Saccharomyces cerevisiae yeast and its use to improve intestinal health in pets.

Abstract

The present invention relates to a Saccharomyces cerevisiae yeast obtained by culturing the Saccharomyces cerevisiae strain filed on March 3, 2021 with the CNCM under number I-5660, which is an active dry yeast exhibiting the following physicochemical characteristics: - a dry matter content greater than 95% by weight and up to 97% by weight; - a water activity ranging from 0.1 to 0.3; - a spherical shape; - a particle size distribution characterized by a Gaussian curve showing that at least 90% of the spherules have a diameter ranging from 0.3 to 0.7 mm. The invention also relates to the use of said yeast to improve the intestinal health of companion animals.The present invention further relates to a solid palatability enhancer for pets or a pet food, characterized in that it comprises a yeast as defined above, said yeast having a lifespan of 12 to 24 months within said palatability enhancer or said food.

Description

Title of the invention: Saccharomyces cerevisiae yeast and its use to improve the intestinal health of pets. Technical field

[0001] This application falls within the field of animal nutrition and intestinal health in companion animals. It relates in particular to a yeast Saccharomyces cerevisiae for its use in improving the intestinal health of companion animals. Previous technique

[0002] The use of probiotic microorganisms, such as yeasts or bacteria, to enhance the intestinal health of pets is known in the prior art. Several documents describe the encapsulation or microencapsulation of microorganisms to ensure their survival in the gastrointestinal tract and thus enable them to exert their health benefits. Document JP2008013534 describes in particular the encapsulation of lactic acid bacteria and yeast in a polymer comprising a mixture of xanthan gum and chitosan. The encapsulated probiotic thus obtained is used to improve gastrointestinal disorders in pets. The article by Bruna M. Rodrigues et al. describes a lyophilization microencapsulation technique for probiotic strains in order to maintain the viability of the probiotics during storage and to evaluate their resistance in the gastrointestinal tract.Experimental results in cats show that taking probiotics or encapsulated probiotics (with fructooligosaccharides) can modulate the gut microbiota.

[0003] The use of probiotic microorganisms in pet food products is also known from the prior art. When a microorganism or mixture of microorganisms is incorporated into a food product such as a kibble coated with said microorganism, it is imperative that the microorganism survive not only the coating phase around the kibble, but also that said microorganism remain viable until the expiration date of the kibble so that it can exert its beneficial effects on the animal's health. Document PCT / US2009 / 056292 describes the use of probiotics (more specifically, bifidobacteria) in a coating of pet kibble. However, this document does not mention a yeast strain or yeast that would be coated around the kibble and have a lifespan long enough in said kibble to be effective on the intestinal health of a pet.

[0004] There remains a need to find new probiotics that are effective in improving the intestinal health of pets and that can be incorporated into a pet food product while having a sufficiently long shelf life within that food product. A sufficiently long shelf life includes, in particular, a shelf life extending until the expiry date of the food product.

[0005] Unexpectedly, the Inventors developed an active dried yeast that proved particularly effective in strengthening the intestinal health of pets and, moreover, exhibits a particularly long shelf life when incorporated into a pet food product. A particularly long shelf life is defined as a period ranging from 12 to 24 months. Summary

[0006] According to a first aspect, the present invention relates to a Saccharomyces cerevisiae yeast obtained by culturing the Saccharomyces cerevisiae strain filed on March 3, 2021 with the CNCM under number 1-5660, which is an active dry yeast having the following physico-chemical characteristics: - a dry matter content greater than 95% by weight and up to 97% by weight; - a water activity ranging from 0.1 to 0.3; - a spherical shape; - a granulometry characterized by a Gaussian size distribution curve showing that at least 90% of the spherules have a diameter ranging from 0.3 to 0.7mm. The said active dry yeast advantageously has a shelf life of 24 to 36 months when stored in vacuum packaging at a temperature of 15 to 25°C, and a shelf life of 12 to 24 months when incorporated into a solid palatability enhancer or pet food, such as kibble, and said palatability enhancer or food is stored at a temperature of 15 to 25°C.

[0007] According to a second aspect, the invention relates to yeast as defined above, for its use 1 / in improving the intestinal immune system in pets and / or 2 / in restoring a normal intestinal microbiota in pets. The invention also relates to the use of yeast as defined above to improve fecal characteristics in pets.

[0008] According to another aspect, the invention relates to a solid palatability factor for pets which comprises the active dry yeast as defined above, said yeast having a lifespan of 12 to 24 months within said palatability factor.

[0009] According to yet another aspect, the invention relates to a pet food which comprises the active dry yeast or the solid palatability factor as defined above, said yeast having a lifespan of 12 to 24 months within said food.

[0010] The invention also relates to a food as defined above, for its use 1 / in improving the intestinal immune system in pets and / or 2 / in restoring a normal intestinal microbiota in pets. The invention further relates to the use of the food as defined above to improve the fecal characteristics of pets. Brief description of the drawings

[0011] Other features, details and advantages will become apparent from reading the detailed description below, and from analyzing the attached drawings. Figures 1 through 6 include, just above the juxtaposed bars in the histograms, the letters a and b (in lowercase) and / or the letters A and B (in uppercase). The letters a and b indicate whether the observed values ​​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 histograms obviously have no relation to the letters A, B, C or D given to differentiate the different patterns within the same figure. The vertical arrows in figures 1 to 6 symbolize an abrupt change in diet in dogs that takes place on day D22, referred to as the "dietary transition" in what follows. Active dry yeast is represented by "Sc50" in figures 1 to 6 below. Fig. 1

