Fermented dairy food that contains live bacteria and is stable at ambient temperature, and method for preparing same

Abstract

The invention relates to a fermented dairy food that does not require refrigeration and contains microencapsulated live microorganisms that are beneficial to health. This product is obtained using a preparation method comprising the following steps: standardising the milk; pasteurising the mixture; fermenting the pasteurised milk mixture via inoculation and incubation of starter microorganisms; adding microencapsulated lactic acid and / or probiotic microorganisms to the fermented milk mixture; and applying a further heat treatment, followed by rapid cooling to ambient temperature.

Description

[0001] FERMENTED DAIRY FOOD WITH LIVE BACTERIA STABLE AT ROOM TEMPERATURE AND METHOD OF PREPARATION

[0002] TECHNICAL FIELD

[0003] This disclosure relates to food products for human consumption. Specifically, it relates to a fermented dairy product containing microencapsulated active microorganisms that enhance the probiotic activity of the food. This product also undergoes two stages of pasteurization, allowing it to be stored and preserved at room temperature.

[0004] DESCRIPTION OF THE STATE OF THE ART

[0005] Fermented dairy beverages have been part of the human diet for centuries. Their consumption has been associated with numerous health benefits, thanks to their unique composition and the fermentation process. These beverages are an excellent source of probiotics, live microorganisms that benefit gut health by helping to restore and maintain the balance of the gut microbiota. This contributes to improved digestion by facilitating nutrient absorption, reducing the symptoms of digestive disorders, helping to strengthen the immune system by stimulating the production of antibodies and immune cells, and reducing inflammation, which is linked to the prevention of chronic diseases.

[0006] Despite their numerous benefits, one of the main characteristics of fermented dairy beverages is their requirement for refrigerated storage. This is primarily because low temperatures help prevent the degradation of essential dairy nutrients, such as vitamins and proteins. Furthermore, refrigeration preserves the food's organoleptic characteristics for longer, and, most importantly, it slows bacterial growth, preventing spoilage. However, due to considerations of convenience, durability, and accessibility, the need for foods that can be stored at room temperature is becoming increasingly evident. Foods that do not require refrigeration play a crucial role in human nutrition and in various other contexts.Their importance lies in several aspects, such as ease of storage and availability under diverse conditions, as they can be transported and stored in places without access to electricity, making them essential in remote areas or during power outages. Their importance, therefore, extends to various areas, from survival to daily sustenance.

[0007] To address this need, various products have developed solutions that allow fermented dairy beverages to be stored at room temperature. For example, patent CN108184999, which discloses a high-fat yogurt that can be stored at room temperature and a method for preparing it, is a current state-of-the-art product. Specifically, this patent provides a yogurt preparation method comprising the steps of mixing raw milk, cream, sweeteners, and stabilizing ingredients; homogenizing; pasteurizing; fermenting; performing secondary homogenization; carrying out secondary pasteurization; cooling; and packaging.

[0008] Furthermore, patent CN109619180 discloses a live bacterial fermented dairy product that can be stored at room temperature and a method for preparing it. The method comprises the steps of mixing pre-sterilized, antibiotic-free milk, sweetener, cream, starch, and a stabilizing agent to obtain a mixed solution; homogenizing, sterilizing, cooling, adding flavoring, inoculating with a starter culture, fermenting to the endpoint to obtain yogurt base material, and demulsifying the yogurt base material, sterilizing, cooling, filling, and mixing with a probiotic culture. Specifically, patent CN109619180 states that the product has a stable probiotic content, a good flavor, and can be stored for 3-4 months at room temperature.

[0009] Considering these developments, it would seem that the need to offer a fermented dairy food that can be preserved at room temperature has been solved mainly by adding a second cycle of pasteurization or sterilization to the production method.

[0010] However, adding this step means that the probiotic bacteria will not be viable in the final product to perform their essential functions for consumer health. This is mainly because the viability, or ability to multiply and metabolize, and metabolic activity of lactic acid bacteria and / or probiotics decline as a result of exposure to industrial heat treatments such as thermization, pasteurization, and ultra-pasteurization, among others. Probiotics are live, heat-sensitive microorganisms. Therefore, when food is subjected to high temperatures during pasteurization, many of these beneficial microorganisms die or lose their viability, significantly reducing their quantity and effectiveness.

[0011] Responding to this opportunity, this development provides a solution that, in addition to offering a fermented dairy product that can be stored at room temperature, guarantees that the probiotic bacteria are live and available in the product, ensuring the consumer receives its beneficial effects. To achieve this, manufacturing processes have been developed that incorporate lactic acid bacteria and / or bacteria with probiotic activity that have undergone a prior microencapsulation process.

