Patent Description:
Probiotics are gaining importance as a dietary supplement owing to their ability to modify the gut microflora yielding increased health benefits. These probiotics can survive the acidic environments of the stomach and small intestine, but are not observed to colonize the gastro-intestinal tract. Thus, there exists a need for constantly supplementing probiotics for the therapeutic benefits to persist.

Beverages are widely consumed food constituents and can be looked upon as a source for co-administering probiotics. It must be noted that beverage compositions are subjected to treatments under extreme stress, temperature and pressure conditions like brewing or aeration wherein the viability of the probiotic spores should be maintained post treatment to result in a successful administration. <NPL> concerns viability of Bacillus coagulans MTCC <NUM> during processing and storage of functional foods. The document discloses a beverage obtained from brewing a composition comprising Bacillus coagulans MTCC <NUM>. As shown on page <NUM>, subsection "Brewed coffee", coffee powder was mixed with Bacillus coagulans MTCC <NUM> spores at room temperature and subsequently added to water preheated at <NUM> for <NUM> followed by incubation at <NUM> for four hours. As shown on page <NUM>, subsection "Brewed coffee" and page <NUM>, Fig. <NUM>, the viability of Bacillus coagulans MTCC <NUM> showed a <NUM> % reduction in viability already <NUM> after brewing. Furthermore, the subsequent incubation at <NUM> resulted in a <NUM> % and <NUM> % viability reduction after three and four hours, respectively. <CIT> describes (i) the growth promotional activity of natural plant based fibres on Bacillus coagulans MTCC <NUM>; (ii) the combination of natural plant based fibres and Bacillus coagulans MTCC <NUM> to inhibit Gram Negative pathogenic bacteria and (iii) the production of short chain fatty acids (SCFA) by Bacillus coagulans MTCC <NUM> using plant based natural fibres. <CIT> relates to a sports nutritional composition comprising a protein source and an isolated Bacillus coagulans spore. <CIT> relates to a composition comprising a non-dairy milk-like product and an isolated Bacillus coagulans spore.

<CIT> discloses a beverage composition comprising Bacillus coagulans strains GBI-<NUM>, GBI-<NUM> and GBI-<NUM>. <CIT> discloses a fruit beverage composition comprising Lactobacillus paracasei. Both these applications do not disclose the method of brewing and the survival of the probiotic bacteria in harsh conditions. Also, it is well known in the scientific art that that biological effects of probiotics or products thereof are strain specific and cannot be generalised among genera, species and strains (<NPL>). Hence, there still exists a need to find a probiotic strain that is more efficient and viable when subjected to extreme stress, temperature and pressure conditions.

The present invention solves the above technical problem by disclosing a probiotic strain that is viable and efficient when co-administered with beverages.

The principle objective of the inventions is to disclose beverage compositions comprising Bacillus coagulans MTCC <NUM> wherein the spore viability is maintained post treatments.

The present invention fulfils aforesaid objectives and provides further related advantages.

The present invention discloses beverage compositions that have been subjected to treatments under extreme stress, temperature and pressure conditions like brewing or aeration and comprise Bacillus coagulans MTCC <NUM> spores and water soluble prebiotic fibers wherein the spore viability is maintained post said treatments.

In a principle embodiment, the present invention discloses a beverage as specified in claim <NUM> which beverage is obtained by brewing of a composition comprising Bacillus coagulans MTCC <NUM> in the form of spores. The beverage is tea. It comprises Bacillus coagulans in the form of spores and viable or heat killed or dead or lysed vegetative cells of Bacillus coagulans MTCC <NUM>.

An embodiment not covered by the invention is a coffee composition comprises Bacillus coagulans MTCC <NUM> in the form of spore and bacterium wherein said bacterial spore is heat resistant having more than <NUM>% viability after brewing coffee. In a related embodiment not covered by the invention, the coffee is selected from the group consisting of decaffeinated coffee, unroasted green coffee and roasted coffee. In another related embodiment not covered by the invention, the spores include viable or heat killed or dead spores of Bacillus coagulans MTCC <NUM>. In yet another related embodiment not covered by the invention, the bacterium includes viable or heat killed or dead or lysed vegetative cells of Bacillus coagulans MTCC <NUM>.

