Saccharomyces cerevisiae and use thereof for improving intestinal health of pets

Abstract

The present invention relates to a Saccharomyces cerevisiae obtained by culturing a strain of Saccharomyces cerevisiae preserved in CNCM at number I 5660 on March 3, 2021, which is an active dry yeast having the following properties: a dry matter content greater than 95 wt% and at most 97 wt%; -a water activity of from 0.1 to 0.3; -a small sphere shape; -a particle size distribution characterized by a Gaussian particle size distribution curve showing that at least 90% of the small spheres have a diameter of 0.3 to 0.7 mm; having a stability of 24 to 36 months when in vacuum packaging and at a temperature of 15 to 25 DEG C. The invention also relates to the use of the yeast for improving intestinal health of pets. The invention also relates to a composition for pets, characterized in that it comprises a yeast as defined above, said yeast having a shelf life of 12 to 24 months in said composition. Such a composition may be a solid palatability enhancer or for example a coarse ground food comprising the palatability enhancer. The invention also relates to the use of the composition for improving intestinal health of pets.

Description

Technical Field

[0001] The present invention relates to the field of animal nutrition and pet intestinal health, and in particular to the use of Saccharomyces cerevisiae in improving pet intestinal health. Background Art

[0002] It is known in the prior art to use probiotic microorganisms (such as yeast or bacteria) to enhance pet intestinal health. Therefore, some documents describe the encapsulation or microencapsulation of microorganisms to ensure that they survive in the gastrointestinal tract, so that they can confer health benefits. Document JP2008013534 more specifically describes encapsulating lactic acid bacteria and yeast in a polymer comprising a mixture of xanthan gum and chitosan. The encapsulated probiotics thus obtained are used to improve the gastrointestinal diseases of pets. The article of Bruna M. Rodrigues et al. describes the technology of microencapsulating probiotic strains by freeze-drying, so as to keep the vitality of probiotics during storage and evaluate their resistance in the gastrointestinal tract. The experimental results of cats show that the use of probiotics or encapsulated probiotics (together with fructooligosaccharides) can regulate intestinal microbiota.

[0003] The use of probiotic microorganisms in pet food is also known in the prior art. When a microorganism or a mixture of microorganisms is incorporated into a food (e.g., a kibble coated with the microorganism), the microorganism must not only survive the kibble coating stage, but must also remain alive before the kibble expiration date in order to be able to impart its beneficial effects on animal health. Document PCT / US2009 / 056292 describes the use of probiotics (more particularly bifidobacteria) in a coating layer on pet kibble. However, the document does not mention yeast strains or yeasts that coat kibble and have a sufficiently long shelf life in the kibble to be effective for pet intestinal health.

[0004] There is still a need to find new probiotics that effectively enhance pet intestinal health and can be incorporated into pet food while having a sufficiently long shelf life in the food. In particular, a sufficiently long shelf life is a shelf life that lasts until the expiration date of the food.

[0005] The present inventors have unexpectedly developed an active dry yeast that has been shown to be particularly effective in enhancing the intestinal health of pets and that also has an exceptionally long shelf life when incorporated into pets' food. The exceptionally long shelf life is 12 to 24 months. SUMMARY OF THE INVENTION

[0007] According to a first aspect, an object of the present invention is Saccharomyces cerevisiae obtained by culturing the Saccharomyces cerevisiae strain deposited at the CNCM under number I-5660 on March 3, 2021. It is an active dry yeast having the following characteristics:

[0008] - a dry matter content of greater than 95% by weight and up to 97% by weight;

[0009] - Water activity from 0.1 to 0.3;

[0010] -Small spherical shape;

[0011] - a particle size distribution characterized by a Gaussian particle size distribution curve showing that at least 90% of said spheroids have a diameter between 0.3 and 0.7 mm;

[0012] - has a stability of 24 to 36 months when in vacuum packaging and at a temperature of 15-25°C.

[0013] The active dry yeast of the present invention also advantageously has a shelf life of 12 to 24 months when incorporated into a composition stored at a temperature of 15 to 25°C.

[0014] Such a composition may be, for example, a solid palatability enhancer or a pet kibble comprising, for example, the palatability enhancer.

[0015] According to a second aspect, the present invention relates to a yeast as defined above for use in pets:

[0016] - Improve the intestinal immune system,

[0017] -Restore normal intestinal microflora,

[0018] - lowering stool pH,

[0019] - Reduce the production of putrefactive metabolites (such as ammonia or biogenic amines) in feces.

[0020] According to another aspect, the present invention relates to a composition for pets, comprising an active dry yeast as defined above, said yeast having a shelf life of 12 to 24 months in said composition. Such a composition for pets may be a solid palatability enhancer or a kibble comprising said palatability enhancer, for example.

[0021] The present invention also relates to a composition as defined above for use in pets:

[0022] - Improve the intestinal immune system,

[0023] -Restore normal intestinal microflora,

[0024] - lowering stool pH,

[0025] - Reduce the production of putrefactive metabolites (such as ammonia or biogenic amines) in feces. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Other features, details and advantages will be apparent from the following detailed description and analysis of the accompanying drawings.

[0027] Figures 1 to 6 Include the letters a and b (lower case) and / or the letters A and B (upper case) just above the adjacent bars in the bar graph. The letters a and b indicate that the values ​​observed between the two groups are statistically different (p≤0.05) or not statistically different (p>0.1). The letters A and B indicate that a statistical trend is observed (p=0.05-0.1).

[0028] The capital letters A and B on the bars of the bar graph are obviously unrelated to the letters A, B, C or D used to distinguish different graphs in the same figure.

[0029] Figures 1 to 6 The vertical arrow in the graph indicates the abrupt change in the dog's diet that occurred on day D22, hereinafter referred to as the “dietary transition”.

[0030] The active dry yeast of the present invention is as follows Figures 1 to 6 It is indicated by “Sc50” in Chinese.

[0031] Figure 1

[0032] [ Figure 1 ] shows fecal pH values ​​(see A) and ammonia concentration values ​​(percent of dry matter) (see B) in beagle dog feces before and after a dietary shift on day 22 (D22) (as described above, the dietary shift is indicated by the vertical arrow).

[0033] The control group (black bars) was fed diet 1 before the diet switch and diet 2 after the diet switch.

[0034] The "Sc50" group (white bars) was fed diet 1 supplemented with active dry yeast Sc50 before the diet switch and diet 2 supplemented with active dry yeast Sc50 after the diet switch.

[0035] Figure 2

[0036] [ Figure 2 ] shows the levels of Blautia bacteria (see A), Bifidobacterium (see B), Lactobacillus (see C) and Faecalibacterium (see D) in beagle feces before and after the diet change on D22. Figure 1 Same as those described above.

[0037] Figure 3

[0038] [ Figure 3 ] shows the E. coli content in beagle feces before and after the diet change on D22. Figure 1 Same as those described above.

[0039] Figure 4

[0040] [ Figure 4 ] shows the dysbiosis index of the intestinal microbiota (see A) and the content of Streptococcus (see B) found in the feces of beagle dogs before and after the dietary change on D22. Figure 1 Same as those described above.

