JP2018538009A - Composition containing probiotics and methods of use thereof - Google Patents

Abstract

The present invention provides a composition comprising probiotics, the composition comprising an effective amount of an amphiphile that enhances the viability of the probiotics in the composition when the composition is subjected to heat. In a mixture with.
[Selection] Figure 1

Description

  The field of the invention relates to compositions and uses of probiotics, including Pediococcus acidilactici.

  Probiotics are beneficial microorganisms that exist naturally in the digestive (GI) tract of humans and animals. In 2001, the World Health Organization announced that probiotics “live microorganisms, which, when administered in adequates, confer a health benfon on the probiotics when they are administered in sufficient amounts. ("Joint FAO / WHO Expert Consultation Report, 2001"). Health effects associated with many probiotics such as antibiotic-induced diarrhea, acute diarrhea, traveler diarrhea, allergies, respiratory and urinary tract infections, inflammatory bowel disease, irritable bowel syndrome, colon and bladder cancer, and A reduction in symptoms from rheumatoid arthritis has been reported. To date, Bifidobacterium and Lactobacillus are commercially available probiotics. However, these bacteria are sensitive to atmospheric exposure, elevated temperature, and gastric acid.

  Pediococcus acidilactici is a plant-derived probiotic that has been widely applied as a sausage cooker for human consumption and as an animal feed additive to improve animal health. Furthermore, P.I. acidilactici has been reported to be able to stimulate antibody production against broiler chicken coccidiosis parasite infection and ovalbumin antibody production in horses vaccinated with ovalbumin (Furr et al., Journal of Equine Science, 34: 1156). -1163 (2014)). The proliferation of both T and B cells is Detected in rats fed Pediococcus probiotics, a mixture of acidilactici and Saccharomyces bouraldiii.

  One of the difficulties of incorporating probiotics into food and pharmaceutical / nutritional supplement products for administration to a subject is the product, and the process of making the product, particularly sterilization under harsh conditions such as heat or Incompatibility with processing. These harsh conditions can cause significant loss of probiotic viability. Failure to effectively sterilize the product in the presence of probiotics without significant loss of viability results in additional processing steps in the manufacturing process, which increases complexity and cost.

  Thus, there is a need for new compositions and methods that include probiotics such as Pediococcus acidilactici, particularly methods for enhancing the viability of probiotics when subjected to heat.

  This preliminary knowledge is provided for informational purposes only. The admission that any of the foregoing information constitutes prior art to the present invention is not necessarily intended and should not be so construed.

  It is understood that both the above summary of the embodiments and the following detailed description are exemplary and therefore do not limit the scope of the embodiments.

  According to a non-limiting embodiment, in one aspect, the present invention is in a mixture with an effective amount of an amphiphile that enhances the viability of probiotics in the composition when the composition is subjected to heat. A composition comprising the probiotics as described above is provided.

  In another aspect, the present invention includes a probiotic that is in a mixture with an effective amount of an amphiphile that enhances the viability of the probiotic in the composition when the composition is subjected to heat. Provided is a container suitable for containing a food product comprising the composition.

  In another aspect, the present invention provides a container suitable for containing food, wherein a portion or surface of the container is coated with an effective amount of Pediococcus acidilactici.

In another aspect, the present invention provides a method for enhancing the viability of probiotics in a composition subjected to heat, the method comprising:
i) adding an effective amount of an amphiphile to the composition comprising probiotics;
ii) subjecting the composition to heat,
Thereby, the viability of the probiotic in the composition subjected to heat is enhanced.

  In another aspect, the present invention provides a method of treating a disease or condition characterized by inflammation in a subject in need thereof, wherein the method comprises an effective amount of the Pediococcus acidilactici pro described herein. Administering a biotic composition to the subject.

  In some embodiments, administration of Pediococcus acidilactici probiotics increases the number of anti-inflammatory M2 macrophage cells in the subject, thereby treating the disease or condition. In some embodiments, administration of Pediococcus acidilactici probiotics increases IL-10 production in the subject, thereby treating the disease or condition. In some embodiments, administration of Pediococcus acidilactici probiotics reduces the level of IL-6 and / or IL-23 in the subject, thereby treating the disease or condition.

  In some embodiments, the disease or condition characterized by inflammation is malignant tumor (cancer), arthritis, cardiovascular disease, hepatitis, infection, wound healing, pancreatitis, gastroesophageal reflux disease, diabetes, inflammatory bowel disease, Peptic ulcer disease, bronchitis, cholecystitis, appendicitis, bursitis, dermatitis, asthma, autoimmune disease, pelvic inflammatory disease, gout, trauma, foreign body infection, burn, dental treatment, tendonitis, rhinitis, Selected from the group consisting of mucositis and exposure to toxins such as chemicals and alcohol.

  In some embodiments, the Pediococcus acidilactici probiotic is the NRRL B-50517 strain. In some embodiments, the subject is a human.

In some embodiments, the subject is administered more than 1.0 × 10 ⁹ cfu probiotics. In some embodiments, the subject is administered probiotics greater than 4.0 × 10 ⁹ cfu.

  In some embodiments, the subject is administered one or more additional therapeutic agents. In some embodiments, the subject is administered one or more chemotherapeutic (anticancer) agents and / or radiation therapy in combination with Pediococcus acidilactici probiotics.

  In some embodiments, the subject is not administered another therapeutic agent. In some embodiments, the subject is not administered another probiotic.

  In another aspect, the present invention provides a composition comprising Pediococcus acidilactici probiotic. In some embodiments, the Pediococcus acidilactici is NRRL B-50517 strain. In some embodiments, the composition is a pharmaceutical composition.

  It is understood that both the foregoing summary and the following detailed description are exemplary and thus do not limit the scope of the invention. Other objects, features and advantages of the present invention will become apparent from the following detailed description. However, while the detailed description and specific examples illustrate specific embodiments of the present invention, various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description. It should be understood that it is given solely for the purpose.

  Those skilled in the art will appreciate that the drawings described below are for illustrative purposes only. The drawings are not intended to limit the scope of the present teachings in any way.

P. This is the effect of acidilactici 5051 probiotics on body fat percentage. P. This is the effect of acidilactici 5051 probiotics on IL-23 activity. P. This is the effect of acidilactici 5051 probiotics on IL-6 activity. This is the effect of Pediococcus probiotics on dogs suffering from pancreatitis. A 14-year-old female contraceptive toy poodle suffering from pancreatitis was treated with 100 mg KAMOSTAAL100 twice a day at a certain time (red dot), and the treatment was stopped approximately 2 and a half months later (red square). Pediococcus probiotics were applied (green dots) and stopped approximately one month later (green squares). After recurrence, Pediococcus probiotics were applied again (green circles) and continued for a period of time (green arrows). It is a flowchart of a manufacturing process outline of Pediococcus acdiactici NRRL B-50517. FIG. 3 is an exemplary container for food containing a probiotic composition. FIG.

  The present invention is based in part on the surprising discovery that the viability of probiotics such as Pediococcus acidilactici can be enhanced when combined with an effective amount of amphiphile when the composition is subjected to heat. The present invention is also based in part on the surprising discovery that an effective amount of Pediococcus acidilactici probiotic can treat a disease or condition characterized by inflammation.

  Reference will now be made in detail to embodiments of the present invention, which together with the drawings and the following examples serve to explain the principles of the invention. These embodiments are described in sufficient detail to enable those skilled in the art to practice the invention, other embodiments may be utilized, and structural, biological, and chemical changes may be made. It will be appreciated that may be made without departing from the spirit and scope of the invention. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

  For purposes of interpreting this specification, the following definitions will apply and where appropriate, terms used in the singular will also include the plural and vice versa. Where any definition set forth below conflicts with the use of the word in any other document, including any document incorporated herein by reference, the definition set forth below clearly indicates the opposite meaning. Unless otherwise intended (eg, in documents where the term is originally used) for the purpose of interpreting the specification and the associated claims. When used in conjunction with the term “comprising” in the claims and / or herein, the use of the word “a” or “an” may mean “one”, but “one” It is also consistent with the meanings of “above”, “at least one”, and “one or more than one”. The use of the term “or” in the claims is used to mean “and / or” unless it is clearly indicated that it refers only to the alternative, or unless the alternative is mutually exclusive. However, this disclosure supports definitions that refer only to alternatives and “and / or”. As used herein and in the claims, the term “comprising” (and any form of comprising such as “comprise” and “comprises”) means “having”. "(And any form of having such as" have "and" has ")," including "(and any form of including such as" includes "and" include "), or" containing " (And any form of containing such as “contains” and “contain”) are inclusive or unconstrained and do not exclude additional unlisted elements or method steps. Further, where the term “comprising” is used to describe one or more embodiments, those skilled in the art will recognize that, in some specific cases, the embodiment (s) may alternatively be “ It can be understood that it may be described using the phrase “consisting essentially of” and / or “consisting of”. As used herein, the term “about” means up to ± 10% of the numerical value used with it.

  Those skilled in the art may refer to general reference documents for a detailed description of the known techniques or equivalent techniques discussed herein. These documents include, for example, Current Protocols in Molecular Biology (Ausubel et.al., eds.John Wiley & Sons, NY and its appendix), Current Protocols in Immunology. Wiley St Sons, NY, and its appendix), Current Protocols in Pharmacology (Enna et al., Eds. John Wiley & Sons, NY, and its appendix), and Remington: The Spicen, The Scicen. Williams & Willicin s, 2Vt edition (2005)).

Heat Resistant Probiotic Compositions and Methods of Making In one embodiment, the present invention provides an effective amount of an amphiphile that enhances the viability of probiotics in the composition when the composition is subjected to heat. A composition comprising probiotics in a mixture of The composition comprises an effective amount of probiotics when suitable for administration to a subject.

  In another embodiment, the invention includes a method of making a probiotic composition with improved viability in response to heat, wherein the method comprises an effective amount of an amphiphile comprising a probiotic. Adding to the product.