[0012] In this regard, it has been disclosed in the state of the art that microencapsulation enables the preservation of the viability and stability of lactic acid and / or probiotic bacteria present in food matrices that have been subjected to stress conditions such as physical, chemical or biochemical treatments. For example, the article “Influence of encapsulation on the survival of probiotics in food matrix under simulated stress conditions” (Afzaal M, Saeed F, Hussain M, Ismail Z, Siddeeg A, Al-Farga A, Aljobair MO. Influence of encapsulation on the survival of probiotics in food matrix under simulated stress conditions. Saudi J Biol Sci. 2022 Sep;29(9): 103394. doi: 10.1016 / j.sjbs.2022.103394. Epub 2022 Jul 25), evaluates the influence of encapsulation on the viability and stability of Bifidobacterium bifidium from yogurt under simulated gastrointestinal conditions.The study found that yogurt containing encapsulated bacteria had decreased acidity, while maintaining lactose levels, viscosity, and syneresis during storage. Furthermore, cell viability in yogurt with free probiotics was shown to be lower than in encapsulated probiotics when exposed to simulated gastric and intestinal juice.

[0013] Similarly, the article “Functional exploration of free and encapsulated probiotic bacteria in yogurt and simulated gastrointestinal conditions” (Afzaal M, Khan AU, Saeed F, et al. Food Sci Nutr. 2019; 7: 3931-3940. https: / / doi.org / 10.1002 / fsn3.1254) reports a study that evaluates the effect of microencapsulation on the viability and stability of probiotic bacteria in yogurt under simulated gastrointestinal conditions, using sodium alginate and carrageenan microcapsules, finding that microencapsulation improved the survival of probiotic bacteria in carrier foods.

[0014] Although these documents reveal the use and advantages of microencapsulation of lactic acid bacteria as a mechanism for preserving the viability of bacteria in dairy products subjected to simulated gastric digestion conditions, no solution is identified that proposes microencapsulated bacteria in heat-treated dairy products after fermentation that can be stored at room temperature and retain cell viability.

[0015] BRIEF DESCRIPTION

[0016] This disclosure refers to a fermented dairy food containing live bacteria, which is stable and microbiologically safe (i.e., poses no health risks or hazards) at room temperature throughout its shelf life, and also refers to the method of preparing this food. The method comprises the steps of modifying milk with additives, stabilizers, vitamins, and minerals; pasteurizing the mixture; fermenting the pasteurized milk mixture by inoculating and incubating starter microorganisms; adding lactic acid bacteria and / or microencapsulated probiotics to the fermented milk mixture; and applying a second heat treatment followed by rapid cooling to room temperature. DETAILED DESCRIPTION

[0017] This disclosure refers to a shelf-stable fermented dairy product containing microencapsulated live microorganisms. It also refers to the preparation method for obtaining this fermented dairy product, which includes the following steps: preparing the milk, pasteurizing the mixture, fermenting the pasteurized milk mixture by inoculating and incubating starter microorganisms, adding microencapsulated lactic acid microorganisms and / or probiotics to the fermented milk mixture, and applying a second heat treatment. Finally, it is cooled to room temperature.

[0018] For the purposes of this document, all percentages expressed, unless expressly stated otherwise, are weight-weight percentages (%w / w). Furthermore, "around" is understood to mean a difference of at least ±10% from the defined value.

[0019] Formulation of fermented dairy food

[0020] Firstly, a shelf-stable fermented dairy product containing microencapsulated live microorganisms was developed. The components of this dairy product include fermented milk, additives, stabilizers, vitamins and minerals, and microencapsulated lactic acid bacteria and / or probiotics. Specifically, these microencapsulated microorganisms are selected from among lactic acid bacteria, probiotics, or both.

[0021] The components included in the developed dairy food include, but are not limited to, fermented milk between 50% w / w and 100% w / w or its equivalent in milk solids; additives up to 50% w / w; stabilizers between 0.5% w / w and 3% w / w up to 10% w / w; vitamins and minerals between 0.001% w / w and 0.5% w / w; and microencapsulated lactic acid microorganisms and / or probiotics with a viable cell count between 100 and 100. 3 UFC / gy IxlO 8CFU / g. For the purposes of this document, "milk solids equivalent" means that, in the formulation of a dairy product, fermented milk in its liquid state (between 50% and 100% of the final product) or, alternatively, an equivalent amount of milk solids may be used. This means that, instead of using liquid milk directly, concentrated ingredients containing the same essential components of milk, such as proteins, fats, and lactose, in powder or solid form, could be used. This alternative is commonly used to adjust the product composition or improve efficiency in the manufacturing process. For example, if a product requires liquid milk, but for logistical or formulation reasons it is more practical to use powdered milk, these milk solids can be added to achieve the same proportion of nutrients and characteristics that would be obtained with liquid milk.

[0022] For the purposes of this development, "fermented milk" refers to milk that meets standard quality parameters and has undergone a fermentation process caused by the action of lactic acid bacteria. For the purposes of this invention, it is understood that the milk provides a nutritious environment for these bacteria and also serves as the basis for the consumable product.

