Login| Sign Up| Help| Contact|

Patent Searching and Data


Title:
PROBIOTIC COMPOSITIONS AND USES THEREOF
Document Type and Number:
WIPO Patent Application WO/2016/179639
Kind Code:
A1
Abstract:
The present invention relates to a probiotic composition comprising: coconut milk; bacteria capable of fermenting saccharides to lactic acid; and saccharides, wherein the saccharides are supplementary to any present in the coconut milk, the bacteria, or both.

Inventors:
MARKSON, Ro (AC Agency, Suite 205 19a Boundary Stree, Rushcutters Bay NSW 2011, AU)
Application Number:
AU2016/000165
Publication Date:
November 17, 2016
Filing Date:
May 13, 2016
Export Citation:
Click for automatic bibliography generation   Help
Assignee:
MARKSON, Ro (AC Agency, Suite 205 19a Boundary Stree, Rushcutters Bay NSW 2011, AU)
International Classes:
A61K36/889; A23C9/133; A23C11/10; C12N1/04; C12N1/20; C12R1/225
Other References:
ABOULFAZLI, F. ET AL.: "Effects of fermentation by Bifidobacterium bifidum on the rheology and physical properties of ice cream mixes made with cow and vegetable milks", INTERNATIONAL JOURNAL OF FOOD SCIENCE & TECHNOLOGY, vol. 50, no. 4, 2015, pages 942 - 949, XP055329492
CORREA, S. ET AL.: "Probiotic potential and sensory properties of coconut flan supplemented with Lactobacillus paracasei and Bifidobacterium lactis", INTERNATIONAL JOURNAL OF FOOD SCIENCE & TECHNOLOGY, vol. 43, no. 9, 2008, pages 1560 - 1568, XP055329493
KOLAPO, A. ET AL.: "Effect of different concentrations of coconut milk on the chemical and sensory properties of soy-coconut milk based yoghurt", FOOD AND PUBLIC HEALTH, vol. 2, no. 4, 2012, pages 85 - 91, XP055329494
LADOKUN, O. ET AL.: "Fermented milk products from different milk types", FOOD AND NUTRITION SCIENCES, vol. 5, no. 13, 2014, pages 1228 - 33, XP055329495
DATABASE WPI Derwent World Patents Index;
DATABASE WPI Derwent World Patents Index;
Attorney, Agent or Firm:
SPRUSON & FERGUSON (GPO Box 3898, Sydney, New South Wales 2001, AU)
Download PDF:
Claims:
CLAIMS

1. A probiotic composition comprising:

coconut milk;

bacteria capable of fermenting saccharides to lactic acid; and

saccharides,

wherein the saccharides are supplementary to any present in the coconut milk, the bacteria, or both.

2. The probiotic composition according to claim 1 , wherein the bacteria are lactobaciUus bacteria, lactococcus bacteria, bifidobacteria, or any combination thereof.

3. The probiotic composition according to claim 2, wherein the lactobaciUus bacteria are selected from the group consisting of: L. acidophilus, L. fermentum, L. paracasei, L. brevis, L. gasseri, L. p/an/arum, L. biiJgaricits, L. helveticus, L. reuteri, L. casei, L. jensenii, L. rhamnosns, L. crispatus, L. johnsonii, L. salivarius, and any combination thereof.

4. The probiotic composition according to claim 3, wherein the lactobaciUus bacteria comprise or consist of L. planlariim strain LM.

5. The probiotic composition according to claim 2 or claim 3, wherein the lactococcus bacteria are selected from the group consisting of: lactis, L. lactis subsp. lactis, L. lactis subsp. cremoris, and combinations thereof.

6. The probiotic composition according to claim 2 or claim 3, wherein the lactococcus bacteria comprise or consist of/,, lactis and L. lactis subsp. Cremoris.

7. The probiotic composition according to any one of claims 2 to 6, wherein the bifidobacteria are selected from the group consisting of: B. adolescentis, B. breve, B. longum, B. animalis, B. infantis, B. thermophilum, B. bifidum, B. lactis, and any combination thereof.

8. The probiotic composition according to any one of claims 1 to 7, further comprising one or more additional components selected from a thickening agent, a prebiotic, a flavouring agent, and citric acid, wherein the one or more additional components are supplementary to the bacteria, coconut milk and saccharides.

9. The probiotic composition according claim 8, wherein the prebiotic is selected from the group consisting of polysaccharides, mucopolysaccharides, oligosaccharides, galactooligosaccharides, fructooligosaccharides, inulin, vitamins, harvested metabolic products of biological organisms, amino acids, nutrient precursors, proteins, lipids, Sunfiber®, Larch Arabinogalactan, Kirkman's Pre-Bio™ Fiber Powder, and any combination thereof.

10. The probiotic composition according claim 8 or claim 9, wherein the thickening agent is selected from the group consisting of pectin, citrus pectin, modified citrus pectin, kuzu, kuzu root starch, starch, polymerised whey protein, agar powder, gelatin, guar gum, arabic gum, Sunfiber and any combination thereof.

1 1 . The probiotic composition according to any one of claims 6 to 10, wherein the thickening agent is citrus pectin, kuzu root starch, or a combination thereof.

12. The probiotic composition according to any one of claims 6 to 1 1 , wherein the flavouring is selected from the group consisting of coconut flavouring, vanilla flavouring, and a combination thereof.

13. The probiotic composition according to any one of claims 1 to 12, wherein the saccharides are a component of fruit juice that included in the composition.

14. The probiotic composition according to claim 13, wherein the fruit juice is pear juice.

15. The probiotic composition according to claim 13 or claim 14, wherein the composition comprises between 10% and 80%, between 20% and 70%, between 30% and 60%, between 40% and 60%, between 45% and 55%, or between 45% and 50% fruit juice.

16. The probiotic composition according to any one of claims 1 to 15, wherein the coconut milk additionally comprises carob bean gum.

17. The probiotic composition according to any one of claims 1 to 16, wherein the composition comprises between 10% and 80%, between 20% and 70%, between 30% and 60%, between 40% and 60%, between 45% and 55%, or between 45% and 50% coconut milk.

18. The probiotic composition according to any one of claims 1 to 17, wherein the composition pH is between 4.0 and 6.5, between 4.5 and 6.0, between 4.0 and 5.5, between 4.5 and 5.5, or between 4.5 and 5.5.

1 . The probiotic composition according to any one of claims 1 to 1 8, wherein the composition comprises more than 50 x 106, more than 60 x lO6, more than 70 x 106, more than 75 x 106, or more than 80 x 106, colony forming units (probiotic units) per millilitre (mL).

20. The probiotic composition according to any one of claims 1 to 1 , wherein the composition is: lactose free, dairy free, vegan, organic, or any combination thereof.

21 . A method for preventing or treating an intestinal infection by pathogenic bacteria in a subject, the method comprising orally administering the composition according to any one of claims 1 to 20 to the subject.

22. The method according to claim 21 , wherein the bacterial infection arises from bacteria selected from the group consisting of: Salmonella, Escherichia, Staphylococcus, Proteus, and any combination thereof.

22. The method according to claim 21 , wherein the bacterial infection is an infection of the large intestine.

23. A method for preventing or treating an intestinal disease or condition in a subject, the method comprising orally administering the composition according to any one of claims 1 to 20 to the subject, wherein the intestinal disease or condition is selected from the group consisting of: diarrhoea, Crohn's disease, ulcerative colitis, inflammatory bowel disease, irritable bowel syndrome, and recurrent pouchitis.

24. The method according to any one of claims 21 to 23, wherein the subject is a human subject.

Description:
PROBIOTIC COMPOSITIONS AND USES THEREOF

Incorporation by Cross-Reference

This application claims priority from Australian provisional patent application number 2015901740 filed on 13 May 2015, the entire content of which is incorporated herein by cross-reference.

Technical Field

The present invention relates generally to the digestive tract and digestive health, More specifically, the present invention relates to probiotic compositions and methods for their use.

Background

A large number of different bacterial species inhabit the human gastrointestinal tract and they are considered to play a number of important roles in metabolism and the prevention of infection by other non-native bacteria. For example, the enterohepatic circulation generally involves conjugation of metabolites to a polar group (e.g. sulfate, taurine, glucuronic acid, glutathione, or glycine) in the liver. Enzymes produced by intestinal bacteria deconjugate the metabolites making them more easily absorbed by intestinal mucosa. Enzymes produced by intestinal bacteria are also important in the metabolism of various vitamins (e.g. vitamin K, vitamin Bj 2 , thiamin, folic acid), and are capable of fermenting indigestible carbohydrates to short chain fatty acids (e.g. butyrate, actetate and propionate). Intestinal bacteria also maintain relatively stable numbers and type over time inhibiting infection by non-native pathogenic bacteria and fungi, for example, by production of antimicrobial substances including short-chain fatty acids and bacteriocins. Intestinal bacteria have also been implicated in the production of anti- carcinogenic chemicals and stimulating the development and/or activity of the immune system.