In another embodiment, the invention includes a method of enhancing the viability of probiotics in a composition subjected to heat, the method comprising:
i) adding an effective amount of an amphiphile to the composition comprising probiotics;
ii) subjecting the composition to heat,
Thereby, the viability of the probiotic in the composition subjected to heat is enhanced.

  The probiotic is not particularly limited. In some embodiments, the probiotic is mixed with an amphiphile as a lyophilized fermentation medium. In some embodiments, the probiotic is Pediococcus acidilactici. Pediococcus acidilactici probiotics that can be used in accordance with the present invention are not limited. In some embodiments, Pediococcus acidilactici is a strain that can survive above 65 ° C. and can grow under aerobic and anaerobic conditions and at pH 1-6.2. In some embodiments, the Pediococcus acidilactici is a strain deposited with the Agricultural Research Service (ARS) Patent Culture Collection as NRRL B-50517. The NRRL B-50517 strain is described in US Application No. 13 / 676,579, which is incorporated herein by reference.

  In some embodiments, Pediococcus acidilactici for use in the present invention may be selected for elevated temperatures, low pH values, and resistance to aerobic and anaerobic conditions.

In some embodiments, the composition comprises greater than about 1.0 × 10 ⁹ cfu probiotics. In some embodiments, the composition comprises greater than 4.0 × 10 ⁹ cfu probiotics.

  Amphiphilic molecules are molecules that have both polar and nonpolar moieties in their structure. Amphiphilic molecules generally have a hydrophobic portion of the molecule that faces the hydrophobic phase and a hydrophilic portion that faces the aqueous layer.

  Some of the chemical compounds that characterize these molecules are essential for acceptance of biological and industrial processes. In some embodiments, the amphiphile comprises lecithin, peanut butter, almond butter, soy butter, or cookie butter as a carrier of probiotics such as Pediococcus acidilactici NRRL B-50517 fermentation culture. In some embodiments, an excess amount of amphiphile is used, which is Acidilactici NRRL B-50517 fermentation culture can be protected from severe dry heat or heat treatment. Lecithin is phosphatidylcholine and may include natural mixtures of neutral and polar fats derived from plants and / or animals. This is an excellent emulsifier with low water solubility. In aqueous solution, the phospholipid can form either liposomes, bilayer sheets, micelles, or lamellar structures, depending on hydration properties and temperature. This produces a type of surfactant that is usually classified as amphiphilic.

  In some embodiments, the amphiphile is a lipid. The hydrophilic character is derived from the presence of polar or charged groups such as carbohydrates, phosphatos, carboxylic acids, sulfatos, aminos, sulfhydryls, nitros, hydroxys, and other similar groups. Hydrophobicity can be imparted by including nonpolar groups, including long chain saturated and unsaturated aliphatic hydrocarbon groups, and one or more aromatic, alicyclic, or heterocyclic groups. Such groups substituted by, but not limited to). Examples of amphiphilic compounds include, but are not limited to, phospholipids, amino lipids, and sphingolipids. Representative examples of phospholipids include phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, palmitoyloleoylphosphatidylcholine, lysophosphatidylcholine, lysophosphatidylethanolamine, dipalmitoylphosphatidylcholine, dioleoylphosphatidylcholine, disteolinyl phosphatidylcholine, Examples include, but are not limited to dilinoleoyl phosphatidylcholine. Other compounds lacking phosphorus, such as sphingolipids, glycosphingolipid families, diacylglycerols, and β-acyloxyacids also belong to the group designated as amphiphilic lipids.

  In some embodiments, the amphiphile comprises lecithin, peanut butter, almond butter, soy butter, or cookie butter. In some embodiments, the amphiphile is sunflower lecithin.

  An “effective amount” of an amphiphile described herein is a measure of the viability of the probiotic in response to heat as compared to the viability of the probiotic in the absence of the amphiphile. The amount that can be strengthened. In some embodiments, the probiotic viability is enhanced by at least about 2-fold compared to a composition lacking an effective amount of amphiphile. In some embodiments, viability is enhanced by at least about 25%, 50%, 75%, 2x, 3x, 4x, 5x, 6x, 7x, 8x, 9x, or 10x. Is done. In some embodiments, viability is enhanced by at least about 10 times compared to a composition lacking an effective amount of amphiphile.

  In some embodiments, the amphiphile to probiotic ratio (w / w) ranges from about 1:10 to about 25: 1. In some embodiments, the amphiphile to probiotic ratio (w / w) ranges from about 1: 1 to about 10: 1. In some embodiments, the amphiphile to probiotic ratio (w / w) is about 5: 1. In some embodiments, the amphiphile to probiotic ratio (w / w) is about 10: 1. In some embodiments, the amphiphile ratio can be greater than 25: 1. For example, in some embodiments, peanut butter flour and / or lecithin can be treated as a sugar carrier. In some embodiments, up to 1 part probiotic can be produced for 25 parts lecithin or peanut butter, but as much probiotics can be blended with the sugar carrier as desired. This means that 1 part probiotic can be successfully mixed with 100 parts (or even more) lecithin or peanut butter flour.

  In some embodiments, the amphiphile is mixed with the probiotic as a dry powder. In some embodiments, the amphiphile acts as a carrier and is combined with probiotics and oils to convert the probiotics to heat, eg, dry or liquid heat treatment such as sterilization processing for food processing. A lipid paste can be formed that makes it more resistant to. The oil is not particularly limited and may include edible oil. In some embodiments, the oil is a plant-derived oil, such as olive oil, coconut oil, soybean oil, fish oil, sunflower oil, canola oil (rapeseed oil), corn oil, peanut oil, and other vegetable oils, and butter and Animal oils such as lard may be included. In some embodiments, the oil is olive oil. In some embodiments, the amphiphile to probiotic to oil ratio (w / w) ranges from about 1: 10: 0.1 to about 10: 1: 25. In some embodiments, the amphiphile to oil ratio (w / w) ranges from about 1: 0.5 to about 0.5: 1. In some embodiments, the amphiphile to oil ratio (w / w) is about 1: 1.

  The nature of the heat is not necessarily limited and can include dry heat, water vapor, and liquid heat treatment. Liquid heat means that the product is immersed in a warm liquid such as water and the heat treatment is performed with the liquid.

  In some embodiments, the composition is subjected to dry heat. Dry heat treatment can be accomplished using an oven or hot plate without water and steam. In some embodiments, heating may be initiated at a first temperature and terminated at a second desired temperature. In some embodiments, the composition is immediately subjected to dry heat at the desired temperature without a heating step. In some embodiments, the desired temperature of dry heat is at least 50 ° C. In some embodiments, the dry heat is at least about 50 ° C, 51 ° C, 52 ° C, 53 ° C, 54 ° C, 55 ° C, 56 ° C, 57 ° C, 58 ° C, 59 ° C, 60 ° C, 61 ° C, 62 ° C. ° C, 63 ° C, 64 ° C, 65 ° C, 70 ° C, 75 ° C, 80 ° C, 85 ° C, 90 ° C, 95 ° C, or 100 ° C. In some embodiments, the dry heat is in the range of 65-100 ° C. The composition can be subjected to dry heat at a first temperature for a period of time and then exposed to one or more heat treatments at one or more different temperatures for a period of time.

  The duration of heat exposure is not particularly limited. In some embodiments, the composition is immediately subjected to heat and immediately removed from the heat. In some embodiments, the composition is subjected to heat for at least about 1 second. In some embodiments, the composition comprises at least about 1 second, 2 seconds, 4 seconds, 5 seconds, 6 seconds, 7 seconds, 8 seconds, 9 seconds, 10 seconds, 20 seconds, 30 seconds, 40 seconds. Second, 50 seconds, 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 35 minutes, Subject to heat for 40 minutes, 45 minutes, 50 minutes, 55 minutes, or at least 2 hours. In some embodiments, the composition is subjected to heat for at least about 10 minutes. In some embodiments, the composition is subjected to a first temperature of dry heat and then subjected to a second temperature of dry heat. In some embodiments, the first temperature is about 50-65 ° C. and the second temperature is about 85 ° C. or higher. In some embodiments, the composition is subjected to heat at a first temperature for at least about 1 second to at least about 2 hours, and is subjected to heat at a second temperature for at least about 1 second to at least about 2 hours. Is done. In some embodiments, the composition is subjected to a first temperature overnight. In some embodiments, the composition is subjected to a second temperature overnight. In some embodiments, the composition is subjected to heat at a first temperature for about 30 minutes and subjected to heat at a second temperature for about 10 minutes. In some embodiments, the composition is subjected to heat at a first temperature for about 30 minutes and subjected to heat at a second temperature for about 30 minutes. In some embodiments, the purpose of multiple heat treatments at different temperatures is to ensure that the sample is at a temperature suitable for processing at the second temperature, such that the material is from room temperature, eg, 85. This is due to the fact that it can take time to warm to ° C. In some embodiments, the composition is pre-treated at 65 ° C. for 30 minutes to overnight and then transferred to 85 ° C. for a treatment time to achieve an appropriate temperature heat treatment.

  In some embodiments, the composition is subjected to liquid heat. In some embodiments, the liquid heat is at least about 50 ° C, 51 ° C, 52 ° C, 53 ° C, 54 ° C, 55 ° C, 56 ° C, 57 ° C, 58 ° C, 59 ° C, 60 ° C, 61 ° C, 62 ° C. ° C, 63 ° C, 64 ° C, 65 ° C, 70 ° C, 75 ° C, 80 ° C, 85 ° C, 90 ° C, or 95 ° C. In some embodiments, the liquid heat is at least about 50-65 ° C. In some embodiments, the liquid heat ranges from about 65 ° C to about 95 ° C. In some embodiments, the liquid heat is at least about 70 ° C, 75 ° C, or 80 ° C. In some embodiments, the liquid heat is at least about 81 ° C, 82 ° C, 83 ° C, 84 ° C, 85 ° C, 86 ° C, 87 ° C, 88 ° C, 89 ° C, 90 ° C, 91 ° C, 92 ° C, 93 ° C. ° C, 94 ° C, or 95 ° C.