[0023] In another embodiment, the fermented dairy food of the present invention may include milk solids, which can be defined as the essential components of milk that remain after water removal. These components include proteins, lactose, fats, minerals, and vitamins, which represent the nutritional value of milk and are also responsible for its organoleptic characteristics, such as flavor and texture. Milk solids are obtained through various technologies, such as vacuum evaporation, spray drying, roller drying, tangential flow filtration, and freeze-drying, among others.

[0024] For the purposes of this invention, lactic acid bacteria are defined as microorganisms capable of metabolizing carbohydrates (primarily lactose) into lactic acid through fermentation processes. This process not only transforms milk into products such as yogurt, kumis, and other fermented foods, but also acidifies the environment, slowing the proliferation of undesirable bacteria, including those responsible for spoilage or pathogens. In addition to their fermentation capacity, lactic acid bacteria produce bioactive compounds, such as bacteriocins and antimicrobial peptides, which contribute to product preservation. These bacteria are also considered technological strains due to their ability to impart essential organoleptic characteristics, such as viscosity, flavor, body, and mouthfeel.

[0025] In one embodiment, the food of the present invention includes one or more strains of lactic acid bacteria, or technological strains. The lactic acid bacteria are selected from, but not limited to, Streptococcus, Lactobacillus, Lactococcus, Lacticaseibacillus, and Lactiplantibacillus, or any other name assigned to them by modifications to international taxonomic classifications. In a preferred embodiment, the lactic acid bacteria strains are Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus, and are present at a concentration between 10 and 100. 3 UFC / gy IxlO 8 UFC / g, in a concentration between IxlO 5 UFC / gy IxlO 8 UFC / g, or in a concentration between IxlO 6 UFC / gy IxlO 7 CFU / g, preferably at a concentration around 1xlO 7 UFC / g.

[0026] In one embodiment, the food of the present invention includes one or more probiotic, or functional, strains. For the purposes of the present invention, functional strains are understood to be strains of probiotic bacteria with documented benefits for digestion, the immune system, cognitive and emotional well-being, skin health, dental health, eye health, vaginal health, and other areas. In some embodiments of the present invention, the functional or probiotic strains are selected from, but are not limited to, bacteria such as Akkermansia muciniphila, Bacillus coagulans, Bacillus subtilis, and Bifidobacterium animalis subsp.lactis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium longum, Clostridium butyricum, Escherichia coli, Faecalibacterium prausnitzii, Lactobacillus acidophilus, Lactobacillus brevis, Lactobacillus gasseri, Lactobacillus helve ti cus, Lactobacillus paracas ei, Lactiplantibacillus plantarum, Lactobacillus rhamnosus, Limosilactobacillus reuteri, Lacticaseibacillus casei, Limosilactobacillus fermentum, Propionibacterium freudenreichii, Streptococcus thermophilus, however, some yeasts with probiotic potential such as Saccharomyces boulardii could also be considered. In a preferred embodiment, the strains of probiotic bacteria are Bifidobacterium bifidium and Lactobacillus plantarum. The probiotic bacteria are at a concentration between 1x10. 3 UFC / gy 1x10 8 UFC / g, preferably in a concentration around 1x10 6 CFU / g.

[0027] Preferably, the lactic acid bacteria and probiotic bacteria are microencapsulated to isolate and protect them from the product components and from pasteurization. For the purposes of the present invention, microencapsulation is understood to occur with mixtures of milk-derived protein (casein, sodium caseinate, whey, milk, or casein protein concentrates and isolates), vegetable protein (pea, lentil, soy, sunflower, among others), gelatin, chitosan, prebiotic fibers (fructooligosaccharides (FOS) and galactooligosaccharides (GOS)), pectin, alginate, gums (arabic, xanthan, guar), agar-agar, carrageenan, microcrystalline cellulose, native and modified starches, and other cellulose derivatives. In a preferred embodiment, the microencapsulation materials include, but are not limited to, a milk or vegetable protein and a gum.

[0028] For the purposes of the present invention, the fermented dairy food is understood to comprise additives, particularly up to 50% additives. The additives included in the developed dairy food include, but are not limited to, a mixture of sugar, proteins, sweeteners, flavor modulators, flavorings, antioxidants, preservatives, and acidity modulators, and combinations thereof.

[0029] In a preferred embodiment of the present invention, the fermented dairy food comprises acidity modulators at concentrations of 0.01% to 5%. Acidity modulators are substances added to food to adjust and maintain its pH within an optimal range, enhancing flavor and maintaining proper consistency. The fermented dairy food of the present invention may include acidity modulators such as citric acid, lactic acid, malic acid, or tartaric acid, among others. The fermented dairy food may also include, as additives, proteins of various types and origins, such as powdered milk proteins, whey protein concentrates or isolates, micellar casein, caseinates, and hydrolyzed collagen, among others.

