Probiotic Fortified Food Products and Methods of Manufacture

Abstract

The invention includes a method of manufacturing a probiotic fortified food product characterised by the step of applying a composition inoculated with at least one probiotic organism to at least one portion of the surface of the food product, wherein the composition includes a base which is fully milk derived and/or includes components inherently found in milk.

Claims

1. A method of manufacturing a probiotic fortified food product characterised by the step of a) applying a composition inoculated with at least one probiotic organism to at least one portion of the surface of the food product, wherein the composition includes a base which is fully milk derived and/or includes components inherently found in milk.

2. The method of claim 1 wherein the probiotic fortified food product is a bread.

3. The method as claimed in any one of the above claims wherein the food product has a water activity (a.sub.w) level between 0.5 to 0.9.

4. The composition as claimed above wherein at least one species of probiotic micro-organism is selected from the group consisting of a lactic acid bacteria, non-lactic acid bacteria and non-pathogenic yeast.

5. The method as claimed in any one of the above claims wherein at least one probiotic micro-organism is selected from the group consisting of Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus bifidus (bifidobacterium), Streptococcus thermophilus, and combinations thereof.

6. The method as claimed in any one of the above claims wherein, prior to step a), the composition is inoculated with the at least one micro-organism and pre-fermented in to achieve a CFU of above 7.0 log CFU ml.sup.1.

7. The method as claimed in any one of the above claims wherein, prior to step a), the composition is inoculated with the at least one micro-organism and pre-fermented to achieve a CFU of above 9.0 log CFU ml.sup.1.

8. The method as claimed in either claim 6 or 7 wherein the pre-fermentation step prior to step a) includes i. inoculating the base of the composition with an amount of a single species of probiotic micro-organism or combination of different probiotic micro-organisms; and ii. incubating the composition during an early stationary growth phase of the probiotic micro-organism until a probiotic culture in the composition is achieved with a cell count above 7 log CFU ml.sup.1.

9. The method of claim 8 wherein after step ii), the composition is homogenised to form a liquid with a viscosity below 4 cP.

10. The method as claimed in any one of claims 1 to 5 wherein at least one micro-organism is added before step a) to the composition to achieve a CFU of above 7.0 log CFU ml.sup.1.

11. The method as claimed in any one of claims 1 to 5 wherein the at least one micro-organism is added to the composition before step a) to achieve a CFU of above 9.0 log CFU ml.sup.1.

12. The method as claimed in any one of the above claims wherein the composition applied in step a) is a liquid with a viscosity below 4 cP.

13. The method as claimed in any one of the above claims wherein the base of the composition applied in step a) includes reconstituted milk.

14. The method as claimed in any one of the above claims wherein the base of the composition applied in step a) includes at least one coagulated component.

15. The method as claimed in claim 14 wherein the at least one coagulated component is protein and a coagulant is lactic acid.

16. The method as claimed in any one of the above claims wherein the base of the composition applied in step a) includes a milk-derived sugar.

17. The method as claimed in claim 16 wherein the milk-derived sugar is selected from the group consisting of lactose, glucose and/or galactose.

18. The method as claimed in claim 16 wherein the milk-derived sugar is lactose.

19. The method as claimed in any one of the above claims wherein the base of the composition applied in step a) has been supplemented with additional amounts of at least one sugar selected from a type of sugar inherently present in milk including lactose, glucose and/or galactose.

20. The method as claimed in any one of the above claims wherein the base of the composition applied at step a) includes a sugar concentration between 3 to 20% (w/v).

21. The method as claimed in any one of the above claims wherein the base of the composition applied at step a) includes a sugar concentration between 7-10% (w/v).

22. The method as claimed in any one of the above claims wherein the base of the composition applied at step a) includes less than 5% fat (w/v).

23. The method as claimed in any one of the above claims wherein the base of the composition applied at step a) includes substantially no fat.

24. The method as claimed in any one of the above claims wherein the the composition applied at step a) is configured to have a viscosity below 4 cP (centipoise).

25. The method as claimed in any one of the above claims wherein the composition applied at step a) is configured to have a viscosity between 1 to 3 cP.