  In some embodiments, the composition is immediately subjected to liquid heat and immediately removed from the heat. In some embodiments, the composition is subjected to liquid heat for at least about 1 second. In some embodiments, the composition is subjected to liquid heat for at least about 1 second to about 1 hour. In some embodiments, the composition comprises at least about 1 second, 2 seconds, 4 seconds, 5 seconds, 6 seconds, 7 seconds, 8 seconds, 9 seconds, 10 seconds, 20 seconds, 30 seconds, 40 seconds. Second, 50 seconds, 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 35 minutes, Subject to liquid heat for 40 minutes, 45 minutes, 50 minutes, 55 minutes, or at least 1 hour. In some embodiments, the composition is subjected to heat for at least about 5 minutes. In some embodiments, the composition is subjected to heat for at least about 10 minutes.

  In some embodiments, the composition is contacted with a food, pharmaceutical, or dietary supplement and subjected to heat with the product. In some embodiments, the heated food, pharmaceutical, or dietary supplement is dispensed into a container containing the composition or otherwise combined with the composition. In some embodiments, the heated food, pharmaceutical, or dietary supplement is contacted immediately after the heat treatment without undergoing a significant cooling step prior to combining the product with the probiotic composition. Thus, in some embodiments, subjecting the composition to the thermal step of part ii) comprises contacting the composition with a food, pharmaceutical, or dietary supplement, wherein the food, pharmaceutical, or The dietary supplement has already been subjected to heat prior to contact with the composition. In some embodiments, the composition will be in a container into which the heated food, pharmaceutical, or dietary supplement is dispensed.

Containers In some embodiments, the present invention further provides containers suitable for containing foods, pharmaceuticals, or dietary supplements that include the compositions described herein. In some embodiments, the container is made from a material comprising glass, plastic, paper / carton, aluminum, or a dried plant fruit shell. The composition and shape of the containers are not limited on the condition that they are suitable for containing foods, pharmaceuticals, or dietary supplements. In some embodiments, the container is coated with a probiotic composition described herein.

  In some embodiments, the container is coated with an effective amount of Pediococcus acidilactici. In some embodiments, the Pediococcus acidilactici is NRRL B-50517 strain. In some embodiments, a composition comprising a probiotic, such as Pediococcus acidilactici, including the NRRL B-50517 strain, and an amphiphile, such as sunflower lecithin, provided herein, Part is coated. In some embodiments, the container further comprises a food, nutritional supplement, or pharmaceutical product. In some embodiments, the food is oil, vinegar, yogurt, fruit product, applesauce, dairy product, beverage, candy, snack confectionery, juice, or dessert food, eg, cookie, cake, JELLO, fruit bar, fruit Includes custard or tiramisu. In some embodiments, after the heated food, dietary supplement, or pharmaceutical is dispensed into the coated container, the container is sealed.

Methods of Treatment In accordance with one non-limiting exemplary embodiment, in one aspect, the invention provides a method of treating a disease or condition characterized by inflammation in a subject in need thereof, the method comprising: Administering to the subject an amount of Pediococcus acidilactici probiotic. In some embodiments, the subject is administered a composition described herein. In some embodiments, the composition comprises a probiotic and an amphiphile described herein.

  As used herein, “treatment” and all its forms and tenses (including, for example, “treating”, “trated”, and “treatment”) refer to therapeutic treatment. In certain embodiments of the invention, a subject in need of treatment includes a subject already suffering from a pathological disease or condition of the invention (eg, including cancer), in which case the treatment comprises a subject (eg, Refers to administering a therapeutically effective amount of the composition (including a human or other mammal in need of treatment) so that the subject has an improvement in the signs or symptoms of the pathological condition of the invention. This improvement may be any observable or measurable improvement. Thus, those skilled in the art will understand that treatment may improve the patient's condition, but may not be a complete cure of the disease or pathological condition.

  The subject to be treated herein is not limited. In some embodiments, the subject is a mammal, bird, reptile, or fish. Mammals that can be treated according to the present invention include humans, dogs, cats, horses, mice, rats, guinea pigs, sheep, cattle, pigs, monkeys, subject to diseases characterized by inflammation or other pathological conditions. Examples include, but are not limited to, apes. The terms “patient” and “subject” are used interchangeably. In some embodiments, the subject is a human.

  Pediococcus acidilactici probiotics can be administered once, twice, three times, four times, five times, six times, seven times, eight times, nine times, ten times, eleven times, or twelve times a day. In some embodiments, Pediococcus acidilactici probiotics are administered four times daily, twice daily, or once daily. In some embodiments, Pediococcus acidilactici probiotics are administered every 2 hours, every 4 hours, every 6 hours, every 8 hours, every 10 hours, every 12 hours, or every 24 hours. In some embodiments, the Pediococcus acidilactici probiotic is administered once daily.

  The duration of administration of Pediococcus acidilactici probiotics can vary from individual to individual being treated / administered, depending on the individual case and the disease or condition being treated. In some embodiments, Pediococcus acidilactici probiotics can be administered continuously over a period of days, weeks, months, or years, or can be administered intermittently, in which case the individual Treats a disease or condition after receiving Pediococcus acidilactici probiotics over a period of time, not receiving treatment for a period of time, and then resuming treatment as needed. For example, in some embodiments, an individual being treated may receive Pedicococcus acidilactici probiotics of the invention daily, every other day, every third day, every fourth day, two days a week, three days a week, four days a week, Administered 5 days a week or 7 days a week. In some embodiments, the individual receives Pediococcus acidilactici probiotics for 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 It is administered for months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year or longer.

  In some embodiments, administration of Pediococcus acidilactici probiotics increases the number of anti-inflammatory M2 macrophage cells in the subject, thereby treating the disease or condition. In some embodiments, the anti-inflammatory M2 macrophage cells are about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, compared to the non-therapeutic level. About 80%, about 90%, about 100%, about 125%, about 150%, about 175%, about 200%, about 250%, about 300%, about 350%, about 400%, or about 450%, about Increase by 500%, about 600%, about 700%, about 800%, about 900%, or about 1000% or more.

  In some embodiments, administration of Pediococcus acidilactici probiotics increases IL-10 production in the subject, thereby treating the disease or condition. In some embodiments, IL-10 production is about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80 compared to non-therapeutic levels. %, About 90%, about 100%, about 125%, about 150%, about 175%, about 200%, about 250%, about 300%, about 350%, about 400%, or about 450%, about 500% , About 600%, about 700%, about 800%, about 900%, or about 1000% or more.

  In some embodiments, administration of Pediococcus acidilactici probiotics reduces the level of IL-6 and / or IL-23 in the subject, thereby treating the disease or condition. In some embodiments, the level of IL-6 and / or IL-23 is about 10%, about 20%, about 30%, about 40%, about 50%, about 60% compared to the non-therapeutic level. , About 70%, about 80%, about 90%, or about 100%.

  In some embodiments, administration of Pediococcus acidilactici probiotics increases the number of anti-inflammatory M2 macrophage cells in the subject, along with increased IL-10 production, and optionally IL-6 in the subject. And / or IL-23 is decreased.

  In some embodiments, the disease or condition characterized by inflammation is malignant tumor (cancer), arthritis, cardiovascular disease, hepatitis, infection, wound healing, pancreatitis, gastroesophageal reflux disease, diabetes, inflammatory bowel disease, Peptic ulcer disease, bronchitis, cholecystitis, appendicitis, bursitis, dermatitis, asthma, autoimmune disease, pelvic inflammatory disease, gout, trauma, foreign body infection, burn, dental treatment, tendonitis, rhinitis, Selected from the group consisting of mucositis and exposure to toxins such as chemicals and alcohol.

  As used herein, “cancer” refers to a pathophysiological condition characterized by unregulated and / or proliferative cell growth and the ability to induce such growth. Epithelial and non-epithelial malignancies such as acute lymphoblastic leukemia, acute myelogenous leukemia, adrenocortical carcinoma, AIDS-related cancer, AIDS-related lymphoma, anal cancer, astrocytoma (eg, cerebellum and Including cerebrum, basal cell carcinoma, cholangiocarcinoma, bladder cancer, bone cancer, brainstem glioma, brain tumor (eg, ependymoma medulloblastoma, medulloblastoma, supratentorial primitive neuroectodermal, visual pathway and hypothalamus Glioma), cerebral astrocytoma / malignant glioma, breast cancer, bronchial adenoma / carcinoid, Burkitt lymphoma, carcinoid tumor (eg, gastrointestinal), cancer of unknown primary site, central nervous system lymphoma, cervical Cancer, chronic lymph Leukemia, chronic myelogenous leukemia, chronic myeloproliferative disorder, colon cancer, colon cancer, cutaneous T cell lymphoma, endometrial cancer, ependymoma, esophageal cancer, Ewing tumor, extrahepatic bile duct cancer, eye cancer (eg, eye) Internal melanoma, including retinoblastoma, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), germ cell tumor (eg, including extracranial, extragonadal, ovary), gestational villus Tumor, glioma, hairy cell leukemia, head and neck cancer, squamous cell carcinoma of the head and neck, hepatocellular carcinoma, Hodgkin lymphoma, hypopharyngeal cancer, islet cell carcinoma (eg, including pancreatic endocrine), Kaposi sarcoma, larynx Cancer, leukemia, lip and oral cancer, liver cancer, lung cancer (eg, including non-small cell carcinoma), lymphoma, macroglobulinemia, malignant fibrous histiocytoma / osteosarcoma of bone, melanoma, Merkel cell carcinoma, Mesothelioma, metastatic squamous cervical cancer of unknown primary origin, Cavity cancer, multiple endocrine tumor syndromes, multiple myeloma / plasmacytoma, mycosis fungoides, myelodysplastic syndrome, myelodysplastic / myeloproliferative disorder, myeloma, nasal and sinus cancer, nasopharyngeal cancer, Neuroblastoma, non-Hodgkin lymphoma, oral cancer, oral cancer, osteosarcoma, oropharyngeal cancer, ovarian cancer (including ovarian, epithelial ovarian cancer, germ cell tumor), ovarian low-grade tumor, pancreatic cancer, Paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, pineoblastoma and supratentorial primitive neuroectodermal tumor, pituitary tumor, plasmacytoma / multiple myeloma, pleura Pulmonary blastoma, pregnancy and breast cancer, primary central nervous system lymphoma, prostate cancer, rectal cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, soft tissue sarcoma, uterine sarcoma, Sezary syndrome, skin cancer (eg, non- Melanoma or melanoma), small intestine cancer, supratentorial primitive neuroectodermal Tumor, T-cell lymphoma, testicular cancer, throat cancer, thymoma, thymoma and thymic carcinoma, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, choriocarcinoma (eg, including pregnancy), rare in children and adults Cancer, urethral cancer, endometrial uterine cancer, uterine sarcoma, vaginal cancer, virus-induced cancer (eg, including HPV-induced cancer), vulvar cancer, Waldenstrom macroglobulinemia, Wilms tumor, and Examples include, but are not limited to, women's cancer.

