Yeast product for reducing or preventing the formation of dental biofilm

Abstract

The present invention relates to the field of oral / dental hygiene and health. The invention relates to a yeast product used for preventing and / or reducing the formation of dental biofilm, particularly in dogs.

Description

[0001] Yeast product used as a postbiotic in the field of oral hygiene and health

[0002] technical field

[0003] The present invention relates to the field of oral hygiene and health. The invention relates to a yeast product obtained from a yeast extract, or a composition comprising it, used for the prevention and / or reduction of dental biofilm formation, particularly in dogs.

[0004] Technical background

[0005] In mammals, the oral cavity is a complex ecosystem, both open to the outside and the inside, inhabited by numerous microorganisms.

[0006] In companion animals, particularly dogs, the oral cavities are also complex ecosystems inhabited by numerous microorganisms (see, for example, the article: Holcombe et al., PlosOne, 2014, 9, 12, e113744). Most of these microorganisms are commensal and necessary to maintain the balance of this ecosystem. However, a disruption of this balance can occur and lead to the development of infectious diseases of the oral cavity.

[0007] The main infectious diseases of the oral cavity are periodontal diseases. Indeed, dental caries are very rare in dogs and cats. Among the bacterial species most frequently implicated in periodontal diseases in pets are bacteria of the species Porphyromonas cangingivalis, Porphyromonas gulae, and Porphyromonas spp.

[0008] To prevent or treat oral infections, it is helpful to maintain or restore the balance between non-pathogenic and pathogenic microorganisms within the oral microbiota. This balance allows for the control of dental biofilm formation, development, and composition. Many pathogenic bacteria can be present in the canine oral microbiota and can stimulate the formation, development, and composition of dental biofilm, potentially contributing to the development of oral infections.

[0009] Therefore, there is a real need to prevent and / or reduce the formation of dental biofilm, particularly in dogs. Summary of the invention

[0010] The invention relates to the non-therapeutic use of a yeast product, or a composition comprising it, to prevent and / or reduce the formation of dental biofilm, preferably canine dental biofilm, the yeast product being a yeast extract having a pH between 6.0 and 7.0; and comprising, by weight by total weight of the yeast product, 35 to 65% protein (nitrogen x 6.25), 10 to 30% ash and 3% or less sodium chloride.

[0011] The invention also relates to the non-therapeutic use to prevent and / or reduce the formation of dental biofilm, preferably canine dental biofilm, of the combination of the yeast product defined above with an additional yeast product comprising at least 20% p-glucans, by weight per total weight of the additional yeast product, or of a composition comprising them.

[0012] The invention also relates to the non-therapeutic use to prevent and / or reduce the formation of dental biofilm, preferably canine dental biofilm, of the combination of the yeast product defined above with a Bacillus inaquosorum strain, and / or cells derived from it.

[0013] The invention also relates to a yeast product, or a composition comprising it, for therapeutic use to prevent and / or reduce the formation of dental biofilm, preferably canine dental biofilm, the yeast product being a yeast extract having a pH between 6.0 and 7.0; and comprising, by weight by total weight of the yeast product, 35 to 65% protein (nitrogen x 6.25), 10 to 30% ash and 3% or less sodium chloride.

[0014] In embodiments, the yeast product is a yeast extract having a pH between 6.1 and 6.9; and comprising, by weight by total weight of the yeast product, 40 to 61% protein (nitrogen x 6.25), 13 to 27% ash and 2% or less sodium chloride.

[0015] In embodiments, the yeast product is a yeast extract having a pH between 6.2 and 6.8; and comprising, by weight by total weight of the yeast product, 45 to 57% protein (nitrogen x 6.25), 16 to 24% ash and 1% or less sodium chloride.

[0016] In some embodiments, the yeast product is obtained by implementing a preparation process comprising the following steps:

[0017] - obtaining a yeast cream by fermentation followed by concentration by centrifugation;

[0018] - washing of the yeast cream obtained by adding water and centrifugal concentration; - heating of the washed yeast cream to a temperature between 60 and 95°C, preferably between 75 and 90°C;

[0019] - incubation under stirring of the yeast cream heated to a temperature of 55 to 90°C, preferably between 55 and 75°C, for a duration of 1 to 5 hours;

[0020] - separation of yeast extract and yeast cell walls by centrifugation;

[0021] - concentration of the yeast extract by evaporation and / or reverse osmosis; and

[0022] - optionally drying of the concentrated yeast extract.