The tea includes green tea, black tea, oolong tea, yellow tea, white tea or decaffeinated tea, herbal tea selected from the group consisting of rosehip tea, chamomile tea, jiaogulan tea, pep permint tea, rooibos tea, ginger tea, ginseng tea, or lemon grass tea.

In another embodiment not covered by the invention, the beverage is fruit juice. The fruit component of the fruit juice is selected from the group consisting of orange, apple, pears, strawberry, raspberry, cranberry, blue berry, apricot, pineapple, peach, banana, mango, lime, grape, tomato, pomegranate, papaya, tender coconut water, kiwi and a fruit mixture.

Another preferred embodiment not covered by the invention is an aerated soda composition comprising Bacillus coagulans MTCC <NUM> in the form of spore and bacterium wherein the said bacterial spore has the ability to survive aeration. In another related embodiment not covered by the invention, the spores include viable or heat killed or dead spores of Bacillus coagulans MTCC <NUM>. In yet another related embodiment not covered by the invention, the bacterium includes viable or heat killed or dead or lysed vegetative cells of Bacillus coagulans MTCC <NUM>.

In another preferred embodiment, the composition comprises a water soluble prebiotic fiber.

Another embodiment not covered by the invention is a synergistic coffee composition comprising a prebiotic water soluble fiber and Bacillus coagulans in the form of spore and bacterium wherein the said bacterial spore is heat resistant having more than <NUM>% viability after brewing coffee. In a related embodiment not covered by the invention, coffee includes decaffeinated coffee, unroasted green coffee and roasted coffee. In another related embodiment not covered by the invention the water soluble prebiotic fiber is selected from the group consisting of fructooligosaccharides (FOS), galacto-oligosaccharide (GOS), inulin and polydextrose. In another related embodiment not covered by the invention, the spores include viable or heat killed or dead spores of Bacillus coagulans. In yet another related embodiment not covered by the invention, the bacterium includes viable or heat killed or dead or lysed vegetative cells of Bacillus coagulans. In a preferable related embodiment not covered by the invention, the Bacillus coagulans strain is preferably Bacillus coagulans MTCC <NUM> and also the strains derived from Bacillus coagulans Hammer strain accession number ATCC <NUM> and ATCC <NUM>.

In another related embodiment of the invention the water soluble probiotic fiber is selected from the group consisting of fructooligosaccharides (FOS), galacto-oligosaccharide (GOS), inulin and polydextrose.

The aforesaid most preferred embodiments incorporating the technical features and technical effects of instant invention, are explained through illustrative examples herein under.

Commercial preparation of Bacillus coagulans MTCC <NUM> spores was standardized to <NUM> × <NUM><NUM> cfu (spores) per gram and added to grounded unroasted green coffee or roasted coffee and blended at room temperature with low r. Further, <NUM> of water soluble prebiotic fibres (fructooligosaccharide, galacto-oligosacharides, inulin and polydextrose) were added to <NUM> gm of unroasted green coffee or roasted coffee and blended at room temperature with low r. for <NUM> (Table <NUM>). Approximately <NUM> of the compositions were brewed using electric brewer and tested for viable spore count as per method described earlier (<NPL>). Average mean of spore viable counts are expressed in log<NUM> CFU.

Commercial preparation of Bacillus coagulans MTCC <NUM> was standardized to <NUM> × <NUM><NUM> cfu (spores) per gram and added to black, green or oolong tea and blended at room temperature with low r. Further, <NUM> of water soluble prebiotic fibres (fructooligosaccharide, galacto-oligosacharides, inulin and polydextrose) were added to <NUM> gm of black, green or oolong tea and blended at room temperature with low r. for <NUM> (Table <NUM>). Approximately <NUM> of these tea compositions were brewed using electric brewer and tested for viable spore count as per method described earlier (<NPL>). Average mean of spore viable counts are expressed in log10 CFU.

