Saccharomyces cerevisiae yeast and its use in improving intestinal health in pets

Abstract

The present invention relates to a Saccharomyces cerevisiae yeast, which is an active dry yeast obtained by culturing the Saccharomyces cerevisiae strain deposited at the CNCM under number I-5660 on March 3, 2021, the active dry yeast having a dry matter content of more than 95% by weight and less than or equal to 97% by weight, a water activity of 0.1 to 0.3, a spheroid shape, a particle size distribution characterized by a Gaussian size distribution curve in which at least 90% of the spheroids have a diameter in the range of 0.3 to 0.7 mm, and a stability of 24 to 36 months when vacuum-packed and at a temperature of 15 to 25°C. The present invention relates to a composition for pets, comprising a yeast as defined above, the yeast having a shelf life of 12 to 24 months in the composition. The present invention further relates to the use of the composition for improving the intestinal health of pets.

Description

[Technical field]

[0001] This application relates to the field of animal nutrition and intestinal health of pets, and in particular to the use of Saccharomyces cerevisiae yeast in improving the intestinal health of pets. [Background technology]

[0002] The use of probiotic microorganisms, such as yeasts or bacteria, to enhance the intestinal health of pets is known from the prior art. Thus, several documents describe the encapsulation or microencapsulation of microorganisms to ensure their survival in the digestive tract and to be able to fulfill their health benefits. JP 2008-013534 more specifically describes the encapsulation of lactic acid bacteria and yeasts in a polymer containing a mixture of xanthan and chitosan. The encapsulated probiotics thus obtained are used to improve gastrointestinal disorders in pets. An article by Bruna M. Rodrigues et al. describes a technique of microencapsulation of probiotic strains by freeze-drying in order to maintain the viability of the probiotics during storage and to evaluate their tolerance in the digestive tract. Experimental results in cats show that the administration of probiotics or encapsulated probiotics (containing fructooligosaccharides) can modulate the gut microbiota.

[0003] The use of probiotic microorganisms in pet food products is also known from the prior art. When incorporating a microorganism or a mixture of microorganisms into a food product such as a kibble coated with said microorganism, it is essential that the microorganism not only survives the coating stage of the kibble, but also maintains viability until the expiration date of the kibble so that it can exert a beneficial effect on the health of the animal. US 2009 / 056292 describes the use of probiotics, in particular bifidobacteria, in the coating of pet kibbles. However, this document does not describe yeast strains or yeasts that coat the kibble and have a sufficiently long shelf life in said kibble to be effective for the intestinal health of the pet.

[0004] Currently, there remains a need to find new probiotics that are effective in enhancing the intestinal health of pets, that can be incorporated into pet food products, and that have a sufficiently long shelf life within said food products, particularly a shelf life that lasts until the expiration date of the food product.

[0005] The inventors have unexpectedly developed an active dry yeast that proves to be particularly effective in enhancing the gut health of pets and has a particularly long shelf life when incorporated into pet food products, the particularly long shelf life being anywhere from 12 to 24 months. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] JP 2008-013534 A [Patent Document 2] US Patent No. 2009 / 056292 [Non-patent literature]

[0007] [Non-Patent Document 1] Bruna M. Rodrigues et al., Hindawi, Int. J. of Microbiology, Vol 2020, ID 1293481, 10 pages

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

Non-Patent Document 5

Non-Patent Document 6

[0008] According to a first aspect, the object of the present invention is to provide a Saccharomyces cerevisiae yeast obtained by culturing the Saccharomyces cerevisiae strain deposited at the CNCM under number I-5660 on March 3, 2021, which is an active dry yeast, The dry matter content is greater than 95% by weight and may be less than or equal to 97% by weight; Water activity is 0.1 to 0.3, It has a spherical shape, having a particle size distribution characterized by a Gaussian size distribution curve showing that at least 90% of the spheroids have diameters in the range of 0.3 to 0.7 mm; It is vacuum-packed and has a stability of 24 to 36 months when kept at a temperature of 15 to 25°C. The active dry yeast of the present invention also advantageously has a shelf life of 12 to 24 months when incorporated into a composition and the composition is stored at a temperature of 15 to 25°C. Such a composition may be, for example, a solid palatability enhancer or a pet kibble, the pet kibble, for example, comprising the palatability enhancer.

[0009] According to a second aspect, the present invention relates to a pet Improves the intestinal immune system, Restoration of normal intestinal flora, Decreased fecal pH, The present invention relates to a yeast as defined above for use in reducing the production of putrefactive catabolic products (such as ammonia or biogenic amines) in faeces.

[0010] According to another aspect, the present invention relates to a pet composition comprising an active dry yeast as defined above, said yeast having a shelf life of 12 to 24 months in said composition. Such a pet composition may be a solid palatability enhancer or, for example, a kibble comprising said palatability enhancer.

[0011] The present invention also relates to a method for treating a pet. Improves the intestinal immune system, Restoration of normal intestinal flora, Decreased fecal pH, and / or The present invention relates to a composition as defined above for use in reducing the production of putrefactive catabolic products (such as ammonia and / or biogenic amines) in faeces. [Brief description of the drawings]

[0012] Other features, details and advantages will become apparent from the following detailed description and an analysis of the accompanying drawings. Figures 1-6 contain the letters a and b (lowercase) and / or the letters A and B (uppercase) immediately above adjacent bars in the histograms. The letters a and b indicate whether the values ​​observed between the two groups are statistically different (p≦0.05) or not (p>0.1). The letters A and B indicate that a statistical trend is observed (p=0.05-0.1). The capital letters A and B above the bars of the histogram clearly bear no relationship to the letters A, B, C or D provided to distinguish different diagrams within the same drawing. The vertical arrows in Figures 1-6 represent the abrupt change in the dog's diet that occurred on D22, hereafter referred to as the "dietary transition." The active dry yeast of the present invention is designated "Sc50" in Figures 1 to 6 below.