[0023] In some embodiments, the additional yeast product comprises, by weight per total weight of the additional yeast product:

[0024] - at least 50%, preferably from 50 to 90%, preferably from 50 to 80%, preferably from 50 to 70%, preferably from 50 to 60%, of p-glucans;

[0025] - 5% or less, preferably between 1 and 5%, of mannans;

[0026] - 10% or less protein; and

[0027] - 10% or less of glycogen.

[0028] In some embodiments, the Bacillus inaquosorum strain is the strain deposited on September 12, 2024 with the Agricultural Research Culture Collection under the Treaty of Budapest and having received the registration number NRRL B-68429.

[0029] In some embodiments, the yeast product, optionally in combination with the additional yeast product or a Bacillus inaquosorum strain, and / or cells derived therefrom, is administered to a subject orally.

[0030] In embodiments (non-therapeutic use), the product, or a composition comprising it, optionally in combination with the additional yeast product or a Bacillus inaquosorum strain, and / or cells derived therefrom, is administered to a healthy subject.

[0031] In some embodiments (therapeutic use), the yeast product, or a composition comprising it, is administered to a subject with a disease of the oral cavity, or at risk of having one.

[0032] In embodiments, the composition comprising the yeast product, optionally in combination with the additional yeast product or a Bacillus inaquosorum strain, and / or cells derived therefrom, is selected from a food supplement or a parapharmaceutical composition or a pharmaceutical composition.

[0033] The inventors have demonstrated, surprisingly, that the yeast product, optionally in combination with an additional yeast product or a Bacillus inaquosorum strain, or the composition containing them, prevents and / or reduces the formation of dental biofilm, particularly canine dental biofilm. The yeast product, optionally in combination with an additional yeast product or a Bacillus inaquosorum strain, or the composition containing them, is particularly interesting because it can be used in healthy individuals (non-therapeutic use) or in individuals with oral cavity disease, or at risk of developing such disease (therapeutic use), these two populations being distinct.

[0034] Detailed description

[0035] The invention is now described in more detail and in a non-limiting manner in the following description.

[0036] Definitions

[0037] By “Saccharomyces cerevisiae yeast strain”, we mean a relatively homogeneous population of Saccharomyces cerevisiae yeast cells obtained by culturing (or multiplying) the starting strain.

[0038] The term "oral microbiota" refers to all the microorganisms, particularly bacteria, present in the oral cavity. The expressions "oral microbiota," "buccal microbiota," "human oral microbiota," "dog and cat oral microbiota," "oral cavity microbiota," "oral flora," and "oral flora" can be used interchangeably. The microorganisms present in the oral microbiota can be non-pathogenic or pathogenic. The balance between non-pathogenic and pathogenic microorganism populations therefore determines, from a clinical and biological perspective, whether the oral microbiota is healthy or not.Des microorganismes présent peuvent être, par exemple, les baccteria appartant au genre Actinomyces, Actinomycetemcomitans, Aggregatibacter, Alloprevotella, Alloscardovia, Anaeroglobus, Atopobium, Bacteroides Bifidobacterium, Capnocytophaga, Catonella, Corynebacterium, Dialister, Eggerthia, Eubacteria, Fusobacterium, Haemophilus, Lachnoanaerobaculum, Leptotrichia, Mogibacterium, Moryella, Neisseriaceae, Olsenella, Oribacterium, Parvimonas, Peptoniphilus, Peptostreptococcus, Propionibacterium, Porphyromonas, Prevotella, Rikenellaceae, Ruminococcaceae, Selenomonas, Shuttleworthia, Slackia, Solobacterium, Stomatobaculum, Streptococcus, Tannerella, Treponema ou Veillonella. Examples of pathogenic microorganisms are: Tannerella forsythia (T forsythia), Porphyromonas gingivalis {P. gingivalis), Treponema denticola (T. denticola), Porphyromonas cangingivalis (P. cangingivalis), Porphyromonas gulae (P. gulae), Porphyromonas endodontalis (P. endodontalis), Prevotella intermedia (P.intermedia), Streptococcus mutans (S. mutans) and Streptococcus sobrinus (S. sobrinus). Non-pathogenic microorganisms include, for example: Streptococcus oralis (S. oralis), and Streptococcus mitis (S. mitis).

[0039] The term "biofilm" refers to the film formed on the surface of teeth and oral mucosa, such as the mucous membranes of the tongue, gums, palate, and cheeks, by the oral microbiota. The terms "biofilm," "dental biofilm," "dental plaque," "polymicrobial biofilm," and "polymicrobial film" can be used interchangeably.

[0040] By "periodontium" we mean all the supporting tissues of the tooth, the gums, the bone tissue, the cementum and the periodontal ligament.

