Saccharomyces cerevisiae yeast and use thereof for improving the intestinal health of pets
Patent Information
- Application Number
- EP2023722439
- Authority / Receiving Office
- EP · EP
- Patent Type
- Applications
- Current Assignee / Owner
- Priority Date
- 2022-04-15
- Filing Date
- 2023-04-14
- Publication Date
- 2025-02-19
Smart Images
Figure 1.1
Abstract
Description
Description Title: Saccharomyces cerevisiae yeast and its use to improve intestinal health in pets. technical field
[0001] This application falls within the field of animal nutrition and intestinal health in companion animals. It specifically concerns the 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 improve the intestinal health of pets is well-established. 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 specifically describes the encapsulation of lactic acid bacteria and yeast in a polymer containing a mixture of xanthan gum and chitosan. The resulting encapsulated probiotic is used to improve gastrointestinal disorders in pets. The article by Bruna M. Rodrigues et al. describes a lyophilized microencapsulation technique for probiotic strains to maintain their viability 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 intestinal 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 kibble coated with said microorganism, it is imperative that the microorganism survive not only the coating process around the kibble, but also remain viable until the kibble's expiration date in order to exert its beneficial effects on the animal's health. Document PCT / US2009 / 056292 describes the use of probiotics (specifically bifidobacteria) in a coating of pet kibble.However, this document does not mention a strain of yeast or a yeast that would be coated around the kibble and that would have a sufficiently long lifespan 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 pet food products while maintaining a sufficiently long shelf life within those products. A sufficiently long shelf life means, in particular, a shelf life extending until the expiration 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, has a particularly long shelf life when incorporated into pet food. This particularly long shelf life is defined as a period of 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 I-5660, which is an active dry yeast with 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 to 0.7mm; - stability, in vacuum packaging and at a temperature of 15 to 25°C, ranging from 24 to 36 months. The active dry yeast of the invention also advantageously has a shelf life of 12 to 24 months when incorporated into a composition, said composition being stored at a temperature of 15 to 25°C. Such a composition could be, for example, a solid palatability enhancer or pet kibble, said kibble including, for example, said palatability enhancer.
[0007] According to a second aspect, the invention relates to yeast as defined above, for its use in companion animals: - in improving the intestinal immune system, - in the restoration of a normal gut microbiota, - in the decrease of fecal pH, - in reducing the production of putrefactive catabolites in feces (such as ammonia or biogenic amines).
[0008] In another aspect, the invention relates to a pet food composition comprising active dry yeast as defined above, said yeast having a shelf life of 12 to 24 months within said composition. Such a pet food composition may be a solid palatability enhancer or a kibble containing, for example, said palatability enhancer.
[0009] The invention also relates to a composition as defined above, for use in companion animals: - in improving the intestinal immune system, - in the restoration of a normal gut microbiota, - in the decrease of fecal pH, and / or - in reducing the production of putrefactive catabolites in feces (such as ammonia and / or biogenic amines). Brief description of the drawings
[0010] Other features, details and advantages will become apparent upon reading the detailed description below, and upon analysis of 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. The active dry yeast of the invention is represented by "Sc50" in Figures 1 to 6 below. Fig. 1
[0011] [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 that 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
[0012] [Fig. 2] illustrates the content of Blautia (see A), Bifidobacterium (see B), Lactobacillus (see C) and Faecalibacterium (see D) bacteria in feces of Beagle breed dogs before and after the dietary transition at day 22. The groups tested are the same as those described for Figure 1. Fig. 3
[0013] [Fig. 3] illustrates the E. coli content in feces of Beagle breed dogs before and after the dietary transition at day 22. The groups tested are the same as those described for Figure 1. Fig. 4
[0014] [Fig. 4] illustrates the dysbiosis index (see A) of the intestinal microbiota and the streptococcal content (see B) found in the feces of Beagle dogs before and after the dietary transition at day 22. The groups tested are the same as those described for Figure 1. Fig. 5
[0015] [Fig. 5] illustrates the fecal immunoglobulin A (IgA) content found in the feces of Beagle dogs before and after the dietary transition at day 22. The groups tested are the same as those described for Figure 1. Fig. 6