[0041] Figure 5

[0042] [ Figure 5 ] shows the fecal immunoglobulin A (IgA) content found in beagle feces before and after the dietary change on D22. Figure 1 Same as those described above.

[0043] Figure 6

[0044] [ Figure 6 ] shows the amount of biogenic amines present in beagle feces (in mg / kg dry matter) before and after the dietary transition on D22. Specifically, Figure A illustrates the amount of total biogenic amines in beagle feces before and after the dietary transition on D22, Figure B illustrates the amount of putrescine, and Figure C illustrates the amount of spermidine. Figure 1 Same as those described above.

[0045] Figure 7

[0046] [ Figure 7 ] shows the stability of active dry yeast Sc50 in two different palatability enhancers for dogs (see A) and for cats (see B).

[0047] Figure 8

[0048] [ Figure 8 ] shows different possible embodiments for coating kibble for dogs or cats.

[0049] In Panel A, a core of kibble is coated with a first coating layer (shown in grey) composed of fat (FAT) and then with a second coating layer (shown in white and bounded by a dotted line) composed of a liquid palatability enhancer.

[0050] In panel B, the kibble core is coated with a first coating layer (shown in grey) consisting of fat (FAT) and then with a second coating layer (represented by small dashed squares) consisting of a solid palatability enhancer mixed with the active dry yeast Sc50 of the present invention.

[0051] Finally, in panel C, the kibble core is coated with a first coating layer consisting of fat (FAT) (shown in grey), a second coating layer consisting of a liquid palatability enhancer (shown in white and demarcated by a dotted line), and then a third coating layer consisting of a solid palatability enhancer mixed with active dry yeast Sc50 (represented by a small dotted square).

[0052] Fig. 9

[0053] [ Fig. 9 ] shows the stability of active dry yeast Sc50 when coated on dog kibble.

[0054] In Panel A, the solid line represents the stability of yeast in coated kibble as a function of the coating method: fat then "solid palatability enhancer + active dry yeast Sc50" then fat then solid palatability enhancer. The dotted line represents the stability of yeast in coated kibble as a function of the coating method: fat then "solid palatability enhancer + active dry yeast Sc50".

[0055] In Panel B, the solid line represents the stability of yeast in coated kibble as a function of the coating method: fat then "solid palatability enhancer + active dry yeast Sc50" then fat then liquid palatability enhancer. The dotted line represents the stability of yeast in coated kibble as a function of the coating method: fat then "solid palatability enhancer + active dry yeast Sc50" then liquid palatability enhancer.

[0056] Fig.10

[0057] [ Fig.10 ] shows the stability of active dry yeast Sc50 when coated on cat kibble. Fig.10 The series shown corresponds to the mean viability values ​​obtained at each time point for the following four coating methods:

[0058] -Fat then "Solid Palatability Enhancer + Active Dry Yeast Sc50" then fat then solid palatability enhancer;

[0059] -Fat then "Solid Palatability Enhancer + Active Dry Yeast Sc50";

[0060] -Fat then "Solid Palatability Enhancer + Active Dry Yeast Sc50" then Vacuum Applied Fat;

[0061] -Fat then "Solid Palatability Enhancer + Active Dry Yeast Sc50" then Vacuum Applied Fat then Solid Palatability Enhancer.

[0062] Error bars represent + / - 1 standard deviation.

[0063] Fig.11

[0064] [ Fig.11 ] shows the stability of two active dry yeasts "Yeast 1" and "Yeast 2" coated on dog kibble. The coating method is:

[0065] -Fat then "Solid Palatability Enhancer + Active Dry Yeast 1" (curve connected by squares);

[0066] - fat then "solid palatability enhancer + active dry yeast 1" then liquid palatability enhancer (curve connected by triangles);

[0067] -Fat then "Solid Palatability Enhancer + Active Dry Yeast 2" (curve connected by circles);

[0068] -Fat then "Solid Palatability Enhancer + Active Dry Yeast 2" then Liquid Palatability Enhancer (curves connected by vertical lines). DETAILED DESCRIPTION

[0069] Therefore, the present invention relates to a Saccharomyces cerevisiae obtained by culturing a Saccharomyces cerevisiae strain deposited in the CNCM under CNCM number I-5660 on March 3, 2021, which is an active dry yeast having the following characteristics:

[0070] - a dry matter content of greater than 95% by weight and up to 97% by weight;

[0071] - Water activity from 0.1 to 0.3;

[0072] -Small spherical shape;

[0073] - a particle size distribution characterized by a Gaussian particle size distribution curve showing that at least 90% of said spheroids have a diameter between 0.3 and 0.7 mm;

[0074] - has a stability of 24 to 36 months when in vacuum packaging and at a temperature of 15-25°C.

[0075] A Saccharomyces cerevisiae strain is a yeast strain. The expression "yeast strain" refers to a relatively homogeneous population of yeast cells.

[0076] Yeast strains are obtained from the isolation of clones, which are cell populations obtained from a single yeast cell. The strain submitted to the CNCM under reference number I-5660 is also identified with the reference "Sc50". In particular, the strain may be named in this application as "strain Sc I-5660", "strain I-5660" or "strain Sc50".

[0077] Saccharomyces cerevisiae obtained by culturing strain I-5660 may be named "yeast Sc I-5660", "yeast I-5660" or "yeast Sc50".

[0078] As an indication, yeast I-5660 can be obtained by culturing strain I-5660 in a culture medium, for example as described in the reference book "Yeast Technology", 1991, G. Reed and TW Nagodawithana, ISBN 0-442-31892-8.

[0079] However, yeast I-5660 can also be obtained by an industrial-scale process consisting in preparing a liquid yeast extract or liquid yeast comprising the following steps:

[0080] - Cultivation of yeast strains in culture medium in several stages, first in a semi-aerobic environment, then in an aerobic environment,

[0081] - The yeast thus produced is separated from its culture medium by centrifugation to obtain a liquid yeast extract or liquid yeast containing a dry matter content of 12% to 25%, or even higher, in particular if the liquid yeast extract or liquid yeast is mixed with a permeate.

[0082] The liquid yeast extract or liquid yeast I-5660 obtained at the end of the above-mentioned culturing process is then subjected to a dehydration and drying process, which comprises the following steps:

[0083] - mixing liquid yeast extract or liquid yeast I-5660 with sodium chloride-based brine to obtain a volume of yeast extract comprising sodium chloride-based brine,

[0084] - dehydrating said volume of yeast extract comprising sodium chloride based saline to obtain dehydrated yeast,

[0085] - extruding the dehydrated yeast to obtain extruded yeast,

[0086] - Drying the extruded yeast so as to obtain a dry yeast having a dry matter content greater than 95% by weight relative to the total weight of the yeast.

[0087] The yeast I-5660 obtained at the end of this dehydration and drying process is called "active dry yeast". It may be referred to in the present application as "active dry yeast Sc I-5660", "active dry yeast I-5660", "active dry yeast Sc50" or simply "active dry yeast".