[0012] [Fig-1] illustrates the values ​​of fecal pH (see A) and ammonia concentration (percentage of dry matter) (see B) in feces of Beagle breed dogs before and after a dietary transition which takes place on day 22 (D22) (as indicated above the dietary transition is signified by a vertical arrow). The control group (black histograms) was subjected to diet 1 before the dietary transition and to 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. Fig. 2

[0013] [Fig.2] illustrates the content of Blautia bacteria (see A), Bifidobacterium (see B), Lactobacillus (see C) and Faecalibacterium (see D) in feces of Beagle dogs before and after dietary transition at day 22. The groups tested are the same as those described for [Fig. 1]. Fig. 3

[0014] [Fig.3] illustrates the E. coli content in the feces of Beagle breed dogs before and after the dietary transition on day 22. The groups tested are the same as those described for [Fig. 1]. Fig. 4

[0015] [Fig.4] illustrates the dysbiosis index (see A) of the intestinal microbiota and the content of Streptococci (see B) were found in the feces of Beagle dogs before and after the dietary transition on day 22. The groups tested are the same as those described for [Fig. 1], Fig. 5

[0016] [Fig.5] illustrates the content of fecal immunoglobulin A (IgA) found in the feces Beagle dogs before and after the dietary transition at day 22. The groups tested are the same as those described for [Fig. 1]. Fig. 6

[0017] [Fig. 6] illustrates the quantity of biogenic amines (in mg / kg of dry matter) present in the feces of Beagle dogs before and after the dietary transition on day 22. Specifically, Figure A illustrates the total amount of biogenic amines, Figure B illustrates the amount of putrescine, and Figure C illustrates the amount of spermidine in the feces of Beagle dogs before and after the dietary transition on day 22. The groups tested are the same as those described for [Fig. 1]. Fig. 7

[0018] [Fig.7] illustrates the stability of the active dry yeast Sc50 within two factors of different palatability for dogs (see A) and for cats (see B). Fig. 8

[0019] [Fig.8] illustrates different possible embodiments for the coating of a dog or cat kibble. In Figure A, the core of the kibble is coated by a first coating layer (shown in gray) made up of fats (FM) and then by a second coating layer (represented in white and delimited by dotted lines) consisting of a liquid palatability enhancer. In Figure B, the core of the kibble is coated by a first coating layer (represented in grey) consisting of fats (FM) and then by a second coating layer (represented by small dotted squares) consisting of a solid palatability factor mixed with the active dry yeast Sc50 of the invention. Finally, in Figure C, the core of the kibble is coated by a first coating layer (represented in grey) consisting of fats (FM), by a second coating layer (represented in white and delimited by dotted lines) consisting of a liquid palatability factor, and then by a third coating layer (represented by small dotted squares) consisting of a solid palatability factor mixed with the active dry yeast Sc50. Fig. 9

[0020] [Fig.9] illustrates the stability of the active dry yeast Sc50 when the latter is coated around dog kibble. In Figure A, the solid line represents the stability of the yeast in the coated kibble according to the coating method: fat followed by "solid palatability enhancer + active dry yeast Sc50" followed by fat followed by solid palatability enhancer. The dashed line represents the stability of the yeast in the coated kibble according to the coating method: fat followed by "solid palatability enhancer + active dry yeast Sc50". In Figure B, the solid line represents the stability of the yeast in the coated kibble according to the coating method: fat, then "solid palatability factor + active dry yeast Sc50", then fat, then liquid palatability factor. The dashed line represents the stability of the yeast in the coated kibble according to the coating method: fat, then "solid palatability factor + active dry yeast Sc50", then liquid palatability factor. Fig. 10

[0021] [Fig.10] illustrates the stability of the active dry yeast Sc50 when the latter is coated around cat kibble. The series shown in [Fig. 10] corresponds to the average viability obtained at each time point for the following 4 coating methods: - fat then "solid palatability factor + active dry yeast Sc50" then fat then solid palatability factor; - fat then "solid palatability factor + active dry yeast Sc50"; - fat then "solid palatability factor + active dry yeast Sc50" then fat under vacuum; - fat then "solid palatability factor + active dry yeast Sc50" then vacuum-packed fat then solid palatability factor. The error bars represent + / - 1 standard deviation. Detailed description