[0013] [Figure 1]Values ​​of fecal pH (see A) and ammonia concentration (percent dry matter) (see B) in faeces of beagle dogs before and after the dietary transition occurring on day 22 (D22) (dietary transitions are indicated by vertical arrows, as above). The control group (black histograms) was fed diet 1 before the dietary transition and diet 2 after the dietary transition. The "Sc50" group (white histograms) was fed diet 1 supplemented with active dry yeast Sc50 before the dietary transition and diet 2 supplemented with active dry yeast Sc50 after the dietary transition. [Diagram 2] The contents of bacteria of the genera Blautia (see A), Bifidobacterium (see B), Lactobacillus (see C) and Faecalibacterium (see D) in the feces of beagle dogs before and after the dietary transition on D22. The tested groups are the same as those described in Figure 1. [Diagram 3] Figure 2 shows the E. coli content in feces of beagle dogs before and after the dietary transition on D22. The groups studied are the same as those described in Figure 1. [Figure 4] Dysbiosis index (see A) and streptococci content (see B) of the intestinal microbiota found in the feces of beagle dogs before and after the dietary transition on D22. The groups tested were the same as those described in Figure 1. [Diagram 5] Figure 1 shows the fecal immunoglobulin A content found in the feces of beagle dogs before and after dietary transition on D22. The groups studied were the same as those described in Figure 1. [Figure 6] The amount of biogenic amines (calculated in mg / kg dry matter) present in the feces of beagles before and after the dietary transition on D22 is shown. Specifically, panel A shows the amount of total biogenic amines, panel B shows the amount of putrescine, and panel C shows the amount of spermidine in the feces of beagles before and after the dietary transition on D22. The groups tested were the same as those described in Figure 1. [Figure 7] The stability of active dry yeast Sc50 in two different palatability enhancers for dogs (see A) and cats (see B). [Figure 8]Various possible embodiments for coating kibbles for dogs or cats are shown. In figure A, the core of the kibble is coated with a first coating layer (shown in grey) consisting of fat (FAT), then coated with a second coating layer (shown in white and separated by a dotted line) consisting of a liquid palatability enhancer. In figure B, the core of the kibble is coated with a first coating layer (shown in grey) consisting of fat (FAT), then coated with a second coating layer (shown in small dotted squares) consisting of a solid palatability enhancer mixed with active dry yeast Sc50 of the present invention. Finally, in figure C, the core of the kibble is coated with a first coating layer (shown in grey) consisting of fat (FAT), then coated with a second coating layer (shown in white and separated by a dotted line) consisting of a liquid palatability enhancer, then coated with a third coating layer (shown in small dotted squares) consisting of a solid palatability enhancer mixed with active dry yeast Sc50. [Figure 9] FIG. 1 shows the stability of active dry yeast Sc50 coated on dog kibble. In FIG. A, the solid line represents the stability of yeast in the coated kibble as a function of coating method (fat, then "solid palatability enhancer + active dry yeast Sc50", then fat, then solid palatability enhancer). The dashed line represents the stability of yeast in the coated kibble as a function of coating method (fat, then "solid palatability enhancer + active dry yeast Sc50"). In FIG. B, the solid line represents the stability of yeast in the coated kibble as a function of coating method (fat, then "solid palatability enhancer + active dry yeast Sc50", then fat, then liquid palatability enhancer). The dashed line represents the stability of yeast in the coated kibble as a function of coating method (fat, then "solid palatability enhancer + active dry yeast Sc50", then liquid palatability enhancer). [Figure 10]Figure 10 shows the stability of active dry yeast Sc50 coated on cat kibble. The series shown in Figure 10 corresponds to the average viability obtained at each time point for the following four coating methods: fat, then "solid palatability enhancer + active dry yeast Sc50", then fat, then solid palatability enhancer; fat, then "solid palatability enhancer + active dry yeast Sc50", then fat, then solid palatability enhancer; fat, then "solid palatability enhancer + active dry yeast Sc50", then vacuum applied fat; fat, then "solid palatability enhancer + active dry yeast Sc50", then vacuum applied fat, then solid palatability enhancer. Error bars represent + / - 1 standard deviation. [Figure 11] The stability of two types of active dry yeast, Yeast 1 and Yeast 2, coated onto dog kibble is shown for the following coating schemes: Fat, then solid palatability enhancer + active dry yeast 1 (curve connected by squares); Fat, then solid palatability enhancer + active dry yeast 1, then liquid palatability enhancer (curve connected by triangles); Fat, then solid palatability enhancer + active dry yeast 2 (curve connected by circles); Fat, then solid palatability enhancer + active dry yeast 2, then liquid palatability enhancer (curve connected by vertical line). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] The present invention therefore relates to an active dry yeast, Saccharomyces cerevisiae yeast, obtained by culturing the Saccharomyces cerevisiae strain deposited at the CNCM under number I-5660 on March 3, 2021, said active dry yeast comprising: The dry matter content is greater than 95% by weight and may be less than or equal to 97% by weight; Water activity is 0.1 to 0.3, It has a spherical shape, having a particle size distribution characterized by a Gaussian size distribution curve showing that at least 90% of the spheroids have diameters in the range of 0.3 to 0.7 mm; The present invention relates to a yeast strain Saccharomyces cerevisiae, which has the characteristic that it has a stability of 24 to 36 months when vacuum-packed and at a temperature of 15 to 25°C. A Saccharomyces cerevisiae strain is a yeast strain. The term "yeast strain" refers to a relatively homogenous population of yeast cells. Yeast strains are obtained by isolation of clones, a clone being a population of cells derived from a single yeast cell. The strain deposited at the CNCM under the number I-5660 is also identified by the reference number "Sc50". In particular, this strain may be referred to in the present application as "strain Sc I-5660", "strain I-5660" or "strain Sc50". The Saccharomyces cerevisiae yeast obtained by culturing the I-5660 strain may be referred to as "Yeast Sc I-5660", "Yeast I-5660" or "Yeast Sc50". By way of example, the yeast I-5660 can be obtained by culturing the strain I-5660 in a medium as described, for example, in the reference book "Yeast Technology", 1991, G. Reed and TW Nagodawithana, ISBN 0-442-31892-8. However, yeast I-5660 Cultivating the yeast strain in a medium in several stages, first under semi-anaerobic conditions and then under aerobic conditions; and separating the yeast produced from the medium by centrifugation, thus obtaining a liquid yeast cream or liquid yeast containing 12-25% dry matter, or even a higher content of dry matter, especially if the yeast cream is mixed with an osmolyte product. And the liquid yeast cream or liquid yeast I-5660 obtained at the end of the above culture process is Mixing the liquid yeast cream or the liquid yeast I-5660 with a sodium chloride-based brine to obtain a quantity of yeast cream containing a sodium chloride-based brine; Dehydrating a quantity of yeast cream containing a sodium chloride-based brine to obtain dehydrated yeast; extruding the dehydrated yeast to obtain extruded yeast; drying the extruded yeast to obtain a dried yeast having a dry matter content of more than 95% by weight relative to the total weight of the yeast. The yeast I-5660 obtained at the end of this dehydration and drying process is called "active dry yeast", which may be indiscriminately 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". The term "active yeast" is synonymous with live yeast and refers to a population of metabolically active yeast cells. "Active dry yeast" for the purposes of this application refers to active yeast having a dry matter content of more than 95% by weight and which may be up to 97% by weight. Dry matter content was assessed using the drying method after placing in an oven at a temperature of 103°C for at least 17 hours. Water activity was assessed using the chilled mirror dew point technique at 25°C using an AquaLab series 3TE instrument (Decagon Devices, Pullman, WA, USA), which is the primary measurement of relative humidity. Active dry yeast is in the form of spherules which have a surface with very low porosity and a very dense internal structure, which gives the spherules a moisture barrier effect, among other effects. These spheroids have a particle size distribution characterized by a Gaussian size distribution curve showing that at least 90% of the spheroids have diameters in the range of 0.3-0.7 mm. The particle size distribution was evaluated by dry measurement using the laser diffraction scattering method with a laser particle size analyzer. The particle size distribution of the spheroids was determined using the ISO 13320:2020 standard. Advantageously, according to the invention, the active dry yeast Sc50 has the property that it has a stability of 24 to 36 months when vacuum packed and at a temperature of 15 to 25°C. The stability of the active dry yeast of the invention is directly related to its viability, 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 viable cells (calculated in CFU / g or log CFU / g). "Viable cells" are viable cells that can grow and form colonies. The term CFU means "colony forming unit". One CFU corresponds to one colony. The viability of the cells was determined by the EN15789 standardized method. For purposes of this application, an active dry yeast that is stable over a period of 24-36 months means that its number of live, metabolically active cells (CFU / g) does not decrease by more than 1 log CFU / g over said period of 24-36 months. This means, for example, that an active dry yeast has a cell count of 1×10 at time t0 (t0 being the time the active dry yeast was obtained, i.e., the time the active dry yeast was prepared). 10 At the end of the 24-36 month period, the yeast has a population of 1 × 10 9 This means that the cells will have a number of live, metabolically active cells equal to or greater than 9 log CFU / g (equivalent to 9 log CFU / g). The stability of the active dry yeast of the present invention for 24 to 36 months has been confirmed when said yeast is stored in a vacuum package at a temperature of 15 to 25°C. Thus, for the purposes of this application, stability of the active dry yeast over a period of 24-36 months means that the active dry yeast has a shelf life of 24-36 months according to the above criteria (viability loss threshold of ≦1 log CFU / g). This shelf life / stability is highly advantageous, since in this way the active dry yeast maintains its properties, namely properties that improve the intestinal health of pets. Thus, active dry yeast Sc50 is characterized by having a shelf life of 24 to 36 months when vacuum packed and at a temperature of 15 to 25°C.