[0088] The term "active yeast" is synonymous with living yeast and refers to a population of metabolically active yeast cells.

[0089] For the purposes of this application, "active dry yeast" refers to active yeast having a dry matter content of greater than 95% by weight and up to 97% by weight.

[0090] The dry matter content was evaluated using the drying method after placing in an oven at a temperature of 103°C for at least 17 hours.

[0091] Water activity was assessed using an AquaLab series 3TE instrument (Decagon Devices, Pullman, WA, USA) using the chilled mirror dew point technique at 25° C. This is the primary measurement of relative humidity.

[0092] Active dry yeast is in the form of small spheres having a very low porosity surface and a very dense internal structure which gives the spheres, among other things, a moisture barrier effect.

[0093] The spheroids have a size distribution characterized by a Gaussian size distribution curve, indicating that at least 90% of the spheroids have a diameter between 0.3 and 0.7 mm.

[0094] The particle size distribution was evaluated by dry measurement using a laser diffraction method with a laser particle size analyzer. The particle size distribution of the spheroids was determined using the ISO 13320:2020 standard.

[0095] Advantageously according to the invention, the active dry yeast Sc50 has a stability of 24 to 36 months in vacuum packaging at a temperature of 15 to 25°C.

[0096] The stability of the active dry yeast of the present invention is directly related to its viability rate, i.e. the number of viable cells contained in the active dry yeast. One method of measuring the viability of yeast cells is to count the number of regenerable cells (in CFU / g or log CFU / g). "Regenerable cells" are viable cells that are able to proliferate and form colonies. The term CFU means "colony forming unit". One CFU corresponds to one colony. The viability of the cells is measured according to the EN15789 standardized method.

[0097] For the purposes of this application, an active dry yeast that is stable over a period of 24 to 36 months is one whose viable cell count and metabolically active cell count (CFU / g) does not decrease by more than 1 log CFU / g over the period of 24 to 36 months. This means, for example, that if the active dry yeast has a CFU / g of 1×10 10 CFU / g (equivalent to 10log CFU / g) of viable cells and metabolically active cells (t0 is the time when the active dry yeast is obtained, that is, the time when the active dry yeast is just prepared), then at the end of 24 to 36 months, the number of viable cells and metabolically active cells will not be less than 1×10 9 CFU / g (equivalent to 9logCFU / g).

[0098] When the active dry yeast of the present invention is stored in a vacuum package at a temperature of 15-25° C., it is confirmed that the active dry yeast of the present invention has a stability of 24 to 36 months.

[0099] Therefore, for the purpose of this application, the stability of active dry yeast over a period of 24 to 36 months means that the active dry yeast has a shelf life of 24 to 36 months according to the above standards (viability loss threshold is less than or equal to 1 log CFU / g). This shelf life / stability is extremely advantageous because in this way the active dry yeast maintains its properties, namely improving the intestinal health of pets.

[0100] Thus, the active dry yeast Sc50 is also characterized in that, when in vacuum packaging and at a temperature of 15 to 25°C, it has a shelf life of 24 to 36 months.

[0101] The inventors were able to determine that the physicochemical characteristics of the active dry yeast of the invention, namely dry matter content, water activity, globule shape and particle size distribution, are important for the shelf life exhibited by the yeast, especially when the latter is incorporated into pet compositions such as kibble. They were also able to determine that these properties are important for good adhesion of the yeast to kibble.

[0102] In fact, they were able to observe that if the dry matter content of the active dry yeast is less than 95%, for example 94 or 93%, the shelf life of said yeast in a composition such as pet food does not exceed 8 months, or even 5 months.

[0103] The inventors have also found that when the yeast of the invention is in the form of small spheres with a diameter of 0.3 to 0.7 mm, this allows good adhesion of the yeast at the periphery of a composition such as a pet food. In fact, when the small spheres of yeast have a diameter of less than 0.3 mm or greater than 0.7 mm, the yeast does not adhere. The adhesion of the yeast at the periphery of the kibble is characterized in particular by the percentage of "fines" produced after the durability / friability test. "Fines" means material that has been separated from the coated kibble and has passed through a sieve with a pre-adjusted mesh size.

[0104] The present invention also relates to the active dry yeast Sc50 as defined above for the following uses:

[0105] - Improve pet's intestinal immune system,

[0106] -Restore your pet's normal intestinal microbiota,

[0107] -Lower pet fecal pH, and / or

[0108] -Reduces the production of putrefactive metabolites in pet feces.

[0109] "Pet" refers to domestic animals such as dogs, cats, rabbits, birds, hamsters, etc. Dogs and cats may be mentioned as more specific examples of pets.

[0110] In humans, as in pets, a large number of immune cells are located in the intestine, which clearly illustrates the importance of the intestinal immune system. The digestive tract is constantly attacked by a variety of antigens and pathogens, which is why the digestive tract has a complex and highly developed immune system.

[0111] According to an advantageous embodiment of the invention, the administration of active dry yeast Sc50 in pets allows to significantly increase the production of fecal immunoglobulin A.

[0112] Therefore, a significant increase in fecal IgA production has the potential to improve the pet's intestinal immune system.

[0113] The gut microbiota refers to all the microorganisms that live in the digestive tract and plays an important role in maintaining good intestinal health. It is also fully involved in the functioning of the intestinal immune system.

[0114] Dysbiosis is a change in the composition and / or diversity of microorganisms in the digestive tract. It can lead to overgrowth of pathogenic bacteria, acute inflammatory responses, and diarrhea. Any change in the intestinal environment will cause changes in the microbiota, thereby causing dysbiosis. The microbiota dysbiosis index provides an overview of overall intestinal health in particular, and generally establishes whether dysbiosis exists. The inventors analyzed the dysbiosis index using a quantitative PCR (polymerase-chain-reaction) method disclosed in the journal FEMS Microbiology Ecology (AlShawaqfeh et al., 2017). Dysbiosis of the fecal microbiota is defined as a dysbiosis index greater than 2. A dysbiosis index of 0 to 2 indicates a moderate change in the fecal microbiota. A normal microbiota is defined as a negative dysbiosis index. Therefore, an improvement in the dysbiosis index means a statistically significant reduction in the dysbiosis index.

[0115] A “normal” gut microbiota is a healthy microbiota that does not show any changes in its digestive tract composition and / or microbial diversity.

[0116] The administration of active dry yeast Sc50 in pets with altered intestinal microflora has the potential to restore normal intestinal microflora. In fact, according to an advantageous embodiment of the invention, the administration of active dry yeast Sc50 in pets with altered intestinal microflora allows:

[0117] - Significant reduction in fecal streptococci and fecal coliforms,

[0118] - A significant effect on improvement of the dysbiosis index, and / or

[0119] - Significant increase in the levels of Blautia, Bifidobacterium, Lactobacillus and Faecalibacterium.

[0120] Blautia, Bifidobacterium, Lactobacillus, and Faecalibacterium are bacteria that contribute to good intestinal health.

