GB2522627A

GB2522627A - Composition

Info

Publication number: GB2522627A
Application number: GB1401510.1A
Authority: GB
Inventors: Nicholas John Larkins
Original assignee: GREENCOAT Ltd
Current assignee: GREENCOAT Ltd
Priority date: 2014-01-29
Filing date: 2014-01-29
Publication date: 2015-08-05
Also published as: GB201401510D0

Abstract

A composition comprises medium chain triglyceride compounds and dietary fibre, and may comprise one or more branched chain amino acid(s), such as leucine, isoleucine, and valine. Preferably, the medium chain triglyceride compounds comprise copra. The dietary fibre may comprise mushroom species, such as mushroom waste, Ganoderma lucidum, Lentinula edodes and Trametes versicolor. The composition may also include biotin. Preferably, the compositions are effective at treating one or more of gastrointestinal tract dysbiosis, diarrhea, gastritis, mega colon, abdominal distension, laminitis, colic, flatus, constipation, gastric ulcer syndrome, and muscle fatigue.

Description

I

Coniposition The present invention provides compositions which may be used in the treatment of, for example, gastrointestinal tract cysbiosis, as wel as conditions that nay be associated therewith, such as mega color, diarrhoea, gastritis and abdominal distension.

Background

The gut mucosa is the largest and most dynamic mmunologca! envronment of the body-It is often the first pont of exposure to pathogens. and many microbes use it as a beachhead for access to the rest of the body. Therefore the gut immune system needs to be ready to respond to pathogens, hut at the same tulle, a must toierate innocuous environfllentai antigens. food partides and commensa! mcrofiora to which it is constantly exposed.

Msdrected mrnune responses to harmless anirgens are the unue lying cause of food aiergies and debilitaUng conditions such as infl9mm-atory bowel disease.

The' mammaian body is home to far more microflora han mammalian cells at least 100 trillion microbial cells (Whitman et al., 1998) and a quadrillion viruses (Haynes and Rohwer, 201 1). Collectively, the microbial associates that reside in and on the mammalian body constitute microbiota, and the genes they encode are the microbiome. This complex community contains taxa from across the tree of life, bacteria, eukaryotes, viruses, and at least one archaeon, that interact with one anothei and with the host, greatly impacti"g health cnd physioogy rhe microbiota plays a major role in health and disease in humans, indeed, it is sometimes referred to as a forgotten organ" (OHara and Shanahan, 2006).

The gut nuicrobiota is integral to development and maturaton of the host immune system, plays a role in the differentiaton and turnover ot gut epithelial tissues and may be an important factor in various diseases The microbiota is involved in energy harvest arid storage, as wefl as in a variety of metabolic functions such as fermenting and absorbing undigested carbohydrates (Gil et al 2006), a trait that has probably acted as a strong evolutionary force toward the establishment of bacteria as symbionts. Perhaps even more importantly, the gut microbiota interacts with the immune system, providing signals to promote the maturation of immune cells and the normal development of immune functions (Chow et aL, 201 0).

Although the nicroorota is generally stable within indiv,duas over time, me composition can be altered due to external perturbations. One of the major factors that can perturb the composition of the n,icrc.biota is anthiotic use. Although the particular taa affected vaiy among individuais, some taxa do not recover eve-n months after treaurienL and in general.

there is a long-term decrease in bacteriai diversity. As the estabflshed gut bacter!ai community eshapes after treatment with antibiotics, there is a redced resstance o colonization, allowing foreign microbes that can outgrow commensal bacteria to cause permanent changes in the structure of the nicrobiota.