  Pediococcus acidilactici probiotics that can be used in accordance with the present invention are not limited. In some embodiments, Pediococcus acidilactici is a strain that can survive above 65 ° C. and can grow under aerobic and anaerobic conditions and at pH 1-6.2. In some embodiments, the Pediococcus acidilactici is a strain deposited with the Agricultural Research Service (ARS) Patent Culture Collection as NRRL B-50517. The NRRL B-50517 strain is described in US Application No. 13 / 676,579, which is incorporated herein by reference.

  In some embodiments, Pediococcus acidilactici for use in the present invention may be selected for elevated temperatures, low pH values, and resistance to aerobic and anaerobic conditions.

In accordance with the treatment methods of the invention, a “therapeutically effective amount” or “effective amount” is administered to the subject. As used herein, a “therapeutically effective amount” or “effective amount” is an amount sufficient to reduce, suppress, or alleviate one or more symptoms associated with a disease or condition. In some embodiments, the subject is administered more than 1.0 × 10 ⁹ cfu probiotics. In some embodiments, the subject is administered probiotics greater than 4.0 × 10 ⁹ cfu.

  In some embodiments, the subject is administered one or more additional therapeutic agents. In some embodiments, the one or more additional therapeutic agents are those commonly used to treat a disease or condition characterized by inflammation.

  In some embodiments, the subject is administered an anti-inflammatory drug in combination. In some embodiments, the administered Pediococcus acidilactici and the anti-inflammatory agent act synergistically. In some embodiments, the anti-inflammatory drug is a non-steroidal anti-inflammatory drug (NSAID). In some embodiments, the anti-inflammatory drug is antazoline, balsalazide, beclomethasone, betamethasone, budesonide, celecoxib, colchicine, deflazacort, dexamethasone, dexbuprofen, diclofenac, etanercept, etodolac, felbinac, fenoprofen, flumethazone, fluoro Metron, flurbiprofen, flurbiprofen, fluticasone, gentamicin, hydrocortisone, ibuprofen, indomethacin, ketoprofen, loteprednol, mefenamic acid, meloxicam, mesalazine, methylprednisolone, mometasone, nabumetone, naproxen, nepafenac, olsalazine, predoxolone Sulfasalazine, sulindac, tenoxicam, thiaprofen It is selected triamcinolone, and combinations thereof.

  In some embodiments, the subject is administered one or more anticancer agents and / or radiation therapy in combination with Pediococcus acidilactici probiotics to treat cancer in the subject.