[0121] Therefore, reductions in fecal Streptococcus and E. coli levels, improvements in the dysbiosis index, and / or increases in the abundance of these bacteria have the potential to restore a pet’s normal intestinal microbiota.

[0122] According to another advantageous embodiment of the invention, the administration of active dry yeast Sc50 in pets allows lowering of fecal pH.

[0123] Fecal pH can be considered a marker of the fermentative activity of the intestinal microbiota, with high pH being associated with proteolytic metabolism. Therefore, lowering fecal pH may help maintain intestinal function by inhibiting the proliferation of potentially pathogenic bacteria. In a publication by Lin and Visek, it was reported that a decrease in fecal pH in rats was associated with a decrease in proliferating cells in the colonic mucosa.

[0124] According to another advantageous embodiment, the administration of the active dry yeast Sc50 of the invention allows to reduce the production of putrefactive catabolites in pet feces, which represents a benefit for their intestinal health. Ammonia and biogenic amines such as spermidine and putrescine can be cited as examples of "putrefactive catabolites".

[0125] In the publication by Celi et al., the authors describe that excessive ammonia production is associated with excessive fermentative activity of bacteria in the intestinal microbiota. They also describe that ammonia can have toxic effects on intestinal cells.

[0126] As for biogenic amines, their excessive concentrations in pet feces may be harmful to the intestinal mucosa and promote the survival of potentially pathogenic bacteria.

[0127] Thus, lowering stool pH and reducing the production of putrefactive catabolites (eg, ammonia and / or biogenic amines) advantageously has the potential to improve a pet's stool characteristics and help improve a pet's intestinal health.

[0128] In addition to the health benefits, the reduction of putrefactive metabolites in pet feces also helps to reduce the bad odor from their feces, which represents a significant advantage because bad odor from feces can be a real nuisance to pet owners.

[0129] According to another advantageous embodiment of the present invention, active dry yeast Sc50 is used at 1×10 8 to 1.5×10 8 The daily dose of CFU / animal is used for one of the above uses.

[0130] The present invention also relates to a probiotic or food supplement for pets, characterized in that it comprises active dry yeast Sc50.

[0131] The terms "probiotics" and "food supplements" used above indicate that the active dry yeast Sc50 acts as a probiotic or a food supplement. The active dry yeast of the present invention may also be referred to as "probiotic yeast".

[0132] In 2001, WHO (World Health Organization) defined probiotics as "live microorganisms which when administered in adequate amounts confer health benefits beyond traditional nutritional effects".

[0133] According to the definition given by Directive 2002 / 46 / EC of the European Parliament, "food supplements are foods intended to supplement the normal diet, which are concentrated sources of nutrients or other substances having a nutritional or physiological effect and are marketed in dosed form, either alone or in combination."

[0134] The probiotic or food supplement of the invention comprises at least 95% by weight, preferably 96-99% by weight, of active dry yeast Sc50. Other ingredients constituting the probiotic or food supplement may be, for example, starch, but also vitamins, dietary minerals, trace elements and / or digestive enzymes.

[0135] According to one embodiment of the present invention, the probiotics or food supplements may be in a form suitable for oral administration, such as tablets, capsules, granules, powders, and the like.

[0136] The administration of the probiotics or food supplements of the invention can be carried out outside of meals or conversely at mealtimes of the pet. In the latter case, the probiotics or food supplements can be mixed, for example, with the animal's meals.

[0137] According to another advantageous embodiment of the invention, the probiotic or food supplement is administered in an amount ranging from 9 to 900 mg / day / animal.

[0138] According to another embodiment, the present invention relates to the use of a probiotic or a food supplement as defined above in a pet food, preferably kibble for dogs or cats.

[0139] The active dry yeast Sc50 of the present invention has the advantage of being easily dispersed in food ingredients, such as palatability enhancers intended to be incorporated into pet food. In fact, when mixed with food ingredients, the active dry yeast does not produce sedimentation or caking effects over time and advantageously has a viability of 12 to 24 months in said food ingredients. According to another advantageous embodiment, the present invention relates to the use of active dry yeast Sc50 in solid palatability enhancers for pets.

[0140] Palatability enhancers are, for example, in powder form and are intended to increase the attractiveness of the food intended for them to pets. Adding palatability enhancers to food allows a significant increase in the consumption of the food by pets. Palatability enhancers can be considered as flavoring agents or other taste factors.

[0141] Poultry offal (e.g. from chicken or turkey), pork or poultry liver, or fats (e.g. butter or cheese) may be mentioned as examples of palatability enhancers.

[0142] The invention also relates to a composition for pets, characterized in that it comprises an active dry yeast as defined above, said yeast having a shelf life in said composition of 12 to 24 months. Such a composition may be a solid palatability enhancer intended to be incorporated into pet food, or may be a pet food, such as kibble.

[0143] When the composition is stored at a temperature of 15 to 25°C, the condition of a 12 to 24 month shelf life of the active dry yeast in the composition is met.

[0144] According to another advantageous embodiment of the invention, the composition as defined above is more particularly characterized in that it is a solid palatability enhancer and that the yeast of the invention is incorporated into said palatability enhancer. According to another advantageous embodiment of the invention, the palatability enhancer has a water content of less than 10%.

[0145] The shelf life of the active dry yeast in the palatability enhancer of 12 to 24 months means that within the period of 12 to 24 months, the threshold of its viability loss is less than or equal to 1 log CFU / g, and preferably within the period of 12 to 24 months, the threshold of its viability loss is 0.1 log CFU / g to 0.8 log CFU / g. This means, for example, that if the active dry yeast is mixed with the palatability enhancer at a concentration of 1×10 7 If the amount of CFU exists (time T0), then 12 to 24 months after time T0 (T0 is the time when the live active dry yeast is mixed with the palatability enhancer), the amount of live active dry yeast per gram of palatability enhancer is not less than 1×10 6 CFU.

[0146] This shelf life is extremely advantageous because in this way, active dry yeast Sc50 retains its biological properties (i.e., improving pet intestinal health) when used in combination with palatability enhancers.

[0147] According to another embodiment of the present invention, the weight of active dry yeast in the solid palatability enhancer is 1 to 40%, more specifically 2 to 35%, relative to the total weight of the palatability enhancer.

[0148] The present invention also relates to the use of active dry yeast Sc50 as defined above or a solid palatability enhancer as defined above in pet food, preferably kibble.

[0149] The term "pet food" is understood to mean a food or food composition intended to be ingested by a pet, such as a dog or cat. As an example of such a food, mention may be made of kibbles for pets, more particularly kibbles for dogs or cats.

[0150] According to one embodiment of the present invention, active dry yeast Sc50 or solid palatability enhancer is incorporated into the food or deposited on the periphery of the food by coating.

[0151] Incorporating active dry yeast or palatability enhancers into food means, for example, that the yeast or palatability enhancer can be mixed directly into the food during its manufacture, so that the food contains the yeast or the palatability enhancer in its composition. However, it may also mean that the yeast or palatability enhancer is mixed into the food when the animal eats it.

[0152] When the yeast is incorporated into the food product during its manufacture, it is preferably active dry yeast as is.

