interaction with certain MMOs (international Patent Publication No. Wt} 2016/065324 incorporated by reference herein). The activated B. infantir is defined herein as the state of the cells, as measured by the tip-regulation or down-regulation of certain genes including, but not limited to, oligosaccharide binding proteins, permeases, and enzymes responsible for the nptalte and internal deconstruction of the hilt/i0. tn the activated form, the B. is becomes the primary consumer of all the MMO and has been shown to se its relative proportion in the gut niicrohiota of hreast-fed infant humans to levels significantly higher than its natural levels and as high as 70% of the total microbial population of the distal colon. When .8. ii/tfiiti/lfis is present in the gut of a baby, and that baby is also provided with ’s milk as a primary or sole source of nutrition, the population of B. informs can increase to levels as high as 90% of the total bacterial population of the gut as measured by the microbial quantification of the stool, When present in this situation, many other genes are also upregulated including those for the production of a number of other metabolites. [9M7] When activated, 8:, iiyfantis is known to bind tightly to the gut mucosa of the baby and facilitate the development of the infant gut (Underwood, et al,, 2,015, Pediatr. 11665,, 77:229-- 235). The proliferation of activated B. infantis in the gut of a newborn infant, triggered and uniquely enabled by the hills/EUs provided in mother’s rnillt, is of significant benefit to the health and long term survival of that infant. B. informs is associated with significant benefits to a newborn infant which include, but are not limited to, a higher binding affinity to the gut mucosa, higher colonization of the Gl tract thereby preventing growth of other ial clades, higher consumption of MMOs, and a greater stimulation of the immune response (Lewis, et al., 20l5, Microbial/nu, 3zl3; Huda, et al., 20%, Pediatrics, l34:2 e362—-e372).

[Quill] Once administered with a sufficient amount of MMOs as a dietary source, the activated 8. s will remain in the gut of a mammal at high concentrations and activated as long as the dietary source of Mths is continuously provided to the mammal, The inventors have discovered that once the source of the MMOS is withdrawn from the diet (tag, at weaning), the B. infantis is no longer ted, and, it can no longer successfully colonize or e with other gut microbiota for nutrients in the gut, and its population rapidly decreases to less than 5% of the total microbiorne. B. infimtis is generally not naturally found in the gut of a weaned infant, child, or adult in levels of more than l%.

Mammalian Milk (lligosaccharide nts [9919} For this invention, the lleOs are lly sourced, from, identical to, or functional lents of those oligosaccharides in ian milks including, but not d to, human, caprine, bovine, equine, or ovinc milk. The term "mammalian mills oligosaccharide" {lvlhltil}, as used , refers to those indigestible glycans, sometimes referred to as "dietary , or the carbohydrate polymers which are not hydrolyzed by the endogenous host enzymes in the digestive tract and remain generally unabsorbed in the intestinal lumen (eg, the small intestine) of the . Although "dietary fiber" usually refers to indigestible plant polysaccharides with degrie of polymerization (DR) of 20 or more carbohydrate residues, WO 56550 MMO includes hranched~chain oligosaecharides and oligosaeeharides between 333 and DP—ZO. aceharides may he free in milk or bound to protein or lipids and are also referred to as glyeanst aeeharides having the chemical structure of the indigestihle oligosaeeharides found in any mammalian mill: are called "M M0" or "milk iiber" herein, whether or not they are actually sourced from ian mill; Ellillel in alternative ments of the instant invention, the MMO (e.g., hovine millt oligosaeeharides (HMO) or human milk oligosaccharides (HMQD may be supplemented with synthetically—produced oligosaceharides ing fueosyllactose (SPF) and/or sialyllaetose {SE38}, or more complicated Structures such as, hut not limited to, 2’—tueosyllactose, 3— fueosyllactose, difueosyllactose, lacto~N—l’ueosylpentaose l, laeto—waucosylpentaose ll, N~ i‘ucosylpentaose lll, laeto-N—fncosylpentaose V, 3’—sialyllaetose, 6’—sialyllaetose, 3’—sialyl~3~ fucosyllaetose, sialyllaeto--N--tetraose, ylgalactosamine, and 6'-—sialyllactosamine, which have been synthetically produced and may be purified to at least 50% purity before addition to the MMOs. The definition of synthetically produced oligosaeeharides in this invention includes those oligosaecharides produced in genetically modified organisms as well as through eherni— synthetic processes that are otherwise identical to MMOS as well as galactooligosaeeharides (G03) that are enriched in DP—4 and Dirt—5 polymers as descrihed in USP 8,425,930 {incorporated here by reference in its entirety) as these structures also provide differential growth of B. infer/iris. in a preferred. embodiment, the synthetically—produced derivatives can he used alone or added to the milk—sourced MMQ and rnalte up from at least 5% to at least 80% of the dry weight of the composition" in some embodiments of the t invention, the mass ratio of MMO:SPF or MMO:SPS is from 20:l to lzfii, in a red embodiment the mass ratio of :8l’F or MMOISPS is from ltlzl to lIZ, and in a most preferred embodiment the mass ratio of MMOzSPF or MMOfil’S is from Szl to l:l., Ratio targets may also he that of human milk wherein one starts with Sialyllactt‘ise—dominant itions such as liovine milk and add one or more of purified 2’~fucosyllaetose., 3vfueosyllactose, difncosyllactosm lacto—N~fiicosylpentaose l, laeto—N-l'ucosylnentaose ll, lacto~N—t‘ueosylpentaose ill, laeto—N-l'ucosylnentaose V, lacto~N~ tetrose, and lactose-N—neotetrose.

Affecting intestinal Mueusa Beyond The Neonatal Stage [Willi] The instant invention can be used to treat a mammalian infant or non-infant patient (heyor‘id 6 months of age), where the patient has a gastrointestinal distress caused by elevated, levels of pathogenic bacteria (dysbiosis) such as, but not limited to, Listeria, dia, Escherichia, Helicobacter, Shigella, ella, Yersinia, Clostridinm, Campylobacrer, and other members of the Proteobacteria which can damage the gut epithelium and mucosa. Such dysbioses include, but are not limited to, irritable Bowel Syndrome (U33), Crohn’s Disease (CD), and Ulcerative colitis (UC); (collectively lBD) Necrotizing colitis (NBC), bacterial oy argrowth (B0), bacterial induced diarrhea (BID), Celiac Disease (CELT), and antibiotic treatment (AT). ln some embodiments of the present invention the dysbiosis may be d by a less complex and/or less abundant microbiome than normal which may be due to causes including, but not limited to, prolonged antibiotic treatments, narrow dietary diversity, and eating disorders, such as, but not limited to, bulimia nervosa, anorexia nervosa, and binge eating disorder.