[0041] An "infectious disease of the oral cavity" is a medical condition located in the oral cavity (mouth) that results from infection by a pathogenic microorganism. Infectious diseases of the oral cavity include, but are not limited to, dental caries and periodontal disease.

[0042] By "dental caries" or "cavity", we mean an infectious disease of the tooth, which causes damage to the enamel, dentin and / or cementum (pulp).

[0043] By "periodontal health" we mean the absence of inflammation or the presence of a low level of inflammation in an intact or reduced but stable periodontium.

[0044] Periodontal disease refers to inflammation of the tissues supporting the teeth, namely the bone and gums. These infections are caused by the accumulation of pathogenic bacteria and their toxins on the gum line surrounding the teeth. It initially manifests as gingivitis and then, if left untreated, progresses to periodontitis. The terms "periodontal disease" and "periodontal disease" can be used interchangeably.

[0045] Gingivitis refers to inflammation of the gums, stage 1 of periodontal disease. The terms gingivitis, gum disease, and gum inflammation can be used interchangeably.

[0046] By "periodontitis" we mean the inflammation of the deep tissues of the periodontium, stage 2 of periodontal disease.

[0047] By "postbiotic" we mean a preparation of inanimate microorganisms and / or their components that confer a health benefit to the host.

[0048] The term "treatment" means a method intended to: (1) delay or prevent the onset of a disease or clinical condition; (2) slow or stop the progression, worsening, or deterioration of disease symptoms; (3) improve disease symptoms; and / or (4) cure the disease. A treatment may be administered before the onset of the disease, for prophylactic action (referred to as "prevention"), or it may be administered after the disease has begun, for therapeutic action. In the context of the invention, the term "treatment" generically refers to the reduction and / or prevention of dental biofilm formation, particularly through the growth of periodontogenic and / or cariogenic bacteria.

[0049] By "physiologically acceptable excipient" is meant any medium or additive which does not interfere with the effectiveness of the biological activity of the active ingredient and which is not excessively toxic to the subject at the concentrations at which it is administered.

[0050] The term “pharmaceutical active ingredient” means any compound or substance whose administration has a therapeutic effect or whose administration has a beneficial effect on the health or general condition of a subject to whom it is administered.

[0051] Unless otherwise stated, all percentages for the quantities indicated are mass percentages. For p-glucan percentages, these are expressed as an equivalent mass of glucose. For mannan percentages, these are expressed as an equivalent mass of mannose.

[0052] Yeast product

[0053] The yeast product is a yeast extract having a pH between 6.0 and 7.0; and comprising, by weight per total weight of the yeast product, 35 to 65% protein (nitrogen x 6.25), 10 to 30% ash and 3% or less sodium chloride.

[0054] Yeast, as a living microorganism, comprises cytoplasm surrounded by a cytoplasmic membrane (also called the inner membrane or cell membrane). The cytoplasm contains intracellular compartments, including the nucleus, mitochondria, and Golgi apparatus. The cytoplasmic membrane is surrounded by a cell wall or cortex (the terms "yeast cell walls" and "yeast cortex" are used interchangeably and refer to the insoluble portion of yeast cells, i.e., the cell wall and the plasma membrane of yeast) composed primarily of p-glucans and mannans. The area between the cell membrane and the cell wall forms the periplasm (also called the periplasmic space).

[0055] Yeast products are obtained from the yeast Saccharomyces cerevisiae, and more specifically from a strain of Saccharomyces cerevisiae. Saccharomyces cerevisiae is also known as brewer's yeast or baker's yeast. Many strains of Saccharomyces cerevisiae are known to the art. They are widely used in the food industry for their role in the production of various foods, including breads and fermented beverages. A yeast strain can be obtained from a clone, a clone being a population of yeast cells derived from a single yeast cell. Culturing a strain of Saccharomyces cerevisiae can be carried out using any suitable method. Yeast culture methods are known in the prior art, and those skilled in the art know how to optimize the culture conditions for each strain according to its specific characteristics.Thus, a Saccharomyces cerevisiae yeast can be obtained by multiplying a strain in an appropriate culture medium, for example, as described in the reference work "Yeast Technology", 2. ème edition, 1991, G. Reed and TW Nagodawithana, published by Van Nostrand Reinhold, ISBN 0-442-31982-8.

[0056] In one embodiment, the yeast product being a yeast extract having a pH between 6.1 and 6.9; and comprising, by weight by total weight of the yeast product, 40 to 61% protein (nitrogen x 6.25), 13 to 27% ash and 2% or less sodium chloride.

[0057] In one embodiment, the yeast product being a yeast extract having a pH between 6.2 and 6.8; and comprising, by weight by total weight of the yeast product, 45 to 57% protein (nitrogen x 6.25), 16 to 24% ash and 1% or less sodium chloride.