[0153] When the yeast is mixed with the food at mealtime of the animal, the yeast is preferably in the form of a probiotic or food supplement.

[0154] According to a preferred embodiment, active dry yeast or a palatability enhancer is deposited on the periphery of a food, such as kibble for dogs or cats, by coating.

[0155] Viability testing of active dry yeast has shown favorably that the yeast has a shelf life / stability of 12 to 24 months when coated on the periphery of the kibble. These results are particularly attractive and promising because they demonstrate an extremely favorable shelf life. The stability of the active dry yeast of the present invention is directly related to its viability rate at the end of the kibble's useful life.

[0156] When used in a food, a palatability enhancer, a probiotic or a food supplement, the stability of the active dry yeast of the present invention is defined according to its viability loss threshold in said food, palatability enhancer, probiotic or food supplement.

[0157] More specifically, the stability of the yeast of the present invention in the food, palatability enhancer, probiotic or food supplement over a period of 12 to 24 months means that its threshold of viability loss is less than or equal to 1 log CFU / g over a period of 12 to 24 months, and preferably its threshold of viability loss is 0.1 to 0.8 log CFU / g over a period of 12 to 24 months. This means, for example, that after the yeast is incorporated into the food, palatability enhancer, probiotic or food supplement (referred to as time T0), if the active dry yeast is present at a concentration of 1×10 7 If the amount of CFU is present, then after a period of 12 to 24 months from time T0 (T0 = the time when the live active dry yeast is incorporated into the food, palatability enhancer, probiotic or food supplement), the amount of live active dry yeast per gram of food, palatability enhancer, probiotic or food supplement is not less than 1×10 6CFU. This shelf life is extremely advantageous because in this way, active dry yeast Sc50 retains its biological properties (i.e. improving pet intestinal health) when incorporated into foods, palatability enhancers, probiotics or food supplements.

[0158] These shelf life conditions for active dry yeast are observed when the food, palatability enhancer, probiotic or food supplement is stored at a temperature between 15 and 25°C.

[0159] According to a particularly advantageous embodiment, the present invention relates to a pet food, characterized in that it comprises an active dry yeast as defined above or a palatability enhancer as defined above, said yeast having a shelf life in said food of 12 to 24 months.

[0160] Another object of the invention is a probiotic or food supplement as defined above, further characterized in that it comprises an active dry yeast as defined above having a shelf life ranging from 12 to 24 months.

[0161] The shelf life of the active dry yeast in the food, probiotic or food supplement is as defined above (ie a threshold of loss of viability less than or equal to 1 log CFU / g, and preferably 0.1 to 0.8 log CFU / g, over a period of 12 to 24 months).

[0162] The variability in the shelf life of active dry yeast in pet food depends inter alia on the water activity of the food.

[0163] Therefore, the shelf life of active dry yeast Sc50 of the present invention is:

[0164] -12 months, when it is present in food with a high water activity, i.e. greater than 0.5;

[0165] - 24 months, when it is present in a food product with a low water activity, ie less than 0.5, preferably less than 0.4.

[0166] According to another embodiment of the present invention, Sc50 yeast is present at 5×10 5 Up to 5×10 9 CFU amount was administered.

[0167] According to another advantageous embodiment, the composition of the invention as defined above is more particularly characterized in that the yeast of the invention is coated on the periphery of said composition. For example, the yeast is more particularly comprised in the coating coating said composition.

[0168] Such a composition may be, for example, a food product such as a kibble comprising a coating comprising the yeast of the invention or a palatability enhancer as defined above.

[0169] Typically, kibbles or coated kibbles for pets consist of a core or center that is coated with at least one coating layer. The kibbles of the present invention more particularly refer to coated kibbles. The terms "kibbles" / "coated kibbles" and the terms "core" / "center" are used interchangeably hereinafter.

[0170] The coating may cover the entire surface of the core or only part of the surface of the core. According to a preferred embodiment of the present invention, the coating covers the entire surface of the core.

[0171] According to an advantageous embodiment of the invention, the weight percentages vary as follows, relative to the total weight of the kibble:

[0172] -50 to 97%, preferably 80 to 95% of the core,

[0173] - 3 to 50%, preferably 5 to 20% coating.

[0174] In this respect, the composition of the invention as defined above is more particularly characterized in that the coating is present in an amount ranging from 3 to 50% by weight, preferably from 5 to 20% by weight relative to the total weight of the composition.

[0175] The composition is, for example, kibble.

[0176] The core of the kibble has a moisture content of less than 10%.

[0177] The core of the kibble may, for example, contain proteins of animal or vegetable origin, carbohydrates, fats of animal or vegetable origin, minerals, vitamins, and the like.

[0178] According to an advantageous embodiment, the coating of the composition according to the invention also comprises fat.

[0179] For example, a kibble of the present invention may consist of a core, a first coating composed of fat, and a second coating comprising a solid palatability enhancer comprising the active dry yeast of the present invention.

[0180] As another example, a kibble may consist of a core and a coating comprising fat and active dry yeast Sc50 or a palatability enhancer comprising active dry yeast.

[0181] The invention also relates to the composition as defined above, for use thereof:

[0182] - Improve pet's intestinal immune system,

[0183] -Restore your pet's normal intestinal microbiota,

[0184] -Lower pet fecal pH, and / or

[0185] -Reduces the production of putrefactive metabolites in pet feces.

[0186] The effects of active dry yeast Sc50 on pet intestinal health as described above are fully applicable to the composition of the present invention comprising active dry yeast Sc50. This is particularly due to the excellent stability / shelf life (12 to 24 months) of the active dry yeast of the present invention in the composition of the present invention, which means that the active dry yeast does not show any loss of activity and is therefore able to exert its benefits on pet intestinal health in the same way as Sc50 yeast alone (not incorporated into a composition such as a palatability enhancer or kibble).

[0187] Example 1: Study on the effect of active dry yeast Sc50 on intestinal health

[0188] refer to Figures 1 to 6 The purpose of this example was to evaluate the effect of daily dietary supplementation with Sc50 active dry yeast on the intestinal health of healthy adult dogs following an abrupt dietary transition. An "abrupt dietary transition" is defined as a sudden change in diet.

[0189] Research plan

[0190] The study was conducted on 40 healthy adult beagle dogs (20 males and 20 females) with an average body weight of 10 to 15 kg.

[0191] The study was repeated twice and was approved by the Comité d'éthique de l'utilisation des animaux (Ethical Committee for the Use of Animals).

[0192] Two different groups were studied: supplemented or not supplemented with active dry yeast Sc50. Active dry yeast Sc50 (1×10 10 CFU of viable cells and metabolically active cells).

[0193] The study was conducted in two phases:

[0194] - Phase 1 lasts 3 weeks (from Day 1 to Day 21 (D1 to D21)), and

[0195] - Phase 2 lasts 4 weeks (from day 22 to day 49 (D22 to D49)).

[0196] On day 1 of phase 2, day 22 (D22), the dogs underwent an abrupt change in diet (hereinafter referred to as the “dietary transition” and Figures 1 to 6 denoted by the vertical arrow in the figure).