[QGEZE in this invention, treatment of the gastrointestinal distress is provided with an oral dose of bacteria such as, but not d to bilidobacteria, and MMGS including, but not limited to milk oligosaccharides from a mammalian , MMOs from other biological sources, or chemically or biologically synthesized MMOs that are the finictional equivalent of those found in mammalian mill: sources, and @053 polymers enriched in DP~4 and EELS. [9923} in some embodiments, any of the compositions described herein can be provided to a non—nursing mammal. The non—nursing mammal can be a human, as well other icated mammalian species such as, but not limited to, an agriculturally~relevant production mammal (sag, cow, pig, rabbit, goat, buffalo, and sheep), a mammalian companion animal tag. cat, dog, rabbit, and , laboratory mammals leg, mice and rats), and performance mammals (eg a thoroughbred race horse, camel, and working dog). [9924} in some embodiments, a composition comprising a Btfidcbacterinm and a MMG is provided. The Bifidohacterizmi can be B. longzmt (eg. B, langnm subsp. infantis, B, longer/n, subsp. longum), B. breve, B. bifidum, B. enimaiis (9.3., B. is snbsp lactis, B. onimalis subsp onin’talis), B, pseudocotermiamm, B. adolescemis, B, crttertttlatnm, B, pseudolongnm, or any ation f. in some embodiments, the composition provides a mucosal healing to a mammal by acting as an anti-inflammatory to sooth intestinal inflammation caused by dysbiosis or other disease and also preventing the growth and y fernoving the unwanted or overgrown bacteria. The composition, when provided to a mammal, may allow for colonization by the bifidobacteria and cement of other bacteria When the composition is administered and, optionally, combined with a regulated diet, the microbiome can have d numbers of nonwhifidohacteria s as compared to a microbiome of one not being administered the composition. in one embodiment, administration of the composition s in a "simple microbiorne" due to the increased tional colonization by the bifidobacteria. [9925} A simple microbiome can he described as the presence of greater than ll.)6 cfu/g stool ol a single genus of bacteria leg, Bifidobacrerinm), more particularly, of a single species or strain of bacteria (eg, B. Zangum subsp. infantis [8. infanrisl). This can be reflected in, for e, up to 80% of the microbiome being dominated by the bacterial genus or, more particularly, by the single subspecies of a bacteria. such as B. refunds in a human breast feeding infant. A simple nucrobiome can also be described. as the presence of greater than l0%, 20%, 230%, 40%, 50%, 60%, 70%, 75%, 80%, or 90% of a single genus of bacteria (e.g., Bifih’obacrerium), more particularly, of a single subspecies as a t of total bacterial cells leg, B. longnm subsp. infer/iris {13. informal) in animals, or in other patient groups [9926} in a preferred embodiment the bifidohacteria is selected from B. Zorigam, B. breve, B. bifldum, B. animaiis subsp testis, B. animafis subsp animaiis, B. pseudocareniridium, B. (:arenulamm, or any combination thereof. in a more preterred embodiment the obacteria is selected from a group of bifidobacteria that internalize mammalian mill: oligosaeeharides prior to their hydrolysis such as, but not limited to B. Magnet, 8. brave, and B. pseudocaimmigrant. in a particularly preferred embodiment of the invention, the bilidobaeteria is B, longnm cies infantry. [fibril Additional embodiments involve the "ceding of a mammal of any age in need of mucosal pment or healing with a composition comprising bacteria (eg, activated hifidohacteria), and a MMG composition. Such a composition can be provided at a dose level of from it) million to l on chi/day of bifidobacteria, and, from l to 60 g/day of MMQ composition for a period of from l—btl days. A mammal leg, human) in need of mucosal healing would include, but would not be limited, to, individuals with signs or symptoms of NEC, lBS, lBD, Crohn’s Disease, leaky gut, auto inflammatory diseases, autism, obesity, , food allergies, eating disorders, or pathogenic bacterial overgrowth, as well as individuals that have had a course of antibiotic therapy and are repopulating their (ll tract.