[0030] Fermented dairy food also includes other additives such as antioxidants, soluble or insoluble fibers, fatty acids, amino acids, enzymes, peptides, polysaccharides, polyphenols, phytosterols, botanical extracts, edible fungi, their parts or extracts, among others.

[0031] In some forms, fermented dairy products may be sweetened with fruit preparations, flavor enhancers, or sweeteners, among other things. For example, some fermented dairy products contain a caloric sweetener, such as sugar, syrup, glucose, dextrose, fructose, galactose, sucrose, or mixtures thereof. In other forms, fermented dairy products are sweetened with a non-caloric sweetener, such as saccharin, aspartame, or sucralose, among others widely known in the field.

[0032] In some varieties, the fermented dairy product may be flavored with natural or artificial flavorings. In one variety, the formulation may have natural flavors, such as blackberry, strawberry, raspberry, cherry, mixed berries, orange, grapefruit, lemon, tangerine, passion fruit, peach, yellow fruits, soursop, dragon fruit, papaya, coconut, gooseberry, dulce de leche, coffee, among others.

[0033] For the purposes of the present invention, stabilizers are understood to be products added to the mixture to maintain the product's consistency and stability throughout its shelf life. The addition of these products aims to prevent phase separation, syneresis, and protein precipitation, thus maintaining the quality and organoleptic properties established for the product stored at room temperature throughout its shelf life. In one embodiment of the present invention, the fermented dairy product comprises stabilizers at concentrations of 0.5% to 3%, which are compatible with the encapsulated bacteria and do not interfere with their release in the digestive system.For the purposes of the present invention, stabilizers include, but are limited to, a mixture of starches, gelatin, pectin and gums, alginates, carrageenans, carboxymethylcellulose, milk solids, calcium, sodium and / or potassium carbonates, potassium and / or sodium orthophosphate, calcium, sodium and / or potassium polyphosphate, alone or in mixtures, and others known to a person reasonably skilled in the art. In a preferred embodiment, the stabilizers are starch, pectin, and gelatin.

[0034] In one embodiment, the fermented dairy food comprises additives that are a mixture of sugar, proteins, sweeteners, flavor modulators, flavorings, antioxidants, preservatives, and acidity modulators at a concentration of between 0.01% w / w and 5% w / w, and stabilizers that are a mixture of starches, gelatin, pectin, and gums at a concentration of between 0.5% w / w and 3% w / w. This mixture is preferred to enable the matrix to withstand the second heat treatment and to preserve the sensory properties of the product.

[0035] Additionally, the developed fermented dairy product contains vitamins and minerals at concentrations between 0.001% and 0.5%. In some formulations, the fermented dairy product includes vitamins such as vitamin A, vitamin D, vitamin E, vitamin K, vitamin C, B vitamins such as B1 (thiamine), B2 (riboflavin), B3 (niacin), B5 (pantothenic acid), B6 ​​(pyridoxine), B7 (biotin), B9 (folic acid), and B12 (cobalamin), and minerals such as calcium, phosphorus, magnesium, zinc, iron, potassium, selenium, and manganese.

[0036] In some forms, the developed fermented dairy product can be presented in different ways. For example, it can be in liquid form, such as a ready-to-drink fermented dairy product like yogurt, kumis, or fermented dairy beverage, among others. The developed fermented dairy product can also be in powder form, meaning dehydrated and ready to be reconstituted with water or other beverages, ideal for transport and long-term storage. Additionally, the developed fermented dairy product can also be in a spoonable form. In some forms, the developed fermented dairy product may be packaged in bottles, cups, boxes, bags with single servings, or in packages for two or more servings, and after manufacturing, it can be stored at room temperature for its shelf life before being opened for the first time.

[0037] For the purposes of the present invention, the developed fermented dairy food may have functional forms, which correspond to mixtures of different strains of technological and / or functional bacteria. Each strain is selected according to the desired benefit. In some forms of the present invention, the fermented dairy food may be in specific formulations for children, young adults, older adults, athletes, or women.

[0038] In one form, the fermented dairy food comprises between 0 and 3.5% fat and between 2 and 15% protein.

[0039] The resulting fermented dairy product is a beverage of dairy origin fermented with live bacteria that remain viable throughout the product's shelf life when stored at room temperature. The shelf life of this fermented dairy product can be up to 12 months or even longer.

[0040] Method of producing fermented dairy food

[0041] Secondly, the present development is directed to the method for obtaining the fermented dairy food that does not require refrigeration and that contains microencapsulated live microorganisms, described in the previous point.

[0042] Adequacy

[0043] The method for obtaining the fermented dairy product described herein preferably begins with the reception and analysis of the milk to ensure compliance with quality parameters, both microbiological and physicochemical. Subsequently, the milk is processed. For the purposes of this invention, the milk processing involves the addition of additives, stabilizers, vitamins, and minerals to standardize the milk solids content in terms of fat and protein. Specifically, to achieve standardized milk fat content according to the desired product designation (whole milk with a minimum of 2.5%, semi-skimmed milk with a minimum of 1.5%, or skimmed milk with a maximum of 0.8%), the fat content is generally expected to be between 0 and 15%, between 0 and 3.5%, or between 10 and 15%, or even the fat-to-protein ratio can be 2:1.As for protein, it is expected to be found in an amount between 2 and 15%.