[0016] [Fig. 6] illustrates the amount of biogenic amines (in mg / kg dry matter) present in the feces of Beagle dogs before and after the dietary transition on day 22. More specifically, Figure A illustrates the amount of total 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 Figure 1. Fig. 7
[0017] [Fig. 7] illustrates the stability of active dry yeast Sc50 within two different palatability factors for dogs (see A) and for cats (see B). Fig. 8
[0018] [Fig. 8] illustrates different possible embodiments for coating a dog or cat kibble. In Figure A, 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 in white and delimited by dotted lines) consisting of a liquid palatability factor. 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
[0019] [Fig. 9] illustrates the stability of the active dry yeast Sc50 when 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
[0020] [Fig. 10] illustrates the stability of active dry yeast Sc50 when coated around cat kibble. The series shown in Figure 10 corresponds to the average viability obtained at each time point for the four coating methods. following: - 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. Fig. 11
[0021] [Fig. 11] illustrates the stability of two active dry yeasts, "yeast 1" and "yeast 2," coated around dog kibble. The coating methods are: - fat then "solid palatability factor + active dry yeast 1" (curve connected by the squares); - fat then "solid palatability factor + active dry yeast 1" then liquid palatability factor (curve connected by the triangles); - fat then "solid palatability factor + active dry yeast 2" (curve connected by circles); - fat then "solid palatability factor + active dry yeast 2" then liquid palatability factor (curve connected by vertical lines). 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 I-5660, which is an active dry yeast exhibiting 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 to 0.7mm; - stability, in vacuum packaging and at a temperature of 15 to 25°C, ranging from 24 to 36 months. 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 I-5660 is also identified by the reference "Sc50". This strain may be referred to, among other things, as "strain Sc I-5660", "strain I-5660", or "strain Sc50" in this application. The Saccharomyces cerevisiae yeast obtained by culture of strain I-5660 may be designated as "Sc I-5660 yeast", "I-5660 yeast" or "Sc50 yeast". As an example, I-5660 yeast can be obtained by culturing the I-5660 strain 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". However, I-5660 yeast can also be obtained by an industrial-scale process that 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% dry matter, or even a higher quantity of dry matter, in particular if the liquid yeast cream or liquid yeast is mixed with osmolyte products. The liquid yeast cream or liquid yeast I-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 i-5660 liquid yeast 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 i-5660 yeast obtained from this dehydration and drying process is called "active dry yeast". It may be referred to interchangeably in this 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 yeast cells that are metabolically active. For the purposes of this application, "active dry yeast" means active yeast. presenting a dry matter content greater than 95% by weight, which can reach 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 at 25°C, with an AquaLab 3TE series instrument (“Decagon Devices, Pullman, WA, USA”). 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 using a dry method with a laser diffraction analyzer. The particle size distribution of the spherules was determined according to ISO 13320:2020. Advantageously according to the invention, the active dry yeast Sc50 exhibits, in a vacuum package and at a temperature of 15 to 25°C, a stability 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 and forming colonies. 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, stable active dry yeast over a period of 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 that period. This means, for example, that if the active dry yeast has a live and metabolically active cell count of 1 x 1010 CFU / g (equivalent to 10 log CFU / g) at time t0 (t0 being the time of obtaining the active dry yeast, i.e., the moment when the active dry yeast has just been prepared), then the yeast will, after a period of 24 to 36 months, have a number of living and metabolically active cells that will not be less than 1x10 9 UFC / g (equivalent to 9 log UFC / g). The stability of the active dry yeast of the invention for 24 to 36 months is verified when said yeast is stored in vacuum packaging at a temperature ranging from 15 to 25°C. Thus, for the purposes of this application, the stability of active dried yeast over a period of 24 to 36 months means that the active dried yeast has a shelf life of 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. Thus, the active dry yeast Sc50 is further characterized in that, in vacuum packaging and at a temperature of 15 to 25°C, it has a shelf life of 24 to 36 months.