26. The method as claimed in any one of the above claims wherein during step a) the composition is spread as the thin layer less than 0.1 mm thick.

27. The method as claimed in any one of the above claims wherein during step a) the composition is spread as the thin layer less than 0.05 mm thick.

28. The method as claimed in any one of the above claims wherein during step a) between 200 l to 2 ml of the composition is applied to a surface of the food product.

29. The method as claimed in any one of the above claims wherein during step a) between 500 l to 1 ml of the composition is applied to a surface of the food product.

30. The method as claimed in any one of the above claims wherein step a) includes spreading the composition across an entire surface of the food product.

31. The method as claimed in any one of the above claims wherein step a) includes spraying the composition on the surface of the food product.

32. The method as claimed in any one of the above claims characterized by the further step of b) drying the composition on the surface of the food product at a temperature below 70 C. after step a).

33. The method as claimed in claim 31 wherein step b) includes drying the composition on the surface of the food product at a temperature between 30 C. to 70 C.

34. The method as claimed in claim 31 wherein step b) includes drying the composition on the surface of the food product at a temperature between 50 C. to 60 C.

35. The method of any one of claims 32 to 34 wherein step b) is performed for between 10 to 120 minutes.

36. The method of any one of claims 32 to 35 wherein step b) is performed until the water activity (a.sub.w) of the composition applied to the surface of the food product and/or surfaces of the food product is between 0.5 to 0.9, and/or closely matches the original water activity (a.sub.w) of the food product before applying the composition to its surface.

37. A probiotic fortified food product characterised in that at least a portion of a surface of the food product includes a composition including at least one probiotic microorganism, wherein the composition includes a base which is fully milk-derived and/or includes components that are inherently found in milk.

38. A probiotic fortified food product prepared by the method as claimed in any one of claims 1 to 36.

39. A use of a fully milk derived composition in the manufacture of a probiotic fortified food product as claimed in claim 37 or 38, for the improvement, treatment or prevention of an intestinal tract dysfunction or disorder, and/or conditions relating to same.

40. A use of a fully milk derived composition in the manufacture of a probiotic fortified food product as claimed in claim 37 or 38 to improve immunity, act as an anti allergenic, treat eczema, and/or aid in cholesterol lowering.

41. A method of treatment, improvement or prevention of an intestinal tract dysfunction or disorder, and/or conditions relating to same by administering a probiotic fortified food product as claimed in claims 37 to 38 to person or other animal in need thereof.

42. A method of treatment by administering a probiotic fortified food product as claimed in claims 37 to 38 to improve immunity, act as an anti allergenic, treat eczema, and/or aid in cholesterol lowering to person or other animal in need thereof.

43. A method of manufacturing a fortified food product as substantially herein described and illustrated with reference to Examples 1 to 7 and FIGS. 1 to 2 in the Best Modes section.

44. A probiotic fortified food product as substantially herein described and illustrated with reference to Examples 1 to 7 and FIGS. 1 to 2 in the Best Modes section.

45. A use of a fully milk derived composition in the manufacture of a probiotic fortified food product as substantially herein described and illustrated with reference to Examples 1 to 7 and FIGS. 1 to 2 in the Best Modes section.

46. A method of treatment as substantially herein described and illustrated with reference to Examples 1 to 7 and FIGS. 1 to 2 in the Best Modes section.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0182] Further aspects of the present invention will become apparent from the ensuing description which is given by way of example only and with reference to the accompanying drawings in which:

[0183] FIG. 1 Analysis of Log CFU following preparation and storage of fortified LC431 probiotic bread; and

[0184] FIG. 2 Analysis of Log CFU following preparation and storage of fortified LC431, BB12 and LA05 combination probiotic bread.

[0185] FIG. 3 Analysis of Log CFU following preparation and storage of a fortified LC431 composition compared to commercially available Yakult and Activate probiotic drinks at various stages of manufacture of probiotic bread.

[0186] FIG. 4 Analysis of Log CFU following preparation and storage of fortified Lactobacillus plantarum 299V.