[QGESE in other embodiments of the t invention the treatment of the intestinal distress is provided by an oral dose of bil‘idobacteria described above that can internalize and consume MMO such as HMO or BMO, along with commensal bacteria that can consume free sugar monomers, Where such commensal ia are preferably Lactobaciilns and/or Pediococcus species that ively consume monomer sugars such as, but no d. to, fucose and/or sialic acid. [9929} in some embodiments of the instant invention, the MMO, bifidobacteria, and optionally lactobacilli are provided to a patient together in a dry powder form or encr psulated in a two-part capsule enrobed with an enteric coating, and provided to a patient in need of such treatment at a dose of from if) million to 1 trillion cfu of bifidobacteria and 10 million to lOO billion cfu of lactobacilli per day and from 1 to (at) g of lvllvtt} per day. in a more preferred embodiment, the MMO and tic bacteria are provided to a patient at a dose of from 4 to 50 Billion cfu of bifidobacteria plus 4—50 Billion cfu of lactobacilli per day and from 2 to 30 g of bib/l0 per day. in a particularly pre ‘erred ment the Btfidohacrerinm is B. longum subsp. injimris, and the Larrmhaciiius is L renter/r7. [9939} in various embodiments of the invention, the composition comprising the bil‘idobacteria and a MMO selected from MMO, SPF and/or SP3 and/or G08, is provided to a mammal (eg, a human) in order to overcome gut~related disorders in obesity including, but not limited to, gut~related metabolic disorders such as hyperphagia and Type l and. Type ll diabetes by any of a number of mechanisms including, but not limited to, the restoration of gut barrier function and the reduction of food intake. This embodiment includes all ages of mammals (e.g., humans) ing newborn infants, children, adolescents, adults, and geriatric mammals (cg, humans). lllll3ll in some embodiments of the instant invention, the combination of (a) hifidohacteria capable of internalizing a MMO prior to hydrolysis and (b) a MMO such as, but not limited to ECO, Bit/l0, HMO, SPF and/or SP3, and/or G03 is provided to a human or mammalian patient exhibiting a dysbiosis—related inal pathology. Such a treatment is maintained on a daily basis until the tration of the bacteria achieves at least a 2—fold se in the numbers of the bifidobacteria in the gut of the mammal. in a preferred embodiment the levels reach at least a ltl-—fold increase. in a more preferred ment the levels reach at least a lOQ—fold increase. in a preferred embodiment of the invention the patient will e essentially no other oligosaccharide or dietary fiber other than the delivered ECO, 8M0, l-lMG, SPF and/or SP8 and/or G03 during the treatment period. Although optional, it may be beneficial to "clean out" the intestine prior to ent, lly by use of laxatives to encourage ion of any residual fiber present prior to treatment. in certain embodiments, the bilidohaeteria capable of internalizing a mammalian milh oligosaecharide prior to hydrolysis is an activated hifidohacteria, An activated hilidohacteria is a bit‘idohaeteria that, through contact with milk glycans has genes of an HMO gene cluster that are upregulated. in certain other embodiments, the hifidohacteria capable of internalizing a MMO prior to hydrolysis is cultured in a manner that is non-activating. [9932} in certain embodiments of the t ion, a "daily ration" of the liifidobacteria and M M0 is provided to the patient. A "daily ration" is an amount provided to the t within the same 24—hour period. A patient can he given a dose of the hifidohacteria and a dose of the hilt/i0 substantially contemporaneously (eg, within six hours, within four hours, within two hours, within one hour, within forty-five minutes, within thirty minutes, within twenty minutes, within filteen minutes, within ten minutes, within five minutes, within three minutes, or within one minute). [$133] in some embodiments of the instant invention, the dosing ol the ohaeteria and MMO is maintained for a period of at least l week to allow mucosal healing. in a more preferred embodiment the dosing of the hifidobaeteria and MMO is maintained. for a period of at least l month to allow full rnucosal healing. in a particularly preferred embodiment the dosing of the hifidohacteria and lleO may be continued through the period of time during which the symptoms of the intestinal pathology are alleviated. Preferably, dosing is discontinued when Gl symptoms have been alleviated, and the patient is able to tion without symptoms to adult dietary s ol fibers using a strategy of weaning away from a single fiber source to le fiber sources supported with commensal organisms adapted to the adult dietary fiber. Such a weaning s is described in US. Patent Application No. ,425, which is incorporated herein by reference in its entirety. Dosing may be continued while the ms are alleviated tor a period, of time (e.g., one hour, two hours, three hours, four hours, six hours, eight hours, ten hours, one day, two days, three days, a week, two weelts, at least l month, at least from l month to 6 months, and at least 6 months to one year).

HMS-4} in various embodiments of the invention, the high levels of bilidohacteria are returned to normal (low) levels by eliminating the dietary supply of M MO, ECO, BMO, HMO, SPS, and/or SPF, and (308, and introducing other conventional food liber sources as part of the daily diet for a period of at least 1 week, in a prel’erred embodiment the levels of bifidobacteria are reduced to normal levels by eliminating the dietary supply of MMO, ECO, EMU, l-llv’ltil, SP3, SPF, and (303 and allowing other conventional food fiber sources as part of the daily for a period of at least l month, ll‘orrnnlating Compositions For This invention [titiSSl 'l‘be hilt/i0 ation may be provided together with red ia or separately. lo a preferred embodiment, the MMO is prepared from a bovine colostrum (ECO), whey permeate. or other dairy streams (Eh/i0), and combined with the bit‘idohacteria at a ratio of from (Mil to ill g of MMG per Billion cfu of bifidobacteria. in a more preferred embodiment, the MMO is combined with the bilidobacteria at a ratio of from (hi to it) g of MMO per Billion cfn of bifidobacteria. [9936} in a more prel’erred embodiment the accharide is provided in a concentrated l‘orm, wherein the concentration of the MMO comprises at least ll)% of the mass of the preparation (on a dry weight basis) delivered. to the human or other mammal in need of the treatment. The ation may be provided in a dry powder lormnlation, a solution, a suspension, or in a tablet or capsule format with or without an enteric coating to allow passage through the stomach and release in the intestine. Such enteric coatings include, but are not limited to, dairy proteins, whey ns fatty acids, waxes, c, plastics, plant , methyl acrylate—methaerylic acid eopolymers, cellulose acetate suecina'te, hydroxy propyl methyl cellulose pbthalate, hydroxy propyl methyl cellulose acetate succinate, polyvinyl acetate plithalate (PVAP), methyl methacrylate—methaerylic acid copolymers, cellulose acetate trimellitate, sodium alginate, and Zein.