[0015] The inventors have been able to determine that the physicochemical properties of the active dry yeast of the present invention, i.e. dry matter content, water activity, spheroid shape and particle size distribution, are important to the shelf life of the yeast, especially when the yeast is incorporated into a pet composition such as a kibble. The inventors have also been able to determine that these properties are important for good adhesion of the yeast to the kibble. In practice, the inventors have been able to observe that when the dry matter content of the active dry yeast is below 95%, for example 94% or 93%, the shelf life of said yeast in a composition such as a pet food does not exceed 8 months, or even 5 months. The inventors have also found that when the yeast of the present invention is in the form of spheres having a diameter of 0.3 to 0.7 mm, the yeast can adhere well to the periphery of a composition such as a pet food. In fact, when the diameter of the yeast spheres is less than 0.3 mm or more than 0.7 mm, the yeast does not adhere well. The adhesion of the yeast to the periphery of the kibble is particularly characterized by the proportion of "fines" generated after durability / friability testing. By "fines" is meant material that has fallen off the coated kibble and passed through the mesh of a sieve having a pre-adjusted mesh size.

[0016] The present invention also provides Improve your pet's intestinal immune system, Restoring normal intestinal flora in pets, A decrease in the pet's fecal pH, and / or The present invention relates to an active dry yeast Sc50 as defined above for use in reducing the production of putrefactive catabolic products in the faeces of pets. "Pets" refers to domestic animals such as dogs, cats, rabbits, birds, hamsters, etc. Dogs and cats are more specific examples of pets. Humans, like pets, have a large number of immune cells in the intestine, which clearly indicates the importance of the intestinal immune system. The digestive tract is constantly under attack by a large number of antigens and pathogens, and therefore has a complex and highly developed immune system. According to one advantageous embodiment of the invention, administration of active dry yeast Sc50 to pets can significantly increase the production of immunoglobulin A in the feces. By significantly increasing the production of immunoglobulin A in the feces, it can improve your pet's intestinal immune system. The gut microbiota, which refers to all the microorganisms living in the digestive tract, plays an essential role in maintaining good gut health and is also integrally involved in the functioning of the gut immune system. Dysbiosis is a change in the composition and / or microbial diversity of the digestive tract. This can lead to an overgrowth of pathogenic bacteria, acute inflammatory responses, and diarrhea. Any change in the intestinal environment can cause a change in the microbiota, resulting in dysbiosis. The dysbiosis index of the microbiota provides an overview of the overall health of the intestine in particular and confirms the presence or absence of dysbiosis in general. The inventors analyzed the dysbiosis index using a quantitative PCR (polymerase chain reaction) method published 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 between 0 and 2 indicates moderate changes in the fecal microbiota. A normal microbiota is defined as a negative dysbiosis index. An improvement in the dysbiosis index therefore means a statistically significant decrease in the dysbiosis index. A "normal" gut microbiota is a healthy microbiota that does not show any alterations in the composition and / or microbial diversity of the digestive tract. Administration of active dry yeast Sc50 to pets with altered intestinal flora can restore normal intestinal flora. Indeed, according to one advantageous embodiment of the invention, administration of active dry yeast Sc50 to pets with altered intestinal flora can: Significantly reduce the content of fecal streptococci and fecal coliform bacteria, has a significant effect on improving the dysbiosis index, and / or The content of Blautia, Bifidobacterium, Lactobacillus and Faecalis bacterium can be significantly increased. Bacteria from the genera Blautia, Bifidobacterium, Lactobacillus and Faecalibacterium are beneficial in maintaining good intestinal health. Thus, by reducing the content of fecal streptococci and Escherichia coli, improving the dysbiosis index and / or increasing the content of the above mentioned bacteria, the intestinal flora of a pet can be restored to normal. According to another advantageous embodiment of the invention, the administration of active dry yeast Sc50 to pets makes it possible to reduce the fecal pH. Fecal pH can be considered as a marker of the fermentation activity of the intestinal microflora, with high pH being associated with proteolytic metabolism. Therefore, lowering fecal pH can inhibit the growth of potential pathogenic bacteria and contribute to maintaining intestinal function. In a publication by Lin and Visek, it is reported that a decrease in fecal pH in rats correlates with a decrease in proliferating cells in the colonic mucosa. According to yet another advantageous embodiment, the administration of the active dry yeast Sc50 of the invention can reduce the production of putrefactive catabolites in the feces of pets, which is beneficial for the intestinal health of pets. Examples of "putrefactive catabolites" include ammonia and biogenic amines such as spermidine and putrescine. In the publication by Celi et al., the authors explain that excessive ammonia production is associated with excessive fermentative activity of bacteria in the gut microbiota, and that ammonia can have a toxic effect on intestinal cells. Regarding biogenic amines, if their concentrations in pet feces are too high, they can be harmful to the intestinal mucosa and promote the survival of potential pathogens. Therefore, lowering fecal pH and reducing the production of putrefactive catabolic products (such as ammonia and / or biogenic amines) can advantageously improve a pet's fecal profile and contribute to improved intestinal health of the pet. In addition to the health benefits, the reduction of putrefactive catabolic products in pet feces also contributes to reducing the odor caused by pet waste, which is a major benefit given that odor caused by waste can be a real nuisance for pet owners.

[0017] According to yet another advantageous embodiment of the invention, the active dry yeast Sc50 is used at a concentration of 1 x 10 per day for one of the uses defined above. 8 ~1.5×10 8 Doses ranging from 0.1 CFU / animal are used.

[0018] The present invention also relates to a probiotic or nutritional supplement for pets, characterized in that it contains active dry yeast Sc50. The terms "probiotic" and "food supplement" used above indicate that the active dry yeast Sc50 functions as a probiotic or food supplement. The active dry yeast of the present invention may also be referred to as "probiotic yeast". In 2001, probiotics were defined by the World Health Organization (WHO) as "live microorganisms that, when administered in adequate amounts, confer a health benefit beyond conventional nutritional benefits." As defined by Directive 2002 / 46 / EC of the European Parliament, "nutritional supplements mean food products marketed in dosage forms which are concentrated sources of nutrients or other substances having a nutritional or physiological effect, either alone or in combination, intended to supplement the normal diet." The probiotic or dietary supplement of the present invention comprises at least 95% by weight, preferably 96-99% by weight, of active dry yeast Sc50. Other components constituting the probiotic or dietary supplement may be, for example, starch, vitamins, dietary minerals, trace elements and / or digestive enzymes. According to one embodiment of the invention, the probiotic or dietary supplement may be in a form suitable for oral administration, such as, for example, tablets, capsules, granules, powders, and the like. The administration of the probiotic or dietary supplement of the present invention may be carried out outside of a meal or conversely at the time of the pet's meal, in which case the probiotic or dietary supplement may, for example, be mixed with the animal's food. According to yet another advantageous embodiment of the invention, the probiotic or dietary supplement is administered in an amount ranging from 9 to 900 mg / day / animal. According to yet another embodiment, the present invention relates to the use of a probiotic or a dietary supplement as defined above in a pet food, said food being preferably a kibble for dogs or cats.

[0019] The active dry yeast Sc50 of the invention has the advantage that it can be easily dispersed in food ingredients, for example in palatability enhancers intended for incorporation in pet food products. In fact, when mixed with a food ingredient, the active dry yeast does not produce any settling or clumping effects over time and advantageously has a viability of 12 to 24 months in said food ingredient. According to another advantageous embodiment, the invention relates to the use of the active dry yeast Sc50 in a solid palatability enhancer for pets. Palatability enhancers, for example in the form of powders, are intended to increase the attractiveness of pet food to pets. Adding a palatability enhancer to a food can significantly increase the consumption of this food by pets. Palatability enhancers can be considered flavorings or other taste factors. Examples of palatability enhancers can include poultry offal (eg, from chicken or turkey), pork or poultry liver, or fat (eg, butter or cheese).

[0020] The present invention also relates to a pet composition, characterized in that it comprises an active dry 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 intended for incorporation into pet food or it may be a pet food, for example a kibble. The requirement of a shelf life of 12 to 24 months for the active dry yeast in the composition is met when the composition is stored at a temperature of 15 to 25°C.