The consequences c-flmbaiances in gut bacteria and ndeed other gut microbiota are manifested n a number of conditions. For example: mmes in gut rncrobiota 0 (dysbiosis can cause a number of conditions affecting both organ systems near and far frorri the gastrointestinal tract. The rrechanisms through which microbiota exerts its beneficial or detrimental influences remain ianjer uncefined, hut include elaboration 01 sign&ling molecules and recognuor1 of hacterai epitopes by both intestinal epithelial and mucosa mmune cefis. Examples 01 conditions which may stem from gut mjcrobota riyshiosis are autoimi une diseases. gastrointestinal conditions, such as abdominal distension and diarrhoea, as WOil as other more remote conditions such-as larninitis Treating gastrointestinal tract dysbiosis has provided a significant chaDenge, not least because of the complexity of the gut microbiota, For exaniple, introducing just one microbial species into a patients gastro intestinal tract is unlikely to provide a therapeutic solution. Probiotic compositions or mixtures that are administered orally often do not lead to a successful treatment of diseases and/cr symptoms associated with gastrointestinal dysbiosis, Faecai microbial transplant is a therapy that is showing potential promise in treat ny conoitions associated with gastrointestinal tract dysbiosis Feca1 microbial transplant therapy involves transplanting taeal matenal from a Healthy donor into a patient This commonly carned oA via an enema, although son'e studies have also administered the transplant v a a iasal gastric tube It will be appreciated that fecal microbial transolant therapy may not appeal to many patients.

Accordingly, new therapeutic compositions are required that lead to good patient compliance and can treat gastrointestinal tract dysbiosis, and/ or treat the conditions associated with gastrontestinal tract dysbiosis for example, diarrhoea, mega colon, laminitis, abdominal distension and constipation.

Statement of invention

According to the present invention in a first aspect there is provided a composition (for example a pharmaceutical or veterinary composition, for example a dietary supplement) comprising medium chain triglyceride compounds (for example a source of medium chain triglyceride compounds) and dietary fibre (for example a source of dietary fibre).

Apphcants have found that a composition comprising medium chain trigycerde compoulFis in addition to detarv fibre (and. possihiy. other components) may be effective ttreating gastrointestnai dysbosis, and/or conditions associated wfth gastrointestnaI dysbioss. In

Medium chain triglyceride compc.'unds MCT) are generally considered a good b!ologicay inert source of energy that are re!atively easy to metaoohse Sources of medium chain triglyceride con:pour:ds include for example, palm kernel oil, coconut oU, camphor tree C! upes and c:opra. Copra is the dried meat or Kernel cf the coconut palm.

Preferably, the medium chain triglyceride compounds comprise copra.

The medium chain triglyceride compounds may be present in an effective amount. The medium chain triglyceride compounds may be present in an amount from around 2.7% by weight to around 98.2% by weight, for example from around 10% to around 95 %, for example from around 10.5% to around 82% by weight of the total amount of the medium chain triglyceride compounds, the dietary fibre, the branched chain amino acid(s) (if present) and the biotin (if present). Applicants have found that the above amounts of the med urn chain triglyceride compounds may be particularly effective for treating gastrointestinal tract dysbiosis and/or corthtions associated therewith Dietary fibre may refer to the indigestible portion of food aerived from plants, rd the waste of animals that eat dietary fibre. Dietary fibre can include a complex mixture of soluble and insoluble fibre and can include, for example, non-starch polysaccharides, cellulose, inulin, ignin and waxes. Another example of a source of dietary fibre is mushroom species. The source of the dietary fibre may be mushroom waste, for example mushroom waste, which includes mushroom stipes and bases of edible mushrooms. The dietary fibre (mushroom waste) may include nonstarch po'ysaccharides, polysacchande-protein and polysaccharide-peptide complexes, ribonucleases, proteases and lectins. Mushrooms also accumulate many secondary metabolites, fur example, polyphenols, polyketides, terpeiles and steroids. Most large indigestible glycans are unable to cross the intestinal epithelium and therefore remain the gut while being used by the colon microbiota or excreted as faeces. Without being bound to any particular theory, these indigestible glycans can enhance colonization of beneficial microbiota, thus providing a prebictic benefit.

Preferably, the detary fibre comp(ses mushrooms species The mushroom species may be (or include) for example Trametes versico or, Lentinula edodes, Pleurotus eryngii, Flarnmulina velutipes arid/or Ganoderma lucidum. The d etary fibre may comprise one or more of mushroom waste, ganodema lucidL m, Lentinula edodes and trametes vers color The apphca its have found that dietary fibre derived from mus rooms is an economica and effectve source of dietary fibre for inclusion in compositions of the invention. In particu a the applicants have found that dietary fibre se ected from at least one of mushroom waste, ganodema ucidum, Lentinula edodes and trarnetes versico or may he particular effective.