Administering The Composition Of This invention lllli37l in some embodiments of the instant invention, the MMQ and bilidobacteria are provided to a patient together in a dry powder form and/or ulated in a two-part capsule enrobed with an enteric coatings, and provided to a patient in need of such treatment at a dose of from ltl million to ltlt) billion cfn of bacteria per day and from l to 60 g of mammalian mi lit oligosaeeliarides per day. in a more preferred embodiment, the mammalian millr oligosaccharide and bacteria are provided to a patient at a dose of from 4 n to 50 billion clip of bacteria per day and from ‘2 to 40 g of MMO per day. [$138] in some embodiments the accharide component can be dissolved in a liquid such as, but not limited to. water, logical saline, mammalian mills, or formulation designed to provide all or part of a daily nutritional ement such as, but not limited to an infant formula or an enteral formula and provided in a liquid form to the patient while the bilidobacteria are provided separately as a powder or suspension in a carrier liquid which may optionally include a solution comprising the MMO. litllfifill in some embodiments of the invention, the patient is maintained on a strictly controlled diet throughout the course of the ent with the bacteria and MMO. Such a diet would contain none or a minimal amount of any other dietary fiber but may contain simple carbohydrates such as monosaccharides and disaccharides in amounts required to in the patients teryention . in a preferred ment of the instant invention, the daily amount of the other dietary fiber is from less than 30 g/day, preferably less than lt) g/day, more pre Terably less that 5 g/clay, and most prefereahly less than l g/day. in a red embodiment of the instant invention, the dietary simple carbohydrates are less than 50 g/d. in a preferred embodiment of the instant invention. the daily amount of the simple carbohydrates is less than 40 g/d, preferably less than 20 g/day, more preferably less than it) g/day, and most preferably less than 5 g/day. [lllléltll in other embodiments the aceharide component and the bil’idobacteria are provided together in a spoonable composition such as, but not limited to, yogurt, hel‘ir, pudding, cream, chocolate, or any edible oil. litllldll in some embodiments of the invention, any oi the itions described herein may be administered to a patient. Patients include mammals suffering from gut-related disorders including, but not limited to, obesity, or gut—related metabolic disorders such as hyperphagia and 'l‘ype l and 'l'ype ll diabetes, by any of a number of mechanisms including, but not limited to, the restoration of gut barrier on and the reduction of food intalre. Mammals may include humans, as well as other domesticated mammalian species ing, but not d to, agrieulturally—releyant production mammals (6.3., cows, pigs, rabbits, goats, and sheep), mammalian companion animals (6.3., cats, dogs, and horses), and performance mammals (tag, thoroughbred race horses. racing camels, and working dogs). Patients may include all ages of s including inlai'it mammals, young mammals, adolescent mammals, adult mammals, and geriatric mammals. [lily-$2} Those who would particularly benefit from the process of this ion include patients with a bacterial bloom that rapidly expands the presence of a particular organism, or patients with reduced diversity Where ltey conunensal s are missing. Both of these cases may present as a microbionie of less ity than expected in a healthy individual, and these patients afe characterized as having a tic microbiome.Shilts in the inicrohionie can he determined using Next Generation Sequencing (see, cg ii at al, "From next-generation sequencing to systematic modeling of the gut rnicrohiome", Front Genet. (June 23, ZOlS), published online at doi.org/l0.3389/fgene20l5002l9) or full nomics approaches (sec, tag, Wang et al, "Application of metagenomics in the human gut niicrohioine", World 3., Gastroenterol. (2015), Vol. 2i No. 3, pp. 803—8l4) to monitor the change in specific sms, or overall shifts in families known to contain members of opportunistic or pathogenic organisms. qPCR can also he used to monitor changes in specific species or subspecies. Typically measurements can be normalized using the amount of DNA per gram of stool. A simple microbioine may be healthy in the case of an infant whose diet is almost entirely composed of a single nutrient source (eg, mother’s millr). However, for an dual consuming a more varied diet, a shift of the nticrohiorne to simpler structu T6. is typically an indication of dysbio sis. [9943} in some embodiments, a patient is administered a composition comprising bilidohacteria and an oligosaccharide component for a period of time, following which the patient is stered a composition comprising an oligosaccharide ent that does not comprise bifidohacteria to keep the hilidobacteria colonized.

[WM-4} in other embodiments, a patient is adrninisteed a ition comprising bifidohacteria for a period of time, following which the patient is administered a composition comprising bit‘idobacteria and an, accharide component. [9945} in some embodiments, a patient is administered a ition comprising hifidohacteria and an oligosaccharide component for a period of time, following which the patient is administered a composition sing hifidohacteria that does not include an oligosaecharide component. [titlddl in some embodiments, a patient is administered an oligosaccharide component for a period of time, following which the patient is administered a composition comprising ohacteria and an accharide component. The initial oligosaccharide component can he provided in an amount that provides at least i g per day of MMO to the patient. For example, the l oligosaccharide component can he provided in an amount that provides at least l g per day, at least 3 g per day. at least 5 g per day, at least 8 g per day, at least it) g per day. at least l5 g per day, at least 20 g per day, at least 25 g per day, at least 30 g per day, at least 35 g per day, at least 40 g per day. at least 50 g per day, or at least 60 g per day of mammalian milk accharides to the patient. [filial-7} in s embodiments, a patient is administered a composition comprising hii‘idohacteria and/or an oligosaccharide component for a period of time. following which the patient is administered a composition comprising bifidohacteria and/or an oligosaccharide component for a period of time in which the amount administered. is tapered (ext... administered at a generally decreasing rate) for a second period of time. [till-48} n embodiments of the invention involve a combination of a composition comprising MMO and/or SPF and/or SP8 and/or GUS and a hii’idohacteria wherein the Bifia’ohacterium is selected from B. longem, B. breve, B. bifidus. B. animalis suhsp lactis. B. animalis snhsp is, B. pseudocatemtlamm and B. (satanic/latent or any combination thereof. in a more preferred embodiment the acterizmi is B. longum snbp infhmz's.

Eitllléllll in other ments of the iiwentioh, the Bifldobacmrimn species is used in combination with a Lacmbacilius species including, but not limited to, L. plantartmi, L. am'ri, L.

Mei/is, L. easel, L. coleohominis, L. fermenmm, L. gasscri, L. johnsoniiy L. pentosm‘. L. sakci, L. salivarius, L. ms (tag, LGG}, L. acidophiius, L. cwvatus, and L. restart. in a preferred embodiment the composition ses MMO and/or SPF and/or SP3, and/or G03, or derivatives thereof, L. rhanmosis and 8. {engine suhsp. infazrztis.

EXAMPLES [llllStll Example l. ation of Human Mill; Gligosaecharide (HMO) Compesitinns that can he used Exclusively by Certain Biiidohacteria. [9951} A concentrated, mixture of HMO is obtained by a process similar to that described by Fournell er a! (US Patent Application l40l75). llnrnan mill; is pasteurized and then centrifugally del’atted. separating it into cream (predominantly fat) and slcim mill: (defatted product). The del’atted skim milk is then nltrafiltered using membranes with a 5—10 We cut off to concentrate a protein fraction (predominantly whey proteins and caseins). The permeate from the nltrafiltration. comprising lactose and the complex l-lMOs. is dried ly by spray , or the lactose is partially eliminated by an additional ultrafiltration using a l icDa cut oil filter helore drying. The composition of this dried traction is typically about 50% lactose and about % mammalian millt. oligosaceharides (HMO) with the remainder of the mass primarily peptides and ash. The HMO traction is predominantly fucosylated. However, these compositions can vary from 20—70% lactose and 10— 50% mammalian mill: oligosaeeharides (HMO) depending on the ultraliltration processes. [llll52} Example 2, Preparation ef Ravine Milk Olignsaeeharide (fill/l0} Compesitiens that can be used Exelusively by Certain haeteria.