[0058] The protein content was measured using the Kjedhl method (6.25 times the total nitrogen). The sodium chloride content was measured using the sodium chloride method. The pH was measured at room temperature (18°C) in a solution containing 8.33% dry matter.

[0059] Yeast products can be in powder form.

[0060] The yeast product may comprise at least 94%, preferably at least 96%, by weight of dry matter per total weight of yeast product. Alternatively, the yeast product may comprise 100% or less, preferably 99% or less, by weight of dry matter per total weight of yeast product. The dry matter content may be measured, for example, by a drying method carried out for 5 hours at 105 °C.

[0061] In one embodiment, the yeast product is soluble. By "soluble" is meant a yeast product which can be homogeneously dissolved in water at a concentration of 50 g / L, under stirring and at a temperature of 50°C, the solution comprising less than 500 mg of solid residues i.e. undissolved.

[0062] The yeast product can be obtained by implementing a preparation process comprising the following steps:

[0063] - obtaining a yeast cream by fermentation followed by concentration by centrifugation; - washing the yeast cream obtained by adding water and centrifugal concentration;

[0064] - heating the washed yeast cream to a temperature between 60 and 95°C, preferably between 75 and 90°C;

[0065] - incubation under stirring of the yeast cream heated to a temperature of 55 to 90°C, preferably between 55 and 75°C, for a duration of 1 to 5 hours;

[0066] - separation of yeast extract and yeast cell walls by centrifugation;

[0067] - concentration of the yeast extract by evaporation and / or reverse osmosis; and

[0068] - optionally drying of the concentrated yeast extract.

[0069] The yeast product is available under the trade name Springer® 4101 ZO-PW-L from the company Biospringer by Lesaffre (Lesaffre group).

[0070] Springer® Yeast Product 4101 ZO-PW-L is in powder form, has a pH between 6.3 and 6.7 (8.33% solution) and comprises at least 94% by weight dry matter, less than 1% by weight sodium chloride, 7.5 to 9.0% by weight total nitrogen, 46.9 to 56.3% by weight protein (nitrogen x 6.25), and 18 to 22% by weight ash (excluding sodium chloride) by total product weight.

[0071] Additional yeast products

[0072] The yeast product according to the invention can be used in combination with at least one additional yeast product.

[0073] The additional yeast product is an extract of p-glucan-rich cell walls of Saccharomyces cerevisiae yeasts, preferentially rich in (3-1,3 / p-1,6-glucans (p-1,3 / [3-1,6-glucans).

[0074] By "rich in p-glucans" is meant a product comprising at least 20% p-glucans by weight of the total weight of the product. The additional yeast product may comprise from 20 to less than 30%, alternatively from 30 to less than 40%, alternatively from 40 to less than 50%, alternatively from 50 to less than 60%, alternatively from 60 to less than 70%, alternatively from 70 to less than 80%, alternatively from 80 to less than 90%, alternatively at least 90%, of p-glucans, by weight of the total weight of the additional yeast product.

[0075] The p-glucans from the yeast cell wall are called "(3-cell wall glucans". These p-glucans are essentially glucose polymers in which the glucose units of the main chain are linked by p-1,3 bonds and the branches are linked by p-1,6 bonds. p-glucans are insoluble and have low viscosity. The mannans from the yeast cell wall are called "cell wall mannans". These mannans are polysaccharides composed mainly of mannose, more precisely of copolymers of neutral or acidic sugars (5 or 6 carbon atoms), linked together by glycosidic bonds and associated with proteins. In the cell wall mannans of Saccharomyces cerevisiae, mannose is present as a skeleton of mannose residues (50 or more) linked by α-(1)-chains. ,6), branched by short chains of mannose linked in a-(1,2) and a-(1,3).

[0076] In one embodiment, the additional yeast product may comprise, by weight per total weight of the additional yeast product:

[0077] - at least 50%, preferably from 50 to 90%, preferably from 50 to 80%, preferably from 50 to 70%, preferably from 50 to 60%, of p-glucans;

[0078] - 5% or less, preferably between 1 and 5%, of mannans;

[0079] - 10% or less protein; and

[0080] - 10% or less of glycogen.

[0081] This additional yeast product may comprise at least 94%, preferably at least 96%, of dry matter, by weight per total weight of the additional yeast product.

[0082] p-glucans and mannans can be present in a (weight / weight) ratio of 12 to 40, preferentially 15 to 30, very preferentially 18 to 22.

[0083] For example, a product of this type is disclosed in application WO 2023194609 A1 published on October 12, 2023.