In some embodiments, the anti-cancer agent is abiraterone acetate, abitrexate (methotrexate), Abraxane (paclitaxel albumin stabilized nanoparticle formulation), ABVD, ABVE, ABVE-PC, AC, AC-T. , ADCETRIS (brentuximab vedotin), ADE, Ado-trastuzumab emtansine, adriamycin (doxorubicin hydrochloride), adolcil (fluorouracil), afatinib dimaleate, affinitol (everolimus), aldara (imiquimod), aldesleukin, alemtumab Pemetrexed disodium), alloxy (palonosetron hydrochloride), ambochlorin (chlorambucil), ambochlorin (chlorambucil), Minolevulinic acid, Anastrozole, Aprepitant, Aredia (Pamidronate disodium), Arimidex (Anastrozole), Aromasin (Exemestane), Alanone (Neralabine), Diarsenic trioxide, Azera (Ofatumumab), Asparaginase Erwinia Chrysanthymy , Avastin (Bevacizumab), Axitinib, Azacitidine, BEACOPP, Becenum (Carmustine), Beleodaq (Belinostat), Belinostat, Bendamustine Hydrochloride, BEP, Bevacizumab Tomobimatomito , BiCNU (Carmustine), Boreomycin, Blinatumomab, Brynsite (Blinatumomab) Bortezomib, Bosrif (Bostinib), Bostinib, Brentuximab vedotin, Busulfan, Busulfex (Busulfan), Cabazitaxel, Cabozantinib-S-malate, CAF, Campus (Alemtuzumab), Camptosar (Irinotepecitabine hydrochloride) , CAPOX, carboplatin, CARBOPLATIN-TAXOL (carboplatin-taxol), carfilzomib, carmbris (carmustine), carmustine, carmustine implant, casodex (bicalutamide), CeeNU (lomustine) ceritinib, cervidin (salbinic acid salt) Replacement HPV bivalent vaccine), cetuximab, chlorambucil, CHLORAMBUCI L-PREDNISONE (chlorambucil-prednisone), CHOP, cisplatin, krafen (cyclophosphamide), clofarabine, clofarex (clofarabine), chloral (clofarabine), CMF, cometric (cabozantinib-S-malate), COPP , COPP-ABV, cosmegen (dactinomycin), crizotinib, CVP, cyclophosphamide, cyfos (ifosfamide), syramza (ramsylmab), cytarabine, cytarabine, liposome, cytosal-U (cytarabine), cytoxan (cyclohexane) Phosphamide), dabrafenib, dacarbazine, dacogen (decitabine), dactinomycin, dasatinib, daunorubicin hydrochloride, decitabine, degarelix, denilo Kindiftitox, denosumab, deposite (cytarabine liposome), depoform (cytarabine liposome), dexrazoxane hydrochloride, docetaxel, doxyl (doxorubicin hydrochloride liposome), doxorubicin hydrochloride, doxorubicin hydrochloride liposome, Dox-SL (doxorubicin hydrochloride liposome) , DTIC-Dome (dacarbazine), EFdex (fluorouracil), Elitek (rasbricase), Elens (epirubicin hydrochloride), eloxatin (oxaliplatin), eltrombopaguolamine, imendo (aprepitant), enzalutamide, epirubicin hydrochloride , EPOCH, Erbitux (cetuximab), Eribulin mesylate, Elevige (Bismodegib), Erlotinib Acid salt, erwinase (asparaginase erwinia chrysanthemi), etopophos (etoposide phosphate), etoposide, etoposide phosphate, evacet (doxorubicin hydrochloride liposome), everolimus, evista (raloxifene hydrochloride) , Exemestane, fairston (toremifene), fasodex (fulvestrant), FEC, femara (retrozole), filgrastim, fludara (fludarabine phosphate), fludarabine phosphate, fluoroplex (fluorouracil), fluorouracil , Forex (methotrexate), Forex PFS (methotrexate), FOLFIRI, FOLFIRI-BEVACIZUMAB, FOLFI RI-CETUXIMAB, FOLFIRINOX, FOLFOX, forotin (pralatrexate), FU-LV, fulvestrant, gardashil (genetically modified HPV tetravalent vaccine), gardashil 9 (genetically recombined HPV 9-valent vaccine), gaziva (Obinutuzumab), gefitinib , Gemcitabine hydrochloride, GEMCITABINE-CISPLATIN (gemcitabine-cisplatin), GEMCITABINE-OXALIPLATIN (gemcitabine-oxaliplatin), gemtuzumab ozogamicin, gemzar (gemcitabine hydrochloride), diolotrifbelaine (fate) Imatinib mesylate), Giliadel (carmustine implant), Gliadelwafer (carmustine implant) ), Glucarpidase, Goserelin acetate, Halaven (Eribulin mesylate), Herceptin (Trastuzumab), Genetically modified HPV bivalent vaccine, Genetically modified HPV 9 valent vaccine, Genetically modified HPV tetravalent vaccine, Hycamtin (Topotecan hydrochloride), Hyper CVAD, ibrans (parvocyclib), ibritumomab tiuxetan, ibrutinib, ICE, iclusig (ponatinib hydrochloride), idamycin (idarubicin hydrochloride), idarubicin hydrochloride, ideralisive, ifex (ifosfamide), ifosfamide Ifosfamidum (ifosfamide), imatinib mesylate, imbrubica (ibrutinib), imiquimod, inwriter (axitinib), intro A (genetically modified interferon α-2b), iodine 131 tositumomab and tositumomab, ipilimumab, iressa (gefitinib), irinotecan hydrochloride, istodax (romidepsin), ixabepilone, ixempura (ixabepilone), jakafi (luxolitinibrate) , Jebutana (cabazitaxel), cadsira (Ado-trastuzumab emtansine), keoxifen (raloxifene hydrochloride), kepivans (palifermine), kytoluda (pembrolizumab), caiprolis (carfilzomib), lanreotide acetate, lapatinibide dosildo Lenvatinib mesylate, lenvima (lenvatinib mesylate), letrozole, leucovorin calcium, leukeran (chlorambucil) Leuprolide acetate, lebrin (aminolevulinic acid), lympholizin (chlorambucil), lipodox (doxorubicin hydrochloride liposome), cytarabine liposome, lomustine, lupron (leuprolide acetate), leupron depot (Lupron deprodot) Acetate), Lupron Depot-Ped (Lupronide acetate), Lupron Depot-3 Month (Lupron Depot-3 Month) (Lupronidepot), Lupron Depot-Lupron Depot- 4 (Month) (leuprolide acetate), Lymphorza (Olaparib), Marquibo (Vincristine sulfate liposome) Matulane (procarbazine hydrochloride), mechlorethamine hydrochloride, megges (megestrol acetate), megestrol acetate, mequinist (tramethinib), mercaptopurine, mesna, mesnex (mesna), metazolaston (temozolomide), Methotrexate, methotrexate LPF (methotrexate), mexate (methotrexate), mexate-AQ (methotrexate), mitomycin C, mitoxantrone hydrochloride, mitosaitrex (mitomycin C), MOPP, mozovir (prerixahol), mustarden (mechloretamine hydrochloride) , Mutamycin (mitomycin C), milleran (busulfan), mylosar (azacytidine), mylotag (Gemtuzumab ozogamicin), nanoparticle paclitaxel (paclitaxel albumin stabilized nanoparticle formulation), navelbine (vinorelbine tartrate), nelarabine, neosal (cyclophosphamide), newpogen (filgrastim), nexavar (sorafenib) Tosylate), nilotinib, nivolumab, norbadex (tamoxifen citrate), N plate (romiplostim), obinutuzumab, OEPA, ofatumumab, OFF, olaparib, omecetaxin mepesuccinate, oncaspar (pegaspargase), Ontac (Denileukin Difty Tox), Opdivo (Nivolumab), OPPA, Oxaliplatin, Paclitaxel, Paclitaxel Albumin Stabilized Nanoparticle Formulation, P D, parvocyclib, parifermine, palonosetron hydrochloride, disodium pamidronate, panitumumab, paraprat (carboplatin), paraplatin (carboplatin), pazopanib hydrochloride, pegase pargase, peginterferon alpha-2b, PEG-intron (peginterferon alpha -2b), pembrolizumab, pemetrexed disodium, pergeta (pertuzumab), pertuzumab, platinol (cisplatin), platinol-AQ (cisplatin), prelixahol, pomalidomide, pomalist (pomaridomide), ponatinib hydrochloride, pralatrexate hydrochloride, prednisone hydrochloride, prednisone hydrochloride Salt, Proleukin (Aldesleukin), Prolia (Denosumab), Promacter (Eltrombopag Lamin), Provenge (Cipleucel-T), Printol (Mercaptopurine), Purixan (Mercaptopurine), Radium dichloride 223, Raloxifene hydrochloride, Ramucirumab, Rasburicase, R-CHOP, R-CVP, Genetic recombination Human papillomavirus (HPV) bivalent vaccine, recombinant human papillomavirus (HPV) 9-valent vaccine, recombinant human papillomavirus (HPV) 4-valent vaccine, recombinant interferon alpha-2b, regorafenib, R-EPOCH, levlimido (lenalidomide) ), Rheumatorex (methotrexate), rituxan (rituximab), rituximab, romidepsin, romiplostim, rubidomycin (daunorubicin hydrochloride), ruxorichi Nibrinate, Sclerosol Intrapleural Aerosol (Talc), Siltuximab, Sipleuxel-T, Somatuline Depot (Lanreotide Acetate), Sorafenib Tosylate, Sprisel (Dasatinib), STANDARD V ), Steritarc (talc), Stiberga (Legorafenib), Sunitinib malate, Sutent (Sunitinib malate), Syratron (Peginterferon alfa-2b), Sylvant (Siltuximab), Sinovir (Sinovir), Thalidomide Synribo (Omacetaxin mepesuccinate), TAC, Taffinlar (Dabrafenib), Talc, Tamoxifen citric acid , Tarabine PFS (cytarabine), tarceva (erlotinib hydrochloride), targretin (bexarotene), tasigna (nilotinib), taxol (paclitaxel), taxotere (docetaxel), temodarl (temozolomide), temozolomide, temilidomide Thiotepa, Toposal (Etoposide), Topotecan hydrochloride, Toremifene, Tricel (Temcilirolimus), Tositumomab and Iodo I 131 Iodine Tositumomab, Tectect (dexrazoxane hydrochloride), TPF, Tramethinib, Trastuzumab, Toreanda (Bendamcetino hydrochloride) Arsenic acid), tie curve (lapatinib ditosylate), vandetanib, VAMP, vetivix (panitumumab), VeIP, Luban (vinblastine sulfate), Velcade (bortezomib), Versal (vinblastine sulfate), Vemurafenib, Bepsid (etoposide), Viadur (leuprolide acetate), Bidaza (azacytidine), Vinblastine sulfate, Vincasartin PFS (vincristine PFS) Salt), vincristine sulfate, vincristine sulfate liposome, vinorelbine tartrate, VIP, bismodegib, boraxase (glucarpidase), vorinostat, botrient (pazopanib hydrochloride), welcovorin (leucovorin calcium), zakori (crizotinib), Capecitabine), XELRI, Xgeva (denosumab), zofigo (radium dichloride 223) Xtandi (Enzalutamide), Yervoy (Ipilimumab), Zartrap (Ziv-Afribercept), Zelboraf (Bemurafenib), Zevalin (Ibritumomab Tiuxetane), Zinecard (Dexrazoxane Hydrochloride), Ziv -Selected from the group consisting of aflibercept, zoladex (goserelin acetate), zoledronic acid, zolinza (vorinostat), zometha (zoledronic acid), zidelig (idelarisib), zikadia (seritinib), and zytiga (aviraterone acetate). In some embodiments, the drug is paclitaxel, curcumin, docetaxel, ixabepilone, vinblastine, colchicine, Y-27632 fasudil, SU6656 dasatinib, HDAC inhibitor, ROCK inhibitor, parthenolide, costunolide, and ML-7 jazzpraquinolide Selected from the group consisting of (Jazplatinolide).

  In some embodiments, the subject is not administered another therapeutic agent and consists of or consists essentially of Pediococcus acidilactici probiotics optionally mixed with amphiphiles and / or food. Is administered.

  In some embodiments, the subject is administered one or more additional probiotics. In some embodiments, the subject is not administered another probiotic.

Compositions In some embodiments, the present invention provides compositions comprising Pediococcus acidilactici probiotics. In some embodiments, the composition comprises Pediococcus acidilactici NRRL B-50517. In some embodiments, the composition comprises an effective amount of Pediococcus acidilactici, including Pediococcus acidilactici NRRL B-50517. In some embodiments, the present invention provides a composition comprising an effective amount of amphiphile and optionally Pediococcus acidilactici probiotics mixed with the oils described herein. In some embodiments, the composition comprises a food, pharmaceutical, or dietary supplement in combination with an effective amount of amphiphile and optionally Pediococcus acidilactici probiotics mixed with oil.

  In some embodiments, the composition is a pharmaceutical composition. In some embodiments, the composition comprises a pharmaceutical composition comprising an effective amount of Pediococcus acidilactici comprising Pediococcus acidilactici NRRL B-50517 capable of treating one or more diseases or conditions characterized by inflammation. It is. In some embodiments, the composition comprises a Pediococcus acidilactici NRRL mixed with an effective amount of an amphiphile and optionally an oil capable of treating one or more diseases or conditions characterized by inflammation. A pharmaceutical composition comprising an effective amount of Pediococcus acidilactici comprising B-50517.

  In some embodiments, the pharmaceutical composition includes one or more pharmaceutically acceptable carriers or additives. Pharmaceutically acceptable carriers and additives are those that are compatible with the other ingredients in the formulation and biologically acceptable. Pediococcus acidilactici can be provided in combination with a pharmaceutically acceptable carrier, additive, or diluent. Suitable carriers, additives, and / or diluents include pharmaceutical grade starch, mannitol, lactose, magnesium stearate, sodium saccharine, talc, cellulose, glucose, sucrose (or other sugar), magnesium carbonate, gelatin, oil , Alcohols, surfactants, emulsifiers, or water (preferably sterilized). The composition may be a mixed preparation of the composition or a combined preparation for simultaneous use, separate use, or continuous use (including administration). The Pediococcus acidilactici can also be administered in sachets that need to be drunk after being added to a glass of water.

  In some embodiments, the composition is suitable for oral administration. In some embodiments, the composition is a tablet, capsule, pill, dragee, suspension, troche, emulsion, aqueous solution, liquid, gel, or syrup. In some embodiments, the composition can be delivered in the form of functional foods and / or beverages, as well as various supplements.

  Formulations of the present invention suitable for oral administration may be presented as separate units such as capsules, cachets, or tablets, each of which separates a predetermined amount of Pediococcus acidilactici in some embodiments. In, as powders or granules, which can be wet, spray dried or lyophilized; as a solution or suspension in an aqueous or non-aqueous liquid; or as an oil-in-water emulsion or a water-in-oil liquid emulsion.

  In some embodiments, tablets may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets are in free-flowing form such as powders or granules, optionally with binders (eg povidone, gelatin, hydroxypropylmethylcellulose), lubricants, inert diluents, preservatives, disintegrants (eg The active ingredient mixed with sodium starch glycolate, cross-linked povidone, cross-linked sodium carboxymethylcellulose), surfactant, or dispersant can be prepared by pressing in a suitable machine. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert diluent. Tablets may optionally be coated or scored and sustained active ingredient in, for example, using hydroxypropyl methylcellulose in different proportions to provide the desired release profile. Or it may be formulated to provide controlled release. Tablets may optionally be provided with an enteric coating to provide release in portions of the gastrointestinal tract other than the stomach.