[0044] For the purposes of the present invention, the mixture may also contain functional ingredients such as antioxidants, prebiotic fibers, fatty acids, amino acids, enzymes, peptides, polysaccharides, polyphenols, phytosterols, botanical extracts, edible fungi, their parts or extracts, among others.

[0045] For the purposes of this development, the mixture of milk with additives, stabilizers, vitamins and minerals is called "dairy base".

[0046] Homogenize and pasteurize

[0047] For the purposes of this development, once the dairy base is ready, the next step is to homogenize and pasteurize the mixture. Homogenization is a mechanical process that reduces the size of the milk fat globules, dispersing them uniformly throughout the liquid. This is achieved by forcing the dairy base at high pressure through small openings. For the purposes of this development, homogenization can be carried out at a temperature between 55 and 65°C and a pressure between 130 and 150 bar. Pasteurization can be performed for a minimum of 5 minutes, or between 5 and 20 minutes, at a temperature between 80 and 86°C, or at the time and temperature deemed necessary by a person with a reasonable level of expertise to ensure the reduction of pathogenic microorganisms to safe levels established by current regulations and the inactivation of enzymes that could affect product quality.In a preferred embodiment, homogenization is performed prior to pasteurization.

[0048] Fermentation

[0049] Following pasteurization, the pasteurized milk mixture must be fermented by inoculating and incubating lactic acid bacteria, which are the starter microorganisms. This fermentation takes place at a temperature between 35 and 45°C for the time necessary to reach a pH between 4.9 and 4. Temperatures below 35°C could result in a less productive and more expensive process, favor the proliferation of microorganisms (mesophytes or pathogens that spoil the product), or a slower metabolic response. Consequently, the necessary amount of lactic acid would not be generated to give the product the desired profile and for the effective formation of the gel that gives the dairy beverage its texture.On the other hand, temperatures above 45°C can significantly reduce the viability of fermenting microorganisms, affecting efficiency. Some lactic acid bacteria may even produce lactic acid at an uncontrolled rate before their activity decreases due to the heat, resulting in an excessively acidic fermented food (which may be unpleasant for consumers or could destabilize proteins), leading to a weak gel or even phase separation (syneresis). Regarding pH, if the value is below 4, the resulting product would be extremely acidic to the consumer's palate, while if it is above 4.9, the necessary conditions for gel formation, which gives the fermented product its texture and stability, would not be present.

[0050] Addition of microencapsulated lactic acid bacteria and / or probiotics

[0051] Next, microencapsulated lactic acid bacteria and / or probiotics are added to the dairy base (or fermented milk mixture). This addition can be done before, during, or after the fermentation stage. If the lactic acid bacteria and / or microencapsulated probiotics are added before fermentation begins, it is necessary to mix them thoroughly at the time of addition to ensure homogeneous distribution throughout the base. Subsequently, the lactic acid bacteria, or bacteria used for technological purposes, ferment the dairy base, while the microencapsulated bacteria remain inactive and homogeneously distributed throughout the product.

[0052] In the alternative where lactic acid microorganisms and / or microencapsulated probiotics are added during fermentation, their incorporation must be carried out through a mixing process that ensures homogeneous distribution throughout the product. This procedure must be performed before the pH reaches the threshold of 5.0 to 5.4, at which point the formation of the protein network responsible for the product's viscosity intensifies. If the bacteria are incorporated after the pH drops below this limit, there is a risk of uneven distribution within the product volume. Furthermore, the agitation required for mixing could alter the structure of the forming gel, compromising the final texture and making it difficult to achieve the desired product viscosity.

[0053] This alternative is preferable in cases where the microcapsule coating material is sensitive or permeable in matrices with pH levels above 6.0. In this scenario, the capsules should be incorporated after fermentation has begun, once the pH has dropped to a level that guarantees the integrity and impermeability of the microcapsules.

[0054] Similarly, this incorporation method is suitable when the ingredient containing the microencapsulated bacteria includes fast-growing or highly fermentative lactic acid bacteria strains. For the purposes of this development, "fast-growing or highly fermentative lactic acid bacteria" refers to strains capable of reducing the pH of the dairy base to 4.5 within 6 hours. Because microencapsulation processes are not completely effective, free cells may remain and compete with the added lactic acid culture for fermentation. This competition can alter the organoleptic characteristics and quality of the final product. Therefore, the microcapsules should be added at a stage where the starter cultures have already established a competitive advantage.