Numerous factors can disrupt the balance of native intestinal bacteria including antibiotics and steroids, other specific medicines, increased acidity in the digestive system (e.g. poor diet, constipation or diarrhoea), fatigue, yeast overgrowth, irritable bowel syndrome, and other intestinal conditions. Supplementing the diet with probiotic bacteria ('probiotics') represent a means of combating the deleterious imbalance of native intestinal bacteria. Apart from inhibiting colonisation of the intestinal tract by non-nati ve pathogenic bacteria, probiotics have been associated with positive outcomes in the treatment of diarrhoea, Crohn's disease, inflammatory bowel disease, irritable bowel syndrome, and recurrent pouchitis. They are also beneficial in urogenital health, the prevention/treatment of allergies (e.g. skin conditions such as eczema), and can assist in reducing adverse effects arising from antibiotics.

In order to exert their beneficial effects probiotic bacteria must be alive when administered to an individual, and they must survive in sufficient numbers within the medium in which they are delivered. They must also retain their genetic and physical stability during storage along and maintain properties responsible for their health benefits upon consumption during manufacture and storage in the delivery medium.

Probiotic bacteria exert their beneficial effects at least in part via production of lactic acid. Lactic acid-producing bacteria are commonly used in the fermentation of dairy products in which they convert lactose to lactic acid. Consequently, dairy products such as yoghurt and kefir are widely used as a medium for the growth and delivery of probiotic bacteria to individuals. However, a number of deficiencies arise from dairy-based probiotic preparations. For example, numerous individuals are lacking in intestinal lactase, a mucosal enzyme responsible for hydrolysing lactose. This means that the lactose is not adequately digested or absorbed, and upon reaching the large intestine it undergoes vigorous bacterial fermentation causing distention, flatus, and diarrhea. Additionally, dairy probiotic fonnulations often suffer from having a relatively short shelf-life and can spoil relatively quickly. The concentration of probiotic bacteria grown in dairy products can also be limited meaning that more medium needs to be utilised and in some cases consumed.

A need exists for improved probiotic products which overcome one or more of the deficienci es of existing products noted above or as noted elsewhere.

Summary of the Invention

The present inventors have identified probiotic formulations that address one or more deficiencies noted in currently available probiotic products.

Accordingly, in a first aspect, the present invention provides a probiotic composition comprising: coconut milk; bacteria capable of fermenting saccharides to lactic acid; and saccharides, wherein the saccharides are supplementary to any present in the coconut milk, the bacteria, or both.

In a second aspect, the present invention provides use of coconut milk; bacteria capable of fermenting saccharides to lactic acid; and saccharides, in the preparation of a medicament for preventing or treating an intestinal infection by pathogenic bacteria in a subject, wherein the saccharides are supplementary to any present in the coconut milk, the bacteria, or both.

In a third aspect, the present invention provides coconut milk; bacteria capable of fermenting saccharides to lactic acid; and saccharides, for use in preventing or treating an intestinal infection by pathogenic bacteria in a subject, wherein the saccharides are supplementary to any present in the coconut milk, the bacteria, or both.

In a fourth aspect, the present invention provides use of coconut milk; bacteria capable of fermenting saccharides to lactic acid; and saccharides, in the preparation of a medicament preventing or treating an intestinal disease or condition in a subject, wherein the saccharides are supplementary to any present in the coconut milk, the bacteria, or both, and wherein the intestinal disease or condition is selected from the group consisting of: diarrhoea, Crohn's disease, ulcerative colitis, inflammatory bowel disease, irritable bowel syndrome, and recurrent pouchitis.

In a fifth aspect, the present invention provides coconut milk; bacteria capable of fermenting saccharides to lactic acid; and saccharides, for use in preventing or treating an intestinal disease or condition in a subject, wherein the saccharides are supplementary to any present in the coconut milk, the bacteria, or both, and wherein the intestinal disease or condition is selected from the group consisting of: diarrhoea, Crohn's disease, ulcerative colitis, inflammatory bowel disease, irritable bowel syndrome, and recurrent pouchitis.

In one embodiment of the first, second, third, fourth or fifth aspects, the bacteria are lactobacillus bacteria, lactococcus bacteria, bifidobacteria, or any combination thereof.

In one embodiment of the first, second, third, fourth or fifth aspects, the lactobacillus bacteria are selected from the group consisting of: L. acidophilus, L. fermentum, L. paracasei, L. brevis, L. gasseri, L. plcmtarum, L. bulgaricus, L. helvelicus, L. renteri, L. casei, L. jensenii, L. rhamnosus, L. crispatus, L. johnsonii, L. salivarius, and any combination thereof.

In one embodiment of the first, second, third, fourth or fifth aspects, the lactobacillus bacteria comprise or consist of L. plcmtarum strain LM.

In one embodiment of the first, second, third, fourth or fifth aspects, the lactococcus bacteria are selected from the group consisting of: L. lactis, L. lactis subsp. lactis, L. lactis subsp. cremoris, and combinations thereof.

In one embodiment of the first, second, third, fourth or fifth aspects, the lactococcus bacteria comprise or consist of L. lactis and L. lactis subsp. Cremoris.

In one embodiment of the first, second, third, fourth or fifth aspects, the bifidobacteria are selected from the group consisting of: B. adolescentis, B. breve, B. longum, B. animalis, B. infantis, B. thermophilum, B. bifidum, B. laciis, and any combination thereof.

In one embodiment of the first, second, third, fourth or fifth aspects, the probiotic composition or medicament further comprises one or more additional components selected from a thickening agent, a prebiotic, a flavouring agent, and citric acid, wherein the one or more additional components are supplementary to the bacteria, coconut milk and saccharides.

In one embodiment of the first, second, third, fourth or fifth aspects, the prebiotic is selected from the group consisting of polysaccharides, mucopolysaccharides, oligosaccharides, galactooligosaccharides, fructooligosaccharides, inulin, vitamins, harvested metabolic products of biological organisms, amino acids, nutrient precursors, proteins, lipids, Sunfiber®, Larch Arabinogalactan, Kirkman's Pre-Bio™ Fiber Powder, and any combination thereof.

In one embodiment of the first, second, third, fourth or fifth aspects, the thickening agent is selected from the group consisting of pectin, citrus pectin, modified citrus pectin, kuzu, kuzu root starch, starch, polymerised whey protein, agar powder, gelatin, guar gum, arabic gum, Sunfiber and any combination thereof.

In one embodiment of the first, second, third, fourth or fifth aspects, the thickening agent is citrus pectin, kuzu root starch, or a combination thereof.

In one embodiment of the first, second, third, fourth or fifth aspects, the flavouring is selected from the group consisting of coconut flavouring, vanilla flavouring, and a combination thereof.

In one embodiment of the first, second, third, fourth or fifth aspects, the saccharides are a component of fruit juice that included in the composition or medicament.

In one embodiment of the first, second, third, fourth or fifth aspects, the fruit juice is pear juice or concentrated pear juice.

In one embodiment of the first, second, third, fourth or fifth aspects, the composition or medicament comprises between 10% and 80%, between 20% and 70%, between 30% and 60%, between 40% and 60%, between 45% and 55%, or between 45% and 50%) fruit juice.

In one embodiment of the first, second, third, fourth or fifth aspects, the coconut milk additionally comprises carob bean gum.

In one embodiment of the first, second, third, fourth or fifth aspects, the composition or medicament comprises between 10% and 80%, between 20% and 70%, between 30% and 60%, between 40% and 60%, between 45% and 55%, or between 45% and 50% coconut milk. In one embodiment of the first, second, third, fourth or fifth aspects, the composition or medicament pH is between 4.0 and 6.5, between 4.5 and 6.0, between 4.0 and 5.5, between 4.5 and 5.5, or between 4.5 and 5.5.

In one embodiment of the first, second, third, fourth or fifth aspects, the composition or medicament comprises more than 50 x 10 6 , more than 60 x 10 6 , more than 70 x 10 6 , more than 75 x 10 6 , more than 80 x 10 6 , more than 1 x 10 7 , more than 1 x 10 8 , more than 1 x 10 9 , more than 1 x 10 10 , more than 1 x 10 1 1 , or more than 1 x 10 12 , colony forming units (probiotic units) per millilitre (mL).