  In some embodiments, the composition comprises tablets, pills, which may contain flavoring or coloring agents for immediate release, delayed release, modified release, sustained release, pulsatile release, or controlled release applications. Contains one or more of capsules, cavity suppositories, solutions, or suspensions.

  For aqueous suspensions and / or elixirs, the compositions of the present invention may be prepared by various sweetening or flavoring agents, coloring materials or pigments, emulsifiers and / or suspending agents, and dilutions such as water, propylene glycol, and glycerin. Can be combined with agents, as well as combinations thereof.

  In some embodiments, the compositions of the invention are formulated in unit dosage form for ease of administration and uniformity of dosage. The term “unit dosage form” as used herein refers to a physically discrete unit suitable as a single dose for the individual to be treated. In some embodiments, the composition comprises a predetermined “unit dose” or “unit dose” of one or more active compounds calculated to produce the desired effect in conjunction with the required pharmaceutical carrier. It is formulated into separate dosage units, each containing a “unit dose”.

  In some embodiments, the composition comprises gelatin capsules. In some embodiments, the gelatin capsules have an effective amount of P.A. with peach fruit powder at a dose of about 1-4 billion CFU. acidilactici NRRL B-50517 fermentation culture.

  While it is possible to administer Pediococcus acidilactici alone in accordance with the present invention, the Pediococcus acidilactici is typically supported as part of a product, particularly as a component of a food, dietary supplement, or pharmaceutical formulation. Administered on or in the body. These products typically contain additional components well known to those skilled in the art. In one embodiment, the composition comprises an effective amount of amphiphile and optionally Pediococcus acidilactici probiotics mixed with oil.

  In one embodiment, the Pediococcus acidilactici is used in accordance with the present description in food products such as supplements, beverages, or milk-based powders. As used herein, the term “food” is used broadly to cover human food and animal food (ie, feed). In one embodiment, the food is for human consumption.

  The food may be in the form of a solution or solid, depending on the use and / or application form and / or dosage form.

  When used as or in the preparation of a food such as a functional food, the composition of the present invention comprises a nutritionally acceptable carrier, a nutritionally acceptable diluent, a nutritionally acceptable additive, a nutritional May be used in combination with one or more of an acceptable adjuvant and a nutritionally active ingredient.

  By way of example, the composition of the present invention comprises a soft drink, a beverage comprising fruit juice or whey protein, a healthy tea, a cocoa beverage, a milk beverage, a yogurt and a drinking yogurt, a cheese, an ice cream, a water ice and a dessert, a confectionery, Biscuits, cakes and cake bases, snack foods, balanced foods and beverages, fruit fillings, care glazes, baking chocolate fillings, cheesecake flavoring fillings, fruity cake fillings, cake and donut sugars Clothing, instant bread filling cream, cookie filling, ready-made bread filling, low calorie filling, adult nutrition drink, acid soy / juice beverage, aseptic / retort chocolate drink, snack snack, beverage powder, calci Beam reinforced soy / plain and chocolate milk, it can be used as a component for calcium-fortified coffee beverage.

  The composition further comprises an anti-caking agent for food products such as American cheese sauce, powdered cheese and shredded cheese, chip dip, cream cheese, dry blended whipped topping fat free cream cream. ), Frozen / thawed fresh whipped cream, frozen / thawed stable whipped topping, low fat and light natural cheddar cheese, low fat Swiss style yogurt, aerated frozen dessert, hardened ice cream, ingredients Easy-to-understand and economical hardened ice cream, low-fat ice cream (soft cream), barbecue sauce, cheese dip sauce, cottage cheese dress Ring, dry mixes Alfred source, mix cheese sauce, it may be used as a component in dry mix tomato sauce and others.

  The term “dairy product” as used herein is meant to include a medium composed of milk of animal and / or plant origin. Examples of animal-derived milk include cow, sheep, goat, or buffalo milk. Plant-derived milk can include any plant-derived fermentable material that can be used according to the present invention, particularly those derived from soybeans, rice, or cereals.

  In some embodiments, the food product used in accordance with the present invention is fermented milk or humanized milk.

  In some embodiments, the composition may be used in connection with yogurt production such as fermented yogurt beverages, yogurt, drinking yogurt, cheese, fermented cream, milk based desserts, and others.

  Suitably, the composition may further be used as a component in one or more of cheese applications, meat applications, or applications involving protective bacteria.

  The present invention provides a method for preparing a food or food ingredient, the method comprising mixing a composition according to the invention with another food ingredient.

  In some embodiments, the present invention relates to a product contacted with a composition of the present invention (and optionally other components / components), wherein the composition comprises the nutrients of the product and / or Used in an amount that can improve health benefits.

  As used herein, the term “contacted” refers to applying the composition of the present invention directly or indirectly to a product. Examples of application methods that can be used include treating the product in a material comprising the composition, applying the composition directly by mixing it with the product, and spraying the composition onto the product surface. Or soaking the product in the preparation of the composition.

  When the product of the present invention is a food product, the composition of the present invention is preferably mixed with the product. Alternatively, the composition may be included in an emulsion or foodstuff ingredient. Still alternatively, the composition may be applied as a seasoning, glaze, colorant mixture, and the like.

  For some applications, it is important that the composition be available on or against the surface of the product to be treated / treated. This allows the composition to confer one or more of the preferred features of nutrients and / or health benefits.

  The composition of the present invention may be applied to the product to disperse, coat with and / or impregnate a controlled amount of live microorganisms.

In some embodiments, the composition is used for the fermentation of milk, or sucrose-enriched milk, or a milk medium having sucrose and / or maltose, where as a result of containing all components of the composition The resulting medium (ie the previously mentioned microorganism according to the invention) can be added as a component to yogurt at a suitable concentration, for example at a concentration in the final product that provides a daily dose of 10 ⁶ to 10 ¹⁰ cfu. . The microorganism according to the present invention may be used before or after the fermentation of yogurt.

  In some embodiments, the microorganisms according to the present invention are used as or in the preparation of animal feeds such as livestock feed, in particular avian (such as chicken) feed, pet food or pet treats.

  In some embodiments where the product is a food product, the Pediococcus acidilactici should remain effective over the normal “sale date” or “expiration date”, which is the period during which the food product is offered for sale by the retailer. is there. Preferably, the shelf life should extend beyond such dates until the end of the normal freshness period when the food spoilage becomes evident. The desired length of time and normal shelf life will vary from one foodstuff to another, and those skilled in the art will understand that the shelf life depends on the type of foodstuff, the size of the foodstuff, the storage temperature, the processing conditions, the packaging material, and You will recognize that it depends on the packaging equipment.

  In some embodiments, the compositions of the present invention may be used as food and / or feed ingredients. As used herein, the term “food ingredient” or “feed ingredient” includes a formulation that can be added or added as a nutritional supplement to a functional food or food product. The food ingredient may be in the form of a solution or solid depending on the use and / or application form and / or dosage form.

  In some embodiments, the compositions of the invention may be supplements (also referred to herein as dietary supplements) or added to them.

  In some embodiments, the compositions of the present invention may be functional foods or added to them. As used herein, the term “functional food” means a food that not only provides a nutritional effect, but can also provide a further beneficial effect to the consumer. Thus, a functional food usually contains components or ingredients (such as those described herein) that confer a specific functional benefit, such as a medical or physiological benefit, to the food in addition to a mere nutritional effect. Of food. Although there is no legal definition of functional food, the majority of people interested in this area are functional foods that are marketed as having specific health benefits beyond basic nutritional effects. I agree that Some functional foods are nutritional supplements. As used herein, the term “nutritional supplement” can not only provide nutritional benefits and / or taste satisfaction, but can also provide a therapeutic (or other beneficial) effect to the consumer. Means. Nutritional supplements cross the traditional border between food and medicine.

  In some embodiments, the present invention provides a composition comprising Pediococcus acidilactici such as Pediococcus acidilactici NRRL B-50517 and an edible oil. In some embodiments, the edible oil is selected from the group consisting of olive oil, corn oil, EVOO, LTOO, peanut oil, and vegetable oil. In some embodiments, the ratio of oil to probiotics (w / w) in the composition ranges from 1: 1 to 10: 1. In some embodiments, the probiotic exhibits heat resistance in the oil composition.

  In some embodiments, the present invention provides a composition comprising a sugar, such as sucrose or lactose, in combination with Pediococcus acidilactici, such as Pediococcus acidilactici NRRL B-50517. In some embodiments, the sugar concentration ranges from 0.1% to 50% of the solution.

  In some embodiments, the present invention provides a composition comprising a salt solution, such as NaCl, in combination with Pediococcus acidilactici, such as Pediococcus acidilactici NRRL B-50517. In some embodiments, the salt concentration ranges from 0.1% to 20% of the solution.

  In some embodiments, the present invention provides a composition comprising peanut butter in combination with Pediococcus acidilactici, such as Pediococcus acidilactici NRRL B-50517. In some embodiments, the ratio of peanut butter to probiotics (w / w) ranges from 1: 1 to 10: 1.

Sample Embodiments This section describes exemplary compositions and methods of the invention that are presented without limitation as a series of paragraphs, some or all of which are specified in alphanumeric order for clarity and efficiency. Can be done. Each of these paragraphs may be combined in any suitable manner with one or more other paragraphs and / or disclosures elsewhere in this application, including materials incorporated by reference. Some of the following paragraphs explicitly refer to other paragraphs, further restrict them, and provide some non-limiting examples of suitable combinations.

  1. A composition comprising an effective amount of probiotic, wherein the composition is mixed with an effective amount of amphiphile that enhances the viability of the probiotic in the composition when subjected to heat A composition within.

  2. The composition of paragraph 1, wherein the probiotic is mixed with the amphiphile as a lyophilized fermentation medium.

  3. The composition of any of paragraphs 1-2, wherein the amphiphile is mixed with the probiotic as a dry powder.

  4). 4. The composition of any of paragraphs 1-3, further comprising an oil mixed with the amphiphile and probiotic.