[0197] Therefore, phase 1 occurred before the dietary transition and phase 2 began on the first day of the dietary transition, i.e. D22.

[0198] Phase 1:

[0199] - Group A: Control group receiving Diet 1 (at Figures 1 to 6 (denoted as “control” in

[0200] - Group B: group receiving diet 1 supplemented with active dry yeast Sc50 (at Figures 1 to 6 denoted as “Sc50” in the literature).

[0201] Diet 1 consisted of a standard and balanced dog food with a low fiber content (ie, approximately 1 to 3%).

[0202] Phase 2 :

[0203] - Group A: Control group receiving Diet 2 (at Figures 1 to 6 denoted as “control” in the

[0204] - Group B: group receiving diet 2 supplemented with active dry yeast Sc50 (at Figures 1 to 6 denoted as “Sc50” in the literature).

[0205] Diet 2 consisted of foods with a high fiber content (ie, approximately 12 to 13%).

[0206] The abrupt switch from a low-fiber to a high-fiber diet is intended to produce intestinal dysbiosis.

[0207] Evaluation parameters

[0208] 1 / Collect stool samples for analysis of stool pH ( Figure 1 A) and fecal ammonia concentration ( Figure 1 B).

[0209] Fecal pH was measured directly in fresh fecal samples using a digital pH meter. Fecal ammonia concentration ([N]) (gr / kg) was determined according to the method described by Brito et al., 2010, and more specifically by calculation according to the following formula and applying a correction factor of 0.9875:

[0210] [Formula.1]

[0211]

[0212] 2 / Bacterial flora present in the intestinal microbiota ( Figure 2 A, B, C, D; Figure 3 B and Figure 4 B) and dysbiosis index ( Figure 4A) The analysis was also performed using the quantitative PCR (polymerase-chain-reaction) method, as published in the journal FEMS Microbiology Ecology (AlShawaqfeh et al., 2017).

[0213] Bacterial populations are expressed as logarithms of DNA.

[0214] The dysbiosis index allows the analysis of fecal microbiota, and more specifically allows the quantitative following bacteria: Faecalibacterium, Fusobacterium, Clostridiumhiranonis, Blautia, Streptococcus, Escherichia coli and Turicibacter. These bacteria are indeed identified in the literature as changing and / or responding to antibiotic therapy in certain intestinal diseases (such as chronic enteropathy). The dysbiosis index allows the reference range of these bacteria to be assessed and the value representing the degree of intestinal dysbiosis. Therefore, when the dysbiosis index is greater than 2, the dysbiosis of the term fecal microbiota is used. The dysbiosis index in the range of 0 to 2 shows the moderate change of fecal microbiota. The term normal biota is used when the dysbiosis index is negative.

[0215] Results

[0216] Active dry yeast Sc50 of the present invention was well accepted and tolerated by dogs in both studies. No occurrence of food refusal, vomiting, diarrhea or weight loss was observed throughout the study, even during the diet transition period. Encouraging and comparable results were reported and are summarized below.

[0217] Effect of active dry yeast Sc50 on fermentation yield ( Figure 1 )

[0218] After the dietary transition, a significant decrease in fecal pH was observed on D35 and D49 ( Figure 1 A) and significant decreases in fecal ammonia were observed at D23, D35 and D49 ( Figure 1 B).

[0219] Effects of active dry yeast Sc50 on intestinal microbiota ( Figure 2 B. Figure 3 B. Figure 4 B) and dysbiosis index ( Figure 4 A) Influence

[0220] Blautia spp. (D49) was observed in the feces of dogs after the diet change ( Figure 2 A), Bifidobacterium (D21 and D49) Figure 2 B), Lactobacillus (D21 and D49) Figure 2 C) and Faecalibacterium (D49) ( Figure 2D) a significant increase in content.

[0221] A significant decrease in fecal Streptococcus levels (D35) was observed after the dietary change ( Figure 4 B). A significant decrease in fecal E. coli levels was also observed after the diet change (D49) and a decreasing trend before the diet change (D21) ( Figure 3 ).

[0222] Similarly, improvements in the dysbiosis index were observed in the gut microbiota of dogs (D21, D23, and D35) before and after the dietary transition. Figure 4 A statistically significant reduction in the dysbiosis index is evident in A. The dysbiosis index allows for the evaluation of how the gut microbiota normalizes in response to a change in treatment, in this case a rapid dietary switch from diet 1 to diet 2.

[0223] Effect of active dry yeast Sc50 on fecal immunoglobulin A (IgA) production Figure 5 )

[0224] Figure 5 It was shown that administration of active dry yeast Sc50 before (D21) and after (D35) the dietary transition resulted in a significant increase in IgA production in dog feces.

[0225] Effects of active dry yeast Sc50 on the production of biogenic amines Figure 6 )

[0226] A significant decrease in total biogenic amine production was observed in dogs before (D21) and after (D49) the dietary transition. In particular, in dogs, a significant decrease in spermidine was observed after the dietary transition (D49), and a significant decrease in putrescine was observed before the dietary transition (D21), and a decreasing trend was observed after the dietary transition (D49).

[0227] in conclusion

[0228] Supplementing the diet of healthy dogs experiencing sudden dietary changes with active dry yeast Sc50 may help to:

[0229] a) Improved fecal properties of the test dogs, due to a reduction in fecal pH and the production of putrefactive catabolites. In fact, it has been reported that reduced fecal ammonia concentrations limit the replication of pathogenic microorganisms in the intestine. In addition, the reduction of biogenic amines contributes to a reduction in fecal odor, an important commercial feature considering the close relationship between dogs and their owners.

[0230] b) Improved local immunity by increasing fecal IgA production, which contributes to greater local resistance to antigen invasion and can limit the absorption of protein antigens and neutralize viruses.

[0231] c) Modulation of the composition and activity of the intestinal microbiota, for example by reducing bacteria such as Streptococcus and Escherichia coli, which are described in the literature as being associated with inflammatory processes in certain intestinal diseases, and by increasing beneficial bacteria in the intestinal microbiota. In addition, supplementation with active dry yeast Sc50 improved the dysbiosis index.

[0232] Example 2: Stability study of active dry yeast Sc50 in dog and cat food

[0233] refer to Figures 7 to 10 The purpose of this example is to evaluate the stability of active dry yeast Sc50 in the following conditions:

[0234] - In two solid palatability enhancers for dogs or cats,

[0235] - In dog or cat kibble, i.e. as a coating around a kibble core.

[0236] A mixture of active dry yeast Sc50 of the present invention and a palatability enhancer in powder form for dogs or cats

[0237] Active dry yeast Sc50 was incorporated at 5% into a palatability enhancer in powder form for dogs or cats, consisting of pork and poultry by-products, protein of vegetable origin, carbohydrates and minerals. The palatability enhancer powders differed in their different water activities, namely:

[0238] - Two palatability enhancers for dogs have a water activity of 0.32 or 0.38,

[0239] - Two palatability enhancers for cats have a water activity of 0.30 or 0.35.