In one embodiment of the first, second, third, fourth or fifth aspects, the composition or medicament comprises more than 50 x l O 6 , more than 60 x 10 6 , more than 70 x 10 6 , more than 75 x 10 6 , more than 80 x 10 6 , more than 1 x 10 7 , more than 1 x 10 8 , more than 1 x 10 9 , more than 1 x 10 10 , more than 1 x 10 11 , or more than 1 x 10 12 , colony forming units (probiotic units) per millilitre (mL) after treatment of the composition or medicament under conditions suitable for the probiotic bacteria to fennent the saccharides to one or more products, wherein the products comprise lactic acid.

In one embodiment of the first, second, third, fourth or fifth aspects, the composition or medicament is: lactose free, dairy free, vegan, organic, or any combination thereof.

In one embodiment of the first, second, third, fourth or fifth aspects, the composition or medicament does not contain any one or more of: skim milk, dairy butter, soy, soy milk, or soy product.

In a sixth aspect, the present invention provides a method for preventing or treating an intestinal infection by pathogenic bacteria in a subject, the method comprising orally administering the composition according to the first aspect to the subject.

In one embodiment of the second, third or sixth aspects, the bacterial infection arises from bacteria selected from the group consisting of: Salmonella, Escherichia, Staphylococcus, Proteus, and any combination thereof.

In one embodiment of the second, third or sixth aspects, the bacterial infection is an infection of the large intestine.

In a seventh aspect, the present invention provides a method for preventing or treating an intestinal disease or condition in a subject, the method comprising orally administering the composition according to the first aspect to the subject, wherein the intestinal disease or condition is selected from the group consisting of: diarrhoea, Crohn's disease, ulcerative colitis, inflammatory bowel disease, irritable bowel syndrome, and recurrent pouchitis. In one embodiment of the second, third, fourth, fifth, sixth or seventh aspects, the subject is a human subject.

Definitions

As used in this application, the singular form "a", "an" and "the" include plural references unless the context clearly dictates otherwise. For example, the phrase "composition" also includes a plurality of compositions.

As used herein, the term "comprising" means "including." Variations of the word "comprising", such as "comprise" and "comprises," have correspondingly varied meanings. Thus, for example, a probiotic composition "comprising" components X and Y may consist exclusively of components X and Y or may include one or more additional components ) (e.g. component Z).

The term "therapeutically effective amount" as used herein, includes within its meaning a non-toxic but sufficient amount of an agent or composition for use in the present invention to provide the desired therapeutic effect. The exact amount required will vary from subject to subject depending on factors such as the species being treated, the age and general condition of the subject, the severity of the condition being treated, the particular agent being administered, the mode of administration and so forth. Thus, it is not possible to specify an exact "effective amount" applicable to all embodiments. However, for any given case, an appropriate "effective amount" may be determined by one of ordinary skill in the art using only routine experimentation.

As used herein, the term "subject" includes any animal of economic, social or research importance including bovine, equine, ovine, primate, avian and rodent species. Hence, a "subject" may be a mammal such as, for example, a human or a non-human mammal.

As used herein, the term "prebiotic" encompasses an ingredient that can be utilised as a nutrient by a given bacterial strain as a source of energy, for example, growth, biomolecule production, replication, and/or other biological activity. Non-limiting examples of prebiotics include polysaccharides, mucopolysaccharides, oligosaccharides, galactooligosaccharides, fructooligosaccharides, polydextrose, inulin, vitamins, harvested metabolic products of biological organisms, amino acids, nutrient precursors, proteins, lipids, Sunfiber®, Larch Arabinogalactan, Kirkman's Pre-Bio™ Fiber Powder, and any combination thereof. The ingredient may stimulate the growth and/or activity of probiotic intestinal bacteria administered to a host organism that is incapable or substantially incapable of digesting the ingredient. The ingredient may be a non-digestible food ingredient that beneficially affects the host by selectively stimulating the growth and/or activity of one or a limited number of bacteria in the colon that can improve the health of the host.

As used herein, the term "probiotic" refers to live microorganisms (e.g. bacteria) which are beneficial to the health of a recipient when administered at appropriate dosages. The exact dosage required will vary from subject to subject depending on factors such as the species being treated, the age and general condition of the subject, the severity of any disease or condition being treated if applicable, and/or the nature and/or quantity of other components administered with the live microorganisms. Thus, it is not possible to specify an exact dosage that would be applicable to all embodiments. However, for any given case, an appropriate dosage may be determined by one of ordinary skill in the art using only routine experimentation.

It will be understood that use of the term "between" herein when referring to a range of numerical values encompasses the numerical values at each endpoint of the range. For example, a polypeptide of between 10 residues and 20 residues in length is inclusive of a polypeptide of 10 residues in length and a polypeptide of 20 residues in length.

Any description of prior art documents herein, or statements herein derived from or based on those documents, is not an admission that the documents or derived statements are part of the common general knowledge of the relevant art.

For the purposes of description all documents referred to herein are hereby incorporated by reference in their entirety unless otherwise stated.

Detailed Description

The following detailed description conveys exemplary embodiments of the present invention in sufficient detail to enable those of ordinary skill in the art to practice the present invention. Features or limitations of the various embodiments described do not necessarily limit other embodiments of the present invention or the present invention as a whole. Hence, the following detailed description does not limit the scope of the present invention, which is defined only by the claims.

A number of deficiencies exist in currently available probiotic formulations. For example, numerous probiotic formulations contain are dairy-based which can be detrimental to individuals who are lactose-intolerant. Additionally, current probiotic formulations often suffer from having a relatively short shelf-life and can spoil relatively quickly. The concentration of probiotic bacteria in existing probiotic formulations can also be limited which can be detrimental in terms of production cost, convenience of use and effectiveness. The present inventors have developed probiotic formulations that address at least one deficiency present in existing probiotic formulations. For example, the formulations of the present invention are lactose free, thus avoiding detrimental outcomes in lactose- intolerant recipients. They exhibit an extended shelf-life in comparison to a number of currently available probiotic preparations. Additionally, the formulations described herein contain a significantly higher number of colony forming units/probiotic units per volume than numerous other currently available products.

Accordingly, the present invention broadly relates to improved probiotic formulations and methods for their use.

Probiotic Formulations

Probiotic formulations according to the present invention comprise probiotic bacteria in coconut milk, saccharides for fermentation by the probiotic bacteria, and optionally additional components. The additional components may include, for example, any one or more of: thickening agent(s), prebiotic(s), flavouring agent(s), citric acid(s), and/or gelling agents(s). These additional components are additive in the sense that they are distinct/separate from the probiotic bacteria in coconut milk, and saccharides.

Probiotic Bacteria

The probiotic compositions of the present invention comprise at least one species of probiotic bacteria. The probiotic bacteria are considered to be bacteria that provide a health benefit when administered to a subject such as a human or an animal . In general, to exert beneficial health effects in the subject, the probiotic bacteria need to be administered in an adequate amount (e.g. a therapeutically effective amount), which can readily be determined by a person of ordinary skill in the art. Without limitation to theory, it is suggested that metabolites produced by the probiotic bacteria (e.g. lactic acid) may account at least in part for the beneficial health effects in a subject to which they have been administered.

Non-limiting examples of probiotic bacteria that may be included in the probiotic compositions described herein include those from any one or more of the following genera: Aerococcus, Bacillus, Bacteroides, Bifidobacterium, Clostridium, Enterococcus, Fusobacterium, Kocuria, Lactobacillus, Lactococcus, Leuconostoc, Melissococcus, Micrococcus, Oenococcus, Pediococcus, Peptostrepococcus, Propionibacterium, Staphylococcus, Streptococcus, and Weissella.

Specific examples of probiotic bacterial species that may be included in the probiotic compositions include, but are not limited to: Bacillus capillosus, Bacillus coagulans, Bacillus lentils, Bacillus licheniformis, Bacillus mesentericus, Bacillus natio, Bacillus pumilus, Bacillus subtilis; Bacteroides amylophilus, Bacteroides ruminocola, Bacteroides suis; Bifidobacterium adolescends, Bifidobacterium animalis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium lac/is, Bifidobacterium longum, Bifidobacterium pseudolongum, Bifidobacterium thermophilum; Enterococcus cremoris, Enterococcus diacetylactis, Enterococcus faechim, Enterococcus intermedins, Enterococcus lactis, Enterococcus muntdi, Enterococcus thermophilic; Escherichia co/i, Lactobacillus acidophilus, Lactobacillus alimentarius, Lactobacillus amvlovorus, Lactobacillus brevis, L. hulgaricus, Lactobacillus casei, Lactobacillus cellobiosus, Lactobacillus crispatus, Lactobacillus curvatus, Lactobacillus delbrueckii, Lactobacillus delbrueckii ss. hulgaricus, Lactobacillus farciminis, Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus helveti ' cus, Lactobacillus jensenii, Lactobacillus johnsonii, Lactobacillus lactis, Lactobacillus lactis subsp. lactis, Lactobacillus lactis subsp. cremoris, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus plantarum strain LM, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus sakei, Lactobacillus salivarius; Leuconostoc mesenteroides; Pediococcus acidilactici; Pediococcus pentosaceus; Propionihacterium freudenreichii, Propionibacterium shermanii; Staphylococcus carnosiis, Staphylococcus xylosus; Streptococcus infantarius, Streptococcus salivarius ss. thermophilus, Streptococcus thermophilus, and Streptococcus lactis.