[0084] In one embodiment, the additional yeast product may comprise, by weight per total weight of the additional yeast product:

[0085] - at least 20%, preferably between 20 and 30%, of p-glucans;

[0086] - at least 20%, preferably 20 to 30%, of mannans;

[0087] - 25% or less, preferably 10 to 25%, of protein; and

[0088] - 10% or less of glycogen.

[0089] This additional yeast product may comprise at least 94%, preferably at least 96%, of dry matter, by weight per total weight of the additional yeast product.

[0090] For example, a product of this type is disclosed in international application WO 2020152229 A1 published on July 30, 2020.

[0091] The additional yeast product can be obtained from any process that yields a product rich in p-glucans. The additional yeast product corresponds to the insoluble fraction of the yeast. The insoluble fraction can be obtained using conventional production processes. The production process can be biochemical and / or mechanical. For example, the mechanical process can be carried out using glass beads, a high-pressure homogenizer, ultrasound, or microwaves. For example, the biochemical process can be carried out by autolysis, thermal plasmolysis, enzymatic hydrolysis, osmotic shock, or repeated freeze-thaw cycles. In particular, the process may include the following steps: autolysis or enzymatic hydrolysis of the whole yeast; then separation of the insoluble fraction and removal of the soluble fraction; and optionally, drying of the soluble fraction.The soluble fraction is conventionally called "yeast extract" and comprises the majority of amino acids, including glutamic acid, peptides, and minerals. The insoluble fraction comprises yeast cell walls, polymers, polysaccharides, nucleotides, and thermocoagulated proteins. Conventional extraction methods are disclosed in the reference work "Yeast Technology," 2. ème edition, 1991, G. Reed and TW Nagodawithana, published by Van Nostrand Reinhold, ISBN 0-442-31982-8.

[0092] The Bacillus inaquosorum strain, the cells derived from it, and their composition

[0093] The yeast product according to the invention can be used in combination with the Bacillus inaquosorum strain, and / or cells derived from it.

[0094] The Bacillus inaquosorum strain is a subspecies of the bacterium Bacillus subtilis (also known as Bacillus subtilis subsp. inaquosorum). The Bacillus inaquosorum strain was documented in the article by AP Rooney et al., "Phylogeny and molecular taxonomy of the Bacillus subtilis species complex and description of Bacillus subtilis subsp. inaquosorum subsp. nov," Int J Syst Evol Microbiol., 2009 Oct, 59, 2429-2436.

[0095] The Bacillus inaquosorum strain was deposited on September 12, 2024 with the International Depository Authority Agricultural Research Culture Collection (1815 N. University Street, Peoria, IL, 61604, USA) on behalf of Envera LIC LLC (220 Garfield Ave. West Chester, PA, 19380 USA) under the Treaty of Budapest and was given the registration number NRRL B-68429.

[0096] Cells can be obtained by culturing the Bacillus inaquosorum strain in a culture medium, according to methods well known to those skilled in the art, for example, those described in the book "Biotechnology, 5th edition, R. Scriban, Edition Tec. & Doc., 1999, ISBN: 2-7430-0309-X". Typically, a cell production method by culturing the strain implemented within the scope of the invention comprises the following steps:

[0097] - inoculation of a culture medium with an inoculum of the strain;

[0098] - culture under aerobic conditions, to obtain cell multiplication; and

[0099] - separation of biomass from its culture medium, to obtain cells.

[0100] The cells thus obtained are predominantly in their vegetative state. The "vegetative state" of a bacterium refers to the state of a bacterium under favorable conditions. The expression "predominantly in its vegetative state" means that at least 70% of the cells are in their vegetative state, preferably at least 80%, and more preferably at least 90%. An example of favorable conditions is a non-limiting culture medium at a temperature and pH conducive to bacterial growth. A non-limiting culture medium contains all the nutrients necessary for cell growth.

[0101] The cell production process may also include an intermediate step, between the aerobic culture stage and the biomass separation stage, of placing the cells under unfavorable conditions. The cells thus obtained are then predominantly in spore form.

[0102] The "spore-forming" form of a bacterium refers to the form of a bacterium in unfavorable conditions. The expression "predominantly in spore form" means that at least 70% of the cells are in spore form, preferably at least 80%, and even more preferably at least 90%. The spore-forming form is a resistant form that allows cells to withstand a hostile environment such as a lack of nutrients (i.e., a nutrient-limiting environment), water stress, significant variations in pH or temperature, or passage through the digestive tract.