[0021] According to another advantageous embodiment of the invention, the composition as defined above is more particularly a solid palatability enhancer, characterized in 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 moisture content of less than 10%. A shelf life of 12-24 months for the active dry yeast in the palatability enhancer means that it has a viability loss threshold of 1 log CFU / g or less over a 12-24 month period, preferably 0.1 log CFU / g to 0.8 log CFU / g over a 12-24 month period. This means, for example, that live active dry yeast has a viability loss threshold of 1×10 immediately after mixing with the palatability enhancer (time T0). 7 CFU / g of the palatability enhancer, after a period of 12–24 months from time T0 (T0 being the time when the live, active dry yeast was mixed with the palatability enhancer), the amount of live, active dry yeast is greater than 1×10 6 This means that the CFU / g palatability enhancer is equal to or greater than 1. This shelf life is highly advantageous since in this way the active dry yeast Sc50 maintains its biological properties (i.e. improves the intestinal health of the pet) when used in combination with a palatability enhancer. According to yet another embodiment of the present invention, the weight of active dry yeast in the solid palatability enhancer is between 1 and 40%, more specifically between 2 and 35%, based on the total weight of the palatability enhancer.

[0022] The present invention also relates to the use of active dry yeast Sc50 as defined above, or of a solid palatability enhancer as defined above, in a pet food, said food being preferably a kibble. The term "pet food" is understood to mean a food product or composition intended for consumption by pets, for example dogs or cats. Examples of such foods include pet kibbles, more particularly dog ​​or cat kibbles.

[0023] According to one embodiment of the invention, active dry yeast Sc50 or a solid palatability enhancer is incorporated into the food or deposited around the food by a coating. Incorporating active dry yeast or a palatability enhancer into a food means that the food contains said yeast or said palatability enhancer in its composition itself, for example by mixing the yeast or palatability enhancer directly into the food during the manufacture of the food, but it may also mean that the yeast or palatability enhancer is mixed into the food at the time the animal eats. The yeast preferably remains as active dry yeast when it is incorporated into the food during manufacture of the food. If the yeast is mixed into the food at the animal's meal time, the yeast is preferably in the form of a probiotic or dietary supplement. According to a preferred embodiment, the active dry yeast or palatability enhancer is deposited by a coating around the food, said food being, for example, a kibble for dogs or cats. When the active dry yeast is coated around a kibble, viability testing of the active dry yeast advantageously demonstrated that the yeast has a shelf life / stability of 12-24 months. These results are particularly attractive and promising as they indicate a very favorable shelf life. The stability of the active dry yeast of the present invention is directly related to its viability at the end of the use life of the kibble. The stability of the active dry yeast of the present invention when used in a food, palatability enhancer, probiotic or probiotic is defined in relation to its viability loss threshold in said food, palatability enhancer, probiotic or dietary supplement. More specifically, the stability of the yeast of the present invention in a food, palatability enhancer, probiotic or dietary supplement for a period of 12-24 months means that it has a viability loss threshold of 1 log CFU / g or less over a period of 12-24 months, and preferably has a viability loss threshold of 0.1-0.8 log CFU / g over a period of 12-24 months. 7 CFU / g of active dry yeast is present in a food, palatability enhancer, probiotic or dietary supplement immediately after incorporation into said food, palatability enhancer, probiotic or dietary supplement (referred to as time T0), after a period of 12-24 months from time T0 (T0 = time the active dry yeast was incorporated into the food, palatability enhancer, probiotic or dietary supplement), the amount of active dry yeast is greater than 1 x 10 6 CFU / g of food, palatability enhancer, probiotic or dietary supplement or more. This shelf life is very advantageous, since the active dry yeast Sc50 thus incorporated into food, palatability enhancer, probiotic or dietary supplement maintains its biological properties (i.e. improves the intestinal health of pets). These shelf life conditions for active dry yeast are observed when the food, palatability enhancer, probiotic or dietary supplement is stored at a temperature between 15 and 25°C.

[0024] According to one particularly advantageous embodiment, the invention relates to a pet food comprising an active dry yeast as defined above or a palatability enhancer as defined above, characterized in that said yeast has a shelf life in said food of between 12 and 24 months. Another object of the invention is a probiotic or dietary supplement as defined above, further characterized in that it comprises an active dry yeast as defined above, the shelf life of which is between 12 and 24 months. The shelf life of the active dry yeast in the food, probiotic or dietary supplement is as defined above (i.e., a viability loss threshold of 1 log CFU / g or less, preferably 0.1-0.8 log CFU / g, over a period of 12-24 months). The variability in shelf life of active dry yeast in pet foods depends, among other things, on the water activity of the food. Therefore, the shelf life of the active dry yeast Sc50 of the present invention is as follows: If high water activity is present in the food, i.e., greater than 0.5, 12 months. 24 months if the water activity present in the food is low, i.e. less than 0.5, preferably less than 0.4. According to yet another embodiment of the present invention, the Sc50 yeast is 5×10 5 ~5×10 9 It is dosed in CFU / g.

[0025] According to yet another advantageous embodiment, the composition of the invention as defined above is more particularly characterized in that the yeast of the invention is coated around said composition, by way of example, the yeast is more particularly comprised in a coating layer coating said composition. Such a composition may for example be a food, such as a kibble, comprising a coating layer comprising the yeast of the invention or a palatability enhancer as defined above. Traditionally, pet kibble or coated kibble is composed of a core or center, and said core or center is coated with at least one coating layer. The kibble of the present invention more specifically refers to a coated kibble. The terms "kibble" / "coated kibble" and "core" / "center" can be used interchangeably below. The coating layer may cover the entire surface of the core or only a portion of the surface of the core. According to a preferred embodiment of the present invention, the coating layer covers the entire surface of the core. According to one advantageous embodiment of the invention, the weight percentages relative to the total weight of the kibble are: For the core, 50 to 97%, preferably 80 to 95%, For the coating layer, it is 3 to 50%, preferably 5 to 20%. In this respect, the composition of the invention as defined above is more particularly characterised in that the coating layer 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. The composition is, for example, a kibble. The moisture content of the kibble core is less than 10%. The kibble core may include, for example, proteins of animal or vegetable origin, carbohydrates, fats of animal or vegetable origin, minerals, vitamins, and the like. According to one advantageous embodiment of the invention, the coating layer of the composition according to the invention further comprises a fat. As an example, a kibble of the present invention may comprise a core, a first coating layer consisting of fat, and a second coating layer comprising a solid palatability enhancer comprising the active dry yeast of the present invention. Also by way of example, the kibble may consist of a core and a coating layer comprising fat and a palatability enhancer comprising active dry yeast Sc50 or active dry yeast.