Probiotic bacteria for inclusion in the compositions of the present invention may be derived from a variety of different sources including commercial sources, including but not limited to international depositories (e.g. America Type Culture Collection - ATCC) and other national equivalents (e.g. Collection Nationale De Cultures De Microorganismes - CNC ; China General Microbiological Culture Collection Center - CGMCC, and the like).

In some embodiments, the probiotic compositions comprise Lactobacillus plantarum (e.g. Lactobacillus plantarum strain LM).

In some embodiments, the probiotic compositions comprise a mixture of Lactococcus and Lactobacillus strains; Lactococcus and Bifidobacterial strains; Bifidobacterial and Lactobacillus strains; or Lactococcus, Lactobacillus and Bifidobacterial strains.

In some embodiments, the probiotic compositions comprise a mixture of Lactococcus and Lactobacillus strains. By way of non-limiting example, the compositions may comprise a mixture of Lactobacillus plantarum (e.g. Lactobacillus plantarum strain LM), Lactobacillus lactis subsp. Lactis {Lactobacillus lactis), and Lactobacillus lactis subsp. cremoris (/,. Cremoris).

Probiotic bacteria included in the compositions of the present invention are preferably alive, although the skilled person will realise that at least a small proportion of bacteria in the compositions may not be. The live bacteria may be replicating or non- replicating, and are preferably capable of metabolising.

According to some embodiments of the present invention, the probiotic bacteria may be present in the compositions at least: 5 x 10 9 colony forming units (CFU) per serving of the composition, 6 x 10 9 CFU per serving, 7 x 10 9 CFU per serving, 8 x 10 9 CFU per serving, 9 x 10 9 CFU per serving, 10 x 10 9 CFU per serving, 1 1 x 10 9 CFU per serving, 12 x 10 9 CFU per serving, 13 x 10 9 CFU per serving, 14 x 10 9 CFU per serving, 15 x 10 9 CFU per serving, 16 x 10 9 CFU per serving, 17 x 10 9 CFU per serving, 18 x 10 9 CFU per serving, 19 x 10 9 CFU per serving, or 20 x 10 9 CFU per serving.

Without limitation, preferred serving sizes include 50mL, 60mL, 70mL, 80mL, 90mL and l OOmL.

Accordingl y, by way of non-limiting example the probiotic bacteria may be present in the compositions at least: 5 x 10 9 colony forming units (CFU) per 80mL serving of the composition, 6 x 10 9 CFU per 80mL serving, 7 x 10 9 CFU per 80mL serving, 8 x 10 CFU per 80mL serving, 9 x l O 9 CFU per serving, 10 x 10 9 CFU per 80mL serving, 1 1 x 10 9 CFU per 80mL serving, 12 x 10 9 CFU per 80mL serving, 13 x 10 9 CFU per 80mL serving, 14 x 10 9 CFU per 80mL serving, 15 x 10 9 CFU per 80mL serving, 16 x 10 CFU per 80m L serving, 17 x 10 9 CFU per 80m L serving, 18 x 10 9 CFU per 80mL serving, 19 x 10 9 CFU per 80mL serving, or 20 x 10 9 CFU per 80mL serving.

Coconut Milk

The probiotic compositions of the present invention comprise coconut milk.

As known to those of ordinary skill in the art, coconut milk comprises a mixture of coconut meat and water. It derives from mature/brown coconut from which meat is grated and mixed with a small amount of water. The fat content of coconut milk is predominantly made up of saturated fats (i.e. triglycerides with only saturated fatty acids), and may vary from, for example 5-10% fat content (thin coconut milk) to for example, 20-25% fat content (thick coconut milk). Coconut milk is available commercially and methods for its preparation are well known in the art.

Compositions according to the present invention may comprise coconut milk with any fat content such as, for example, at least: 1% (v/v), 2% (v/v), 3% (v/v), 4% (v/v), 5% (v/v), 6% (v/v). 7% (v/v), 8% (v/v), 9% (v/v), 10% (v/v), 1 1% (v/v), 12% (v/v), 13% (v/v), 14% (v/v), 15% (v/v), 16% (v/v), 17% (v/v), 18% (v/v), 19% (v/v), 20% (v/v), 21% (v/v), 22% (v/v), 23% (v/v), 24% (v/v), 25% (v/v), 26% (v/v), 27% (v/v), 28% (v/v), 29% (v/v), or 30% (v/v) saturated fat.

Optionally, the coconut milk may comprise a gelling agent. Non-limiting examples of gelling agents include carob bean gum, locust bean gum, and the like. The gelling agent may be used in an amount suitable to achieve a desired texture of the compositions, as can be readily optimised by persons of ordinary skill in the art.

The coconut may be included in the compositions of the present invention in any suitable amount. For example, the coconut milk may constitute at least: 10% (v/v) of the total composition, 20% ( v/v) of the total composition, 30% (v/v) of the total composition, 40% (v/v) of the total composition, 50% (v/v) of the total composition, 60% (v/v) of the total composition, 70% (v/v) of the total composition, or 80% (v/v) of the composition. In some embodiments, the compositions comprise between 35% (v/v) and 65% (v/v) coconut milk, between 35% (v/v) and 60% (v/v) coconut milk, between 40% (v/v) and 65% (v/v) coconut milk, between 40% (v/v) and 60% (v/v) coconut milk, between 45% (v/v) and 60% (v/v) coconut milk, between 45% (v/v) and 55% (v/v) coconut milk, or between 45% (v/v) and 50% (v/v) coconut milk.

Saccharides

The probiotic compositions of the present invention comprise saccharides. The saccharides may be additional to any saccharides that may be or are present in the probiotic bacteria and/or the coconut milk and/or any other component of the compositions.

The saccharides may serve as feed material for the probiotic bacteria to produce metabolites such as, for example, lactic acid, acetic acid, lineolic acid and the like, by fermentation.

Any suitable saccharide or mixtures thereof capable of fermentation by probiotic bacteria may be included in the compositions. Non-limiting examples of suitable saccharides include fermentable monosaccharides, oligosaccharides (e.g. disaccharides), polysaccharides, and mixtures thereof.

The monosaccharide may, for example, be a hexose or a pentose. Suitable examples of pentose saccharides include, but are not limited to, arabinose, lyxose, ribose, ribulose, xylose, xylulose, and any combination thereof. Suitable examples of hexose saccharides include, but are not limited to, allose, altrose, fructose, galactose, glucose, idose, mannose, psicose, sorbose, tagatose, talose, and any combination thereof. Additional examples of suitable monosaccharides include: trioses including aldotrioses (e.g. glyceraldehyde) and ketotrioses (e.g. dihydroxyacetone), tetroses including aldotetroses (e.g. threose and erythrose) and ketotetroses (e.g. erythrulose), pentoses including aldopentoses (e.g. lyxose, ribose, arabinose, deoxyribose and xylose) and ketopentoses (e.g. xylulose and ribulose), hexoses including aldohexoses (e.g. glucose, mannose, altrose, idose, galactose, allose, talose and gulose) and ketohexoses (e.g. fructose, psicose, tagatose and sorbose), heptoses including keto-heptoses (e.g. sedoheptulose and mannoheptulose), octoses including octolose and 2-keto-3-deoxy-manno-octonate, and nonoses including sialose.

Non-limiting examples of disaccharide fragments that may be produced by the methods described herein include, but are not limited to, sucrose, lactose, maltose, trehalose, cellobiose, laminaribiose, xylobiose, gentiobiose, isomaltose, mannobiose, kojibiose, rutinose, nigerose, and melibiose.

The oligosaccharides may, for example, include cellobiose, fructo-oligosaccharides (FOS), galactooligosaccharides (GOS), glucooligosaccharides, lactulose, lactose, maltose, mannooligosaccharides, mannanoligosaccharides, sucrose, raffinose, trehalose, xylooligosaccharides (XOS), and any combination thereof.