[0055] Finally, in the alternative where lactic acid microorganisms and / or microencapsulated probiotics are added after fermentation begins or at the end of the fermentation process, especially in low-viscosity fermented dairy beverages, the addition of the microencapsulated microorganisms should be carried out in later stages of mixing and / or recirculation of the already fermented base. This procedure ensures the homogeneous distribution of the microencapsulated microorganisms in the final product, preserving the quality and desired characteristics of the food.

[0056] In other formulations, microencapsulated lactic acid microorganisms and / or probiotics are added during fermentation. For the purposes of the present invention, the selection of microencapsulated lactic acid strains, as well as microencapsulated probiotic strains, shall be made in accordance with the preferred functional characteristics of the fermented dairy product.

[0057] Once the desired pH values ​​are reached, a cooling stage of the fermented mixture is carried out at a temperature between 10 and 20°C or between 18 and 25°C to decrease the metabolic activity of the bacteria and therefore the acidification of the product.

[0058] Heat treatment

[0059] The method culminates in a second heat treatment applied to the mixture obtained in the previous stage. In a preferred embodiment, this second heat treatment is performed at a temperature between 90 and 120°C for 45–120 seconds, more specifically between 90 and 95°C for 45–60 seconds, followed by cooling to room temperature in no more than 5 seconds to ensure the physicochemical and organoleptic stability of the finished product. This second heat treatment eliminates free or non-microencapsulated microorganisms, allowing the finished product to be transported and / or stored at room temperature or without refrigeration while maintaining the defined microbiological, physicochemical, and organoleptic quality characteristics throughout its shelf life.

[0060] Once the product has completed the second heat treatment stage, it can be packaged using standard (aseptic) techniques to prevent contamination by environmental microorganisms. The fermented dairy product can be stored in its packaging at room temperature for up to 12 months or even longer.

[0061] At the end of the process, a fermented dairy product (yogurt, dairy product, spoonable dairy product, dairy beverage, kumis, powder, etc.) is obtained that has undergone at least two heat treatments (for example, two pasteurization treatments), one before and one after fermentation. This fermented dairy product can be stored and preserved at room temperature and contains live bacteria that may have probiotic activity. To ensure that the probiotic bacteria remain viable after the second heat treatment, they have undergone a prior microencapsulation process.

[0062] Microencapsulated bacteria are coated with a harmless material that isolates them from the product matrix and protects them from heat treatments, allowing them to remain viable throughout their shelf life at room temperature. The microencapsulated bacteria present in the fermented dairy product are released from the microcapsules in the digestive system as a result of the interaction between gastric enzymes and the pH of bile salts with the coating material. The formulation of the fermented product is designed so that, despite heat treatments, it remains stable; that is, there is no agglomeration or precipitation of the protein, phase separation, or syneresis during the product's shelf life when stored at room temperature. EXAMPLES

[0063] Example 1. Industrial test of one ton of fermented dairy beverage with the addition of microencapsulated probiotics

[0064] An industrial trial was conducted in which one ton of fermented dairy beverage was manufactured using traditional methods without microencapsulated bacteria as a reference standard, and one ton of fermented dairy beverage was manufactured with the addition of microencapsulated probiotics. After manufacturing, microbiological, physicochemical, and sensory analyses were performed to determine the impact of the addition of microencapsulated bacteria on the established quality parameters for the product during its shelf life under storage conditions at ambient temperature in Sopó, 25°C, and 35°C.

[0065] The manufacturing method is described below:

[0066] Adequacy

[0067] • Standardize the milk for a fat content (2.7 to 3.1%) and protein content (1.6 to 1.9%).

[0068] Mixture of ingredients

[0069] The mixture consists of milk, additives, stabilizers, vitamins, and minerals. The mixture should be recirculated for a minimum of 10 minutes to ensure homogeneity.

[0070] Homogenization

[0071] • Perform homogenization at a pressure between 130 and 150 bar.

[0072] • The homogenization temperature should be kept between 55 and 65 °C.

[0073] Pasteurization (First heat treatment)

[0074] • Pasteurize the mixture at a temperature between 80 and 86°C.

[0075] • Maintain a holding time of around 6 minutes in the pasteurization equipment.

[0076] • The product outlet temperature must be between 40 and 45°C. Fermentation • Add the starter cultures (promoting lactic acid bacteria) at a rate of 500U per 2500L of dairy base (U being the units defined by the culture manufacturer) and the microencapsulated probiotic bacteria (in the modality that is before fermentation).

[0077] • Add the microencapsulated probiotic bacteria at a rate of 9x10 6 UFC / g.

[0078] • Mix for 15 minutes.

[0079] • Carry out the acidification by incubating at a temperature between 40 and 42°C until a pH of 4.9 - 4.0 is reached.

[0080] Cooling

[0081] • Cool the promoted mixture to a temperature between 18 and 25 °C after completing the acidification.

[0082] Second thermal treatment

[0083] • Apply a heat treatment at a temperature between 90 and 93 °C for 45-60 seconds.