[0022] Thus, the present invention relates to a Saccharomyces cerevisiae yeast obtained by culturing the Saccharomyces cerevisiae strain filed on March 3, 2021 with the CNCM under number 1-5660, which is an active dry yeast with the following physico-chemical characteristics: - a dry matter content greater than 95% by weight and up to 97% by weight; - a water activity ranging from 0.1 to 0.3; - a spherical shape; - a granulometry characterized by a Gaussian size distribution curve showing that at least 90% of the spherules have a diameter ranging from 0.3 to 0.7mm. The Saccharomyces cerevisiae strain is a yeast strain. The term "yeast strain" refers to a relatively homogeneous population of yeast cells. A yeast strain is obtained from the isolation of a clone, a clone being a cell population derived from a single yeast cell. The strain filed with the CNCM under number 1-5660 is also identified by the reference "Sc50". This strain may be referred to, among other things, as "strain Sc 1-5660", "strain 1-5660", or "strain Sc50" in this application. The Saccharomyces cerevisiae yeast obtained by culture of strain 1-5660 may be designated as “Sc 1-5660 yeast”, “1-5660 yeast” or “Sc50 yeast”. As an example, yeast 1-5660 can be obtained by culturing strain 1-5660 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". Yeast 1-5660 can also be obtained by an industrial-scale process which involves preparing a liquid yeast cream or liquid yeast which includes the following steps: - Culturing the yeast strain in a multi-stage culture medium, first under semi-anaerobic conditions, then under aerobic conditions, - separation by centrifugation of the yeast thus produced from its culture medium, to obtain a liquid yeast cream or liquid yeast containing 12 to 25% of dry matter, or even a higher quantity of dry matter, particularly if the liquid yeast cream or liquid yeast is mixed with osmolyte products. The liquid yeast cream or liquid yeast 1-5660 obtained from the culture process described above is then subjected to a dehydration and drying process which includes the following steps: - mixing of liquid yeast cream or liquid yeast 1-5660 with a sodium chloride-based brine to obtain a volume of yeast cream comprising a sodium chloride-based brine, - dehydration of the volume of yeast cream including a sodium chloride-based brine to obtain dehydrated yeast, - extrusion of dehydrated yeast to obtain extruded yeast, - drying of 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. The yeast 1-5660 obtained as a result of this dehydration and drying process is designated "active dry yeast". It may be referred to interchangeably in this application as "active dry yeast Sc 1-5660", "active dry yeast 1-5660", "active dry yeast Sc50" or simply "active dry yeast". The term "active yeast" is synonymous with live yeast and refers to a population of yeast cells that are metabolically active. For the purposes of this application, "active dry yeast" means active yeast with a dry matter content exceeding 95% by weight and potentially reaching up to 97% by weight. The dry matter content was assessed using the drying method, after passing through an oven for a minimum of 17 hours at a temperature of 103°C. Water activity was assessed using the cooled mirror dew point technique with an AquaLab. This is a primary measurement of relative humidity. Active dry yeast comes in the form of spherules which have a very low porosity surface and a very dense internal structure, which gives these spherules, among other things, a moisture barrier effect. These spherules exhibit a granulometry characterized by a Gaussian size distribution curve showing that at least 90% of the spherules have a diameter ranging from 0.3 to 0.7mm. Particle size distribution was assessed by dry method using a laser particle size analyzer.

[0023] According to a particularly advantageous embodiment of the invention, the active dry yeast Sc50 is further characterized in that it exhibits a stability, in a vacuum package and at a temperature ranging from 15 to 25°C, of ​​at least 24 months, and preferably ranging from 24 to 36 months. The stability of the active dried yeast of the invention is directly related to its viability rate, that is, the number of viable cells it contains. One way to measure yeast cell viability is to count the number of recoverable cells (in CFU / g or log CFU / g). "Recoverable cells" are viable cells capable of multiplying. The term CFU stands for "colony-forming unit." One CFU corresponds to one colony. Cell viability was measured according to the standardized method EN15789. For the purposes of this application, a stable active dry yeast over a period of at least 24 months, and preferably from 24 to 36 months, means that its number of live and metabolically active cells (CFU / g) does not decrease by more than one log CFU / g over a period of at least 24 months, and preferably from 24 to 36 months. This means, for example, that if the active dry yeast has a number of live and metabolically active cells of 1xlO10 CFU / g (equivalent to 10 log CFU / g) at time tO (tO being the time at which the active dry yeast is obtained, i.e., the moment when the active dry yeast has just been prepared), then the yeast will have, at the end of a period of at least 24 months, and preferably from 24 to 36 months, a number of live and metabolically active cells that will not be less than 1xlO9 CFU / g (equivalent to 9 log CFU / g). The stability of at least 24 months, and preferably from 24 to 36 months, of the active dry yeast of the invention is verified when said yeast is stored in a vacuum package and at a temperature of 15 to 25°C. Thus, for the purposes of this application, the stability of active dried yeast over a period of at least 24 months, and preferably from 24 to 36 months, means that the active dried yeast has a shelf life of at least 24 months, and preferably from 24 to 36 months, according to the criteria indicated above (loss of viability threshold less than or equal to 1 log CFU / g). This shelf life / stability is extremely advantageous since it allows the active dried yeast to retain its properties, namely improving the intestinal health of pets.

[0024] According to an advantageous embodiment, the invention relates to an active dry yeast Sc50 which has the following characteristics: - a dry matter content greater than 95% by weight and up to 97% by weight; - a water activity ranging from 0.1 to 0.3; - a spherical shape; - a granulometry characterized by a Gaussian size distribution curve showing that at least 90% of the spherules have a diameter ranging from 0.3 at 0.7mm.- a dry matter content greater than 95% by weight and up to 97% by weight; - stability, in vacuum packaging and at a temperature ranging from 15 to 25°C, of ​​at least 24 months, and preferably ranging from 24 to 36 months.

[0025] The invention also relates to the active dry yeast Sc50 as defined above for its use: - in improving the intestinal immune system in pets and / or, - in the restoration of a normal intestinal microbiota in pets. Pets refer to domestic animals such as dogs, cats, rabbits, birds, hamsters, etc. Dogs and cats are more specific examples of pets. In both humans and pets, a large number of immune cells are located in the gut, clearly illustrating the importance of the intestinal immune system. The digestive tract is constantly attacked by a multitude of antigens and pathogens, which is why it possesses a complex and highly developed immune system. According to an advantageous embodiment of the invention, the administration of active dry yeast Sc50 in pets makes it possible to significantly increase the production of fecal immunoglobulins. The gut microbiota, which refers to all the microorganisms that live in the digestive tract, plays a vital role in maintaining good intestinal health. It is also fully involved in the functioning of the intestinal immune system. Dysbiosis is an alteration in the composition and / or microbial diversity of the digestive tract. It can lead to an overgrowth of pathogenic bacteria, an acute inflammatory response, and diarrhea. Any alteration of the intestinal environment can cause a change in the microbiota and thus lead to dysbiosis. The microbiota dysbiosis index provides an overview of overall intestinal health and generally indicates the presence or absence of dysbiosis. A "normal" gut microbiota is a healthy microbiota, showing no alteration in the composition and / or microbial diversity of the digestive tract. Administering active dry yeast Sc50 to pets with an altered gut microbiota helps restore a normal gut microbiota. Indeed, according to an advantageous embodiment of the invention, administering active dry yeast Sc50 to pets with an altered gut microbiota allows: - a significant decrease in the levels of fecal streptococci and fecal Escherichia coli, - a significant effect on improving the dysbiosis index, - a significant increase in the levels of Blautia, Bifidobacterium, Lactobacillus and Faecalibacterium bacteria. Blautia, Bifidobacterium, Lactobacillus and Faecalibacterium bacteria are beneficial for maintaining good intestinal health.