The polysaccharides may, for example, include arabinoxylans, cellulose, chitin cyclodextrin, glycogen, guar gum, gum arabic, pectins and starches.

The saccharides may be provided in an amount suitable for the probiotic bacteria to produce sufficient levels of metabolites (e.g. lactic acid, acetic acid, Hneolic acid and the like) and so assist in providing a beneficial health effect to a subject. Additionally or alternatively, the saccharides may be provided in an amount suitable for the probiotic bacteria to survive during manufacture of the compositions and/or storage of the compositions. The particular amount of saccharides included will depend on the identity and concentration of the probiotic bacteria in the compositions, and other related factors. Such factors can be tested and the amount of saccharides optimised without difficulty by persons of ordinary skill in the art.

In certain embodiments, the saccharides are provided in the form of a fruit juice (e.g. pear blend) which is added to the probiotic composition. The fruit juice may be concentrated.

The fruit juice may be included in the compositions of the present invention in any suitable amount, as influenced by the concentration of saccharides desired and their abundance in the particular fruit juice selected. For example, the fruit juice may constitute at least: 10% (v/v) of the total compositi on, 20% (v/v) of the total composition, 30% (v/v) of the total composition, 40% (v/v) of the total composition, 50% (v/v) of the total composition, 60% (v/v) of the total composition, 70% (v/v) of the total composition, or 80%) (v/v) of the composition. In some embodiments, the compositions comprise between 35% (v/v) and 65% (v/v) fruit juice, between 35% (v/v) and 60 fruit juice, between 40% (v/v) and 65% (v/v) fruit juice, between 40%o (v/v) and 60%o (v/v) fruit juice, between 45% (v/v) and 60% (v/v) fruit juice, between 45% (v/v) and 55% (v/v) fruit juice, or between 45% (v/v) and 50% (v/v) fruit juice.

Prebiotics

The probiotic compositions of the present invention may optionally comprise one or more prebiotics.

In general, the prebiotic can be utilised as a nutrient by a given bacterial strain as a source of energy, for example, growth, biomolecule production, replication, and/or other biological activity. The prebiotic may, for example, be an oligosaccharide produced from glucose, galactose, hemicellulose, inulin, lactose, maltose, sucrose, starch, xylan, xylose, or any combination thereof. Non-limiting examples of suitable prebiotics include polysaccharides, polydextrose, mucopolysaccharides, oligosaccharides, galactooligosaccharides, fructooligosaccharides, inulin, vitamins, harvested metabolic products of biological organisms, amino acids, nutrient precursors, proteins, lipids, Sunfiber®, Larch Arabinogalactan, Kirkman's Pre-Bio™ Fiber Powder, and any combination thereof.

In some embodiments, the prebiotic may selectively stimulate the growth and/or activity of probiotic intestinal bacteria administered to a subject that is incapable or substantially incapable of digesting the ingredient.

The prebiotic(s) may be included in the compositions at any suitable level to achieve the desired growth and/or activity of probiotic bacteria as can be readily determined by persons of ordinary skill in the art. By way of non-limiting example only, the prebiotic(s) may constitute at least: 0.1 %) (v/v) of the total composition, 0.2% (v/v) of the total composition, 0.3% (v/v) of the total composition, 0.4% (v/v) of the total composition, 0.5% (v/v) of the total composition, 0.6% (v/v) of the total composition, 0.7% (v/v) of the total composition, 0.8% (v/v) of the total composition, 0.9% (v/v) of the composition, 1 .0% (v/v) of the total composition, 1 .2% (v/v) of the total composition, 1 .4% (v/v) of the total composition, 1.6% (v/v) of the total composition, 1 .8% (v/v) of the total composition, 2.0% (v/v) of the total composition, 2.5% (v/v) of the total composition, 3.0% (v/v) of the total composition, 3.5% (v/v) of the total composition, 4.0% (v/v) of the total composition, 4.5% (v/v) of the total composition, or 5% (v/v) of the total composition. In some embodiments, the compositions comprise between 0.1% (v/v) and 5% (v/v) prebiotic, between 0.5% (v/v) and 5% (v/v) prebiotic, between 0.5% (v/v) and 4.5% (v/v) prebiotic, between 1% (v/v) and 5% (v/v) prebiotic, between 1 % (v/v) and 4% (v/v) prebiotic, between 2% (v/v) and 5% (v/v) prebiotic, or between 2% (v/v) and 4% (v/v) prebiotic.

Thickening Agents

The probiotic compositions of the present invention may optionally comprise one or more thickening agents. Thickening agents as contemplated herein include at least thickeners and gelling agents. The thickening agents may be used to increase the viscosity of the compositions without substantially modifying the efficacy of other ingredient(s) (e.g. probiotic bacteria) within the compositions. The thickening agents may also increase the stability of the compositions of the present invention.

Non-limiting examples of suitable thickening agents include carboxylic acid polymers, polysaccharides, hydrogenated polyisobutene, crosslinked polyacrylate polymers, trihydroxystearin, polyacrylamide polymers, gums, and any combination thereof.

Non-limiting examples of suitable gums include acacia, agar, hydroxypropyl guar, algin, alginic acid, amylopectin, calcium carrageenan, sodium carrageenan, sodium carboxymethyl dextran, carrageenan, dextrin, gelatin, gellan gum, hydrated silica, guar gum, propylene glycol alginate, hyaluronic acid, carnitine, sclerotium gum, kelp, guar hydroxypropyltrimonium chloride, potassium carrageenan, hectorite, hydroxypropyl chitosan, karaya gum, calcium alginate, locust bean gum, natto gum, potassium alginate, ammonium alginate, tragacanth gum, xanthan gum, and any combination thereof.

In some embodiments, the thickening agent may be pectin, citrus pectin, modified citrus pectin, kuzu, kuzu root starch, starch, polymerised whey protein, agar powder, gelatin, guar gum, arabic gum, Sunfiber, or any combination thereof.

By way of non-limiting example only, the thickening agent(s) may be included in the compositions at any suitable level to achieve the desired consistency and texture as can be readily determined by persons of ordinary skill in the art. By way of non-limiting example only, the thickening agent(s) may constitute at least: 0.1 % (v/v) of the total composition, 0.2% (v/v) of the total composition, 0.3% (v/v) of the total composition, 0.4% (v/v) of the total composition, 0.5% (v/v) of the total composition, 0.6% (v/v) of the total composition, 0.7% (v/v) of the total composition, 0.8% (v/v) of the total composition, 0.9% (v/v) of the composition, 1 .0% (v/v) of the total composition, 1.2% (v/v) of the total composition, 1 .4% (v/v) of the total composition, 1.6% (v/v) of the total composition, 1.8% (v/v) of the total composition, 2.0% (v/v) of the total composition, 2.5% (v/v) of the total composition, 3.0% (v/v) of the total composition, 3.5% (v/v) of the total composition, 4.0% (v/v) of the total composition, 4.5% (v/v) of the total composition, or 5% (v/v) of the total composition.

In some embodiments, the compositions comprise between 0.1 % (v/v) and 5% (v/v) thickening agent, between 0.5% (v/v) and 5% (v/v) thickening agent, between 0.5% (v/v) and 4.5% (v/v) thickening agent, between 1% (v/v) and 5% (v/v) thickening agent, between 1 % (v/v) and 4% (v/v) thickening agent, between 2% (v/v) and 5% (v/v) thickening agent, or between 2% (v/v) and 4 % (v/v) thickening agent. pH

The probiotic compositions of the present invention may be prepared at a final pH suitable for the replication and/or activity of probiotic bacteria within the compositions. The pH may be selectively suitable for the probiotic bacteria over other forms of bacteria.

By way of non-limiting example only, probiotic compositions of the present invention may be provided at a pH of: between 2.0 and 6.5, between 2.5 and 6.0, between 3.0 and 5.5, between 3.0 and 5.0, between 3.0 and 4.5, between 3.0 and 4.0, between 3.2 and 4.0, between 3.4 and 3.8, or between 3.4 and 3.6.

In some embodiments, the probiotic compositions may comprise an additional acidifying agent which may be used to achieve the desired pH. For example, an appropriate volume of acid (e.g. citric acid) may be included to reach a targeted pH.

Colouring and Flavouring

The probiotic compositions of the present invention may optionally comprise colouring agent(s) and/or flavouring agent(s). Any suitable flavouring and/or colouring agent(s) may be used, provided that the reproduction, growth and/or activity of probiotic bacteria in the composition is not hindered or not substantially hindered.