[0023] The inventors determined that the physicochemical characteristics of the active dried yeast of the invention, namely its dry matter content, water activity, spherical shape, and particle size, were important for the yeast's shelf life, particularly when incorporated into a pet food composition such as kibble. They also determined that these characteristics were important for the yeast's good adhesion to the kibble. Indeed, they were able to observe that if the active dry yeast had a dry matter content of less than 95%, namely for example 94 or 93%, then the lifespan of said yeast in a composition such as pet food did not exceed 8 months, or even 5 months. The inventors also observed that when the yeast of the invention is in the form of spherules with a diameter ranging from 0.3 to 0.7 mm, it allows for good adhesion of the yeast around a composition such as pet food. Indeed, when the yeast spherules have a diameter less than 0.3 mm or greater than 0.7 mm, the yeast adhesion is less effective. The adhesion of the yeast around the pet food is characterized in particular by the percentage of "fines" generated after a durability / friability test. "Fines" refers to the material that has detached from the coated pet food and passed through the mesh of a sieve whose mesh size has been pre-adjusted.
[0024] The invention also relates to the active dry yeast Sc50 as defined above for its use: - in improving the intestinal immune system in pets, - in restoring a normal gut microbiota in pets, - in the decrease of fecal pH in pets, and / or - in the reduction of the production of putrefactive catabolites in animal feces company. 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, administration of active dry yeast Sc50 to pets significantly increases the production of fecal immunoglobulin A. The significant increase in fecal immunoglobulin A production thus improves the intestinal immune system in pets. The gut microbiota, which refers to all the microorganisms that live in the digestive tract, plays an essential role in maintaining good intestinal health. It also plays a key part 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 establishes the presence or absence of dysbiosis. The Inventors analyzed the dysbiosis index using a quantitative PCR (polymerase chain reaction) methodology published in the journal FEMS Microbiology Ecology (AlShawaqfeh et al., 2017). 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.Thus, an improvement in the dysbiosis index means a statistically significant decrease in the dysbiosis index. A "normal" gut microbiota is a healthy microbiota, showing no changes 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, and / or - 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. Thus, reducing the levels of fecal streptococci and E. coli, improving the dysbiosis index and / or increasing the levels of the aforementioned bacteria helps to restore a normal intestinal microbiota in pets. According to another advantageous embodiment of the invention, the administration of active dry yeast Sc50 in pets makes it possible to decrease fecal pH. Fecal pH can be considered a marker of the fermentative activity of the gut microbiota, with a high pH associated with proteolytic metabolism. Therefore, a decrease in fecal pH helps maintain intestinal function by inhibiting the proliferation of potentially pathogenic bacteria. In the publication by Lin and Visek, it is reported that a decrease in fecal pH in rats is correlated with a decrease in proliferative cells in the colonic mucosa. According to yet another advantageous embodiment, administering the active dry yeast Sc50 of the invention reduces the production of putrefactive catabolites in the feces of pets, thereby benefiting their intestinal health. Examples of "putrefactive catabolites" include ammonia and biogenic amines such as spermidine and putrescine. The publication by Celi et al. describes, in particular, that excessive ammonia production is associated with excessive fermentative activity of gut microbiota bacteria. It also describes that ammonia can have toxic effects on enterocytes. As for biogenic amines, an excessive concentration of these in the feces of pets can be detrimental to the intestinal mucosa and promote the survival of potentially pathogenic bacteria. Thus, the decrease in fecal pH, the decrease in the production of putrefactive catabolites (such as ammonia and / or biogenic amines) advantageously improves the fecal characteristics of pets and contributes to improving the intestinal health of pets. In addition to the health benefits, reducing putrefactive catabolites in pet feces also helps reduce unpleasant odors in their stools, which is a significant advantage given that Bad odors from feces are a real nuisance for pet owners.
[0025] 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 1x10 8 at 1.5x10 8 UFC / animal.
[0026] 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. The active dry yeast of the invention may also be called "probiotic yeast". 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 food 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 either between meals or, conversely, during the pets' mealtimes. 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 dietary 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 dietary supplement as defined above, in a feed for pets, the food in question preferably being dog or cat kibble.