[0084] • Cool the product to a temperature between 20 and 25 °C after heat treatment.

[0085] Temporary storage

[0086] • The product must be stored for a maximum of 24 hours before moving on to the final stages.

[0087] Packed

[0088] • The product must be aseptically packaged in bottles, cups, bags, or any other format compatible with the dosing and packaging technology available on the manufacturing line.

[0089] Details of composition and parameters of fermented dairy beverage with viable microencapsulated probiotic bacteria for storage at room temperature

[0090] Example 2. Analysis of the finished product after manufacturing

[0091] After the manufacturing process of Example 1, the finished product (FP) was analyzed according to established quality parameters. The results of the physicochemical, microbiological, and sensory analyses are shown below:

[0092] Table 1. Initial results of the product, physicochemical parameters

[0093] Pasteurized mixture = before fermentation; Dairy base = the fermented product that could then be flavored and that mixture would be called the finished product.

[0094] Table 2. Initial results of the product: microbiological parameters

[0095] Table 3. Initial results of the finished product: physicochemical parameters.

[0096] Table 4. Initial results of finished product microbiological parameters.

[0097] Regarding the initial sensory analysis (immediately after manufacturing) of the standard fermented dairy beverage (without microencapsulated bacteria) and the fermented dairy beverage with microencapsulated probiotic bacteria for storage at room temperature, the following results were mostly (greater than 80%) for both beverages:

[0098] Color: suitable;

[0099] - Aroma: fermented dairy, characteristic aroma, fermented dairy flavor, characteristic flavor, sweetness level, acidity level: a good balance;

[0100] Consistency (liquids): suitable liquid; and

[0101] Texture: homogeneous and creamy.

[0102] Example 3. Digestion process to release lactic acid microorganisms and / or microencapsulated probiotics

[0103] After manufacturing, samples were taken of the control product without added microencapsulated bacteria and of the test product with microencapsulated bacteria. These samples underwent a digestion process to break down the microcapsules and release the probiotic bacteria, allowing for a CFU / g count of product to be performed and compared with the count obtained before digestion and with the counts of the control product. The results are shown below:

[0104] Table 5. Viability analysis of microencapsulated bacteria after the second heat treatment.

[0105] Subsequently, counter-samples were taken from both the control batch without microencapsulated bacteria (25240115A) and the batch containing microencapsulated bacteria (71006316). Their organoleptic characteristics and physicochemical and microbiological parameters were monitored periodically for 150 days from the date of manufacture. The results of this shelf-life monitoring are shown below:

[0106] Regarding the microbiological parameters, it was confirmed that for the control group (without microcapsules), from 30 to 150 days, the following parameters were defined: total coliforms <10, total count (CFU / ml) <10, molds (CFU / ml) <10, yeasts (CFU / ml) <10, and E. coli absent. For the assay with microencapsulated bacteria – stored at room temperature, from 30 to 150 days, the following parameters were defined: total coliforms <10, total count (CFU / ml) <10, molds (CFU / ml) <10, yeasts (CFU / ml) <10, and E. coli absent. For testing with microencapsulated bacteria - storage at 25 °C and 35 °C from 30 to 150 days, a total coliform count <10, total count (CFU / ml) <10, molds (CFU / ml) <10, yeasts (CFU / ml) and E. coli absence were defined.

[0107] Regarding the sensory properties during the shelf life of a standard dairy beverage (without microencapsulated bacteria), from day 30 to day 150 the following results were mostly observed (greater than 80%):

[0108] Color: suitable;

[0109] - Fermented dairy aroma, characteristic aroma, fermented dairy flavor, characteristic flavor, sweetness level, acidity level: a good balance; Consistency (liquids): suitable liquid (40%) and viscous (50%);

[0110] Texture: homogeneous and creamy;

[0111] Product preference: I like it (70%).

[0112] Regarding the dairy beverage corresponding to the trial with microencapsulated bacteria at room temperature, from day 30 to day 150 the following results were mostly (greater than 80%):

[0113] Color: suitable;

[0114] - Fermented dairy aroma, characteristic aroma, fermented dairy flavor, characteristic flavor, sweetness level, acidity level: a good balance;

[0115] Consistency (liquids): suitable liquid (50%) and viscous (50%);

[0116] Texture: homogeneous and creamy;

[0117] Product preference: I like it (90%).

[0118] Finally, a physicochemical analysis was also performed in terms of viscosity (cP), pH, acidity (°TH), and visual inspection of the control (without microencapsulated bacteria), the dairy beverage with microencapsulated bacteria stored at room temperature, the dairy beverage with microencapsulated bacteria stored at 25°C, and the dairy beverage with microencapsulated bacteria stored at 35°C. These analyses confirmed that the same parameters were obtained, demonstrating that the addition of microcapsules does not alter the final product. Although after day 30, visual analysis showed syneresis in the dairy beverages containing microencapsulated bacteria stored at 25°C and 35°C, it was evident that standard stirring homogenized the base again, and therefore the product quality was not affected.