BEST MODES FOR CARRYING OUT THE INVENTION

Example 1: Preparation of the Growth Media

[0187] A growth media for Lactobacillus casei LC431 was made according to the following steps. [0188] 1. Prepare a media including 8.0% w/w reconstituted skim milk and 4.0% w/w glucose or lactose. [0189] 2. Heat the media to 90 C. for 15 minutes. [0190] 3. Cool to room temperature.

Example 2: Preparation of the Pre-Fermented Priobiotic Composition

[0191] The Lactobacillus casei LC431 probiotic composition was prepared as follows: [0192] 1. Add 0.05% w/w inoculant of freeze dried LC431 culture to the growth media as pre-prepared in Example 1. [0193] 2. Incubate the inoculated growth media for 16 hours at 37 C. [0194] 3. Homogenize the resulting soft curd to form a spreadable/sprayable liquid.

Example 3: Preparation of the Probiotic Fortified Bread

[0195] A fortified Lactobacillus casei LC431 probiotic bread was prepared as follows: [0196] a) Spray 500 l to 1 ml of composition (see Example 2) to a surface of a pre-baked bread loaf, ensuring a thin layer is evenly applied to the surface [0197] b) Dry the thin layer onto the bread's surface in a convention type forced air oven for 15 minutes at 50 C. [0198] c) Store the probiotic fortified bread at 25 C. or lower.

Example 4: Analysis of Log CFU Following Preparation and Storage of Fortified L. casei 431 (LC431) Probiotic Bread

[0199] A probiotic fortified bread was prepared according to Example 3. The inventors tested the viability of the LC431 cells (Log CFU) in three different trials. Log CFU was recorded for the control LC431, after application/drying, as well as following storage for five days at 25 C. (standard storage testing conditions for bread).

[0200] As can be seen in Table 1 below (and subsequently in FIG. 1), the Log CFU, the process of applying and then drying the composition on the top crust and bottom base of the bread resulted in very little cell viability loss. After storage, minor losses in cell viability were recorded in both samples, but were still well above the commercially and therapeutic level of 7.0 Log CFU required. It should be noted that various types of breads were tested using this method with similar probiotic viability results observed. The water activity of the breads before application of probiotics generally ranged between 0.75 to 0.94, and after drying, water activity ranged from 0.66 to 0.74.

TABLE-US-00001 TABLE 1 Analysis of Log CFU Sample Trial 1 Trial 2 Trial 3 Avg Fermented LC431 (control) 9.98 9.7 9.82 9.83 Bread top crust (after drying) 9.7 9.65 9.5 9.62 Bread bottom base (after drying) 9.43 9.3 9.32 9.35 Bread top crust (after storage) 8.42 8.26 8.38 8.35 Bread bottom base (after storage) 8.3 8.22 8.21 8.24

Example 5: Analysis of Log CFU Following Preparation and Storage of Fortified LC431, B. Lactis 12 (BB12) and L. acidophilus 05 (LA05) Combination Probiotic Bread

[0201] A probiotic LC431, BB12 and LA05 combination fortified bread was prepared according to Example 3, although using all three micro-organisms as the inoculant, not just LC431. The results are shown in Table 2 (and also FIG. 2).

[0202] Similar to Example 4, the inventors then tested the viability of the fortified combination probiotic cells. Similar beneficial and surprising results were seen in this trial showing that the inventive concept is not limited to specific probiotic types and may similarly also include combinations of probiotics.

TABLE-US-00002 TABLE 2 Analysis of Log CFU Sample Trial 1 Trial 2 Avg Fermented LC431 + BB12 + LA05 9.42 9.6 9.5 Bread top crust (after drying) 9.1 9.3 9.2 Bread bottom base (after drying) 9.3 9.45 9.4 Bread top crust (after storage) 8.02 8.23 8.1 Bread bottom base (after storage) 8.3 8.25 8.3

Example 6: Analysis of Log CFU Following Preparation and Storage of Fortified Lactobacillus plantarum 299V

[0203] A similar study was performed to Example 5, but this time using Lactobacillus plantarum 299V. As discussed previously, this probiotic micro-organism is an attractive target owing to its usefulness in treating IBS. The results are shown in FIG. 4.