[0103] Unfavorable cell conditions can be achieved, for example, by: not renewing the bacterial culture medium, stopping feeding into the culture medium, using a limiting culture medium, changing the temperature and / or pH, and controlling aeration and / or agitation to maintain a chlorophyll pressure above 20% in the culture medium and a CO2 pressure below 1.5% in the outgoing gases or their combination. It is also possible to obtain predominantly spore-forming cells by adding glutamic acid to the culture medium in an amount of approximately 0.75 g / L.

[0104] In a preferred embodiment, the cells are predominantly in spore form.

[0105] The cell production process may also include a subsequent step of drying the cells to obtain them in a dry form. Drying methods include, for example, freeze-drying, fluidized bed drying, or spray drying.

[0106] In a particular embodiment, the cells are in dry form. The expression "cells in dry form" advantageously means that the biomass obtained at the end of the cell production process advantageously comprises more than 90% dry matter, preferably more than 95% dry matter.

[0107] The Bacillus inaquosorum strain, and / or cells derived from it, can be formulated in any form suitable for administration to mammals; preferably to dogs.

[0108] Uses

[0109] The present invention relates to the use of a yeast product as a postbiotic in the field of oral hygiene and health.

[0110] The yeast product is used to prevent and / or reduce the formation of dental biofilm, particularly in dogs.

[0111] The action of the yeast product on the prevention and / or reduction of the formation of dental biofilm helps to maintain or restore the good health of the oral microbiota and therefore helps to prevent or treat infectious diseases of the oral cavity, in particular periodontal diseases.

[0112] The use of the yeast product, including its therapeutic or non-therapeutic use, depends on the subject to whom it is administered.

[0113] If the individual is in good health, the yeast product may be administered to maintain their well-being, including maintaining satisfactory oral hygiene. This is a non-therapeutic use of the yeast product. The yeast product may be formulated as a food supplement, a parapharmaceutical composition, or any other suitable non-therapeutic form. If the individual has an infectious disease of the oral cavity, or is at risk of developing one, the yeast product may be administered to prevent, limit, or treat the disease. This is a therapeutic use of the yeast product. The yeast product may be formulated as a pharmaceutical composition or any other suitable therapeutic form.

[0114] The method according to the invention can be used to treat a first episode of infectious disease of the oral cavity, in particular gingivitis or periodontitis, or a recurrence.

[0115] The yeast product can be administered alone. Alternatively, the product can be administered with another yeast product as defined above. Alternatively, the yeast product can be administered with another therapy, for example, an antibiotic or an antiseptic. Alternatively, the yeast product can be administered with another prebiotic, probiotic, and / or postbiotic product. Alternatively, the yeast product can be administered in combination with a mechanical or surgical procedure, including scaling, root planing, curettage, filling, restoration, or root canal treatment.

[0116] Topics

[0117] The yeast product may be administered to a subject. The subject may already have an infectious disease of the oral cavity, or be at risk of developing one. When the subject has such a disease, the term "patient" may be used interchangeably.

[0118] The yeast product can be administered to an animal, preferably to a pet, very preferably to a mammal, more preferably to a dog.

[0119] Effective quantity

[0120] Use to prevent and / or reduce dental biofilm formation (and the corresponding method of prevention and / or treatment) involves administering an effective amount of the yeast product to an animal, optionally in combination with the additional yeast product or a Bacillus inaquosorum strain, and / or cells derived from it. The effective amount, which may be administered in one or more doses, may be determined by the veterinarian, the breeder, or any other appropriate person. The exact amount to be administered may vary from animal to animal, depending on the species, age, weight, general condition of the animal, the absence or presence of an infectious disease of the oral cavity, and if so, its nature, severity, and / or extent. The effective amount may also vary depending on the desired therapeutic effect (prevention and / or reduction of dental biofilm formation).

[0121] Form of administration

[0122] The yeast product can be administered as such or in the form of a preparation or composition.

[0123] The yeast product can be administered as a dietary supplement to prevent and / or reduce the formation of dental biofilm, particularly in dogs. The dietary supplement may be in the form of a powder, sachet, capsule (encapsulated form), biscuit, treat, gummies, paste, sticks, strips, granules, or may be incorporated directly into food during its manufacture, especially pet food such as kibble. The yeast product can also be administered as a parapharmaceutical composition to prevent and / or reduce the formation of dental biofilm, particularly in dogs. The parapharmaceutical composition may be in the form of sachets, powders, capsules, softgels, dental pastes, gummies, or gel.

[0124] The yeast product can be administered as a pharmaceutical formulation to prevent and / or reduce dental biofilm formation, particularly in dogs. The pharmaceutical formulation may be available by prescription or over the counter. It can be administered using any combination of dosage and route of administration effective in achieving the desired therapeutic or prophylactic effect. The effective amount to be administered may vary from one individual to another, depending on the species, age, weight, general condition, the presence or absence of an infectious disease of the oral cavity, and, if applicable, its nature, severity, and / or extent, etc.