  5. The composition according to paragraph 4, wherein the oil is olive oil.

  6). Paragraph 4-6, wherein the ratio of the amphiphile to the probiotic to the oil (w / w) ranges from about 1: 10: 0.1 to about 10: 1: 25. The composition as described.

  7). The composition according to any of paragraphs 1-6, wherein the amphiphile forms a lipid paste and acts as a carrier.

  8). The composition according to any of paragraphs 1-7, wherein the probiotic is Pediococcus acidilactici.

  9. The composition according to any of paragraphs 1-8, wherein the probiotic is Pediococcus acidilactici NRRL B-50517.

  10. 10. The composition of any of paragraphs 1-9, wherein the amphiphile comprises lecithin, peanut butter, almond butter, soy butter, or cookie butter.

  11. The composition according to any one of paragraphs 1 to 10, wherein the heat is dry heat.

  12 The composition of paragraph 11, wherein the dry heat is at least 65 ° C.

  13. The composition according to any of paragraphs 11 to 12, wherein the dry heat is in the range of 65 ° C to 95 ° C.

  14 14. The composition of any of paragraphs 1-13, wherein the composition is subjected to heat for at least 1 second.

  15. The composition of any of paragraphs 1 through 14, wherein the composition is subjected to heat for at least about 10 minutes.

  16. 16. The composition of any of paragraphs 1-15, wherein the composition is subjected to a second temperature of dry heat after the composition has been subjected to a first temperature of dry heat.

  17. The composition of paragraph 16, wherein the first temperature is about 65 ° C and the second temperature is about 85 ° C.

  18. Paragraph 16 wherein the composition is subjected to heat at the first temperature for at least about 1 second to at least about 30 minutes and is subjected to heat at the second temperature for at least about 1 second to at least about 30 minutes. The composition in any one of -17.

  19. 18. The composition of any of paragraphs 16 through 17, wherein the composition is subjected to the first temperature heat for about 30 minutes and is subjected to the second temperature heat for about 10 minutes.

  20. 18. The composition of any of paragraphs 16-17, wherein the composition is subjected to heat at the first temperature for about 30 minutes and is subjected to heat at the second temperature for about 30 minutes.

  21. The composition according to any one of paragraphs 1 to 10, wherein the heat is liquid heat.

  22. The composition of paragraph 21, wherein the liquid heat is at least about 65 ° C.

  23. The composition of paragraph 21, wherein the liquid heat is at least about 80 ° C.

  24. The composition of paragraph 21, wherein the liquid heat is at least about 82 ° C.

  25. 25. The composition of any of paragraphs 21-24, wherein the composition is subjected to the liquid heat for at least about 1 second.

  26. 25. The composition of any of paragraphs 21-24, wherein the composition is subjected to the heat for at least about 5 minutes.

  27. 25. The composition of any of paragraphs 21-24, wherein the composition is subjected to the heat for at least about 10 minutes.

  28. 28. The composition of any of paragraphs 1-27, wherein the viability is enhanced at least about 2-fold compared to a composition lacking the effective amount of amphiphile.

  29. 28. The composition of any of paragraphs 1-27, wherein the viability is enhanced at least about 10-fold compared to a composition lacking the effective amount of amphiphile.

  30. 30. The composition according to any of paragraphs 1-29, wherein the amphiphile is sunflower lecithin.

  31. 31. The composition of any of paragraphs 1-30, wherein the ratio of the amphiphile to the probiotic (w / w) ranges from about 1:10 to about 25: 1.

  32. 32. The composition of any of paragraphs 1-31, wherein the ratio of amphiphile to the probiotic (w / w) ranges from about 1: 1 to about 10: 1.

  33. 33. The composition of any of paragraphs 1-32, wherein the ratio of the amphiphile to the probiotic (w / w) is about 5: 1.

  34. 33. The composition of any of paragraphs 1-32, wherein the ratio of the amphiphile to the probiotic (w / w) is about 10: 1.

  35. The container suitable for accommodating the foodstuff containing the composition in any one of Paragraphs 1-34.

  36. The container of paragraph 35, wherein the container is made from a material comprising glass, plastic, paper / carton, or aluminum.

  37. 37. The container of any of paragraphs 35-36, wherein the composition is coated on the surface or part of the container.

  38. The container according to any of paragraphs 35 to 37, further comprising a food.

  39. 39. The container of paragraph 38, wherein the food product comprises oil, vinegar, yogurt, fruit product, apple sauce, dairy product, beverage, candy, snack confectionery, and juice.

  40. A container suitable for containing food, wherein a part or surface of the container is coated with an effective amount of Pediococcus acidilactici.

  41. 41. The container of paragraph 40, wherein the Pediococcus acidilactici is NRRL B-50517 strain.

  42. 42. The container of any of paragraphs 40 or 41, wherein the container is made from a material that includes glass, plastic, paper / carton, and aluminum.

  43. 44. The container of any of paragraphs 40-43, wherein the Pediococcus acidilactici is coated on the inner surface of the container.

  44. The container according to any of paragraphs 40 to 43, further comprising a food.

  45. 45. The container of paragraph 44, wherein the food product comprises oil, vinegar, yogurt, fruit product, applesauce, dairy product, beverage, candy, snack confectionery, and juice.

46. A method for enhancing the viability of probiotics in a composition subjected to heat comprising:
iii) adding an effective amount of an amphiphile to the composition comprising probiotics;
iv) subjecting the composition to heat,
A method whereby the viability of probiotics in a composition subjected to heat is enhanced.

  47. 47. The method of paragraph 46, wherein the amphiphile is mixed with the probiotic lyophilized fermentation medium.

  48. 48. A method according to any of paragraphs 46-47, wherein the amphiphile is in the form of a dry powder.

  49. 49. The method of any of paragraphs 46-48, wherein the amphiphile is mixed with the probiotic and oil.

  50. 50. The method of paragraph 49, wherein the oil is olive oil.

  51. Paragraph 49-50, wherein the ratio of the amphiphile to the probiotic to the oil (w / w) ranges from about 1: 10: 0.1 to about 10: 1: 25. The method described.

  52. 51. The method of any of paragraphs 46-50, wherein the amphiphile forms a lipid paste and acts as a carrier.

  53. 53. The method of any of paragraphs 46 through 52, wherein the probiotic is Pediococcus acidilactici.

  54. The method according to paragraph 2, wherein the probiotic is Pediococcus acidilactici NRRL B-50517.

  55. 55. The method of any of paragraphs 46-54, wherein the amphiphile comprises lecithin, peanut butter, almond butter, soy butter, or cookie butter.

  56. 56. The method according to any of paragraphs 46-55, wherein the heat is dry heat.

  57. The method according to paragraph 56, wherein the dry heat is at least 50 ° C.

  58. 57. A method according to any of claims 55 to 56, wherein the dry heat is in the range of 65 to 95 [deg.] C.

  59. 59. The method of any of paragraphs 46-58, wherein the composition is subjected to heat for at least 1 second.

  60. 60. The method of any of paragraphs 46 through 59, wherein the composition is subjected to heat for at least about 10 minutes.

  61. 61. The method of any of paragraphs 46-60, wherein the composition is subjected to a second temperature of dry heat after the composition is subjected to a first temperature of dry heat.

  62. 62. The method of paragraph 61, wherein the first temperature is about 65 ° C and the second temperature is about 85 ° C.

  63. Paragraphs 61-62, wherein the composition is subjected to heat at the first temperature for at least about 1 to at least about 30 minutes and is subjected to heat at the second temperature for at least about 1 to at least about 30 minutes. The method in any one of.

  64. 64. The method of any of paragraphs 61-63, wherein the composition is subjected to heat at the first temperature for about 30 minutes and subjected to heat at the second temperature for about 10 minutes.

  65. 64. The method of any of paragraphs 61-63, wherein the composition is subjected to heat at the first temperature for about 30 minutes and is subjected to heat at the second temperature for about 30 minutes.

  66. 56. The method according to any of paragraphs 46-55, wherein the heat is liquid heat.

  67. 68. The method of paragraph 66, wherein the liquid heat is at least about 50 ° C.

  68. The process of paragraph 66, wherein the liquid heat is at least about 80 ° C.

  69. 68. The method of paragraph 66, wherein the liquid heat is at least about 82 ° C.

  70. 70. The method of any of paragraphs 66 through 69, wherein the composition is subjected to the heat for at least about 1 second.

  71. 71. The method of any of paragraphs 66-70, wherein the composition is subjected to the heat for at least about 5 minutes.

  72. 71. The method of any of paragraphs 66-70, wherein the composition is subjected to the heat for at least about 10 minutes.

  73. 73. The method of any of paragraphs 46-72, wherein the viability is enhanced at least about 2-fold compared to a composition lacking the effective amount of amphiphile.

  74. 73. The method of any of paragraphs 46-72, wherein the viability is enhanced at least about 10 times compared to a composition lacking the effective amount of amphiphile.

  75. 75. The method according to any of paragraphs 46-74, wherein the amphiphile is sunflower lecithin.

  76. 76. The method of any of paragraphs 46-75, wherein the ratio of the amphiphile to the probiotic (w / w) ranges from about 1:10 to about 25: 1.

  77. 77. The method of any of paragraphs 46-76, wherein the ratio of the amphiphile to the probiotic (w / w) ranges from about 1: 1 to about 10: 1.

  78. 78. The method of any of paragraphs 46-77, wherein the ratio of the amphiphile to the probiotic (w / w) is about 5: 1.

  79. 78. The method of any of paragraphs 46 through 77, wherein the ratio of the amphiphile to the probiotic (w / w) is about 10: 1.

  80. 80. The method of any of paragraphs 46 through 79, wherein the composition is contacted with food and subjected to heat with the food.

  81. Subjecting the composition to the thermal step of part ii) comprises contacting the composition with food, wherein the food is already subjected to heat prior to contacting the composition, paragraphs 46- 79. The method according to 79.