[0026] The invention further relates to the use of active dry yeast Sc50 to improve fecal characteristics in pets. "Improving fecal characteristics" means, in particular, a decrease in fecal pH, a decrease in the production of putrefactive catabolites such as ammonia, and a decrease in the production of biogenic amines (such as spermidine and putrescine). Reducing putrefactive catabolites in pet feces helps to reduce bad stool odors, which are a real nuisance for pet owners.

[0027] According to yet another advantageous embodiment of the invention, the active dry yeast Sc50, for one of the uses as defined above, is used at a daily dose ranging from 1xlO8 to 1.5xlO8 CFU / animal.

[0028] The invention further relates to a probiotic or food supplement for pets, characterized in that it comprises the active dry yeast Sc50. The terms "probiotic" and "food supplement" used above mean that the active dry yeast Sc50 acts as a probiotic or a food supplement. In 2001, probiotics were defined by the WHO (World Health Organization) as "live microorganisms which, when administered in adequate amounts, confer a health benefit beyond traditional nutritional effects." According to the definition given by Directive 2002 / 46 / EC of the European Parliament, "food supplements are foodstuffs the purpose of which is to supplement the normal diet and which constitute a concentrated source of nutrients or other substances with a nutritional or physiological effect, alone or in combination, marketed in dose form." The probiotic or food supplement of the invention comprises at least 95% by weight of active dry yeast Sc50, and preferably 96 to 99% by weight. The other ingredients constituting the probiotic or food supplement may, for example, be starch, but also vitamins, dietary minerals, trace elements, and / or dietary enzymes. According to one embodiment of the invention, the probiotic or dietary supplement may be in a form suitable for oral administration, such as tablets, capsules, granules, powders, etc. The probiotic or dietary supplement of the invention may be administered between meals or, conversely, during the pets' meals. In the latter 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 food supplement is administered in an amount ranging from 9 to 900 mg / day / animal According to yet another embodiment, the invention relates to the use of the probiotic or food supplement as defined above, in a pet food, said food preferably being a kibble for dogs or cats.

[0029] The active dry yeast Sc50 of the invention has the advantage of being easily dispersed in a food ingredient, such as a palatability enhancer intended for use in pet food products. Indeed, when mixed with a food ingredient, the active dry yeast does not undergo settling or clumping over time and advantageously exhibits a viability of 12 to 24 months within 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, often available in powder form, are designed to increase the appeal of pet food to pets. Adding a palatability enhancer to food significantly increases pet consumption. Palatability enhancers can be considered flavorings or other taste factors. Examples of palatability factors include poultry offal (chicken or turkey, for example), pork or poultry livers, and fats (butter or cheese, for example).

[0030] In this regard the invention also relates to a solid palatability factor intended to be integrated into pet food, characterized in that it comprises an active dry yeast as defined above, said yeast having a lifespan of 12 to 24 months within said palatability factor. These conditions for the shelf life of active dry yeast are observed when the palatability factor is kept at a temperature ranging from 15 to 25°C. According to an advantageous embodiment of the invention, the palatability factor has a water content of less than 10%. The 12- to 24-month lifespan of the active dry yeast within the palatability factor means that it has a viability loss threshold of less than or equal to 1 log CFU / g over a period of 12 to 24 months, and preferably a viability loss threshold of 0.1 to 0.8 log CFU / g over the same period. This means, for example, that if the live active dry yeast is present in an amount of 1xlO7 CFU / g of palatability factor immediately after being mixed with said palatability factor (referred to as time T0), then after a period of 12 to 24 months from time T0 (T0 being the moment when the live active dry yeast is mixed with the palatability factor), the amount of live active dry yeast is not less than 1xlO6 CFU / g of palatability factor. This shelf life is extremely advantageous since in this way the active dry yeast Sc50 retains its biological properties when used in association with a palatability factor (namely to improve the intestinal health of pets). According to yet another embodiment of the invention, the weight of the active dry yeast in the solid palatability factor varies from 1 to 40%, and more particularly from 2 to 35% relative to the total weight of the palatability factor.

[0031] The present invention also relates to the use of active dry yeast Sc50 as defined above or of a solid palatability factor as defined above, in a pet food, said food preferably being a kibble. The term "pet food" refers to any food product or food composition intended to be ingested by a pet, such as a dog or cat. An example of such food is pet kibble, and more specifically, kibble for dogs or cats.