The detary fibre may be present in an effectve amount. The dietary fibre may be present in an amount from around 0. 9% to 75% by weight, for examp e from around 7% to around 72% by weight, for examp e from around 8% to a ound 27% by weight of the total amount of the med urn chain triglyce ide compounds, the dietary fibre, the branched chain amino acid(s) if present) and the biotin (it present). App icants have found that the above amounts of the dietary fibre may be particu any effective for treating gastrointestinal tract dysbiosis and/or cond tons associated therewith, The composition may further comprise one or more branched chain amino acid(si: Branched chain amino acids (BCAA) include amino acids having branched aliphatic side chains! for example eucine, isoleucine and valine. Without being bound by any particular theory, it has been found that certain branched chain amino acids may act as substrates and as key regulators for various nutrient metabolism and therefore enhance the activity of the composition The branched chain amino acid(s) may be selected Iron' leucine, isoleucine and valine.

The composition may include all of leucine isoleucine and valine Preferably, the branched cha'n amino acid is leucine The amount of leucine to isoleucine to valine may be from around 1 0 0 to arouna 8 1 1 For example, the amount of leucine to isoleucine to valine may be around 4:1:1.

The branched chain amino acid(s) (f present) may be present n an effective amount The branched chain amino acid(s) may be present in an amount from around 0% to around 83.9%, for exampie from around 8% to around 70% for example from around 9% to around 64% by weight of the total amount of the medium chai trg yceride compounds, the dietary fibre, the branched chain amino acid(s) (if present) an the biotin (if present). The appUcants have found that the above amounts of the branched chain amino acrd(s) U present) may be particu arty effective for treating gastrointestinal tract dysbiosis and/or conditions assocated therewith The composition may include biotin. Biotin is also sometimes referred to as Vitamin B7 or Vitamin H. Without being bound to any particular theory, it has been found that biotn acts to fully catabohse isoleucine and leucine (if present). The compositions of the invention may include biotin. The biotin if present) may be present in an effective amount. The compositions of the invention may include biotin in an amount from around 0 % to around 1%, for examole from around 00006% to around 0 9%, for example from around 0 0025% to around 0.5%, for example around 0.01% by weight of the total amount of the medium chain triglyceride compounds, the dietary fibre, the branched chain amino acid(s) (if present) and the biotin (if present). The applicants have found that the above amounts of the biotin (if present) may be particularly effectIve for treating gastrointestinal tract dysbiosis and/or conditions associated therewith.

The compositions of the invention may be used for the treatment of, for example gastrointestinal tract dysbiosis, for example the treatment of one or more of diarrhoea, mega colon, laminitis, colic, muscle fatigLe infreciuert bowel movements, frequent bowel movements, abdominal distension, fatus, constipation, gastritis and gastric ulcer syndrome.

According to the present invention in a further aspect there s a provided a composition comprising medium chain triglyceride compounds and dietary fibre, for, or for use in, the treatment of gastro intestinal tract dysbiosis. The composition may further comprise one or more branched chain amino acid(s), and/or biotin. The composition may comprise medium chaIn triglyceride compounds, dietary fibre1 one or more branched chain amino acid(s) and biotin.

The compositions of the invention may be for. or for use in, the prevention and/or the treatment of lower gastrointestinal tract dysbiosis. for example large intestine dysbosis The cOrnpos!tions of the invention may he for, or for use in. the prevention and/cl the treatmer1t 01 one or more of diarrhoea, mega colon, !amuiitis, cohc, muscle fatigue, infrequent bowel moverr.ents. frequent bowel movements. abdominal dstenson, flatus.

constipation, oastritis and gastrc ulcer syndrome The applicants have found that compositions acconiing to the nvention may be remarkabiy effective at treating diarrhoea, mega coion laminitis. colic. muscle fatigue. infrequent bowel movements, frequent bowei movements, abdominal distension, fiatus, corisripaUon gastritis and gastric ulcer syndrome. Without being bound b'y any particular tneorv, tne applicants hypothes!se that the ccmoostions of the invention Freat the above diseases by address!nçl uriderlying gastrcintestnai tracL dvsbiosis. Ft wiil be appi eciated that the composit:OnS of the invention may be used to treat a number of other conditions that may be associat d with g'astrcintest!nai tract dysbiosis, such as nflammatory bowe! disease. Chrohns disease.

ulcerative colitis, irritable bowel disease and chronic fat!gue syn.c:rorn According to the present invention in a stili further aspect there is provided the use of medurn chain trglyceride compounds and dietary fibre (e g specific dietary fibres described herein) in the manufacture of a medicament for the treatment of gastrointestinal tract dysbiosis.