[0119] Compatibility with the infrastructure and developed formulation:

[0120] The fermented dairy beverage product with added microencapsulated probiotic bacteria at the start of fermentation can be manufactured using the company's existing industrial infrastructure without significant modifications, thanks to formulations specifically developed for this process. Physicochemical compatibility:

[0121] The addition of microencapsulated probiotic bacteria does not alter the performance of the formulated fermented dairy base, demonstrating physicochemical compatibility with the dairy matrix. This allows the product to remain stable at room temperature even after undergoing the second heat treatment.

[0122] Viability of microencapsulated bacteria:

[0123] The microencapsulated probiotic bacteria survive and remain viable after the second heat treatment applied after fermentation, validating the effectiveness of the microencapsulation process and their resistance to the industrial conditions used.

[0124] Stability during storage:

[0125] The microbiological, physicochemical, and sensory parameters of the fermented dairy beverage with microencapsulated probiotics were stable and met the defined standards for the product for a period of at least 150 days under storage conditions at room temperature, 25°C, and 35°C. The results are comparable to those obtained for the control product (without the addition of microencapsulated bacteria). Although slight syneresis was observed after day 30 of storage, this does not represent a significant impact on product quality, as syneresis is easily corrected by stirring.

[0126] Product viability and functionality:

[0127] The industrial trial confirms that it is possible to manufacture a fermented dairy beverage that, even when stored at room temperature for at least 150 days, retains its microbiological, physicochemical, and sensory properties, guaranteeing a safe product that meets established quality standards. Furthermore, it was found that the microencapsulated probiotic bacteria remain viable and in adequate quantities even after the second heat treatment, ensuring their functionality as a probiotic ingredient.

Claims

CLAIMS 1. A method for obtaining a fermented dairy food comprising the steps of: i. preparing the milk by adding additives, stabilizers, vitamins and minerals; ii. pasteurizing the milk mixture obtained in step i); iii. fermenting the pasteurized milk mixture by inoculating and incubating starter microorganisms; iv. adding lactic acid microorganisms and / or microencapsulated probiotics to the fermented milk mixture; and v. heat-treating the mixture obtained in step iv) at a temperature between 90 and 120°C for 15-120 seconds and subsequently cooling to room temperature.

2. The method according to Claim 1 wherein in step i) the adaptation process corresponds to a standardization of the milk in terms of fat between 0 and 15% and protein between 2 and 15%.

3. The method according to Claim 1 wherein in step i) functional ingredients such as antioxidants, prebiotic fibers, fatty acids, amino acids, enzymes, peptides, polysaccharides, polyphenols, phytosterols, botanical extracts, edible fungi, their parts or extracts may also be added.

4. The method according to Claim 1, wherein step ii) includes a homogenization step.

5. The method according to Claim 1, wherein step ii) is performed under conditions of a minimum time of 5 min and a temperature between 80 and 86°C.

6. The method according to Claim 1, wherein the fermentation of step iii) is carried out at a temperature between 35 and 45°C and the time required to reach a pH between 4.9 and 4.

7. The method according to Claim 1, wherein step iv) can be performed before, during, and after step iii).

8. The method according to Claim 1, wherein in step iv) the lactic acid microorganisms and / or microencapsulated probiotics are added before the fermentation begins.

9. The method according to Claim 1, wherein in step iv) the lactic acid microorganisms and / or microencapsulated probiotics are added during fermentation.

10. The method according to Claim 1, wherein the microencapsulated microorganisms of step iv) are lactic acid, probiotic, or lactic acid and probiotic.

11. The method according to Claim 1, wherein the heat treatment of step v) allows the elimination of free or non-microencapsulated microorganisms, so that the finished product can be transported and / or stored at room temperature or without refrigeration, preserving the microbiological, physicochemical and organoleptic quality characteristics defined for the product throughout its shelf life.

12. A fermented dairy food obtained by the method described in Claim 1, which is a dairy beverage fermented with live bacteria that remain alive throughout the shelf life of the product stored at room temperature.

13. The fermented dairy food according to Claim 12, wherein the entire shelf life is up to 12 months.

14. A fermented dairy food comprising a content of: fermented milk between 50% and 100% or its equivalent in milk solids; additives up to 50%; stabilizers between 0.5% and 3%; vitamins and minerals between 0.001% and 0.5%; microencapsulated lactic acid microorganisms and / or probiotics with a viable cell count between 100,000 and 1100. 3 UFC / gy IxlO 8UFC / g.

15. The fermented dairy food of Claim 14, wherein the additives are a mixture of sugar, proteins, sweeteners, flavor modulators, flavors, antioxidants, preservatives and acidity modulators that are in concentrations from 0.01 to 5%, and the stabilizers include a mixture of starches, gelatin, pectin and gums, which are in concentrations from 0.5% to 3%.

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