In some embodiments, coconut and/or vanilla flavouring agent(s) may be used. Where the flavouring agent(s) are representative of food materials that differ in colour from coconut milk, colouring agent(s) can be utilised to match the colour of the compositions to that of the representative food material. Additionally or alternatively, the flavouring agent(s) may provide the desired colouring effect.

Preparation of Probiotic Compositions

The probiotic compositions of the present invention may be fermented during their production. For example, coconut milk may be mixed with an inoculating dose of probiotic bacteria and saccharides (including, but not limited to fruit juice, pear juice) as described herein, and the suspension incubated under suitable conditions in order for the probiotic bacteria to ferment at least a portion of the saccharides. Suitable conditions for the fermentation are well known to those of ordinary skill in the field (see, for example, in Heller KJ, Am J Clin Nutr. 2001 Feb;73(2 Suppl):374S-379S). By way of non-limiting example, the fermentation may comprise incubating the suspension at a temperature of between 20°C and 60°C, 25°C and 55°C, 30°C and 50°C, 35°C and 50°C, 35°C and 45°C, 37°C and 45°C, or 40°C and 45°C. The suspension may be incubated for a time period of more than: 1 , 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 36 or 48 hours. The suspension may be incubated for a time period of between about: 1 hour and 48 hours, 1 hour and 36 hours, 1 hour and 24 hours, 1 hour and 12 hours, 6 hours and 48 hours, 6 hours and 36 hours, 6 hours and 24 hours, 6 hours and 12 hours, 12 hours and 48 hours, 12 hours and 36 hours, 12 hours and 24 hours, 18 hours and 48 hours, 18 hours and 36 hours, 18 hours and 24 hours, 6 hours and 18 hours, 8 hours and 14 hours, 10 hours and 12 hours.

In some embodiments, the inoculating dose of probiotic bacteria may be less than the number of probiotic bacteria present in the composition following the fermentation. By way of non-limiting example, there may be: 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, 250%, 300%, 400%, 500%, more probiotic bacteria in the fermented composition as compared to an equivalent volume of the composition prior to the fermentation.

In some embodiments, the concentration of probiotic bacteria in the composition prior to fermentation may be: less than 1 x 10 6 CFU/mL, less than 1 x 10 7 CFU/mL, less than 1 x 10 8 CFU/mL, less than 1 x 10 9 CFU/mL, less than 1 x 10 10 CFU/mL, less than 1 x 10 11 CFU/mL, less than 1 x 10 12 CFU/mL.

In some embodiments, the concentration of probiotic bacteria in the composition after fermentation may be: more than 1 x 10 6 CFU/mL, more than 1 x 10 7 CFU/mL, more than 1 x 10 8 CFU/mL, more than 1 x 10 9 CFU/mL, more than 1 x 10 10 CFU/mL, more than 1 x 10 1 1 CFU/mL, more than 1 x 10 12 CFU/mL, more than 1 x 10 13 CFU/mL.

In some embodiments:

(i) the concentration of probiotic bacteria in the composition prior to fermentation may be less than 0.1 x lO 9 CFU/mL and the concentration of probiotic bacteria in the composition after fennentation may be more than 1 x 10 9 CFU/mL;

(ii) the concentration of probiotic bacteria in the composition prior to fermentation may be less than 1 x 10 9 CFU/mL and the concentration of probiotic bacteria in the composition after fermentation may be more than 1 x 10 9 CFU/mL: (iii) the concentration of probiotic bacteria in the composition prior to fermentation may be less than 0.1 x 10 9 CFU/mL and the concentration of probiotic bacteria in the composition after fermentation may be more than 1 x 10 10 CFU/mL;

(iv) the concentration of probiotic bacteria in the composition prior to fermentation may be less than 1 x 10 9 CFU/mL and the concentration of probiotic bacteria in the composition after fermentation may be more than 1 x 10 10 CFU/mL:

(v) the concentration of probiotic bacteria in the composition prior to fermentation may be less than 0.1 x 10 9 CFU/mL and the concentration of probiotic bacteria in the composition after fermentation may be more than 1 x 10 1 1 CFU/mL:

(vi) the concentration of probiotic bacteria in the composition prior to fermentation may be less than 1 x 10 9 CFU/mL and the concentration of probiotic bacteria in the composition after fermentation may be more than 1 x 10 11 CFU/mL;

(vii) the concentration of probiotic bacteria in the composition prior to fermentation may be less than 0.1 x 10 9 CFU/mL and the concentration of probiotic bacteria in the composition after fermentation may be more than 1 x 10 12 CFU/mL;

(viii) the concentration of probiotic bacteria in the composition prior to fermentation may be less than 1 x 10 9 CFU/mL and the concentration of probiotic

1 ^

bacteria in the composition after fermentation may be more than 1 x 10 CFU/mL;

In some embodiments, the pH of the composition after the fermentation is between: 2.0 and 6.0, 2.0 and 5.0, 2.0 and 4.0, 2.5 and 6.0, 2.5 and 5.0, 2.5 and 4.0, 3.0 and 6.0, 3.0 and 5.0, 3.0 and 4.5, 3.0 and 4.0, 3.0 and 3.5, 3.5 and 6.0, 3.5 and 5.5, 3.5 and 5.0, 3.5 and 4.5, 3.5 and 4.0. 4.0 and 6.0, 4.0 and 5.5, 4.0 and 4.5.

Exemplary Probiotic Compositions

The following formulations are representative non-limiting examples of suitable probiotic compositions according to the present invention. It will be appreciated by persons of ordinary skill in the art that numerous variations and/or modifications can be made to the compositions disclosed in these specific embodiments without extending beyond the scope of the invention as broadly described herein.

Exemplary Formula #1

Coconut Milk (Coconut meat, water) 30%-60% (v/v)

• Probiotic bacteria (e.g. Lactococci, Lactobacilli and/or Bifidobacteria): at least: 8 x 10 9 , 8 x 10 10 , 8 x 10 u , 8 x l O 12 , 8 x 10 13 , 8 x 10 14 , or 8 x 10 15 CFU (per 80mL serve)

Fruit juice (e.g. pear juice) 30%-60% (v/v) (pH 3.0-5.5)

Exemplary Formula #2

Coconut Milk (Coconut meat, water, carob bean gum) 30%-60% (v/v)

• Probiotic bacteria (e.g. Lactococci, Lactobacilli and/or Bifidobacteria): at least: 8 x 10 9 , 8 x l O 10 , 8 x 10 11 , 8 x 10 12 , 8 x 10 13 , 8 x 10 14 , or 8 x 10 15 CFU (per 80mL serve)

Fruit juice (e.g. pear juice) 30%-60% (v/v)

Thickening agent (e.g. Citrus Pectin and/or Kuzu root starch) 0. l%-2%

(pH 3.0-5.5)

Exemplary Formula #3

Coconut Milk (Coconut meat, water) 30%-60% (v/v)

Probiotic bacteria (e.g. Lactococci, Lactobacilli and/or Bifidobacteria): at least: 8 x 10 9 , 8 x 10 10 , 8 x 10 u , 8 x 10 12 , 8 x 10 13 , 8 x 10 14 , or 8 x 10 15 CFU (per 80mL serve)

• Fruit juice (e.g. pear juice) 30%-60% (v/v)

• Prebiotic (e.g. Sunfiber®) 2-5% (v/v)

(pH 3.0-5.5)

Exemplary Fonnula #4

• Coconut Milk (Coconut meat, water) 30%-60% (v/v)

• Probiotic bacteria (e.g. Lactococci, Lactobacilli and/or Bifidobacteria): at least: 8 x 10 9 , 8 x 10 10 , 8 x 10 11 , 8 x 10 12 , 8 x 10 13 , 8 x 10 14 , or 8 x 10 15 CFU (per 80mL serve)

Fruit juice (e.g. pear juice) 30%-60% (v/v)

• Prebiotic (e.g. Sunfiber©) 2-5% (v/v)

• Thickening agent (e.g. Citrus Pectin and/or Kuzu root starch) 0.1 %-2% (v/v) (pH 3.0-5.5)

Exemplary Formula #5

Coconut Milk (Coconut meat, water) 30%-60% (v/v)

• Probiotic bacteria (e.g. Lactococci, Lactobacilli and/or Bifidobacteria): at least: 8 x 10 9 , 8 x 10 10 , 8 x 10 u , 8 x l O 12 , 8 x 10 13 , 8 x 10 14 , or 8 x 10 15 CFU (per 80mL serve)

Fruit juice (e.g. pear juice) 30%-60% (v/v) • Prebiotic (e.g. Sunfiber®) 2-5% (v/v)