The medicarnent may further comprise one or more branched chain amino acid(s) and/or biotin.

The treatment of gastrointestinal tract dysbiosis may be lower gastrointestinal tract dysbiosis, for exarrpte large intestine dysbiosis According to the present invention in a still further aspect there is provided the use of medium chain triglyceride compounds and dietary fibre in the manufacture of a medicament for the treatment of one or more of diarrhoea mega colon, laminitis, colic, muscle fatigue, infrequent bowel movements, frequent bowel movements, abdominal distension, flatus, constipation, gastritis and gastric ulcer syndrome.

The medicament may Lrther comprise one or more branched chain amino acid(s) and/or biotin According to the present invention in a further aspect there is provided a method of prevention and/or treatment of gastrointestinal tract dysbiosis comprising a step of administering to a human or a non-human animal subject in ne'd theteof a composition S comprising medium chain triglyceride compounds and dietary fibre.

The composition may further comprise one or more branched chain ammo acid(s) anc:/or hiotin. The method or treatment may be a method of treatment of lower gastrointestinal tract dysbosis. for exampie large intestine dysbiosis.

According to the present invention n a sUM further aspect there is provided a method c1 prevention and/or treatment of one or more ot da toes. mnega coion. laminitis. colic:.

muscle fatigue, infrequent tc"jvei moverr:&hts. fteuent bowel:poveme.nts abdominal diatension. Ilatus constipation, gastritis and gastric ulcer syndrome comprising a step of administering to a human or a non-humar anin'.ai subject in need thereof a composition comprising medium chain triglyceriøe CompoUnds and c:ietary fibre.

The composition may further comprise one or more branched chain amino acid(s) and/or biotin, In addition to the components described above, the compositions and medicaments of the invention may further comprise other components such as excipients, diluents, binders and the like.

The compositions and medicarnents of the invention may be suitable for oral adninistration The methods of formulation of the compositions for oral adminisVation are well known in the art. For example, the composition for administration may be prepared usng a pharmaceutically acceptable carrier rn a form suitable for administration Such a carrier can be prepared as a tablet, a pill, a sugar-coated agent, a capsule, a liquid, a gel, a syrup, a slurry, a suspension, a cachet etc. The carrier may be one or more pharmaceutically acceptable carriers such as liposomes, lactose, trehalose, sucrose, marinitol, xyiitol, crystalline cellulose, ohitosan, calcium carbonate, talc, titanium oxide, or silica (silicon oxide) or the like.

The composition and rnedicaments may be obtained, for example, by combining the active ingredients with a solid excipient, pulverizing the mixture (if necessary) and inserting into a capsule, for example, a soft sealed capsule consisting of a gelatin capsule, gelatin and coating (e.g., giycerol or sorbitol) or a capsule composition suitable for vegetarians. In the soft capsule, the composthon may be dissolved or suspended in an appropriate liquid, such as a fatty oil, Uquid paraffin or liquid polyethylene glycol, with or without a stabilizer.

The compositions and medicaments of the invention. may he suitable for!ncorporator into food products for both humans and non human animals. For example, the compostons may be ncorporated into animal feed.

The composiUons and medicarients of the invention. may he for use in treating (or for use in the manufacture of a medcament for the prevention of and/or treatment of) both humans and non human animals, for example equine species such as horses. dogs, cats. pigs.

tum!nants such as cattle, sheep. goats, camels, deer, poultry and game hftds.

The present invention will now be described in detail with reference to the following

examples.