The effect of gastric stress on the Bacillus coagulans MTCC <NUM> spores was studied by treating brewed coffee (not according to the invention) or tea (as described in above paragraph) in acidic conditions. The pH of brewed coffee or tea was adjusted aseptically to <NUM> using <NUM> N HCl and incubated at <NUM> for <NUM> with low r. After <NUM> of incubation, again pH was adjusted aseptically back to <NUM> using <NUM> N NaOH and <NUM>% (w/v) ox bile was added to tea or coffee flask. Oxygen reducing enzyme Oxyrase (Oxyrase® for Broth, Oxyrase, Inc, Mansfield, OH, USA) was also added to each flask. Further flasks were incubated at <NUM> for <NUM> with low r. pH values at <NUM> of incubation and after <NUM> of incubation were also recorded. Optical density (OD) at <NUM> was recorded at <NUM> and after <NUM> of incubation by diluting the sample into sterilized demineralized water to <NUM>:<NUM>.

The thermo-stability and compatibility of Bacillus coagulans MTCC <NUM> spores in coffee was evaluated during brewing using electric brewer. Stainless steel and paper filter were used in brewer to understand the feasibility of probiotic brewing. There was <NUM> Log<NUM> cfu (spores) reduction after brewing which remained almost same even after <NUM> of incubation at room temperature (Table <NUM>). The data suggested that Bacillus coagulans MTCC <NUM> spores remained <NUM>% viable after brewing using Stainless steel filter. There was no significant difference observed when coffee was brewed using stainless steel or paper filters.

Black tea, green tea and oolong tea were supplemented with Bacillus coagulans MTCC <NUM> spores and brewed to determine the spore survival during brewing. Bacillus coagulans MTCC <NUM> spores were found to be highly stable and the viability remained above <NUM>% after brewing (Table <NUM> & <NUM>).

Same brewed tea samples were chilled for <NUM> and then spore viability was determined. The viability of spores in black, green and oolong tea after chilling for <NUM> was95. <NUM>%, <NUM>% and <NUM> % respectively (Table <NUM> & <NUM>).

To deliver the right amount of viable Bacillus coagulans MTCC <NUM> spores in the human gut, the effect of brewing conditions and gastric stress on the germination of Bacillus coagulans MTCC <NUM> spores was conducted. Tea and coffee along with the Bacillus coagulans MTCC <NUM> spores were brewed and then subjected to gastric stress (acid and bile) followed by anaerobic incubation at <NUM> with bile salts to mimic human conditions. This set of experiment was conducted to understand the spore germination when Bacillus coagulans MTCC <NUM> is brewed and digested by human. Additionally, along with tea and coffee B. coagulans MTCC <NUM> was brewed with water soluble prebiotic fibres (fructooligosaccharide, galacto-oligosacharides, inulin and polydextrose). Results of the experiment revealed that B. coagulans MTCC <NUM> spores could germinate after brewing and surviving in harsh condition of gastric stress by using tea or coffee as sole nutritional source (Table <NUM>). Additionally, there was further increase in the germination by the addition of water soluble prebiotic fibres (Table <NUM>). This suggested that soluble prebiotic fibres have synergistic effect along with tea and coffee on the germination of B. coagulans MTCC <NUM> spores.

The viability of Bacillus coagulans MTCC <NUM> was also tested in other beverages like aerated soda/cola, fruit juices, mineral water, chocolate, cocoa, fruit drinks, smoothies, milk shakes, cheese spread, honey, tomato ketchup, jam, mayonnaise, protein drink and ice cream. The results are tabulated in tables <NUM> & <NUM>.

Claim 1:
A beverage comprising Bacillus coagulans in the form of spores having more than <NUM> % viability, wherein the beverage is obtained by brewing of a composition comprising Bacillus coagulans MTCC <NUM> in the form of spores, and viable or heat killed or dead or lysed vegetative cells of Bacillus coagulans MTCC <NUM>, wherein said bacterial spores have the ability to survive stress, temperature and aeration treatments of said beverage composition, wherein the composition comprises tea and the viability is viability <NUM> minutes after brewing, wherein the tea is selected from the group consisting of green tea, black tea, oolong tea, yellow tea, white tea or decaffeinated tea, herbal tea selected from the group consisting of rosehip tea, chamomile tea, jiaogulan tea, peppermint tea, rooibos tea, ginger tea, ginseng tea, or lemon grass tea.