[0240] The "palatability enhancer + active dry yeast Sc50" mixture was mixed in a mixer sold under the name Forberg (twin-shaft paddle mixer) by first introducing all the palatability enhancer powder into the mixer and then introducing the active dry yeast Sc50. The mixer was then run at a speed of 50 Hz for 5 minutes.

[0241] Study on the stability of active dry yeast Sc50 in palatability enhancer

[0242] The palatability enhancers supplemented with active dry yeast Sc50 were then packaged in plastic bags (for palatability enhancers for dogs) or aluminum bags (for palatability enhancers for cats) and stored at room temperature (20°C + / - 5°C).

[0243] The viability of active dry yeast Sc50 was measured at different time points (at least 4) throughout the study period, i.e. 0 to 12 months. The stability marker monitored during storage of active dry yeast Sc50 was the level of revivable yeast cells, which was measured according to the standardized method NF EN 15789. The results of the viability of active dry yeast Sc50 in various palatability enhancers are shown in Figure 7 .

[0244] More specifically, Figure 7 A illustrates the stability of active dry yeast Sc50 in two palatability enhancers for dogs, the solid line represents the stability of the palatability enhancer with a water activity of 0.32, and the dashed line represents the stability of the palatability enhancer with a water activity of 0.38.

[0245] Figure 7 B illustrates the stability of active dry yeast Sc50 in two palatability enhancers for cats, the solid line represents the stability of the palatability enhancer with a water activity of 0.30, and the dashed line represents the stability of the palatability enhancer with a water activity of 0.35.

[0246] Comments and Conclusions

[0247] from Figure 7 It can be clearly seen from Figures A and 7B that when the active dry yeast Sc50 of the present invention is incorporated into a palatability enhancer, it exhibits excellent stability.

[0248] The results show that the activity of active dry yeast Sc50 is stable in the four tested dog and cat palatability enhancers (which have a water activity of 0.30 to 0.38) under the tested storage conditions (20+ / -5°C) over a 12 month period. Compared to T0 (the time when the active dry yeast of the present invention is incorporated into the palatability enhancer), the activity of the four palatability enhancers after 12 months of contact differed by less than 1 log CFU / g.

[0249] Coating kibble cores with a palatability enhancer comprising active dry yeast Sc50

[0250] The various "palatability enhancer + active dry yeast Sc50" mixtures prepared in the previous step were then applied to dog or cat kibble in different coating sequences. Once these mixtures were prepared, they were applied to the kibble. The stability study of active dry yeast Sc50 was conducted over a period of 24 months.

[0251] Figure 8 Three possible embodiments for coating the core of a kibble are shown. This figure is provided as an example only, as other methods exist for coating the core of a kibble.

[0252] As an example, the main ingredients of cat and dog kibble can be summarized as follows:

[0253] -For dogs: barley, wheat and its derivatives, corn, poultry meal, whole soy beans, poultry fat;

[0254] -For cats: poultry meal, barley, corn and its derivatives, poultry fat, wheat, soy and its derivatives.

[0255] The percentage of active dry yeast Sc50 incorporated into the kibble was 0.05%.

[0256] Active dry yeast Sc50 pre-incorporated into palatability enhancer powder was applied to dog or cat kibble according to the different coating methods shown in Table 1 below. The abbreviations used in Table 1 have the following meanings:

[0257] -FAT: encapsulated fat;

[0258] -PEP: palatability enhancer powder;

[0259] -PEP+Sc50: palatability enhancer powder plus active dry yeast Sc50;

[0260] -LEP: Liquid Palatability Enhancer.

[0261] Vacuum applied FAT means that the coating fat has been deposited on the kibble while it is under a vacuum within the production equipment (which promotes penetration of the fat into the bulk of the kibble).

[0262] [Table 1]

[0263]

[0264]

[0265] Kibbles coated with active dry yeast Sc50 in this manner were packaged in aluminum bags and then stored at room temperature (20°C + / - 5°C).

[0266] Study on the stability of active dry yeast Sc50 coated on kibble

[0267] The stability marker monitored during storage of the coated kibbles was the level of regenerable yeast cells (standardized method NF EN 15789).

[0268] Fig. 9 The stability results of active dry yeast Sc50 coated dog kibble according to coating numbers 1-4 are shown.

[0269] More specifically, Fig. 9A illustrates the stability of active dry yeast Sc50 in kibble with a water activity of 0.45-0.49. The solid line represents the stability of yeast in kibble coated according to coating number 2, while the dotted line represents the stability of yeast in kibble coated according to coating number 1.

[0270] Fig. 9 B illustrates the stability of active dry yeast Sc50 in kibble with a water activity of 0.53-0.58. The solid line represents the stability of yeast in kibble coated according to coating number 4, while the dotted line represents the stability of yeast in kibble coated according to coating number 3.

[0271] Fig.10 Stability results for active dry yeast Sc50 coated on cat kibble with a water activity of 0.32-0.35 are presented. Fig.10 Shown in FIG. 5 are the average values ​​of the viabilities obtained at each time point for the four coating methods corresponding to coating numbers 1-2 and 5-6.

[0272] Comments and Conclusions

[0273] from Fig. 9 and 10 It can be clearly seen that the active dry yeast Sc50 of the present invention has excellent stability once coated around the coarse grind, which can achieve:

[0274] - When the coating layer contains only the palatability enhancer powder (Coating Nos. 1 and 2, Fig. 9 A) on dog kibble up to 15 months,

[0275] - When the coating contains a liquid palatability enhancer specific to certain dog kibbles (Coating Nos. 3 and 4, Fig. 9 B) reaches 12 months,

[0276] - Regardless of the coating method tested (No. 1-2, 5-6, Fig.10 ), up to 24 months on cat kibble.

[0277] Example 3: Comparative study with Example 2

[0278] The aim of this study was to demonstrate that the physicochemical properties of active dry yeast are critical for:

[0279] - their shelf life within the kibble, and / or

[0280] -Their adhesion within the kibble.

[0281] Shelf Life of Active Dry Yeast in Dog Kibble

[0282] The active dry yeast tested in this example has the following physicochemical properties.

[0283] Yeast "1" :

[0284] - Particle size distribution: The diameter of the small spheres is 0.3 to 0.7 mm;

[0285] - Dry matter content: 93% by weight.

[0286] Yeast "2" :

[0287] - Particle size distribution: The diameter of the small spheres is 0.05 to 0.2 mm;

[0288] - Dry matter content: 94.5% by weight.

[0289] Therefore, these active dry yeasts do not have all the physicochemical properties of the active dry yeast of the present invention.

[0290] The yeasts were incorporated into a palatability enhancer for dogs having a water activity of 0.32 according to the method described in Example 2. Similarly, the yeasts were coated on the periphery of dog kibble according to the method described in Example 2.

[0291] More specifically, yeast 1 is coated according to coating numbers 1 and 3, namely:

[0292] -FAT then "PEP+yeast 1" (coating number 1);

[0293] - FAT then "PEP+Yeast 1" then LPE (coating number 3).