[QGSE A concentrated mixture of bovine mill; oligosaccharide (BMO) was obtained from Whole mill: which was pasteurized by heating tn 145 degrees F fer 30 minutes, eeoled and centrifugally defatted, separating it into cream ‘loininantly fat) and sltim millt (defatted product). The ed skim milk was then ultravfiltered using membranes with a S—lO hDa cut off to concentrate a protein fraction (predominantly whey, proteins and caseins). The lactose in the permeate was partially ated by an additional nanofiltration using a lkDa cut off. The composition was then, spray dried. This composition of dried BMOS comprised about 15% lactose and about 10% Eli/l0 with the remainder of the mass primarily peptides, ash and other components. Twenty grams of this ition was combined with 5 g of GOS (Vivinal GEES) as the daily ration for treatment. [9954} Example 3. Preparation of Bovine Celestrum Ollgesaeeharide (ECO) Ceiripositieus that can be used lixelusiyely by Certain llii’idohaeteria. [9955} A concentrated e of ECG is obtained by a process such as that described by iansen er al (2010) International Dairy Journal, 20:630—636. Bovine colostrum (preferably from the first milking) is pasteurized by heating to lélS s F for 30 minutes. cooled and centrifugally defatted, separating it into cream, (predominantly fat) and skim milk (defatted product). The defatted skim milk is then ultra-filtered using nes with a fill) hDa cut eff to concentrate a protein fraction (predominantly whey. proteins and caseins). The permeate, comprising the e and mammalian niilk oligosaccharides, is dried directly by spray drying. Alternatively, the lactose is partially ated by an additional nanofiltration using a lkDa cut off. The composition of this dried oligosaccharide fraction is about 40% lactose and about 40% bovine colostrum oligosaccharides (ECO) with the remainder of the mass primarily es and ash. The ECG fraction, is inantly ated.

[QGSQ Example 4. Preparation iii an Activated Bilidebaeteria Cempesitien that can Exclusively use Certain Mammalian milk elignsaceharides. {995?} Bifidobncrerium longum snbsp infamis was isolated and, ed from the feces of a vaginally delivered, hreast fed human infant, and its identification was confirmed by DNA analysis that reflected the presence of a gene set that is specifically associated. with this sm {Sela at at, 2008, PNAS, 105:18964-l8969). A seed culture of this organism was added to a standard growth medium comprising glucose and the ECG of Example 3 as carbon sources in a 500 L ed t‘ermenter. Following 3 days of growth under anaerobic ions, a, sample of the culture was tested for the presence of activated Bifidohacreriztm $072ng subsp. infantis.

Activated, Be infantis was identified by the presence of gene transcripts for sialidase, The terrnenter was harvested by centrifugation, the concentrated cell mass was mixed with a cryopreservative (trehalose plus mills proteins) and freeze dried, The final dry product was 5.5 kg ot‘haeterial mass with a live cell count of l3t) )1 l09 ctu/g. [@1581 Example 5. Preparation and use of Therapeutic Compositions for the Treatment of Digestive ogies, [9959} The activated 8. inflmtis product of Example 4 was blended with pharmaceutical grade lactose to provide a minimum dose of 30 Billion chi of B. longer/n snhsp. infimtis per gram. 0.625 g of this diluted activated B. infamis product was then packaged in oxygen~and moisture~ resistant sachets, to provide doses of l5 Billion cfu of 1:?" iongnm suhsp, irtfrznris per sachet, One sachet of 18 hillion efu of B. lwigam subsp. infra/iris was consumed with a g 'ast and one with an evening meal. lithititll Twenty grams of the BMG preparation of Example 2 was combined with live g of €303, packaged in separate bags and administered in a daily ration of 20 g Bit/l0 + 5 g G03. This preparation provided the carbon source (Eh/l0 and SOS) to support the ‘ie growth of the supplemented B. longzmi subsp. infmilis in the colon of the patient, thereby providing a gut environment favoring mncosal g, [9961} The 8M0 preparation was consumed 5 times per day (5 x 5g EMU/(30$ mixture of Example 2), approximately every 3—4 hr by blending the 5 g of powder with a meal replacer (Boost, Nestle Nutrition) containing 240 Cal/drink with ng/protein and 6 g of fat and t) g of dietary fiber. The subject was allowed to e 2-3 eggs each morning, and one serving of fish or meat with lunch and dinner Any dietary fiber consumption e the therapeutic formulation of BMQ was kept at less than 1 g per day. [lititiZl As a step to accelerate the switch trom a microhiome ing adult dietary fiber to a microbiome consuming milk—based fiber, the subject completed a colonoscopy preparation involving a clear liquid diet and laxatives to clear out the bowels of fiber in preparation for the diet change. Once this was completed. the subject followed. the specific diet n that limited the non—mills dietary fiber to less than l gram per day and ensured the t was still eating a diet with sufficient protein, fat and carbohydrate to maintain a constant weight. lilllltifll Fecal samples were talren the day before the colonoscopy prep (pretreatment) and on a daily hasis for the 7 days on the dietary regimen of consumption of the B. infimiis and EMS.

The subject also filled out questionnaire forms regarding a self—assessment of the subject’s gastrointestinal responses or indicators of the palliative ell'ect of the composition on symptoms of gastrointestinal distress. Following the seven days of the dietary regimen. the subject was allowed to return to his pretreatment standard diet and post treatment fecal samples were talren during a l weel: post-treatment phase. DNA was extracted and ted to qFClt is and NextGen sequencing for niicrohiom't analysis. B. infantis was specific; lly measured using qPCR (Figure 1). At baseline, 8. infantis was helow the limit of detection in an adult gut. ahle levels were observed with supplementation and diet changes. Figure l shows that the 76. was at least a l,tlt)t)—fold difference in levels of colonic B. infimtis between ne and treatment. The NGS data ed a means of Visualizing the relative changes in different clades and families of bacteria. Samples were also prepared for other measurements including l3 M0 content by Mass Spectrometry in the stool to monitor in vivo consumption, short chain fatty acid and lactate. pH inations, ements of cytolrines and a full metabolomics determination. 13.3.5 mix; fifyéy .m§ mwgmwwxfifim .r .............................................................................................................. mgfi§32>§% wze%mam :wfi. .mmafi wwcmw ER £meme Amwfiy‘mfié $4:chme N ........................................ wmwfimamwmsw x x, M V 5m x ‘x _.. ................ " mm nnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn Example ti. Use of a composition of B: lorrgttm snhsp. irtfantis with Lactobacilius plarrt‘amm to reduce Clostridinm species in newborn foals. [9964} Newborn foals born to mares at a large horse breeding barn were monitored during an ak of severe hemorrhagic diarrhea among the foals The foals were found to he culture- and toxin-positive for Clostria’inm difficile. Seventeen foals were horn during the initial phase of the onthreale, ol which fifteen animals became ill and required intervention according to the standard of care as described in the lvlerclt nary . rd of care involved metronidazole treatment given at a dose of l5-—-20 mg/leg, PO, tid—qid. and may also involve administration of large volumes of eneons polyionie llnids, with supplemental electrolytes (potassium, magnesium, and calcium), plasma or synthetic ds for low oneotic pressure, anti~inflammatories such as tlunixin meglnmine, and spectrunr antibiotics if the horse is leukopenic and at risk of bacterial translocation across the compromised Gl tract. l’olyinyxin B may aid in binding ic endotoxin. [9965} Of these seventeen foals, fifteen developed loose stool or diarrhea lasting 3-4 days, and 2 died as a result of the infection. After observing the outhrealr, the next loals were provided a formulation of of Stall)" CFU Bifidobaeterium Iongum suhsp. s EVEMKHand Spill? CFU of antobociilus plantamm EVLPGGl every 12 hours, The two foals that were provided with the formulation at l2 hours of age developed a mild diarrhea, hut recovered within 8 hours compared to 3—4 days with standard of care. None of the foals provided with this dose starting at birth developed diarrhea (r126). [dildo] Recovery time for the two d animals that eventually developed the infection was approximately eight hours, which was significantly shorter than the normal recovery time of at least 3-4 days for animals given the standard care regimen. No adverse events were recorded among the treated animals and the dosages were well tolerated. A Fisher°s exact test of the two populations (Standard of Care and Prohiotic treated) yields a significant dill'erence in incidences of C. iie ion (p x 0.00%?) (Table 2).