[0204] The absolute values after five days were 9.12 and 9.07 log CFU per bread (top and bottom crust after storage, respectively), which translates into 1.3 billion and 1.17 billion of viable cells delivered per bread. This is an important commercial achievement, because the Applicant has been able to apply the invention using a wide range of different probiotic organisms, and in the case of Lactobacillus plantarum 299V, the results are showing extremely good probiotic cell viability counts after a five day storage trial.

[0205] Therefore the delivery of the clinically validated and popular probiotic strain Lactobacillus plantarum 299v via fortified breads may be a very attractive option to the targeted consumers suffering from a common gastro-intestinal disorder called Irritable Bowel Syndrome.

Example 7: Comparison of Pre-Fermented Composition Freshly Applied to Bread to Commercially Available Probiotic Drinks

[0206] To test the stability of different probiotic strains and the benefit of pre-fermenting the microbes in the composition, a test composition with LC431 (fermented in 8.0% (w/w) reconstituted skim milk and 4.0% (w/w) dextrose monohydrate as described previously) was compared to two commercially available probiotic drinks (Yakult and Activate).

[0207] For each group, 1 ml of the composition was spread on each piece of bread and dried at 50 C. for 15 min in a convection type forced air oven. A micro-pipette was used to distribute 1 ml of the liquid in mini droplet forms over the bread surfaces and spread evenly with the help of a spatula. The samples were then dried at 50 C. for 15 min in a convection type forced air oven. The bread samples were then packed in LDPE sachets and stored in the laboratory incubator maintained at a constant temperature of 25 C. for 5 days. Probiotic viability was tested before and after application to the top and bottom crust of the bread's surfaces, and also after the storage for 5 days.

[0208] The results are shown in FIG. 3. The fermented curd containing single strain of L. casei 431 yielded very high concentrations of viable cells to the tune of 9.9 log CFU. The drying losses in viability were minimal and recorded as 9.7 and 9.43 respectively for bread tops and bottoms. Moreover, only 1.3 and 1.1 log cell reductions were observed on bread top and bottom respectively, after the storage period. This was compared to the commercially available products which lost cell viability much more rapidly. For the test composition, the absolute values after 5 days were 8.4 and 8.3 log CFU, which translates into 250 million and 200 million of viable cells delivered per bread. These were more than 10 folds higher than the benchmark of delivering, at least 10 million cells per bread.

Example 8: Sensory Analysis of Probiotic Bread

[0209] A sensory trial was performed to determine the acceptability of the taste, feel and appearance of a fortified probiotic bread sample 1 (as used in Example 4) and a probiotic bread sample 2 (as used in Example 5) compared to the control (normal, non probiotic bread). Scores were provided for acceptability from a scale starting at 1 (poor acceptability) to 9 (excellent acceptability).

[0210] As can be seen in Table 3 below, the probiotic bread samples 1 and 2 are very comparable in terms of each specific test, and overall acceptability, to the control bread sample.

TABLE-US-00003 TABLE 3 Sensory analysis Probiotic Probiotic Bread Bread Test Control Sample 1 Sample 2 Crust appearance 6 6 6.5 Odour 7.5 7 6.7 Crust colour 6.5 6.85 6.55 Crispiness 7 7.2 6.8 Crust hardness 8 7 6.7 Taste 7.6 7.2 7.8 Overall acceptability 7.4 7.1 6.9

Example 9: Moisture Analysis of Probiotic Bread

[0211] The crust moisture content (%) and water activity of the probiotic bread samples 1 and 2 are only slightly higher than the control. These values are acceptable for commercial purposes, and it is unlikely that consumers will be able to notice any significant differences, as exemplified by the acceptability analysis shown in Example 6.

[0212] The slightly higher moisture content in the test samples will not adversely affect the shelf life of the bread, and it is still expected that the probiotic fortified bread will retain the full five day shelf life requirements.

TABLE-US-00004 TABLE 4 Moisture analysis Probiotic Probiotic Bread Bread Test Control Sample 1 Sample 2 Crust moisture content % 7.2 8.1 7.85 Crust water activity 0.75 0.74 0.72

[0213] Aspects of the present invention have been described by way of example only and it should be appreciated that modifications and additions may be made thereto without departing from the scope thereof as defined in the appended claims.