[0125] The pharmaceutical composition may be intended for topical administration or oral administration.

[0126] The pharmaceutical composition may include a physiologically acceptable excipient. A physiologically acceptable excipient may be one suitable for administration to mammals, particularly dogs.

[0127] The pharmaceutical composition may include at least one additional active pharmaceutical ingredient having soothing, anti-irritant, analgesic, pain-relieving, anti-inflammatory, healing, antibiotic, antipyretic or antifungal activity.

[0128] The pharmaceutical composition may include at least one additive, for example an additive chosen from the group consisting of preservatives, sweeteners, flavorings, thickening agents, colorings, humectants, disintegrating agents, absorption accelerators, lubricating agents, and mixtures thereof.

[0129] The pharmaceutical composition may be in any form suitable for administration to a subject, preferably a mammal, most preferably a dog.

[0130] The pharmaceutical composition may be in the form of tablets, pills, dragees, capsules, pearls, syrups, emulsions, ointments, pastes, gels, powders, sachets or injectable solutions.

[0131] The yeast product, optionally in combination with the additional yeast product or a Bacillus inaquosorum strain, and / or cells derived therefrom, or the composition comprising it(s), may be administered by incorporation into a dental medical device comprising the products, as defined above, to prevent or reduce the formation of dental biofilm, particularly in dogs.

[0132] Example

[0133] The following examples illustrate the invention without limiting it.

[0134] Products tested

[0135] Product 1: product in powder form having a pH between 6.3 and 6.7 (8.33% solution) and comprising less than 1% by weight of sodium chloride, 7.5 to 9.0% by weight of total nitrogen, 46.9 to 56.3% by weight of protein (nitrogen x 6.25) and 18 to 22% by weight of ash (excluding sodium chloride by total weight of the product, and comprising at least 94% by weight of dry matter (Springer® 4101 / 0-PW-L product marketed by Biospringer by Lesaffre, Lesaffre Group).

[0136] Product 2: product comprising 50 to 60% p-glucans, 1 and 5% mannans, 10% or less protein and 10% or less glycogen, by weight per total weight of product (Safglucan® product, Phileo by Lesaffre, Lesaffre group).

[0137] Product 3: Suspension of Bacillus inaquosorum bacteria filed on September 12, 2024 with Agricultural Research Culture Collection and having the registration number NRRL B-68429.

[0138] Study model

[0139] In vitro model of a polymicrobial biofilm comprising five different bacterial species mimicking a canine dental biofilm, namely Neisseria zoodegmatis CCUG 52598T, Corynebacterium canis CCUG 58627T, Porphyromonas cangingivalis DSMZ VPB 4874, Peptostreptococcus canis CCUG 57081 and an isolate of Enterococcus faecalis.

[0140] Analysis

[0141] The tests were performed in triplicate. A dental plaque biofilm was artificially created from five microorganisms described in the literature as being involved in periodontal disease: early plaque colonizers such as Neisseria zoodegmatis and Corynebacterium canis, other anaerobic bacteria such as Peptostreptococcus canis and Porphyromonas cangingivalis, and Enterococcus faecalis, which is involved in the development of periodontal disease in dogs. A bacterial suspension of each strain was deposited into the wells of a 96-well microplate placed on a Calgary device. A lid with pegs, previously incubated for two hours in artificial canine saliva, was then added to the microplate.After 48 hours of incubation at 37°C under microaerophilic conditions, biofilm formed on the surface of the PEGs. Following incubation, the PEGs were washed three times in 0.9% NaCl and transferred to a new microplate supplemented with product 1 at a concentration of 10% of each product. The plate was then incubated for 24 hours at 37°C under microaerophilic conditions, after which the percentage of biofilm formation inhibition was determined by Crystal Violet staining (by determining optical density).

[0142] Results regarding the inhibition of dental biofilm - product 1 only

[0143] The results (average of the three trials) concerning the inhibition of dental biofilm formation in the presence of product 1 are reported in Table 1.

[0144] Table 1

[0145] The results reported in Table 1 demonstrate that product 1 enables inhibition of dental biofilm formation in dogs.

[0146] Results regarding the inhibition of dental biofilm - products 1 and 2 in combination

[0147] The results concerning the inhibition of dental biofilm formation in the presence of product 1 (concentration: 250 g / L) and product 2 (concentration: 25 g / L) are reported in Table 2.

[0148] Table 2

[0149] The results reported in Table 2 demonstrate that products 1 and 2, used in combination, enable inhibition of dental biofilm formation in dogs.