Examples

Amount of Amount of Total medium medium Total amount of Total chain chain amount of branched Amount amount Total glyceride glyceride branched chain Amount Amount of of amount compounds compounds chain amino of of Amount Amount Bioun I dietary of dietary (Copra) / (Copra) 1 % amino acids 1 % leucine! isoleucine of valine I of Biotm % by fibre! fibre t°,: _ExamplQ jg_ by weight acids! my by weight my 1mg my w*3ght.9!9 by weight 103 600 63.15 4C0 00 100 0.1 0.01 250 630 400 00 100,.l 0.01 250 2631 S!Q. 600 63.15 00 100 0.1 0.31 250 2631 100 28.57. . --250 7L43 4 10000 90 91. .. . . woo jog 13000 8197 200 934 803 20:1 203 01 00008 1000 3.20 10000 90.9. . . . 1000 909 7 13000 81 97 1200 934 330 2C0 200 0 1 00006.000 8.20 8 10000 8.97 123.t'4 00 200 200 0: :30003. 000 820 9 50 20.30 150 6000 100 25 25 000 50 20.00 100 30.77 ____ 153 46.15 100 25 26. 0.09 75 flOE ii 50 20.00 50 6000 100 25 25. . 0.00 50 20.00 CO 2000 150 noo:oo 25 25: 0.00 50 20.00 1:3 50 2000 150 6000 00 25 25. 3.thJ. 50 2000 Table 1. surir.ary of the compos:tions usec: Examples Ito 13 c In the following examples the copra was obtained from Worlee Naturprodukte GmbH, of Germany; the eucine was obtained from Cambridge Commodities Ltd of Cambridge United Kingdom (Product cede P1204), the Eiiotin was obtained from Cambridge Comrnooities Ltd of Cambridge United Kingdom (Product code P0207), the m ishroom species and mushroom waste were obtained from Worlée NaturProdukte GmbH, of Germany. Human

Example I

year old male with "massive gastrointestinal dssenUon and discomfort alter eating rataj fish (cod) at a local restaurant. Although there were frequent niassivi fiatus eruptons, which were relieving, the patent recorted that stool nroduction continued to be normal f somewhat reduced ri quantity. The patient complained that he found it more difficult to urinate when hs "tummy' wr.s bloated. The p9Uent reported that probiotcs were not hep!ng. The consulting gastroenteroioist considered that that the problem might due an anaerobic. infection and accordingly prescribed metronidazole 500 mg taken two times per day for 5 days, which provided minimal symptomatic reef The Patent observed that when he did not consume ?ny da!ry prodi:cts. he was syrnptomaticafly much improved.

FoHowV a dairvp roduct free diet aflcwed the oatient a much: inproved quality of life.

Whenever any dairy product was wicluaed into the patients diet, the patient experienced a return of the' qastrontestina bloating and excessive flatus production. However, this was much more manageable than in the past. copra (MCI) 100mg; BCPA --leucine:isoleucine:viine (400mg.100mg.10Ong: b!ot!n 100 pg; ganoderma ucidum 250 iig was then added to this dairy-product-free diet regime At 5 days, the patient reported that he had started consuming dairy products again, and now he did not suffer any of the previous synptornatic discomforts/problems This Copra (MCT) 100mg, BCAA-leucine isoleucinevalre (400mg 100mg 100mg) biotin 100 pg, ganoderma ucidum 250 mg regime continued over the foflowing months with the patient reporting continued good health.

Example 2

32 year old male with a history of "unstable" gastrointestinal activity that was "sometimes normal, sometimes runny: sometimes gassy doesn't matter what eat". Copra (MCT) 100mg; BCAA ((eucine: isoleucine: valine (400mg:lOOmg:lOOmg)); biotin 100 pg; ganoderma lucidum 250 mg was advised to be taken in the morning and the evening or 5 days. At day 101 the patient reported consistent problem-free and norma bowel movements The patien' then connnued this daily regime of Copra (MCT) 100mg, BCAA leucineisoleucine:vallne (400mg: 100mg: 100mg); biotin 100 jig; ganoderma lucidum 250 rng and reported after 3 months that aD was perfect".

Example 3

year old ma:e patient wth a long history of dsrLlpti'de bowel movements that frequently dsturbed him durir:g the rugt. Over the years. the patient had tried various produors incluthng an assomtnlent of prohotics. lt was advised that Copra (MCT) 100mg: BCAA eucne:scleucine.vane (400rng.100mg:100mg, b!otin IOU pg; g9noderma lucdum 250 mc: was taken twice per day miqht help reEolve this prohiem. At 48 hours, the patient reported that he now had d!arrhoea -"iOOSC hut not runny stools He wished tO 5O taking the Cc'pra (MCI) 100mg; BCAA -eucne:.so;eucne'vahne (4O0mg100ng.100mg). biotin jig' ganodenna lucidum 250 mg. After expaining the possible longer term oenefits of this mx. the patient agreed to continue wit' a mix variation of copra MCT'i 100mg.