Healthy Diarrhea Total Control 2 15 1 "F Treated E 2 3 Tetal 9 "17' 25 Fisher’s Exact Test The twevtailed P value annals asses Table 2.. A 2x2. Contingency table analyzed by " 5 Exact test indicates a significant reduction in sick animals among those treated with the probiotie mixture (Treated), relative to the rd of care (Control). {new} Two treatrnent options were attempted. in the first, animals were dosed at l2 hours of life. but this fails to significantly reduce incidence of diarrhea (given the small n), though the severity (duration) was dramatically shortened to l2, hours or less (p x 0.0074; Fisher exact test, comparing populations of diarrheal foals segregated by on of ea). The second option, dosing at birth, was significant at redueing incidences of diarrhea (p = 0.0025).

All animals were dosed at birth with fining/kg of ceftiofur (Excede). and this did not affect health outcome, related to diarrhea. Additionally, the treated population did not develop foal heat diarrhea, which typically affects >5l)% of anirnals, and requires treatment in approximately % of cases (Weese and Rousseau 2005). if a >50% risk is extrapolated. to a hypothetical population of 8 anirnals to match the 8 observed; this yields a significant reduction in foal heat diarrhea (p x 0.0256).

Elliltiiil The results described above demonstrate that administration of a composition that includes Bifidohaeteria (eg. 8. longzmi subspecies infhniis) with a acillus (e.g., Lg plantamm) that was chosen to consume the free sugar monomers that a known pathogen (e.g., a idiztm species) preferred to consume was ef‘ective at ng the dysbiotic episodes for the newborn foals. This example is not limited to newborn foals. but demonstrates that administration of the compositions described herein can be effective to reduce or eliminate dysbiotic es in mammals. While this example provides experimental support for the concept underlying this invention, it should be noted that these foals were nursing animals where MMO was ed by ’s mills. Therefore, a supplemental MMO was not provided.

Similar results would be expected, were the Mir/if) to be provided as a supplement, rather than mother" s niilk. [9969} Example 7. ing B. infantis in nursing piglets. {997%} in untreated young nursing pigs, populations el‘linterohacteriaceae in the gut were feund tn correlate with the abundance of Bactemidcs (r2 : 0.66li p < 0,001), it was also ieund that these pepnlatiens of Ei’iterehacteriaceae t, by themselves, censume sialylated pig milk eligesaccharides, hut Bacter‘oidss pessess enzymes capable cf releasing sialic acid from pig milk alignsaccharides, which is asseciated with sed abundances of sialic acid in ices ehacteriaceae can censume the sialic acid released by Bactcmides. The treatment cf pigs with Bifidohacterinm and/0r Lactabacillns reduced. the anthem of sialic acid available and resulted in a reductien in scours (See W0 ZOl 6/094836 & W0 2m {mild-9149, the disclesnres of which are incerperated herein in their entirety). [film’ll Example 8. increase in Weight Gain in Nursing Piglets fed a tic Cnn‘tpesitien. [tlll'lZl Pig litters are typically given antihietics prephylactically at birth te prevent early infectiens during nursing including scours" Sceurs can he infectieus from viral or hacterial causes (meat are viral) or can he ated with early pest—weaning. Sceurs is ental tn the Overall performance and health (if the pig. [9973} Several s were ized intn nne of three groups, One grnup received a standard «of care dose ef henzylpenicillin at birth, anether gro up received ne henzylpenicillin, and another group received nn henzylpenicillin but was given 18 billion CPU cf activated B. infer/iris EVCQOl {Example 4) and i hillinn CFU of L. piamamm EVLPOOl daily by nral gavage fer seven days", from day 14-21 of life. L. piamarztm EVLPGOl was iselated frem the feces cf a nursing piglet and cultured in a feed-grade sterile milk medium, t stirring, at 37C and enumerated on MRS medium to cenfirm desing. All animals were weighed at 28 days cf li ‘e and compared acrnss ent grnups. As shown in Figure 2, the piglets that received 8. infantis at l4-Zl days had the highest weight gain.

Claims (9)

1.A medicament fer ng a patient with intestinal distress ct‘nnprising a mammalian mi llt nligosaccharide and a hacteria that internalizes said mammalian milk nligcsaccharide prior in its hydrclysis.

2.The medicament or" claim l, wherein the intestinal distress represents a microbial

3.The medicament of claim 1, wherein the intestinal distress is the result of irritable Bowel e, Crohn9s Disease, Ulceratiye cnlitis, Necrotizing Enterocnlitis, bacterial nr Viral overgrowth, bacterial induced diarrhea, antibiotic treatment, eating disnrders, autism, obesity, or lnw diversity in dietary intake.

4.The medicament of any one cf claims l—3, wherein the hil‘idnhacteria is selected from B. longum B. is, B. caiemtl’amm, B. pseudolangzmi, B. pseudocaraniridium, and B. breve.