[0027] 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).
[0028] The invention further relates to a pet food composition, characterized in that it comprises an active dry yeast as defined above, said yeast having a shelf life of 12 to 24 months within said composition. Such a composition may be a solid palatability enhancer intended for incorporation into pet food, or may be a pet food itself, such as kibble. The shelf life of 12 to 24 months of the active dry yeast within the composition is observed when the composition is stored at a temperature ranging from 15 to 25°C.
[0029] According to another advantageous embodiment of the invention, the composition as defined above is more particularly characterized in that it is a solid palatability enhancer and in that the yeast of the invention is integrated into said palatability enhancer. According to yet another advantageous embodiment of the invention, the palatability enhancer 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 log CFU / g to 0.8 log CFU / g over a period of 12 to 24 months. This means, for example, that if the live active dry yeast is present in an amount of 1 x 10 7CFU / g of palatability factor, immediately after mixing with said palatability factor (called TO time), then at the end of a period ranging from 12 to 24 months from TO time (TO being the time when the live active dry yeast is mixed with the palatability factor), the quantity of live active dry yeast is not less than 1x10 6 UFC / g of palatability factor. This shelf life is extremely advantageous because it allows the active dry yeast Sc50 to retain its biological properties when used in combination with a palatability enhancer (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 enhancer varies from 1 to 40%, and more particularly from 2 to 35% relative to the total weight of the palatability enhancer.
[0030] 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.
[0031] 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 food during the manufacturing process, it will preferably be active dry yeast. When yeast is mixed with the animal's food at mealtimes, then said yeast is preferably in the form of a probiotic or a food 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, palatability enhancer, probiotic, or food supplement, over a period of 12 to 24 months, means that it exhibits 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 the live active dry yeast is present in an amount of 1 x 10 7CFU / g of feed, palatability enhancer, probiotic, or food supplement, immediately after said yeast has been incorporated into said feed, palatability enhancer, probiotic, or food supplement (referred to as the TO time), then, after a period of 12 to 24 months from the TO time (TO = time when the active dry yeast is incorporated into the feed, palatability enhancer, probiotic, or food supplement), the quantity of live active dry yeast is not less than 1 x 10 6 UFC / g of food, palatability factor, 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.
[0032] 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 enhancer as defined above, said yeast with 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 dry yeast as defined above, having a shelf life of 12 to 24 months. The lifespan of the active dry yeast within said food, probiotic or food supplement is as defined above (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 shelf life: - 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, the Sc50 yeast is administered in a quantity ranging from 5x10 5at 5x10 9 UFC / g of food.
[0033] 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 contained within a coating layer that encases said composition. Such a composition may, for example, be a food such as a kibble comprising a coating layer, said coating layer comprising the yeast of the invention or the 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 nucleus, preferably 80 to 95%, - 3 to 50% for the coating layer, preferably 5 to 20%. In this respect, the composition of the invention as defined above is more particularly characterized in that the coating layer is present in an amount by weight ranging from 3 to 50%, and preferably from 5 to 20%, relative to the total weight of the composition. Said composition is, for example, a kibble. 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 of the composition of the invention further comprises a fat. As an 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 factor, said palatability factor 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.
[0034] The invention also relates to a composition as defined above, for its use: - in improving the intestinal immune system in pets, - in restoring a normal gut microbiota in pets, - in the decrease of fecal pH in pets, and / or - in reducing the production of putrefactive catabolites in the feces of pets. The effects of Sc50 active dry yeast on the intestinal health of pets described above are fully applicable to the compositions of the invention that include Sc50 active dry yeast. This is due in particular to the fact that the active dry yeast of the invention exhibits excellent stability / shelf life within the composition of the invention (ranging from 12 to 24 months), which means that said active dry yeast does not exhibit any loss of activity and is therefore able to exert its benefits on the intestinal health of pets in the same way as if Sc50 yeast were used alone (without being incorporated into a composition such as a palatability enhancer or kibble). Example 1: Study of the effectiveness of Sc50 active dried yeast on intestinal health
[0035] 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 with good intestinal health after a sudden food transition. A sudden food transition is defined as an abrupt change in diet.