[0032] According to one embodiment of the invention, the active dry yeast Sc50 or the solid palatability factor is integrated within the food or is deposited by coating around the food. The integration of active dry yeast or a palatability enhancer into the feed means, for example, that the yeast or palatability enhancer can be mixed directly into the feed during its manufacture, so that the feed contains said yeast or palatability enhancer in its very composition. However, it can also mean that the yeast or palatability enhancer is mixed into the feed at the time the animal eats. When yeast is incorporated into the feed during manufacturing, it should preferably be active dried yeast. When yeast is mixed into the feed at mealtimes, it should preferably be in the form of a probiotic or a dietary supplement. According to a preferred embodiment, the active dry yeast or palatability factor is deposited by coating around the food, said food being for example dog or cat kibble. Viability tests of the active dry yeast, when coated around the kibble, have advantageously shown that the yeast exhibits a shelf life / stability ranging from 12 to 24 months. These results are particularly interesting and promising because they represent an extremely advantageous shelf life. The stability of the active dry yeast of the invention is directly related to its viability rate at the end of the kibble's usage period. The stability of the active dry yeast of the invention, when used in a food, in a palatability enhancer, in a probiotic or in a food supplement, is defined with regard to its threshold of loss of viability in said food, palatability enhancer, probiotic or food supplement. More specifically, the stability of the yeast of the invention, in a food, a palatability factor, a probiotic or a food supplement, over a period of 12 to 24 months, means that it has a loss of viability threshold less than or equal to 1 log CFU / g over a period of 12 to 24 months, and preferably a loss of viability threshold of 0.1 to 0.8 log CFU / g over a period of 12 to 24 months.This means, for example, that if live active dry yeast is present in a quantity of 1xlO7 CFU / g of food, palatability enhancer, probiotic or food supplement immediately after said yeast has been incorporated into said food, palatability enhancer, probiotic or food supplement (called time T0), then after a period of 12 to 24 months from time T0 (T0 = moment when the active dry yeast is incorporated into the food, palatability enhancer, probiotic or food supplement), the quantity of live active dry yeast is not less than 1xlO6 CFU / g of food, palatability enhancer, probiotic or food supplement. This lifespan is extremely advantageous since in this way the active dry yeast Sc50 retains its biological properties when incorporated into food, as a palatability enhancer, probiotic or food supplement (namely to improve the intestinal health of pets). These conditions for the shelf life of active dry yeast are observed when the food, palatability factor, probiotic or food supplement is kept at a temperature ranging from 15 to 25°C.

[0033] According to a particularly advantageous embodiment, the invention relates to a pet food, characterized in that it comprises an active dry yeast as defined above or a palatability factor as defined above, said yeast having a lifespan of 12 to 24 months within said food. Another object of the invention lies in a probiotic or food supplement as defined above, which is further characterized in that it comprises an active dried yeast as defined above, having a shelf life of 12 to 24 months. The shelf life of the active dried yeast within said food, probiotic, or food supplement is as defined previously (i.e., a loss of viability threshold less than or equal to 1 log CFU / g, and preferably ranging from 0.1 to 0.8 log CFU / g over a period of 12 to 24 months). The variability in the lifespan of active dry yeast in pet food depends in particular on the water activity of said food. Thus the active dry yeast Sc50 of the invention has a lifespan of: - 12 months when present in a food with a high water activity, namely greater than 0.5; - of 24 months when present in a food with low water activity, namely less than 0.5, and preferably less than 0.4. According to yet another embodiment of the invention, Sc50 yeast is administered in an amount ranging from 5xl05 to 5xl09 CFU / g of food.

[0034] According to an advantageous embodiment, the food of the invention is more particularly a kibble coated by at least one coating layer, said layer comprising an active dry yeast as defined above or a palatability factor as defined above. Traditionally, a pet food kibble or coated kibble consists of a core, which is coated by at least one layer of coating. The kibble of this invention refers more specifically to a coated kibble. The terms "kibble" / "coated kibble" and "core" / "nail" may be used interchangeably in the following. The coating layer may cover the entire surface of the core or only a portion of it. According to a preferred embodiment of the invention, the coating layer covers the entire surface of the core. According to an advantageous embodiment of the invention, the percentages by weight, relative to the total weight of the kibble, vary from: - 50 to 97% for the core, preferably 80 to 95%, - 3 to 50% for the coating layer, preferably 5 to 20%. In this respect, the feed of the invention is more particularly characterized in that the coating layer is present in a quantity by weight ranging from 3 to 50%, and preferably from 5 to 20% relative to the total weight of the feed. The core of the kibble has a water content of less than 10%. The core of the kibble may, for example, include proteins of animal or vegetable origin, carbohydrates, fats of animal or vegetable origin, minerals, vitamins, etc. According to an advantageous embodiment, the coating layer comprises, in addition to the active dry yeast of the invention or the solid palatability enhancer, a fat. By way of example, a croquette of the invention may consist of a core, a first coating layer made of a fat, and a second coating layer comprising a solid palatability enhancer, said palatability enhancer comprising the active dry yeast of the invention. As another example, the kibble may consist of a core and a coating layer including fat and active dry yeast Sc50 or a palatability factor including active dry yeast.

[0035] The invention also relates to a food product as defined above, for its use: - in improving the intestinal immune system in pets and / or, - in the restoration of a normal intestinal microbiota in pets. Finally, another object of the invention is the use of a food such as defined above to improve the fecal characteristics of pets. The improvements noted for the use of Sc50 active dry yeast alone with regard to the intestinal health of pets (improved intestinal immune system, restoration of a normal gut microbiota, improved fecal characteristics) apply to the use of foods that include the active dry yeast of the invention, and in particular to kibble coated with a coating layer comprising the active dry yeast of the invention or the palatability enhancer of the invention. This is notably due to the fact that the active dry yeast of the invention exhibits excellent stability / shelf life within said food (ranging from 12 to 24 months), which means that it does not lose any activity and that said active dry yeast is therefore able to exert its benefits on the intestinal health of pets.