[0150] Results concerning the inhibition of dental biofilm - products 1 and 3 in combination

[0151] The results concerning the inhibition of dental biofilm formation in the presence of product 1 (concentration: 250 g / L) and product 3 (concentration:

[0152] 10 9 CFU / mL) are reported in Table 3.

[0153] Table 3

[0154] The results reported in Table 3 demonstrate that products 1 and 3, used in combination, enable inhibition of dental biofilm formation in dogs.

Claims

DEMANDS 1. Non-therapeutic use of a yeast product, or a composition comprising it, to prevent and / or reduce the formation of dental biofilm, preferably canine dental biofilm, the yeast product being a yeast extract having a pH between 6.0 and 7.0; and comprising, by weight by total weight of the yeast product, 35 to 65% protein (nitrogen x 6.25), 10 to 30% ash and 3% or less sodium chloride.

2. Non-therapeutic use according to any one of the preceding claims, the yeast product being a yeast extract having a pH between 6.1 and 6.9; and comprising, by weight by total weight of the yeast product, 40 to 61% protein (nitrogen x 6.25), 13 to 27% ash and 2% or less sodium chloride.

3. Non-therapeutic use according to any one of the preceding claims, the yeast product being a yeast extract having a pH between 6.2 and 6.8; and comprising, by weight by total weight of the yeast product, 45 to 57% protein (nitrogen x 6.25), 16 to 24% ash and 1% or less sodium chloride.

4. Non-therapeutic use according to any one of the preceding claims, the yeast product being obtained by implementing a preparation process comprising the following steps: - obtaining a yeast cream by fermentation followed by concentration by centrifugation; - washing of the yeast cream obtained by adding water and centrifugal concentration; - heating the washed yeast cream to a temperature between 60 and 95°C, preferably between 75 and 90°C; - incubation under stirring of the yeast cream heated to a temperature of 55 to 90°C, preferably between 55 and 75°C, for a duration of 1 to 5 hours; - separation of yeast extract and yeast cell walls by centrifugation; - concentration of the yeast extract by evaporation and / or osmosis inverse; and - optionally drying of the concentrated yeast extract.

5. Non-therapeutic use according to any of the preceding claims, the product, or a composition comprising it, being administered to a healthy subject.

6. Non-therapeutic use to prevent and / or reduce the formation of dental biofilm, preferably canine dental biofilm, of the combination of the yeast product according to any of the preceding claims with an additional yeast product comprising at least 20% p-glucans, by weight per total weight of the additional yeast product, or of a composition comprising them.

7. Non-therapeutic use according to claim 6, the additional yeast product comprising, by weight per total weight of the additional yeast product: - at least 50%, preferably from 50 to 90%, preferably from 50 to 80%, preferably from 50 to 70%, preferably from 50 to 60%, of p-glucans; - 5% or less, preferably between 1 and 5%, of mannans; - 10% or less protein; and - 10% or less of glycogen.

8. Non-therapeutic use to prevent and / or reduce the formation of dental biofilm, preferably canine dental biofilm, of the combination of the yeast product according to any one of claims 1 to 5 with a Bacillus inaquosorum strain, and / or cells derived therefrom.

9. Non-therapeutic use according to claim 8, the Bacillus inaquosorum strain being the strain filed on September 12, 2024 with Agricultural Research Culture Collection under the Treaty of Budapest and having received the registration number NRRL B-68429.

10. Non-therapeutic use according to any of the preceding claims, the product, optionally in combination with the additional yeast product or a Bacillus inaquosorum strain, and / or cells derived therefrom, being administered to a subject orally.

11. Non-therapeutic use according to any of the preceding claims, the composition comprising the yeast product, optionally in combination with the additional yeast product or a Bacillus inaquosorum strain, and / or cells derived therefrom, being selected from a food supplement or a parapharmaceutical composition.

12. Yeast product, or a composition comprising it, for therapeutic use to prevent and / or reduce the formation of dental biofilm, preferably canine dental biofilm, the yeast product being a yeast extract having a pH between 6.0 and 7.0; and comprising, by weight by total weight of the yeast product, 35 to 65% protein (nitrogen x 6.25), 10 to 30% ash and 3% or less sodium chloride.

13. The yeast product, or a composition comprising it, for use according to claim 12, the product being administered to a subject having a disease of the oral cavity, or at risk of having one.

14. The composition comprising the yeast product, optionally in combination with the additional yeast product or a Bacillus inaquosorum strain, and / or cells derived therefrom, for use according to any one of claims 12 or 13, the composition being a pharmaceutical composition.

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