[0036] 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 were studied: with or without supplementation of active dry yeast Sc50. The active dry yeast Sc50 (comprising 1x10 10(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 (D1 to D21)) and, - a phase 2 which lasts 4 weeks (from day 22 to day 49 (J22 to J49)). On day 1 of phase 2, namely day 22 (D22) the dogs undergo a sudden 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. 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%. 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.
[0037] Parameters evaluated 1 / Fecal samples were collected for analysis of fecal pH (Figure 1, A) and fecal ammonia concentration (Figure 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 a correction factor of 0.9875 according to the formula below: [Math. 1] 2J 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.
[0038] 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.
[0039] Effects of active dry yeast Sc50 on fermentation production (figure 1) A significant decrease in fecal pH (Figure 1, A) at days 35 and 49 and in fecal ammonia (Figure 1, B) at days 23, 35 and 49 was observed after the dietary transition.
[0040] Effect of active dry yeast Sc50 on the gut microbiota (figure 2, figure 3, figure 4, B) and the dvsbiosis index (figure 4, A) A significant increase in the content of Blautia (J49) (figure 2, A), Bifidobacterium (J21 and J49) (figure 2, B), Lactobacillus (J21 and J49) (figure 2, C) and Faecalibacterium (J49) (figure 2, D) was observed after the dietary transition in the feces of the dogs. A significant decrease in fecal streptococcus content (day 35) (Figure 4, B) was observed after the dietary transition. A significant decrease in fecal Escherichia coli content after the dietary transition (day 49) and a downward trend before the dietary transition (day 21) (Figure 3) were also observed. 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). Indeed, Figure 4, A, shows a statistically significant decrease in the dysbiosis index. 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.
[0041] Effect of active dry yeast Sc50 on fecal immunoglobulin A production Figure 5 shows that administration of active dry yeast Sc50 leads to a significant increase in IgA production in the feces of dogs before (J21) and after (J35) the dietary transition.
[0042] Effect of active dry yeast Sc50 on the production of biogenic amines (Figure 6). 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 toward a decrease after the dietary transition (Day 49) in dogs.
[0043] Conclusions Supplementation with Sc50 active dry yeast in the diet of healthy dogs undergoing an abrupt dietary change contributes to: a) Improved fecal characteristics in tested dogs through 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, the reduction in biogenic amines contributes to a decrease in fecal odor, which is an important commercial characteristic given the close relationship between dogs and their owners. b) Improved local immunity through an increase in fecal IgA production. 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 bacteria in the gut microbiota. Furthermore, supplementation with active dry yeast Sc50 has been shown to improve the dysbiosis index. Example 2: Study of the stability of active dry yeast Sc50 in dog and cat food
[0044] 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.
[0045] Mixture of the active dry yeast Sc50 of the invention with palatability enhancers 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 factors for dogs, - a water activity of 0.30 or 0.35 for the 2 palatability factors for cats. The "palatability factor + Sc50 active dry yeast" mixture is prepared in a mixer marketed under the Forberg name (double shaft paddle mixer) by first introducing all of the powdered palatability factor into the mixer, then the Sc50 active dry yeast. The mixture is then run for 5 minutes at a speed of 50Hz.
[0046] Study of the stability of 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 the active dry yeast Sc50 is measured at different time points. (at least 4) over the entire duration of the study, i.e., from 0 to 12 months. The stability marker monitored during storage of the active dry yeast Sc50 is the percentage of viable yeast cells, this percentage being measured according to the standardized method NF EN 15789. The viability results of the active dry yeast Sc50 within the various palatability factors are shown in Figure 7. In particular, Figure 7 A illustrates the stability of active dry yeast Sc50 in two canine palatability factors, the solid line representing stability in the palatability factor having a water activity of 0.32 and the dashed line representing stability in the palatability factor having a water activity of 0.38. Figure 7B illustrates the stability of active dry yeast Sc50 in two cat palatability factors, the solid line representing stability in the palatability factor having a water activity of 0.30 and the dashed line representing stability in the palatability factor having a water activity of 0.35. 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).