ganoderma ucidurn 250mg at a dose regime of once per day. At 10 days, tne patient reported that bowel activity had mproved. he was no lonc.ar having darrhoea reiated problems, and over the last days he had had a c:ecert rnghts sleep'. The patient reported that he a now ab;e to take the onginal Gopra UV1CT) 100mg, BCAA leucine:isoleucr.e,vahrie &iOOmg. 100mg: 100mg); biotin 100 ig; qanoderma licioum 250 mg combination wthout any further problenis Equine Some of these equine studies relate to laniinitis provoked by gut microbiota dysbiosis.

Without beirg bound by any partcular theory it a believed that endotoxins are produced by gut dgsbiosis, which are then reeased nto the blood, targeting and provoking pathological cestrjcticn of the sensitive/fragile hoof Iaminae Laminitis induced by gut microbiota dysbiosis is known to be ti ggered inter alia, b' causes such as crbohydrate overload; it can develop after colic; and lush pastures due to sugars which cause increase in insulin levels, which is knowr to tigger lamin tis Wrthout being bound by any particular theory, it is hypothesised that fructan causes larninitis by causing an imbalance of the normal bowel flora, leading to endotoxin production.

The mix comprising MCT, Mushroom, and BCAA may help restore gut microbicta to normal/healthy homeostatic levels. The addition of the above Mix as a supplement inclusion may provide a prophylactic benefit by maintaining a healthy gut homeostasis,

Example 4

Lam! ntis year oid mare with a recent history of a min.itis. Two months c4 on gong veterinary care and supervision involving rem edial shoeing; daily non steroidal anti nflammatory drug NSAID) medication1 and dietary and supplement management provided some benefit A combination of Copra (MCI) 10000 my. muthroom waste 1 000mg was added to the FRS supplement already being used. An irnprovc-meffl was jbsered within. 7 days. At 21 days, the owner reported the mare was much improved and no longer requhina NSAILJ ned!CatiorI. 4.-' U)

Example 5

Darrhoea year Old mare with a hstory of sudden onset diarrhoea dag nosed as a possible food related ailergic response. Along w!ti poLio s. a non-cereal controlled diet yeas prescribed. W#hn 7 days, the owner reported some mprovemenl and the fec.a! droppngs were more ftc loose pats. A comhnaticn of Copra (MCT) 10.000 mg: BCAA le.uclneisoleuc!ne.vaiine (800mg.20Orng.2.00mg Potin lOG: mcg: and mushroom waste 1000mg was added to the problotic inclusion. Within 48 hours a further mprovement was observed. At 10 days. the owner reported normal healthy leca! dropp:.ngs.

Example &

Laminitis 7 year old gelding Witk a history of "winter' laminitis (potentially due to excessively high sugar (fructari) levels in frosted grasses). A management programme that involved NSAID medication along with stable confinement and a controlled diet was advised. After 7 days the horse was stable but progressing at slower rate than hoped (there was still some increased digital palmar arterial pulsations n both forefeet) A combination of Copra (MCT) 101000 mg; mushroom waste I.000mg was added to the FRS and probiotic supplement already being used Within 24 hours, the digital pulses on both forefee were norma and the horse was able to move freely

Example 7

Laminitis 12 year old gelding with a history of acute onset of severe laminitis. The gelding was unable to bare weight, and had pounding pulses in the palmar digital arteries of all four feet. The horse was placed/bedded in a deep sand litter box. Veterinary care and superv!sic.'n nvo!ved remedial hoof re-sectiorung: h!gh close NSAID medication; 24 hours fasting (adlib water only). followed by slur rv feeding of lucerne provciecJ some benefit over the oi!o'.viriq dy.s However the proQncsis con.tinuecJ o he very ooor. Lamnase. which cota rs RS was fl: ded nto -e ucetmo slury di 1V th n 5 hcLa s, the nnrs able to sell-support ris Own WeigMt wtriout assistau. althc'U_.*h al iCt 0tifl nan mcrku paimar c:igitais pu:ses After 5 days, no further mprovements were dscern!ble A combnation of Copra (MCT) 10.000 mg: BCAA -!eucinelsoleucinevaline 800.m200mg:200mg Biotin. 100 pg: Lentnua edodes (Shitake n!ycehffl 1.000mg was added to the Laminase supplement. W!thln 48 hours, aU dgita puLses on both forefoot were barely detectable nd the horse was abJe to siowly move around the stable At 5 days, the horse was able c walk out from -he stabe wuh some discomfort, which was to be expected.