[0036] Example 1: Study of the effectiveness of Sc50 active dry yeast on intestinal health

[0037] The objective of this example, which refers to Figures 1 to 6, is to evaluate the effect of daily dietary supplementation with active dry yeast Sc50 in healthy adult dogs on their intestinal health after an abrupt dietary transition. An "abrupt dietary transition" is defined as a sudden change in diet.

[0038] Protocol of the studies carried out The study was conducted on forty healthy adult Beagle dogs (twenty males and twenty females), with an average body weight ranging from 10 to 15 kg. The study was repeated twice and approved by the Animal Use Ethics Committee. Two different groups are studied: with or without supplementation of active dry yeast Sc50. Active dry yeast Sc50 (comprising 1xlO10 CFU of live and metabolically active cells per gram of active dry yeast) is administered at a dose of 150 mg per day per dog. The study is conducted in two stages: - a phase 1 which lasts 3 weeks (from day 1 to day 21 (D 1 to D21)) and, - a phase 2 which lasts 4 weeks (from day 22 to day 49 (D22 to D49)). On day 1 of phase 2, namely day 22 (D22) the dogs undergo an abrupt change in diet (referred to as "dietary transition" in what follows and represented by the vertical arrow in figures 1 to 6). Phase 1 therefore takes place before the dietary transition and phase 2 starts on the first day of the dietary transition, at day 22.

[0039] Phase 1: - Group A: control group receiving diet 1 (represented by "Control" in figures 1 to 6), - group B: group receiving diet 1 supplemented with active dry yeast Sc50 (represented by "Sc50" in figures 1 to 6). Diet 1 consists of a standard and balanced food for a dog, which has a low fiber content, namely in the range of 1 to 3%.

[0040] Phase 2: - Group A: control group receiving diet 2 (represented by "Control" in figures 1 to 6), - group B: group receiving diet 2 supplemented with active dry yeast Sc50 (represented by "Sc50" in figures 1 to 6). Diet 2 consists of a food with a high fiber content, namely around 12 to 13%. The abrupt transition from a low-fiber diet to a high-fiber diet is carried out to create intestinal dysbiosis.

[0041] Evaluated parameters 1 / Fecal samples were collected for analysis of fecal pH ([Fig. 1], A) and fecal ammonia concentration ([Fig. 1], B). Fecal pH was measured directly on fresh fecal samples using a digital pH meter. Fecal ammonia concentration ([N]) (g / kg) was determined according to the method described by Brito et al., 2010 and was specifically calculated by applying the correction factor 0.9875 according to the formula below:

[0042] [Math.l] Vacil in the tested sample — Vacid in the control sample = A' x correction factor x!7 x -------------------;—;—- ;----------------------------------- sample loaves in grams 2 / The bacterial population present in the intestinal microbiota (figure 2, A, B, C, D, figure 3 and figure 4, B) as well as the dysbiosis index (figure 4, A) were also analyzed using a quantitative PCR (polymerase-chain-reaction) methodology published in the journal FEMS Microbiology Ecology (AlShawaqfeh et al., 2017). The bacterial population is expressed as a logarithm of the DNA. The dysbiosis index allows for the analysis of the fecal microbiota, and more specifically, the quantification of the following bacteria: Faecalibacterium, Fusobacterium, Clostridium hiranonis, Blautia, Streptococcus, Escherichia coli, and Turicibacter. These bacteria have been identified in the literature as being altered in certain digestive pathologies (e.g., chronic enteropathies) and / or in response to antibiotic treatment. The dysbiosis index allows for the evaluation of reference ranges for these bacteria as well as values ​​expressing the extent of intestinal dysbiosis. Thus, fecal microbiota dysbiosis is defined as a dysbiosis index greater than 2. A dysbiosis index ranging from 0 to 2 indicates a moderate alteration in the fecal microbiota. A normal microbiota is defined as a negative dysbiosis index.

[0043] Results obtained The active dry yeast of the invention, Sc50, was well accepted and tolerated by dogs in both studies. No episodes of food refusal, vomiting, diarrhea, or weight loss were observed throughout the study, even during the dietary transition. Encouraging and comparable results were reported and are summarized below.

[0044] Effects of active dry yeast Sc50 on fermentation production ([Fig. 1]) A significant decrease in fecal pH ([Fig. 1], A) at J35 and J49 and in fecal ammonia ([Fig. 1], B) at J23, J35 and J49 was observed after the dietary transition.

[0045] Effect of active dry yeast Sc50 on the intestinal microbiota ([Fig.2], JTigJL [Fig.4], B) and the dysbiosis index ([Fig.4], A) A significant increase in the content of Blautia (J49) ([Fig.2], A), Bifidobacterium (J21 and J49) ([Fig.2], B), Lactobacillus (J21 and J49) ([Fig.2], C) and Faecalibacterium (J49) ([Fig.2], D) was observed after the dietary transition in the feces of the dogs. A significant decrease in fecal streptococcus content (day 35) (Fig. 4, B) was observed after the dietary transition. A significant decrease in fecal Escherichia coli content was also observed after the dietary transition (day 49), as well as a downward trend before the dietary transition (day 21) (Fig. 3). Similarly, an improvement in the dysbiosis index before and after the dietary transition was observed in the gut microbiota of the dogs (days 21, 23, and 35) (Fig. 4, A). The dysbiosis index allows us to assess how the gut microbiota normalizes in response to a change in treatment, in this case, the rapid dietary transition from diet 1 to diet 2.