[0047] Coating of the kibble core with the palatability enhancer including 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, fat poultry; - for cats: poultry meal, barley, corn and derivatives, poultry fat, wheat, soy and derivatives. In the kibble, the incorporation rate of active dry yeast Sc50 is 0.05%. Active dry yeast Sc50, previously incorporated into the powdered palatability enhancers, is applied to 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).
[0048] [Table 1]
[0049] The kibble coated with the active dry yeast Sc50 is packaged in aluminum bags and then stored at room temperature (20°C + / - 5°C).
[0050] 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 (standardized 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. Specifically, Figure 9A illustrates the stability of active dry yeast Sc50 in kibble exhibiting water activity ranging from 0.45 to 0.49. The solid line represents the stability of the yeast in the coated kibble according to coating code 2. while the dashed line represents the stability of the yeast in the coated kibble according to coating code 1. Figure 9B illustrates the stability of active dry yeast Sc50 in kibble with water activity ranging from 0.53 to 0.58. The solid line represents the stability of the yeast in the coated kibble according to coating code 4, while the dashed line represents the stability of the yeast in the 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. 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 enhancers (coating codes 1 and 2, figure 9 A), - 12 months when the coating layer includes a liquid palatability factor, specific to certain dog kibble (coating codes 3 and 4, figure 9 B), - 24 months in cat kibble regardless of the coating methods tested (codes 1-2, 5-6, figure 10). Example 3: Comparative study to example 2
[0051] The objective of this study is to demonstrate that the physicochemical characteristics of active dry yeasts are essential with regard to: - their lifespan within a kibble, and / or - to their membership within a kibble.
[0052] Shelf life of active dry yeasts in dog kibble. The active dry yeasts tested in this example exhibit the physico-chemical characteristics described below. Yeast "1": - granulometry: diameter of the spherules ranging from 0.3 to 0.7 mm; - dry matter content: 93% by weight. Yeast "2": - granulometry: diameter of spherules ranging from 0.05 to 0.2 mm; - dry matter content: 94.5% by weight. These active dry yeasts therefore do not possess all the characteristics physico-chemical properties of the active dry yeast of the invention. These yeasts are incorporated into palatability factors for dogs with a water activity of 0.32 according to the methods described in example 2. Similarly, these yeasts are coated around a dog kibble according to the methods described in example 2. More specifically, yeast 1 is coated according to coating codes 1 and 3, namely: - MG then "FAP + Yeast 1" (coating code 1); - MG then "FAP + Yeast 1" then FAL (coating code 3). Similarly, yeast 2 is coated according to coating codes 1 and 3: - MG then "FAP + Yeast 2"; - MG then "FAP + Yeast 2" then FAL. The solid palatability factors (PPF) and liquid palatability factors (ALF) are those described in example 2. Dog kibble coated in this way is stored under the same conditions as those described in example 2. Viability measurement is assessed monthly according to the protocol described in example 2. The results obtained are illustrated in figure 11. It appears from this figure that yeast 1 does not exceed 5 months of viability and yeast 2 does not exceed 8 months of viability.