Thickening agent (e.g. Citrus Pectin and/or Kuzu root starch) 0.1 %-2% (v/v) Flavouring (e.g. coconut and/or vanilla) 0.05%-0.5% (v/v)

(pH 3.0-5.5)

Exemplary Formula #6

• Coconut Milk (Coconut meat, water) 30%-60% (v/v)

Probiotic bacteria (e.g. Lactococci, Lactobacilli and/or Bifidobacteria): at least: 8 x 10 9 , 8 x 10 10 , 8 x 10 u , 8 x 10 12 , 8 x 10 13 , 8 x 10 14 , or 8 x 10 15 CFU (per 80mL serve)

• Fruit juice (e.g. pear juice) 30%-60% (v/v)

• Prebiotic (e.g. Sunfiber®) 2-5% (v/v)

Citric Acid (e.g. citric acid) 0.01%-1% (v/v)

(pH 3.0-5.5)

Exemplary Formula #7

• Coconut Milk (Coconut meat, water) 30%-60% (v/v)

• Probiotic bacteria (e.g. Lactococci, Lactobacilli and/or Bifidobacteria): at least: 8 x 10 9 , 8 x l O 10 , 8 x 10 11 , 8 x 10 12 , 8 x 10 13 , 8 x 10 14 , or 8 x 10 15 CFU (per 80mL serve)

Fruit juice (e.g. pear juice) 30%-60% (v/v)

• Prebiotic (e.g. Sunfiber®) 2-5% (v/v)

• Thickening agent (e.g. Citrus Pectin and/or Kuzu root starch) 0.1 %-2% (v/v)

• Flavouring (e.g. coconut and/or vanilla) 0.05%-0.5% (v/v)

Citric Acid (e.g. citric acid) 0.01%-1% (v/v)

(pH 3.0-5.5)

Exemplary Formula #8

• Coconut Milk (Coconut meat, water, carob bean gum) 30%-60% (v/v)

Probiotic bacteria (e.g. Lactococci, Lactobacilli and/or Bifidobacteria): at least: 8 x 10 9 , 8 x l O 10 , 8 x 10 11 , 8 x 10 12 , 8 x 10 13 , 8 x 10 14 , or 8 x 10 15 CFU (per 80mL serve)

• Fruit juice (e.g. pear juice) 30%-60% (v/v)

• Prebiotic (e.g. Sunfiber®) 2-5% (v/v)

• Thickening agent (e.g. Citrus Pectin and/or Kuzu root starch) 0.1 %-2% (v/v) Flavouring (e.g. coconut and/or vanilla) 0.05%-0.5% (v/v) • Citric Acid (e.g. citric acid) 0.01%-1% (v/v)

(pH 3.0-5.5)

Exemplary Formula #9

• Coconut Milk (Coconut meat, water) 30%-60% (v/v)

• Probiotic bacteria (e.g. Lactococci, Lactobacilli and/or Bifidobacteria): at least: 8 x 10 9 , 8 x l O 10 , 8 x 10 11 , 8 x 10 12 , 8 x 10 13 , 8 x 10 14 , or 8 x 10 15 CFU (per 80mL serve)

Fruit juice (e.g. pear juice) 30%-60% (v/v)

(pH 3.0-5.5)

Exemplary Formula # 10

Coconut Milk (Coconut meat, water, carob bean gum) 30%-60% (v/v)

Probiotic bacteria (e.g. L. Plantar m LM™, L. lactis subsp. lactis, L. laciis subsp. cremoris) at least: 8 x 10 9 , 8 x 10 10 , 8 x 10 11 , 8 x 10 12 , 8 x 10 13 , 8 x 10 14 , or 8 x 10 15 CFU (per 80mL serve)

• Fruit juice (e.g. pear juice) 30%-60% (v/v)

• Thickening agent (e.g. Citrus Pectin and/or Kuzu root starch) 0.1 %-2%

(pH 3.0-4.5)

Exemplary Fonnula #1 1

• Coconut Milk (Coconut meat, water) 30%-60% (v/v)

• Probiotic bacteria (e.g. L. Plantarum LM™, L. lactis subsp. lactis, L. lactis subsp. cremoris): at least: 8 x 10 9 , 8 x 10 10 , 8 x 10 11 , 8 x 10 12 , 8 x 10 13 , 8 x 10 14 , or 8 x lO 15 CFU (per 80mL serve)

Fruit juice (e.g. pear juice) 30%-60% (v/v)

• Prebiotic (e.g. Sunfiber©) 2-5% (v/v)

(pH 3.0-4.5)

Exemplary Formula #12

Coconut Milk (Coconut meat, water) 30%-60% (v/v)

Probiotic bacteria (e.g. L. Plantarum LM™, Λ. lactis subsp. lactis, L. lactis subsp. cremoris): at least: 8 x lO 9 , 8 x 10 1 , 8 x 10 11 , 8 x 10 12 , 8 x 10 13 , 8 x 10 14 , or 8 x 10 15 CFU (per 80mL serve)

• Fruit juice (e.g. pear juice) 30%-60% (v/v)

Prebiotic (e.g. Sunfiber®) 2-5% (v/v) • Thickening agent (e.g. Citrus Pectin and/or Kuzu root starch) 0.1 %-2% (v/v) (pH 3.0-4.5)

Exemplary Formula #13

• Coconut Milk (Coconut meat, water) 30%-60% (v/v)

• Probiotic bacteria (e.g. L. Planlarum LM™, L. lactis subsp. lactis, L. lactis subsp. cremoris): at least: 8 x 10 9 , 8 x 10 10 , 8 x 10 n , 8 x 10 12 , 8 x 10 13 , 8 x 10 14 , or 8 x lO 15 CFU (per 80mL serve)

Fruit juice (e.g. pear j uice) 30%-60% (v/v)

• Prebiotic (e.g. Sunfiber®) 2-5% (v/v)

• Thickening agent (e.g. Citrus Pectin and/or Kuzu root starch) 0.1 %-2% (v/v)

• Flavouring (e.g. coconut and/or vanilla) 0.05%-0.5% (v/v)

(pH 3.0-4.5)

Exemplary Formula # 14

Coconut Milk (Coconut meat, water) 30%-60% (v/v)

• Probiotic bacteria (e.g. L. Plantarum LM™, L. lactis subsp. lactis, L. lactis subsp. cremoris): at least: 8 x lO 9 , 8 x 10 10 , 8 x 10 1 1 , 8 x 10 12 , 8 x l O 13 , 8 x 10 14 , or 8 x 10 15 CFU (per 80mL serve)

Fruit juice (e.g. pear juice) 30%-60% (v/v)

• Prebiotic (e.g. Sunfiber®) 2-5% (v/v)

• Citric Acid (e.g. citric acid) 0.01 %-l % (v/v)

(pH 3.0-4.5)

Exemplary Formula #15

Coconut Milk (Coconut meat, water) 30%-60% (v/v)

• Probiotic bacteria (e.g. L. Plantarum LM™, L. lactis subsp. lactis, L. lactis subsp. cremoris): at least: 8 x 10 9 , 8 x 10 10 , 8 x 10 11 , 8 x 10 12 , 8 x l O 13 , 8 x 10 14 , or 8 x 10 15 CFU (per 80mL serve)

Fruit juice (e.g. pear juice) 30%-60% (v/v)

Prebiotic (e.g. Sunfiber®) 2-5% (v/v)

Thickening agent (e.g. Citrus Pectin and/or Kuzu root starch) 0.1 %-2% (v/v)

• Flavouring (e.g. coconut and/or vanilla) 0.05%-0.5% (v/v)

• Citric Acid (e.g. citric acid) 0.01 %-l% (v/v)

(pH 3.0-4.5) Exemplary Formula #16

Coconut Milk (Coconut meat, water, carob bean gum) 30%-60% (v/v)

Probiotic bacteria (e.g. L. Plantation LM™, L. lactis stibsp. lactis, L. lactis subsp. cremoris): at least: 8 x 10 9 , 8 x 10 10 , 8 x 10 n , 8 x 10 12 , 8 x l O 13 , 8 x l O 14 , or 8 x 10 15 CFU (per 80mL serve)

• Fruit juice (e.g. pear juice) 30%-60% (v/v)

• Prebiotic (e.g. Sunfiber®) 2-5% (v/v)

Thickening agent (e.g. Citrus Pectin and/or Kuzu root starch) 0.1 %-2% (v/v) Flavouring (e.g. coconut and/or vanilla) 0.05%-0.5% (v/v)

· Citric Acid (e.g. citric acid) 0.01 %-l% (v/v)