[0026] The present invention also provides Improve your pet's intestinal immune system, Restoring normal intestinal flora in pets, A decrease in the pet's fecal pH, and / or The present invention relates to a composition as defined above for use in reducing the production of putrefactive catabolic products in the faeces of pets. The above effects of the active dry yeast Sc50 on the intestinal health of pets are fully applicable to the compositions of the invention comprising the active dry yeast Sc50. This is in particular due to the fact that the active dry yeast of the invention has an excellent stability / shelf life (12-24 months) in the compositions of the invention, which means that said active dry yeast does not show any loss of activity and can therefore exert its benefits on the intestinal health of pets in the same way as when the Sc50 yeast is used alone (without being incorporated into a composition such as a palatability enhancer or kibble).

[0027] Example 1: Study of the effectiveness of active dry yeast Sc50 on intestinal health This example aims to evaluate the impact of daily dietary supplementation with Sc50-active dry yeast on the intestinal health of healthy adult dogs following an abrupt dietary transition, as shown in Figures 1 to 6. An "abrupt dietary transition" is defined as a sudden change in diet.

[0028] Protocol of the study carried out The study was conducted on 40 healthy adult beagle dogs (20 males, 20 females) with an average weight of 10–15 kg. This study was repeated twice and approved by the Animal Use Ethics Committee. Two different groups were studied, one with and one without supplementation of active dry yeast Sc50 (1 × 10 per gram of active dry yeast). 10 CFU of live metabolically active cells) was administered at a dose of 150 mg per dog per day. The study was conducted in two stages. Phase 1 lasts for 3 weeks (days 1-21 (D1-D21)). Phase 2 lasts for 4 weeks (days 22 to 49 (D22-D49)). On day 1 of phase 2, i.e., day 22 (D22), dogs underwent a sudden change in diet (hereafter referred to as “dietary transition” and represented by a vertical arrow in Figures 1 – 6 ). Thus, phase 1 takes place before the eclipse transition and phase 2 begins on D22, the first day of the eclipse transition. Phase 1 Group A was the control group consuming diet 1 (represented as "Control" in Figures 1 to 6). Group B was the group that consumed diet 1 supplemented with active dry yeast Sc50 (represented as "Sc50" in Figures 1 to 6). Diet 1 consisted of a balanced standard dog food with a low fiber content, i.e., approximately 1-3%. Phase 2 Group A was the control group consuming diet 2 (represented as "Control" in Figures 1 to 6). Group B was the group that consumed diet 2 supplemented with active dry yeast Sc50 (represented as "Sc50" in Figures 1 to 6). Diet 2 consisted of foods with a high fiber content, ie, about 12-13%. The sudden transition from a low-fiber to a high-fiber diet is done to induce intestinal dysbiosis.

[0029] Evaluated parameters 1. Fecal samples were collected and analyzed for fecal pH (Fig. 1A) and fecal ammonia concentration (Fig. 1B). 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 by Brito et al. (2010) and, more specifically, calculated by applying a correction factor of 0.9875 according to the following formula:

number

[0030] Results obtained The active dry yeast Sc50 of the present invention was well accepted and tolerated by dogs in both studies. No refusal to eat, vomiting, diarrhea or weight loss was observed throughout the study or during diet transition. Promising and comparable results were reported and are summarized below.

[0031] Effects of active dry yeast Sc50 on fermentation production (Figure 1) After dietary transition, a significant decrease in fecal pH at D35 and D49 (Fig. 1A) and fecal ammonia at D23, D35, and D49 (Fig. 1B) was observed.

[0032] Effects of active dry yeast Sc50 on gut microbiota (Figures 2B, 3B, and 4B) and dysbiosis index (Figure 4A). After the dietary transition, a significant increase in the contents of Blautia (D49) (Figure 2A), Bifidobacterium (D21 and D49) (Figure 2B), Lactobacillus (D21 and D49) (Figure 2C) and Faecalibacterium (D49) (Figure 2D) in the feces of dogs was observed. After the dietary transition, a significant decrease in fecal Streptococcus content (D35) (Figure 4B) was observed. After the dietary transition (D49), a significant decrease in fecal E. coli content was observed, and before the dietary transition (D21), a decreasing trend was observed (Figure 3). Similarly, an improvement in the dysbiosis index was observed in the gut microbiota of dogs before and after the dietary transition (D21, D23 and D35). Indeed, a statistically significant decrease in the dysbiosis index is evident from FIG. 4A. The dysbiosis index allows us to evaluate how to normalize the gut microbiota in response to a treatment change, in this case a rapid dietary transition from diet 1 to diet 2.

[0033] Effect of active dry yeast Sc50 on fecal immunoglobulin A (IgA) production (Figure 5) FIG. 5 shows that administration of active dry yeast Sc50 significantly increases IgA production in the faeces of dogs before (D21) and after (D35) the dietary transition.

[0034] Effect of active dry yeast Sc50 on biogenic amine production (Figure 6) In dogs, a significant decrease in total biogenic amine production was observed before (D21) and after (D49) the dietary transition. In particular, a significant decrease in spermidine was observed after (D49) the dietary transition, and a significant decrease in putrescine was observed before (D21) and with a tendency to decrease after (D49) the dietary transition in dogs.

[0035] conclusion Supplementing the diet of healthy dogs undergoing an acute dietary transition with active dried yeast Sc50 achieves the following effects: a) Improve fecal characteristics of the test dogs by reducing fecal pH and generating putrefactive catabolic products. In fact, a reduction in fecal ammonia concentration has been reported to limit the replication of pathogenic microorganisms in the intestine. Also, the reduction in biogenic amines contributes to a reduction in fecal malodor, which is an important commercial attribute given the close relationship between dogs and their owners. b) improving local immunity by increasing fecal IgA production, which may contribute to greater local resistance to antigenic invasion, limit the absorption of protein antigens, and subdue viruses. c) modulating the composition and activity of the intestinal microbiota, for example by decreasing bacteria such as Streptococcus and E. coli, which are described in the literature as being associated with inflammatory processes in certain intestinal diseases, and increasing beneficial bacteria in the intestinal microbiota. Also, supplementation with active dry yeast Sc50 improved the dysbiosis index.