[0053] Yeast adhesion within a kibble The active dry yeasts tested in this example exhibit the physico-chemical characteristics described below. Yeast "3": - granulometry: diameter of the spherules ranging from 0.1 to 0.2 mm; - dry matter content: 94% by weight. Yeast "4": - granulometry: diameter of spherules ranging from 0.01 to 0.1 mm; - dry matter content: 94% by weight. These active dry yeasts therefore do not exhibit all the physico-chemical characteristics of the active dry yeast of the invention. These yeasts are incorporated into palatability enhancers for dogs with a water activity of 0.32, as described in Example 2. Similarly, these yeasts are coated around a dog kibble, as described in Example 2. More specifically, yeasts 3 and 4 are each coated according to coating code 1, namely: - MG then "FAP + Yeast 3"; - MG then "FAP + Yeast 4". The solid palatability factor (SPF) is that described in example 2. The coated dog kibble is subjected to a friability / durability test. The durability of the coated kibble is analyzed using a durability tester marketed under the name "Holmen NHP 100". One hundred grams of kibble are placed in the durability tester, and each analysis is performed in duplicate. The tester is equipped with a 1 mm grid, and the test lasts 120 seconds. During the durability test, the kibble is subjected to standardized mechanical stresses. At the end of the mechanical test, the kibble is sieved through a sieve with a mesh size corresponding to 80% of the kibble's diameter. The proportion of material from the kibble with a size smaller than the sieve mesh is called "fines." The higher the fines content, the less effective the yeast adhesion to the kibble. The results show that kibble coated with yeast 3 and subjected to the durability test generated an average fines content of 1.79%, while kibble coated with yeast 4 and subjected to the durability test generated an average fines content of 1.55%. The differences observed after durability testing are mainly due to the physicochemical characteristics of the yeasts, particularly their particle size. Larger yeasts will generate a higher percentage of fines than smaller yeasts. The spherical shape of yeast is important because it has very low surface porosity. Smaller spherules therefore have a larger surface area and are thus more exposed to environmental moisture. However, moisture is critical for the long-term stability of yeast. Therefore, a compromise must be found between spherules that are small enough to adhere well to the surface of the kibble but not so small as to compromise long-term stability by being more resistant to moisture, while also being industrially feasible.
[0054] This disclosure is not limited to the examples described above, which are merely examples, but encompasses all the variations that a person skilled in the art may consider in the context of the protection sought. Reference to deposited biological material
[0055] This application refers to the following biological material and derived variant or mutant strains: - identification reference “Sc50” and registration number “I-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
[0056] For the record, the following patent documents are cited: - patcitl: JP2008013534; and - patcit2: PCT / US2009 / 056292. Littérature non-brevet
[0057] À toute fin utile, l’élément non-brevet suivant est cité : - nplcitl : Bruna M. Rodrigues et al., Hindawi, Int. J. of Microbiology, Vol 2020, ID 1293481 , 10 pages ; - nplcit2 : Yeast Technology, 2ème édition, 1991 , G. Reed et T.W. Nagodawithana, publié par Van Nostrand Reinhold, ISBN 0-442-31892-8 ; - nplcit3 : Journal FEMS Microbiology Ecology, « A dysbiosis index to assess microbial changes in fecal samples of dogs with chronic inflammatory enteropathy. AlShawaqfeh et al., 2017. - nplcit4 : Large intestinal pH and ammonia in rats: dietary fat and protein interactions 1-2. Hsi-Chiang Lin and Villard J. Visek. American Institute of Nutrition. 1991 , 832-843 ; - nplcit5 : Cell, P; Verlhac, V; Calvo, EP; Schmeisser, J; Kluenter, AM. Biomarkers of gastrointestinal functionality in animal nutrition and health. Anim. Feed. Sci. Technol. 2019, 250, 9-31 ; - nplcit6: Brito, C.B.M., Félix, A.P., Jesus, R.M., França, M.I., 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.
Claims
Demands
1. 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 with 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 to 0.7mm; - 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, - in restoring a normal gut microbiota in pets, - in the decrease of fecal pH in pets, and / or - in reducing the production of putrefactive catabolites in the feces of pets.
3. Pet composition, characterized in that it comprises a yeast according to claim 1, said yeast having a shelf life of 12 to 24 months within said composition.
4. Composition according to claim 3, characterized in that the yeast is coated around said composition.
5. Composition according to claim 3 or 4, characterized in that the yeast is included in a coating layer which coats said composition.
6. Composition 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 composition.
7. Composition according to claim 5 or 6, characterized in that the coating layer further comprises a fat.
8. Composition according to any one of claims 3 to 7, for its use: - in improving the intestinal immune system in pets, - in restoring a normal gut microbiota in pets, - in the decrease of fecal pH in pets, and / or - in reducing the production of putrefactive catabolites in the feces of pets.