5.): The medicament of Claim 4, wherein the B. longzmi is B. longnm snhs. infhniis. The medicament at any nne of claims l—S, wherein the ian lillllh nlignsaccharid e is from mammalian mi llt. The ment of claim 6, wherein the mammalian milk is frem human, bovine, er caprine sources. The ment til Claim 7, wherein the bovine source is frnm bovine celestrnm. The medicament of Claim 6, wherein the mammalian milk nligcsaccharide is frnm whey permeate. ll}. The medicament cf any one cf claims 1—9, wherein the mammalian mill; accharide ses fncesyllactose, sialyllactose or tives thereof. ll. The medicament cf any one at claims l-—-l0, wherein the mammalian milk nligcsaecharide comprises one er more carbohydrates selected frem synthetically produced and purified, 2’—fncosyllactnse, 3—incnsyllact0se, difnensyllactose, lactn~N— fncosylpentaose l, lactc~N—fncnsylpentaose ll, lactc~N~fnc0sylpentaose lll, lacto~N~ fncosylpentaose V, 3’--sialyllact0se, 6“--sialyllactnse, 3'--sialyl--t"ncnsyllact0se, sialyllactn-N—tetranse, ylgalactnsamine, and 6’—sialyllactesamine l2, The medicament of any one of claims lull, wherein the medicament further comprises l..actnhacillns and/er Pedicccccus. 13. The medicament of claim 12.. where in the Lactohacillns is ed from L. plai'zmmm, L. amri, L. brat/is, L. coleohominis, L. fermenttmz, L. gasseri, L. joizm‘onii, L. penlmtts, L. sakei, L. salit’rzrinsy L. casei, L. rhamnosus (

6.5

7.. LGG}, L. acidopizilns. L. cumming, L. renteri, L. marinade, and L. criapal‘us Ed. The medicament or" claim l3, wherein the Lactohacillns is L. rental". 15. The medicament of any one of claims l2—lr-l, wherein the l..a.ctohacillns is provided in a daily dose of from l0 million to l trillion cfn. Ltd. The medicament of any one of claims l2~-—l5, wherein the Lactohacilins is provided in a daily dose of from ll) billion to 50 billion cfn. 1’7. The medicament of any one of claims l---16.. wherein the mammalian milk oligosaccliaride is in the form of a powder. 1