(pH 3.0-4.5)

Exemplary Formula #17

Coconut Milk (Coconut meat, water, carob bean gum) 40-50% (v/v)

Pear Juice Blend from Concentrate 40-50% (v/v)

• Sunfiber® Prebiotic 3-4% (v/v)

• Citrus Pectin (Kosher) 0.1 %-0.3% (v/v)

• L. Plantarum LM™, L. lactis subsp. lactis, L. lactis subsp. cremoris: at least: 8 x 10 9 , 8 x 10 1 () , 8 x lO 1 1 , 8 x 10 12 , 8 x 10 13 , 8 x 10 14 , or 8 x 10 15 CFU (per 80mL serve)

Organic Coconut Flavour 0.7% - 1% (v/v)

• Organic Vanilla Flavour 0.05%-2% (v/v)

• Citric Acid 0.05%-0.25% (v/v)

(pH 3.0-4.0)

Exemplary Formula #18

• L. Plantarum LM™, L. lactis subsp. lactis, L. lactis subsp. cremoris: at least: 8 x 10 9 , 8 x 10 10 , 8 x 10 11 , 8 x 10 12 , 8 x 10 13 , 8 x 10 14 , or 8 x 10 15 CFU (per 80mL serve)

Coconut Milk (Coconut meat, water, carob bean gum) 40-50% (v/v)

Pear Juice Blend from Concentrate 40-50% (v/v)

Organic Kuzu Root Starch 2.0%-4.0% (v/v)

• Organic Coconut Flavour 0.7% - 1% (v/v)

• Organic Vanilla Flavour 0.05%-2% (v/v)

• Citric Acid 0.05%-0.25% (v/v)

(pH 3.0-4.0) Methods of Treatment

Probiotic compositions according to the present invention may be administered to subjects for health purposes. The subjects may be human subjects.

At a general level, administration of the probiotic compositions described herein may hinder or prevent intestinal colonisation by non-native microorganisms (e.g. pathogenic bacteria) which may be pathogenic or otherwise deD imental to recipients of the compositions. Consequently, ill-effects arising from harmful microorganisms and their undesirable metabolites can be avoided.

Accordingly, certain embodiments of the present invention relate to methods for preventing or treating an intestinal infection by pathogenic microorganisms in a subject by orally administering a composition according to the present invention. The intestinal infection may be of the large intestine.

By way of non-limiting example, the infection may arise from colonisation of the intestine by pathogenic bacteria. The bacteria may, for example, be selected from the group consisting of Salmonella, Escherichia, Staphylococcus, Proteus, and any combination thereof.

Additional embodiments of the present invention relate to methods for preventing or treating an intestinal disease or condition in a subject by orally administering composition according to the present invention to the subject. By way of non-limiting example, wherein the intestinal disease or condition may be selected from the group consisting of: diarrhoea, Crohn's disease, ulcerative colitis, inflammatory bowel disease, irritable bowel syndrome, and recurrent pouchitis.

Other diseases and/or conditions that may be treated and/or prevented by orally administering composition according to the present invention include, but are not limited to, autism, autoimmune diseases, renal disease including chronic renal disease, pancreatitis, and allergy.

In some embodiments compositions according to the present invention may be administered to improve energy levels, heart health, and/or cognitive function, and/or to relieve stress.

The present invention also contemplates that components within compositions according to the present invention can be used in the preparation of medicaments for the prevention or treatment of the aforementioned diseases and conditions to which the methods of the present invention apply. Examples

The present invention will now be described with reference to specific example(s), which should not be construed as in any way limiting.

Example 1: Preparation and characterisation of probiotic formulations

/. / Materials and Methods

The following formulations were prepared:

Fonnula one:

• Coconut Milk (Coconut meat, water, carob bean gum) 47.368%

Pear Juice Blend from Concentrate 48.163%

• Sunfiber® Prebiotic 3.645%

· Citrus Pectin (Kosher) .197%

• L. Plantarum LM™, L. Lactis, L Cremoris 100B CFU/g .099% (use 1 g/L)

• Organic Coconut Flavour .862%

• Organic Vanilla Flavour .123%)

• Citric Acid .197%

NB: target pH 3.4-3.6

Formula two:

• pH 3.5 L. Plantarum LM™, L. Lactis, L Cremoris 80mL/serv.

• Coconut Milk (Coconut meat, water, carob bean gum) 47.368%

Pear Juice Blend from Concentrate 48.163%

Organic Kuzu Root Starch 3.178%

Organic Coconut Flavour .869%)

• Organic Vanilla Flavour .123%)

• Citric Acid .197%

NB: for preparation use Ig/L of dry culture L. Plantarum LM™, L. Lactis, L Cr 100B CFU/g .099%

NB: target pH 3.4-3.6

Briefly, the coconut milk and juice component were mixed. The mix was then flash pasteurized to 185° F. The kuzu root ingredient was then mixed in until dissolved, followed by agitation continually while cooking the mixture to 1 10° F. The cultures were then mixed in until dissolved and the mixture held at 105° F for 24 hours (alternatively the mixture can be held at 105° F until the desired acidification/pH is achieved). Flavour components were then mixed in followed by cooling to commercial refrigeration temperature. The preparation was down packed to 80mL (alternatively to whatever size desired) HDPE or PP bottles/containers.

Where cold filling is not feasible, it is acceptable to warm fill straight from fermentation temperature (105° F). Steps should be taken to ensure the shortest time possible from completion of fermentation to refrigeration.

The colony forming unit (CFU)/ 80mL serving noted (16 x 10 6 CFU) assumes a doubling of CFU during fermentation (substrate was started with 8 x 10 6 CFU/serving). 16 x 10 6 CFU is an estimate only, from Vivolac (culture supplier), based on the substrate content, fermentation time, and excellent acidification the cultures produced in coco shot. The estimated finished product is between 12-16 x l O 6 CFU.

1.2 Results

Each formula was used to successfully produce viable probiotic preparations. Based on an inoculating amount of 8 x 10 6 CFU/80mL, post-fermentation CFU is estimated to be in the region of 12-16 x 10 6 CFU/80mL. Safety profile testing revealed low/insignificant counts of contaminating bacteria/fungi. The shelf-life of each formulation is at least 90-120 days.

Example 2: preparation of additional probiotic formulation

An additional formulation was prepared according to the parameters set out in Table 1

The culture starter packets' CFU from Vivolac were at 100 billion per packet. Each packet was used to ferment 1 Litre, thus 8 billion CFU/80mL serving at time of manufacturing. Significant multiplication is expected upon 24 hours fermentation, taking the CFU potentially as high as 16 billion/80mL serving. The finished product can be fortified with additional probiotic culture if desired.

Example 3: preparation of probiotic formulation with and without prebiotics

The following two formulations were prepared according to the methodology and specifications below.

Combine and heat coconut milk, pear juice, and Sunfiber and pectin (Formula #1) or kudzu root (Formula #2) to 185°F, stirring constantly. Remove heat. Let cool to 1 10° F then stir in probiotic cultures until thoroughly dissolved and distributed. Ferment inoculated substrate at 105° F for 24 hours. After 24 hours, pH should be at 3.5. Distribute flavourings and citric acid into the finished fermentation, and refrigerate.

Formula #1 - Coconut 1 pH3.5 Plantarum™, L. Lactis, L. Cremoris (80mL/serve)

Coconut Milk (Coconut meat, water, carob bean gum) 47.361 %

Pear Juice Blend from Concentrate 48.163%

Sunfiber® Prebiotic 3.099%

Citrus Pectin (Kosher) .197%

L. Plantarum LM™, L. Lactis, L Cremoris l OOB CFU/g .001% (use 1 g/L)

Organic Coconut Flavour .862%

Organic Vanilla Flavour .120%

Citric Acid .197%

100.000%

Formula #2 - Coconut 2 pH3.5 Plantarum ' , L. Lactis, L. Cremoris (SQmlJserv)

Coconut Milk (Coconut meat, water, carob bean gum) 47.361 %

Pear Juice Blend from Concentrate 48.163%

Organic Kuzu Root Starch 3.286% L. Plantarum LM™, L. Lactis, L Cremoris l OOB CFU/g .001 % (use l g/L)

Organic Coconut Flavour .869%

Organic Vanilla Flavour .123%

Citric Acid .197%

100.000%

Characteristi cs of Formulas 1 and 2

Type: Creamy-textured Liquid

Appearance: Cream Beige

Taste: Slightly Sweet, Sour, Tart, Rich, Natural. Named flavours perceptible

Odour: Subtle, Identifiable with named flavour, Natural