[0036] Example 2. Stability study of active dry yeast Sc50 in dog and cat food In this embodiment, as shown in FIGS. Two types of solid palatability enhancers for dogs and cats: The aim is to evaluate the stability of active dry yeast Sc50 within dog or cat kibbles, i.e. as a coating around the kibble core.

[0037] A mixture of the active dry yeast Sc50 of the present invention and a palatability enhancer in powder form for dogs or cats. The active dry yeast Sc50 is incorporated at 5% into a powdered palatability enhancer for dogs or cats, which is made of pork and chicken by-products, plant-derived proteins, carbohydrates and minerals. The powdered palatability enhancers are differentiated by their water activity, specifically as follows: The water activity of the two palatability enhancers for dogs is 0.32 or 0.38. The two palatability enhancers for cats have a water activity of 0.30 or 0.35. The mixture of "palatability enhancers + active dry yeast Sc50" is combined in a mixer sold under the name Forberg (two-shaft paddle mixer), firstly all the palatability enhancer powders are added to the mixer, followed by the active dry yeast Sc50, and the mixer is run at a speed of 50 Hz for 5 min.

[0038] Study of the stability of active dry yeast Sc50 in palatability enhancers The palatability enhancers containing the active dry yeast Sc50 are packed in plastic bags (for dog palatability enhancers) or aluminium bags (for cat palatability enhancers) and stored at room temperature (20°C + / - 5°C). Measurements of the viability of the active dry yeast Sc50 are carried out at different time points (at least four) over the entire study period, i.e. from 0 to 12 months. The stability marker monitored during storage of the active dry yeast Sc50 is the level of resurrectable yeast cells, said level being measured according to the standardized method NF EN 15789. The results of the viability of the active dry yeast Sc50 in the various palatability enhancers are shown in Figure 7. More specifically, FIG. 7A shows the stability of active dry yeast Sc50 in two palatability enhancers for dogs, with the solid line showing stability in a palatability enhancer with a water activity of 0.32 and the dashed line showing stability in a palatability enhancer with a water activity of 0.38. FIG. 7B shows the stability of active dry yeast Sc50 in two palatability enhancers for cats, where the solid line shows stability in a palatability enhancer with a water activity of 0.30 and the dashed line shows stability in a palatability enhancer with a water activity of 0.35. Opinions and conclusions From Figures 7A and 7B it is clear that the active dry yeast Sc50 of the present invention exhibits excellent stability when incorporated into a palatability enhancer. The results show that over a 12 month period under the storage conditions tested (20+ / -5°C), the viability of active dry yeast Sc50 is stable in the four tested palatability enhancers for dogs and cats with water activities ranging from 0.30 to 0.38. The difference in viability of these four palatability enhancers after 12 months of contact is less than 1 log CFU / g compared to TO (the time when the active dry yeast of the present invention was incorporated into the palatability enhancer).

[0039] Coating kibble cores with a palatability enhancer containing active dry yeast Sc50 The various "palatability enhancer + active dry yeast Sc50" mixtures prepared in the previous step are applied to dog or cat kibbles in different coating sequences. These mixtures are applied to the kibbles as soon as they are prepared. A stability study of active dry yeast Sc50 was conducted over a period of 24 months. Figure 8 shows three possible embodiments for coating the kibble core. This figure is provided as an example only, as there are other ways to coat the kibble core. By way of example, the main ingredients of cat and dog kibbles can be summarized as follows: For dogs, these include barley, wheat and derivatives, corn, poultry meal, whole soybeans, and poultry fat. For cats, there are poultry meal, barley, corn and its derivatives, poultry fat, wheat, soybean and its derivatives. The proportion of active dry yeast Sc50 incorporated into the kibble is 0.05%. The active dry yeast Sc50, previously incorporated into the palatability enhancer powder, is applied to the dog or cat kibble according to the various coating methods shown below in Table 1. The abbreviations used in Table 1 have the following meanings: FAT is a coating fat. PEP is a palatability enhancer powder. PEP+Sc50 is a palatability enhancer powder and active dry yeast Sc50. LPE is a liquid palatability enhancer. Vacuum applied FAT means that the coating fat was applied to the kibble while it was under vacuum in the manufacturing equipment (this helps the fat penetrate into the kibble body).

[0040] [Table 1]

[0041] The kibbles thus coated with active dry yeast Sc50 are packed in aluminium bags and stored at room temperature (20°C + / - 5°C).

[0042] Stability study of active dry yeast Sc50 coated on kibble A stability marker monitored during storage of coated kibbles is the level of resurrectable yeast cells (standardization method NF EN 15789). FIG. 9 shows the stability results of dog kibbles coated with active dry yeast Sc50 according to coating codes 1-4. More specifically, Figure 9A shows the stability of active dry yeast Sc50 in kibbles with water activities between 0.45 and 0.49. The solid line represents the stability of the yeast in kibbles coated according to Coating Code 2, and the dashed line represents the stability of the yeast in kibbles coated according to Coating Code 1. 9B shows the stability of active dry yeast Sc50 in kibbles with water activities between 0.53 and 0.58. The solid line represents the stability of the yeast in kibbles coated according to coating code 4, and the dashed line represents the stability of the yeast in kibbles coated according to coating code 3. Figure 10 shows the stability results for cat kibbles coated with active dry yeast Sc50 at water activities between 0.32 and 0.35. The series shown in Figure 10 correspond to the average viability values ​​obtained at each time point for the four coating methods, coating codes 1-2 and 5-6. Opinions and conclusions From Figures 9 and 10, it is clear that the active dry yeast Sc50 of the present invention, when coated around a kibble, has excellent stability performance up to: For dog kibbles, the shelf life is 15 months when the coating layer contains only the palatability enhancer powder (coating codes 1 and 2, FIG. 9A). 12 months if the coating layer contains a liquid palatability enhancer specific to that particular dog kibble (Coating Codes 3 and 4, FIG. 9B). For cat kibbles, regardless of the coating method tested, it is 24 months (Codes 1-2, 5-6, Figure 10).