[0046] Effect of active dry yeast Sc50 on the production of fecal immunoglobulin A (IgA L ([Fig.5]) It appears from [Fig.5] that the administration of active dry yeast Sc50 leads to a significant increase in the production of IgA in the feces of dogs before (J21) and after (J35) the dietary transition.

[0047] Effect of active dry yeast Sc50 on the production of biogenic amines ([Fig. 611] A significant decrease in the production of total biogenic amines before (Day 21) and after the dietary transition (Day 49) was observed in dogs. More specifically, a significant decrease in spermidine was observed after the dietary transition (Day 49), as well as a significant decrease in putrescine before the dietary transition (Day 21) and a trend towards a decrease after the dietary transition (Day 49) in dogs.

[0048] Conclusions Supplementing the diet of healthy dogs undergoing a sudden dietary change with active dry yeast Sc50 helps to: a) Improvement in the fecal characteristics of the tested dogs due to a decrease in fecal pH and the production of putrefactive catabolites. Indeed, a decrease in fecal ammonia concentration has been reported to limit the replication of pathogenic microorganisms in the gut. Furthermore, a decrease in biogenic amines contributes to a reduction in fecal odor, which is a characteristic important commercial activity given the close relationship between dogs and their owners. b) Improvement of local immunity by an increase in the production of fecal IgA which contributes to greater local resistance to antigen invasion and can limit the absorption of protein antigens and the neutralization of viruses. c) Modulation of the composition and activity of the gut microbiota, for example by reducing bacteria such as streptococci and Escherichia coli, which are described in the literature as being associated with inflammatory processes in certain digestive pathologies, and increasing beneficial gut microbiota bacteria. Furthermore, supplementation with active dry yeast Sc50 improved the dysbiosis index.

[0049] Example 2: Study of the stability of active dry yeast Sc50 in dog and cat food

[0050] The objective of this example, which refers to Figures 7 to 10, is to evaluate the stability of the active dry yeast Sc50: - within two solid palatability factors for dogs or cats, - within a dog or cat kibble, namely as a coating around the core of the kibble.

[0051] Mixture of the active dry yeast Sc50 of the invention with palatability factors in powder form for dogs or cats Active dry yeast Sc50 is incorporated at 5% in powdered palatability enhancers for dogs or cats, these palatability enhancers being composed of pork and poultry by-products, vegetable proteins, carbohydrates, and minerals. The powdered palatability enhancers differ in their water activity, namely: - a water activity of 0.32 or 0.38 for the two palatability enhancers for dogs, - a water activity of 0.30 or 0.35 for the two palatability enhancers for cats. The "palatability enhancer + Sc50 active dry yeast" mixture is prepared in a mixer marketed under the Forberg brand (double-shaft paddle mixer) by first introducing all of the powdered palatability enhancer into the mixer, followed by the Sc50 active dry yeast. The mixture is then run for 5 minutes at a speed of 50 Hz.

[0052] Study of the stability of the active dry yeast Sc50 within the palatability factor The palatability factor with added active dry yeast Sc50 is then packaged in plastic bags (for palatability factors for dogs) or in aluminum bags (for palatability factors for cats) and is stored at room temperature (20°C + / - 5°C). The viability of active dry yeast Sc50 was measured at various time points (at least 4) throughout the study period, from 0 to 12 months. The stability marker monitored during storage of active dry yeast Sc50 was the percentage of viable yeast cells, measured according to the standardized method NF EN 15789. The viability results of active dry yeast Sc50 with respect to various palatability factors are shown in [Fig. 7]. Specifically, [Fig. 7] A illustrates the stability of active dry yeast Sc50 in two palatability factors for dogs, the solid line representing stability in the palatability factor with a water activity of 0.32 and the dashed line representing stability in the palatability factor with a water activity of 0.38. [Fig. 7] B illustrates the stability of active dry yeast Sc50 in two palatability factors for cats, the solid line representing stability in the palatability factor with a water activity of 0.30 and the dashed line representing stability in the palatability factor with a water activity of 0.35.

[0053] Remarks and conclusion Figures 7A and 7B show that the active dry yeast Sc50 of the invention exhibits excellent stability performance when incorporated with palatability factors. The results show that over the 12-month period under the tested storage conditions (20 + / -5°C), the viability of the active dry yeast Sc50 is stable in the 4 tested dog and cat palatability factors with water activities ranging from 0.30 to 0.38. Thus, the viability difference in these 4 palatability factors after 12 months of contact is less than 1 log CFU / g compared to T0 (the moment when the active dry yeast of the invention is incorporated into the palatability factor).

[0054] Coating the core of a kibble with the palatability factor comprising active dry yeast Sc50 The various "palatability enhancer + active dry yeast Sc50" mixtures prepared in the previous step are then applied to dog or cat kibble, according to different coating sequences. These mixtures are applied to the kibble immediately after preparation. Stability studies of the active dry yeast Sc50 were conducted over a 24-month period. Figure 8 illustrates three possible embodiments for coating the core of a kibble. This figure is only given as an example, as other methods of coating the core of a kibble exist. As an example, the main components of cat and dog kibble can be summarized as follows: - for dogs: barley, wheat and derivatives, corn, poultry meal, whole soy, poultry fat; - for cats: poultry meal, barley, corn and derivatives, poultry fat, wheat, soy and derivatives. The incorporation rate of active dry yeast Sc50 in the kibble is 0.05%. The active dry yeast Sc50, previously mixed with the powdered palatability enhancers, is applied to the dog or cat kibble according to the different coating methods illustrated in Table 1 below. The abbreviations used in Table 1 have the following meanings: - MG: coating fat; - FAP: powder palatability factor; - FAP + Sc50: palatability factor powder with added active dry yeast Sc50; - FAL: liquid palatability factor. MG under vacuum means that the coating fat has been deposited on the kibble while a vacuum has been applied in the equipment (this helps to promote the penetration of the fat into the body of the kibble).