ExampteB Mild colic/gastrointestinal bloating year old mare with a long term history of mild colic along with gastrointestinal bloating that frequently required veterinary attention and medications. including antispasmodics and probiotics. Along with a controfled dietary management in conjunction to being supplemented with NAF Pink Powder Probiotic (available from NAF of Monmouth, United Kingdom) the mare Improved wah ninimal cohc-like discomfort episodes that no longer required veterinary attention. She was much improved and able to be ridden without any apparent discomfort. A combination of Copra (MCT) 10,000 mg: BCAA leucine:isoleucine;valine (SOOrng:200nig:ZOOnig); Biotin 100 mcg; Lentinula edodes (shutake rrycelia)1,000mg was added to the NA Pink Powder Probiotic supplement Over the following 3 months the owner reported that the horse has been coic free and had not shown any noticeable gastrointestinal discomfort.

Canine

Example 9

7 year old male dog with a clinical history of megacolon with constipation. Over a period of months, a varied and well balanced wet and dry food twice daiiy dietary regime was unable to control the repeated straining when attempting to defecate. A combination of Copra (MCT) 50mg; BCEkA -leucine;isoleucine:valine (lOOmg:25mg: 25mg); Trametes versicolor 1 4 mg was added to th:s twice daily dietary regime. Over the 1ol!owng iC cays. the owner repoited that that te repeated straining when attempting to defecate seemed to be somewhat IeSS At the end of three weeks, the owner reported that that the steain!n.g when.

attemphng to defecate was no longer observable and the bowel movements were "healthy normal'. At 4 months of daily supplementatior: of the Copra (MCI) 5.0mg: BCAA -.

ieucine:!soleucine'valine (1 OOrno'25rng: 25mg1: rrametes versicolor 51) fig, the owner reported conunued good health with nc tunner problems whatsoever'.

Example IC

8 year old male dog presented with a reoccurrence of gastritis. Over the past two months there had been repeated gastric episodes.)f vomiting and riappetance. After undegong an extensive examination. detaiy management changes were acvised which a!so included probiotc combinations. This dietary management regime rroved not to be successfuL Symptomatic relief was obtained by prescribing Dc'xycycflne 100mg twice per da over 5 days: however the relief was temporary.A:ombination of Copra (MCI) 100mg; BCAA eucine:so!eucne.va!ine (100mg 25ma: 25mgL Shitake (1.entinuia edodes) 75 mg was aced to this tw!ce aany dietary regime At me end or three weeks, me owner reported that there had be-en no further problems The owner was advised to continue wth the Copra (MC[) 1UQnic: BCPA -leucinelsoleucne:va;i no (iOOmg:25.mg: 25mg); suifitake cLertinula edodes) 75 mq. At two months, the owne rreported fu ther pro-bems Examples 11,12 and 13 For fnancia reasons three dogs all from the same housing environment (3 year old male, 5 year old male and 7 year old male) had their wet-food daily dietary regimes changed to a more cost effective commercialy available tinned dog food The owner reported that within 24 hours al dogs were producing a lot of really stinky noisy farts (flatulence)" As they were Indoor dogs" I ving with them was not a very oleasant experience -as you might imagine". The owner was provided with a premixed combination of Copra (MCT) 50mg; BCAA -leucine soleucine valino (100mg 25mg 25mg), Shutake (Lentinula edodes) 50 mg, which was to be added to the diet at each feedirg The owner reported that at 45 hours "al is back to normal -thank you" The owner was advised to continue adding the Copra (MCT) 50mg BCAA -eucine soleu3ne valine (100mg 25mg 25mg), Sh take (Lentinula edodes) 50 mg supplementation.

Some of the above exarnpes nc ude branch chan am'rio acids whch nclude leucine, soeuoine and vahne. Other examples of the compos tbns of the nvenUon ray nclude euc ne (as the on y BCAA

S