[0043] Example 3 Comparative Example to Example 2 The aim of this study was to determine whether the physicochemical properties of active dry yeast are their shelf life in the kibble, and / or The aim is to show that their attachment within the kibble is essential.

[0044] Shelf life of active dry yeast in dog kibble The active dry yeast tested in this example has the following physicochemical properties: Yeast "1" The particle size distribution is such that the diameter of the spheres is 0.3 to 0.7 mm. The dry matter content is 93% by weight. Yeast "2" The particle size distribution is such that the diameter of the spheres is 0.05 to 0.2 mm. The dry matter content is 94.5% by weight. Therefore, these active dry yeasts do not have all the physicochemical properties of the active dry yeasts of the present invention. These yeasts are incorporated into a palatability enhancer for dogs having a water activity of 0.32 according to the method described in Example 2. Similarly, these yeasts are coated around dog kibbles according to the method described in Example 2. More specifically, yeast 1 has coating codes 1 and 3, i.e. FAT, then "PEP+Yeast 1" (coating code 1), and FAT, then "PEP+yeast 1", and then coated according to LPE (coating code 3). Similarly, yeast 2 has coating codes 1 and 3, i.e. FAT, then "PEP+Yeast2", and FAT, then "PEP+yeast2", and then coated with LPE. The solid palatability enhancer (PEP) and liquid palatability enhancer (LPE) were as described in Example 2. The dog kibbles thus coated are stored under the same conditions as those described in Example 2. Viability measurements are assessed monthly according to the protocol described in Example 2. The results obtained are shown in Figure 11. It is clear from this figure that the viability of yeast 1 does not exceed 5 months and that of yeast 2 does not exceed 8 months.

[0045] Yeast adhesion in kibble The active dry yeast tested in this example has the following physicochemical properties: Yeast "3" The particle size distribution is such that the diameter of the spheres is 0.1 to 0.2 mm. The dry matter content is 94% by weight. Yeast "4" The particle size distribution is such that the diameter of the spheres is 0.01 to 0.1 mm. The dry matter content is 94% by weight. Therefore, these active dry yeasts do not have all the physicochemical properties of the active dry yeasts of the present invention. These yeasts are incorporated into palatability enhancers for dogs having a water activity of 0.32 according to the method described in Example 2. Similarly, these yeasts are coated around dog kibbles according to the method described in Example 2. More specifically, yeasts 3 and 4 are each coated with coating code 1, i.e. FAT, then "PEP+Yeast3", and FAT, then coated according to "PEP+Yeast 4". The solid palatability enhancer (PEP) was as described in Example 2. The dog kibbles thus coated are subjected to a friability / durability test. The durability of the coated kibbles is analyzed using a pellet durability tester sold under the name "Holmen NHP 100". 100 grams of kibbles are placed in the durability tester and each analysis is carried out in duplicate. The tester is equipped with a 1 mm screen and the test lasts for 120 seconds. During the durability test, the kibble is subjected to a standard mechanical stress. At the end of the mechanical test, the kibble is sieved through a sieve having a mesh size corresponding to 80% of the diameter of the kibble. The proportion of material originating from the kibble and smaller than the size of the mesh of the sieve corresponds to "fines". The higher the percentage of fines, the worse the yeast will adhere to the kibble. The results obtained show that the average fines percentage for kibbles coated with yeast 3 and subjected to durability testing was 1.79%, and the average fines percentage for kibbles coated with yeast 4 and subjected to durability testing was 1.55%. The differences observed after the durability test are explained in particular by the physicochemical properties of the yeasts, especially their particle size distribution: larger yeasts have a higher fines fraction than smaller ones. The spheroid shape of yeast is important because it has a very low surface porosity. Small spheroids have a larger net surface area and therefore provide a larger surface area to environmental humidity. However, humidity is important for the stability of yeast over time. Therefore, a compromise must be found between spheroids having a size small enough to adhere well to the surface of the kibble, and spheroids that are not too small to increase resistance to humidity and be stable over time and industrially feasible.

[0046] The present disclosure is not limited to the above embodiments merely as examples, but encompasses all variants that may occur to those skilled in the art within the scope of the intended protection.

[0047] Reference to deposited biological material In this application, reference is made to the following biological agents and variants or mutants derived therefrom: The identification reference "Sc50" and the accession number "I-5660" were deposited on March 3, 2021 at the Collection Nationale de Cultures de Micro-organismes (CNCM), 25, rue du Docteur Roux, 75724 Paris Cedex 15, by Lesaffre et Compagnie, 41 rue Etienne Marcel, 75009 Paris.

Claims

1. A Saccharomyces cerevisiae yeast, which is an active dry yeast, obtained by culturing the Saccharomyces cerevisiae strain deposited at the CNCM under the number I-5660 on March 3, 2021, wherein the active dry yeast is The dry matter content is more than 95% by weight and less than or equal to 97% by weight, The water activity is 0.1 to 0.3, It has a spherical shape, having a particle size distribution characterized by a Gaussian size distribution curve showing that at least 90% of the spheroids have diameters in the range of 0.3 to 0.7 mm; A yeast strain of Saccharomyces cerevisiae having a stability characteristic of 24 to 36 months when vacuum-packaged and at a temperature of 15 to 25°C.

2. Improve your pet's intestinal immune system, Restoring normal intestinal flora in pets, A decrease in the pet's fecal pH, and / or The yeast according to claim 1 for use in reducing the production of putrefactive catabolic products in the faeces of pets.

3. 13. A pet composition comprising the yeast of claim 1, said yeast having a shelf life of 12 to 24 months within said composition.

4. The composition of claim 3 , wherein the yeast is coated around the composition.

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

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

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

8. Improve your pet's intestinal immune system, Restoring normal intestinal flora in pets, A decrease in the pet's fecal pH, and / or A composition according to any one of claims 3 to 7 for use in reducing the production of putrefactive catabolic products in the faeces of pets.

Images