[0294] In the same way, encapsulate yeast 2 according to encapsulation numbers 1 and 3:

[0295] -FAT then "PEP+yeast2";

[0296] -FAT then "PEP+yeast2" then LPE.

[0297] The solid (PEP) and liquid (LPE) palatability enhancers were those described in Example 2.

[0298] Dog kibble coated in this manner was stored under the same conditions as described in Example 2.

[0299] Viability measurements were assessed monthly according to the protocol described in Example 2.

[0300] The results are as follows Fig.11 It can be clearly seen from the figure that the viability of yeast 1 does not exceed 5 months, and the viability of yeast 2 does not exceed 8 months.

[0301] Yeast adhesion in kibble

[0302] The active dry yeast tested in this example has the following physicochemical properties.

[0303] Yeast "3" :

[0304] - Particle size distribution: The diameter of the small spheres is 0.1 to 0.2 mm;

[0305] - Dry matter content: 94% by weight.

[0306] Yeast "4" :

[0307] - Particle size distribution: The diameter of the small spheres is 0.01 to 0.1 mm;

[0308] - Dry matter content: 94% by weight.

[0309] Therefore, these active dry yeasts do not have all the physicochemical properties of the active dry yeast of the present invention.

[0310] The yeasts were incorporated into a palatability enhancer for dogs having a water activity of 0.32 according to the method described in Example 2. Similarly, the yeasts were coated on the periphery of dog kibble according to the method described in Example 2. More specifically, yeasts 3 and 4 were each coated according to coating number 1, namely:

[0311] -FAT then "PEP+yeast 3";

[0312] -FAT then "PEP+yeast 4".

[0313] The solid palatability enhancer (PEP) was the one described in Example 2.

[0314] The dog kibble coated in this way was subjected to a friability / durability test. The durability of the coated kibble was analyzed using a pellet durability tester sold under the name "Holmen NHP100". 100 grams of kibble were placed in the durability tester and each analysis was repeated twice. The tester was equipped with a 1 mm screen and the test lasted 120 seconds.

[0315] During the durability test, the kibble is subjected to standardized mechanical stresses. At the end of the mechanical test, the kibble is sieved through a sieve with a mesh size corresponding to 80% of the diameter of the kibble. The proportion of material originating from the kibble and having a size smaller than the mesh size of the sieve corresponds to "fines".

[0316] The higher the percentage of fines, the worse the yeast will adhere to the kibble.

[0317] The results obtained showed that the kibbles coated with Yeast 3 and tested for durability produced an average fines percentage of 1.79%, while the kibbles coated with Yeast 4 and tested for durability produced an average fines percentage of 1.55%.

[0318] The differences observed after the durability test are explained in particular by the physicochemical properties of the yeasts, in particular their particle size distribution. Larger yeasts will produce a higher percentage of fines compared to smaller yeasts.

[0319] The spherical shape of the yeast is important because it has very low surface porosity. Small spherical shapes have a large net surface area, thus providing a large surface area for ambient humidity. However, humidity is critical for the stability of the yeast over time. Therefore, a compromise is made between a spherical size that is small enough to adhere well to the kibble surface but not too small so as to be more stable over time through better moisture resistance while being industrially feasible.

[0320] The present disclosure is not limited to the embodiments described above merely as examples, but comprises all variants that are imaginable to a person skilled in the art within the framework of the protection sought.

[0321] References for depositing biological material

[0322] In this application, reference is made to the following biological materials and variants or mutant strains derived therefrom:

[0323] - Identified with reference "Sc50" and deposited on March 3, 2021 by Lesaffre SA, 41 rue Étienne Marcel 75009 Paris, under the reference "I-5660" with the Collection National des Microorganisms (CNCM), 25 rue Docteur Roux, 15th arrondissement of Paris, 75724, France.

[0324] Citation list

[0325] Patent Documents

[0326] The following patent documents are cited for the purpose of this article:

[0327] -patcit1: JP 2008013534; and

[0328] -patcit2: PCT / US2009 / 056292.

[0329] Non-patent literature

[0330] The following non-patent elements are cited for the purposes of this article:

[0331] -nplcit1:Bruna M.Rodrigues et al.,Hindawi,Int.J.of Microbiology,Vol2020,ID 1293481,10pages;

[0332] -nplcit2:Yeast Technology,2 nd edition,1991,G.Reed andT.W.Nagodawithana,published by Van Nostrand Reinhold,ISBN 0-442-31892-8;

[0333] -nplcit3:Journal FEMS Microbiology Ecology,"A dysbiosis index toassess microbial changes in fecal samples of dogs with chronic inflammatoryenteropathy."AlShawaqfeh et al.,2017.

[0334] -nplcit4:Large intestinal pH and ammonia in rats:dietary fat andprotein interactions 1-2.Hsi-Chiang Lin and Villard J.Visek.AmericanInstitute of Nutrition.1991,832-843;

[0335] -nplcit5:Celi,P;Verlhac,V;Calvo,EP;Schmeisser,J;Kluenter,AM.Biomarkers of gastrointestinal functionality in animal nutrition and health.Anim.Feed.Sci.Technol.2019,250,9-31;

[0336] -nplcit6:Brito,C.B.M.,Félix,A.P.,Jesus,R.M., M.I.,Oliveira,S.G.,Krabbe,E.L.,Maiorka,A.,2010.Digestibility and palatability of dog foodscontaining different moisture levels,and the inclusion of a mouldinhibitor.Anim.Feed Sci.Technol.159,150-155。

Claims

1. Saccharomyces cerevisiae obtained by culturing the strain of Saccharomyces cerevisiae submitted to the CNCM on March 3, 2021 under the number I 5660, which is an active dry yeast having the following characteristics: - a dry matter content of greater than 95% by weight and at most 97% by weight; - Water activity from 0.1 to 0.3; -Small spherical shape; - a particle size distribution characterized by a Gaussian particle size distribution curve showing that at least 90% of said spheroids have a diameter between 0.3 and 0.7 mm; - has a stability of 24 to 36 months when in vacuum packaging and at a temperature of 15-25°C.

2. The yeast according to claim 1, for use in: - Improve pet's intestinal immune system, -Restore your pet's normal intestinal microbiota, -Lower pet fecal pH, and / or -Reduces the production of putrefactive metabolites in pet feces.

3. A composition for pets, characterized in that It comprises the yeast according to claim 1, wherein the yeast has a shelf life of 12 to 24 months in the composition. The composition according to claim 3 , wherein the yeast is coated on the periphery of the composition.

5. The composition according to claim 3 or 4, wherein the yeast is contained in a coating layer coating the composition.

6. The composition according to claim 5, wherein the coating layer is present in an amount of 3 to 50 wt%, preferably 5 to 20 wt%, relative to the total weight of the composition.

7. The composition according to claim 5 or 6, wherein the coating further comprises fat.

8. The composition according to any one of claims 3 to 7, for use in: - Improve pet's intestinal immune system, -Restore your pet's normal intestinal microbiota, -Lower pet fecal pH, and / or -Reduces the production of putrefactive metabolites in pet feces.

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