[0055] [Tables] Coating Code Coating Method 1 MG then "FAP + Sc50" 2 MG then "FAP + Sc50" then MG then FAP 3 MG then "FAP + Sc50" then FAL 4 MG then "FAP + Sc50" then MG then FAL 5 MG then "FAP + Sc50" then MG under vacuum 6 MG then "FAP + Sc50" then MG under vacuum then FAP

[0056] The kibble thus coated with the active dry yeast Sc50 is packaged in aluminium bags, then stored at room temperature (20°C + / - 5°C).

[0057] Study of the stability of the active dry yeast Sc50 coated around the kibble The stability marker monitored during the storage of the coated kibble is the rate of revivable yeast cells (standard method NF EN 15789). Figure 9 illustrates the stability results of active dry yeast Sc50 around dog kibble according to coating codes 1 to 4. In particular, [Fig.9] A illustrates the stability of active dry yeast Sc50 in kibble that exhibits water activity ranging from 0.45 to 0.49. The solid line represents the stability of the yeast in the kibble coated according to coating code 2, while the dashed line represents the stability of the yeast in the kibble coated according to coating code 1. Figure 9 B illustrates the stability of active dry yeast Sc50 in kibble exhibiting water activity ranging from 0.53 to 0.58. The solid line represents the stability of yeast in coated kibble according to coating code 4 while the dashed line represents the stability of yeast in coated kibble according to coating code 3. Figure 10 illustrates the results of the stability of the active dry yeast Sc50 around cat kibble which has an activity in water ranging from 0.32 to 0.35. The series shown in Figure 10 corresponds to the average of the viabilities obtained at each time point for the 4 coating methods according to coating codes 1-2 and 5-6.

[0058] Remarks and conclusions Figures 9 and 10 show that the active dry yeast Sc50 of the invention exhibits excellent stability performance once coated around the kibble, up to: - 15 months in dog kibble when the coating layer only includes powdered palatability factors (coating codes 1 and 2, [Fig.9] A), - 12 months when the coating layer includes a liquid palatability factor, specific to certain dog kibble (coating codes 3 and 4, [Fig.9] B), - 24 months in cat kibble regardless of the coating methods tested (codes 1-2, 5-6, [Fig. 10]).

[0059] This disclosure is not limited to the examples described above, which are only examples, but encompasses all the variants that a person skilled in the art may consider in the context of the protection sought. Reference to deposited biological material

[0060] In this application, reference is made to the following biological material and to the derived variant or mutant strains: - identification reference “Sc50” and registration number “1-5660” filed with the National Collection of Microorganism Cultures (CNCM) (25, rue du Docteur Roux, 75724 Paris Cedex 15), in France, on March 3, 2021 by Lesaffre et Compagnie whose address is 41 rue Etienne Marcel, 75009 Paris. List of documents cited Patent documents

[0061] For the avoidance of doubt, the following patent documents are cited: - patcitl: JP2008013534; and - patcit2: PCT / US2009 / 056292. Non-patent literature

[0062] For the sake of clarity, the following non-patent element is cited: - nplcitl: Bruna M. Rodrigues et al., Hindawi, Int. J. of Microbiology, Vol 2020, ID 1293481, 10 pages; - nplcit2: Yeast Technology, 2nd edition, 1991, G. Reed and TW Nagodawithana, published by Van Nostrand Reinhold, ISBN 0-442-31892-8; - nplcit3 : Brito, C.B.M., Félix, A.P., Jésus, R.M., França, M.L, Oliveira, S.G., Krabbe, E.L., Maiorka, A., 2010. Digestibility and palatability of dog foods containing different moisture levels, and the inclusion of a mould inhibitor. Anim. Feed Sci. Technol. 159, 150-155 ; - nplcit4 : Journal FEMS Microbiology Ecology, « A dysbiosis index to assess microbial changes in fecal samples of dogs with chronic inflammatory enteropathy. AlShawaqfeh et al., 2017.».

Claims

Demands

1. Saccharomyces cerevisiae yeast obtained by culture of the Saccharomyces cerevisiae strain filed on March 3, 2021 with the CNCM under number 1-5660, which is an active dry yeast which has the following characteristics: - a dry matter content greater than 95% by weight and up to 97% by weight; - a water activity ranging from 0.1 to 0.3; - a spherule shape; - a particle size distribution characterized by a Gaussian curve showing that at least 90% of the spherules have a diameter ranging from 0.3 to 0.7mm; - a stability, in vacuum packaging and at a temperature of 15 to 25°C, ranging from 24 to 36 months.

2. Yeast according to claim 1 for its use: - in improving the intestinal immune system in pets, and / or - in restoring a normal intestinal microbiota in pets.

3. Solid palatability enhancer for pets, characterized in that it comprises a yeast according to claim 1, said yeast having a lifespan of 12 to 24 months within said palatability enhancer.

4. Pet food characterized in that it comprises a yeast according to claim 1 or a palatability factor according to claim 3, said yeast having a lifespan of 12 to 24 months within said food.

5. Food according to claim 4, characterized in that it is a kibble coated by at least one coating layer, said coating layer comprising a yeast according to claim 1 or a palatability factor according to claim 3.

6. Food according to claim 5, characterized in that the coating layer is present in an amount by weight from 3 to 50%, and preferably from 5 to 20% relative to the total weight of the food.

7. Food according to claim 5 or 6, characterized in that the coating layer further comprises a fat.

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