8. The medicament of any one of claims l»---l7, wherein the medicament is in the form of a powder. 1

9., The medicament of any of claims l»——l8, wherein the oligosaccharide is formulated to e dietary liher in an amount suitable for a non—infant patient. 2%. The medicament of claim l9, wherein the non—infant t is at least 6 months of age. 21. The medicament of any one of claims lv---2tl, n the medicament is formulated. to provide a daily ration sufficient to support the inal microhioine of a patient with a body weight r than l0 kg. 22, A method of t feating gastrointestinal dyshiosis by providing a, patient with a composition comprising a mammalian milk oligosaccharide from a mammalian rnilk source and hifidohacteria that alizes said mammalian milk aecharide prior to its hydrolysis for at least 5 days. 23., The , of claim 22. wherein the gastrointestinal dyshiosis is associated with irritable Bowel Disease, Crolin’s Disease, Ulceratiye colitis, Necrotizing Enterocolitis, hacterial or viral overgrowth, bacterial induced diarrhea, antibiotic treatment, eating disorders, autism, obesity, or low diversity in dietary intake. 24., The method of any one of claims 22 or 23., n the hifidohacteria is selected from B. lngum, B. pscudocairmiridium, and B. breve. 25. The method of any one of claims 24, wherein the B. e is B. lat/23ml snhs. infimris. 26. The method ol any one of claims 22—25, wherein the mammalian milk oligosaecharide is WO 56550 from a mammalian miljk sonrced, 27. The method of any one of Claims 22—26, wherein the mammalian milk is from human, hoyine, equine, or caprine sources, 28. The method of claim 27, wherein the bovine sonrce is from bovine colostrum, 29., The method of any one of claims 22-28“, wherein the ian milk oligosaceharide is from whey permeate 3th The method or" any one of claims 22-»29, wherein the mammalian mill; oligosaecharide is selected from fncosyllaetose, sialyllactose, combinations thereof, and derivatives thereof, Ill. The method of any one of claims 22—30, n the mammalian milk oligosaceharide is selected from synthetically produced and purified 2’~fucosyllactose., 3—lneosyllactose, difneosyllactose, lacto~N—i‘ncosylpentaose l, lacto—N—l‘ueosylpentaose ll, lacto—N— fneosylpentaose lll, lacto-N-fneosylpentaose V, 3’--sialyllaetose, 6”--sialyllactose, 3-- sialyl—3—fneosyllactose, sialyllacto—N—tetraose, N—aeetylgalaetosamine, 6'— sialyllactosamine, and combinations thereof. 32, The method of Claim 22—27, wherein the mammalian mill: oligosaeeharide is floor heman so nrces. 33. The method of any one of claims 22-25., wherein the mammalian milk oligosaccharide comprises GGS enriched in DP4 and DPfi. 34., The method of any one of claims 22-—-25 or 33“, wherein the ian milk aceharide is lifom a recombinant microorganism. 35, The method of any one of claims 23-25 or 33, wherein the mammalian milk oligosaecharide is produced by ehemiet l synthesis. 36., The method of any one of claims ZZZ-35, wherein the hifidohacteria is provided in a daily dose of from l n to lth billion Clo, 37. The method of any one of Claims 22—36, wherein the hil‘idohaeteria is ed in a daily dose of from l0 billion to 50 billion elu, 38. The method of any one of claims 22—37, wherein the mammalian milk oligosaceharide is ed in a daily dose of from l to 20 g. 39, The method of any one of claims 22—38, whe fein the ian mill; oligosaceharide is provided in a daily dose of from i to ill gt fill}. The method ol any one of claims 22—39, wherein the mammalian milk oligosaccharide and the hifidohacteria are present in a dry form and enrohed. in a material that would provide enteric protection. 41. The method of any one of claims 22-39., wherein the mammalian mill< oligosaccharide and the hil‘idohaeteria are encapsulated and the capsule has an enteric coating. 42, The method of any one of claims 22-89., wherein the mammalian milk oligosaccharide is provided as a solution and the hit‘idohacteria, is provided as an enterie—coated tahlet or capsule. 43. The method of any one of claims 22-42., wherein the composition further comprises Laetohacillus and/or Pediococcns. 44., The method. of claim 43, wherein the Lactobacillns is selected from L. plantamm, L. casei, L. rhamnosus {as}, LGG}, L. acidophilns, L. curt/rains, L. renteri, L. e, and L. crispams 45. The method of claim 44-, wherein the l..aetohacillns is L. rented sit-ti. The method of any one of claims 43»—--45, wherein the Laetohacillns is provided in a daily dose of from ll) million to l, on cfn. 47. The method of any one of claims 4-3—46, wherein the lactohaeillos is provided in a daily dose of from l0 to 50 Billion chi. 48. The method of any one of claims 22—4-7, wherein the mammalian mi llr oligosaeeharide does not e casein, 49, The method of any one of claims 22—48, wherein the patient is not an infant. St}. The method of any one of claims 22-49, wherein the patient is on a strict diet restricting all other dietary Tiber when said composition is provided to said patient 51., The method of any one of claims 22-50., wherein the mammalian milk oligosaccharide is in the form of a . 52. The method of any one of claims 22—51, wherein the ition is in the form of a. 53. The method of any of claims 22—52, wherein the oligosaceharide is formulated to provide dietary fiber in an amount suitable for a non— infant patient. 54, The method of claim 53, whe fein the fant t is at least 6 rnontl'is of age. 55. The method of any one of claims 22—54, wherein the composition is formulated to provide a daily ration sufficient to t the intestinal niicrohiome of a patient with a body weight greater than l0 kg 56. A composition comprising human mi Eh oiigosaccharides ti—iMO) and t‘ncosyiated glycans (SPF) anti/or siaiylated. glyeans (SP3). wherein the SPF and/or SPS are not of millc origin. 57. The conipositimi of claim 56, wherein the ratio ofl-lMOzSPF is from 20:1 to l:5. 58., The composition of any one of claims 56 or 57, wherein the ition comprises human milk oligosaccharitles and syntheticalily—produced and purified sialyl glycans (SPS), and. wherein the ratio of HMO:SPS is from 5:1 to 1:1. 59. The composition of any one of claims sass. wherein the SPF or SPS comprises one or more of 2'~fncosyllaetose, 3'—fncosyilactose, riifncosyllaetose, iacto—N-f’oeosylpentose l, lacto—N~fncosylpentose ll, iaeto—N~fncosylpentose lll, iaeto—N—fncosyipentose V, 3'~ sialyllactose, iyllaetose, 3'-sialyl—3~fncosyllactose, sialyllacto-N—tetraose, and 6'- sialyllaetosamine {it}. The composition of any of claims 56—59, wherein the composition is formulated to provide dietary fiber in an amount suitable for a non-infant patient. 61, The composition of claim 60, wherein the rimi—infant patient is at least 6 months of age. 62. The composition of any one of claims So—ol n the composition is formulated to provide a daily ration sufficient to support the intestinal mierohiome of a t with a body weight greater than li) leg. 63. A composition comprising bovine mill: oiigosaceharides (BMO) and, fucosylated. giycans (SPF), wherein the SPF is not of mill; origin. 64, The ition of claim 63, wherein the ratio of BMO:SPF is from QUIT to 1:5. 65. The composition of any one of claims 63 or 64, wherein the composition comprises bovine mill: oligosaccharicles and synthetically-produeed and. purified siaiyl glycans (SP8), anti wherein the ratio of BMOzSPS is from 5 :l to lzi. an. The composition of any one of claims 63—65, wherein the SPF or SPS comprises of one or more of 2’—focosyiiaetose, ifucosyilactose, difucosyllactose. lacto—N—fucosylpentaose l, N~fucosylpentaose ll, iacto—N-fncosylpentaose ill, N-fncosylpentaose V, 3’~ sialyllactose. 6”--sialyllactose, 3‘--sialyl-—3~fncosyllactose. sialyllacto--N-tetraose, N-— galactosamine, and 6?—sialyllactosaniine. 67, The composition of any of claims 63—66, wherein the composition is formulated to provide dietary fiber in an amount suitable for a non—infant patient. 68. The ition of claim 67, wherein the non—infant patient is at least 6 months of age. 69. The composition of any one of claims 63—68, wherein the composition is formulated to e a daily ration sufficient to t the intestinal microbiome of a patient with a hody weight greater than 10 kg. 7t}. A method of treating gastrointestinal dyshiosis comprising administering a mammalian milk oligosaccharide to a patient in need thereoi‘, wherein the patient in need thereof is a patient with a dyshiotic niicrohiome. 71. The method, of claim 70, wherein the dyshiotie microbiome is a bacterial overgrowth of opportunistic ens and/or overt pathogens. 72., The method. of claim 70 or 7L wherein the dyshiotic microhiome has a reduced ial diversity. ’73, The method or" any one of claims 70-72, wherein the mammalian milk is from human, hovihe, or caprine sources. 74., The method of claim 73., wherein the bovine source is from bovine colostrum. 75, The method of any one of claims 70—74, whe fein the mammalian mill; oligosaccharide is from whey permeate. 76. The method of any one of claims ’70-—-—75, wherein the mammalian millg oiigosaccharide is selected from focosyllactose, sialyilactose, combinations thereof, and derivatives thereof. 77., The method of any one of claims 70-376., wherein the mammalian mill: oligosaceharide is sel acted trom synthetically produced and purified 2’-t‘ncosyllactose, 3—fucosyllactose, difucosyllactose, Nnt'ucosylpentaose l, lacto-N--t"ucosylpentaose ll: lacto-—N-- lpentaose ill, lacto—N—fncosylpehtaose V, 3’—sialyllactose, 6’—sialyllactose, 3‘— sialyl-—3"focosyllactose, sialyllacto-N-tetraose, tylgaiactosamine, 6-- sialyllactosamine, and combinations f, 78. The method of claim '73, wherein the mammalian milk is from human sources. 79. The method of any one of claims 70-378, wherein the patient in need thereof is not an lit}. The method of any one of claims 70-79., wherein the mammalian milk accharide is in the form of a powder. 81, The method of any of claims 70-807 wherein the mammalian mill: oligosaccharide is formulated to provide dietary fiber in an, amount suitable for a non—intant patient. 82., The methed 0f claim 8L wherein the non—infant t is at leagt 6 mtmths 0f age, 83. The methed at any one Of Ciaimg 70—82, wherein the mammaiian milk Oiigosaceharide is farmuiated to previde a daily ration sufficient to support the intestinai aficmbmme Of a patient with a body weight greater than it) kg. WO 56550 PEG” ”E