  82. A method of treating a disease or condition characterized by inflammation in a subject in need thereof, comprising administering to the subject an effective amount of Pediococcus acidilactici probiotic.

  83. 85. The method of paragraph 82, wherein the method comprises administering to the subject a composition according to any of claims 1-34.

  84. 84. The method of paragraph 83, wherein the composition further comprises a food, a dietary supplement, or a pharmaceutical.

  85. The disease or condition is malignant tumor (cancer), arthritis, cardiovascular disease, hepatitis, infection, wound healing, pancreatitis, gastroesophageal reflux disease, diabetes, inflammatory bowel disease, peptic ulcer disease, bronchitis, cholecystitis, Appendicitis, bursitis, dermatitis, asthma, autoimmune disease, pelvic inflammatory disease, gout, trauma, foreign body infection, burn, dental treatment, tendinitis, rhinitis, mucositis, and toxins such as chemicals and alcohol 85. The method according to any of paragraphs 82-84, selected from the group consisting of exposure to:

  86. 86. The method of any of paragraphs 82-85, wherein the Pediococcus acidilactici probiotic is NRRL B-50517 strain.

  87. The method according to any of paragraphs 82-86, wherein the subject is a human.

88. 88. The method of any of paragraphs 82-87, wherein the subject is administered more than 1.0 × 10 ⁹ cfu probiotic.

89. 89. The method of any of paragraphs 82-88, wherein the subject is administered more than 4.0 × 10 ⁹ cfu probiotics.

  90. 90. The method of any of paragraphs 82-89, wherein the subject is administered one or more additional therapeutic agents.

  91. 90. The method of any of paragraphs 82-89, wherein the subject is not administered another therapeutic agent.

  92. 92. The method of any of paragraphs 82-91, wherein the Pediococcus acidilactici probiotic increases the number of anti-inflammatory M2 macrophage cells in the subject.

  93. 93. The method of any of paragraphs 82-92, wherein the subject exhibits increased IL-10 production.

  94. 94. The method of any of paragraphs 82-93, wherein the subject exhibits a reduced level of IL-6 and / or IL-23.

  95. 95. The method of any of paragraphs 82-94, wherein the disease is cancer.

  96. 96. The method of paragraph 95, wherein the subject is administered one or more chemotherapeutic agents and / or radiation therapy in combination with Pediococcus acidilactici probiotics.

  97. 95. The method of any of paragraphs 82-94, wherein the disease is pancreatitis.

  98. 98. A composition comprising an effective amount of Pediococcus acidilactici probiotic for use in the method of any of paragraphs 82-97.

  99. 99. The composition of paragraph 98, wherein the Pediococcus acidilactici probiotic is formulated as a tablet.

  100. 99. The composition of paragraph 98, wherein the Pediococcus acidilactici probiotic is formulated as a capsule.

  101. The composition according to any of paragraphs 98-100, comprising peach fruit powder as a flavoring agent.

  102. 102. The composition of any of paragraphs 98-101, wherein the Pediococcus acidilactici probiotic is NRRL B-50517 strain.

  The invention is further illustrated by the following examples. These examples are provided to aid the understanding of the present invention and should not be construed as limiting thereof.

＊：Ｓｐｒａｇｕｅ Ｄａｗｌｅｙラット（９週）に、Ｈａｒｌａｎ＃７０１２ラット餌を不断給餌した。プロバイオティクスは、Ｐｅｄｉｏｃｏｃｃｕｓ系プロバイオティクスであった（Ｉｍａｇｉｌｉｎ，Ｆｒｅｄｅｒｉｃｋ，ＭＤ）。群は、対照群（プロバイオティクスなし）、低用量（１×１０^９ｃｆｕ）、中用量（２×１０^９）、及び高用量（１０×１０^９）の４つであった（ｎ＝１０）。動物に、プロバイオティクスと混合した２グラムの食物を午前１１時に与えた。その後、午後８時〜午前１１時まで、餌を不断給餌した。水は２４時間不断給餌した。尾血試料を全血球算定のために分析した。 Example 1-P. administration of acidilacticis stimulates innate immune responses in animals. The effect of administration of acidilactici on the innate immune response in rats will be described.

*: Sprague Dawley rats (9 weeks) were fed the Harlan # 7012 rat diet ad libitum. Probiotics were Pediococcus-based probiotics (Imagilin, Frederick, MD). There were four groups (n = 10): control group (no probiotics), low dose (1 × 10 ⁹ cfu), medium dose (2 × 10 ⁹ ), and high dose (10 × 10 ⁹ ). ). The animals were fed 2 grams of food mixed with probiotics at 11:00 am. Thereafter, food was fed continuously from 8 pm to 11 am. Water was fed 24 hours a day. Tail blood samples were analyzed for complete blood count.

When rats were administered 2.0 × 10 ⁹ to 10 × 10 ⁹ cfu of Pediococcus probiotics per day for 15 days, the amount of macrophages was compared to that of rats not receiving probiotics Increased by 150% to 180%. The increase in macrophages indicates that Pediococcus probiotics can stimulate rat innate immune responses. Interestingly, when rats were fed a small amount (1.0 × 10 ⁹ ) of Pediococcus probiotics, the number of macrophages was similar to that of control-derived macrophages. This indicates that a sufficient amount of Pediococcus probiotics is required for stimulating the innate immune response, such as increasing the amount of macrophage cells.

Example 2-P. Administration of acidilactici stimulates cytokine production in human subjects.
This example is shown in P.A. The effect of administration of acidilactici on cytokine production in human subjects will be described.

＊：血清試料を、Ｐｅｄｉｏｃｏｃｃｕｓ ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７プロバイオティクスを投与する前と、１日あたり４０億ｃｆｕのＰｅｄｉｏｃｏｃｃｕｓプロバイオティクスを４５日間投与した後とで５人の有志から収集した。血清試料を、炎症促進性ＩＬ−６及び抗炎症性ＩＬ−１０と関連付けられるバイオマーカーを測定するように設計されたＬｕｍｉｎｅｘ系多重アッセイ（ＥＭＤ Ｍｉｌｌｉｐｏｒｅ；Ｍｉｌｌｉｐｌｅｘ）を使用して分析した。 With respect to the innate immune response, macrophages are roughly divided into two groups: pro-inflammatory M1 macrophages and anti-inflammatory M2 macrophages. M2 macrophages also function as macrophages that function in construction processes such as wound healing and tissue repair, as well as macrophages that abolish harmful immune system activation through the production of anti-inflammatory cytokines such as interleukin-10 (IL-10). Point to.

*: Serum samples were collected from 5 volunteers before administration of Pediococcus acidilactici NRRL B-50517 probiotic and after administration of 4 billion cfu of Pediococcus probiotic for 45 days per day. Serum samples were analyzed using a Luminex-based multiplex assay (EMD Millipore; Milliplex) designed to measure biomarkers associated with pro-inflammatory IL-6 and anti-inflammatory IL-10.

  All five volunteers exhibited significantly increased anti-inflammatory IL-10 activity (158% -422% increase) after 45 days of Pediococcus probiotic administration. In contrast, the effect on pro-inflammatory IL-6 showed inconsistent results, with reduced activity in 3 volunteers and increased activity in 2 volunteers. These results indicate that administration of Pediococcus probiotics in human subjects can enhance anti-inflammatory IL-10 activity more than 2-fold. These results, in conjunction with the results showing increased macrophages in rats treated with Pediococcus probiotics, indicate that innate immunity of humans and animals can be enhanced by Pediococcus probiotics. The innate immune response of humans and animals treated with Pediococcus exhibits an increase in M2 macrophages and anti-inflammatory IL-10.

Example 3 P. for use in weight management. Effect of supplementation with acidilactici NRRL B-50517: a subject randomized double-blind This weight management study is a Pediococcus acidilactici NRRL B-50517 probiotic strain for 30 adult participants in a control randomized double-blind Of 12 weeks of supplementation. Body fat percentage was measured at the start and end of the test by bioelectrical impedance analysis (BIA). The levels of the pro-inflammatory biomarkers interleukin-6 (IL-6) and interleukin-23 (IL-23) were determined using blood samples collected before and after the start of the study. Appetite, vitality level, defecation, stool quality, bloating, and gas were monitored throughout the study using weekly questionnaires. P. The specific weight loss and anti-inflammatory effects of acidilactici are described here for the first time. 4 billion CFU Daily supplementation of acidilactici resulted in an average reduction in body fat percentage of 0.86 ± 0.42% in the probiotic group and an increase of 0.28% ± 0.19 in the control group (p = 0.0264). The ratio of pro-inflammatory IL-6 differs by 0.61 ± 0.22 and 3.06 ± 0.87 (p = 0.0295) in the probiotic group and the control group, respectively. The 23 ratio was 0.65 ± 0.14 in the probiotic group and 1.71 ± 0.38 in the control group (p = 0.068).

Methods and Materials Participants in this study were selected on a volunteer basis and age, gender, and BMI were equivalent across treatment groups. Volunteers were advised not to change their normal dietary patterns or exercise habits during the study. Probiotics were tested in subject groups divided as follows: normal body weight status 20% (18.5 to 24.99), 47% overweight (25 to 29.99), and 33% obesity (30 Super).

  Prior to the start of the replenishment period, the participant conducted a large-scale physical examination, including bioelectrical impedance analysis to determine body fat percentage, and blood tests to quantify IL-6 and IL-23 levels. I received it. The same inspection procedure was repeated at the end of the test. Over a 12-week period, 30 participants were given P. pylori in combination with peach fruit powder reaching a dose of 4 billion CFU of Pediococcus probiotics / day. Either two gelatin capsules containing the compound of acidilactici NRRL B-50517 fermentation culture or twice a placebo treatment containing only peach powder was administered. The safety of probiotics was analyzed in terms of appetite, vitality level, defecation, stool quality, bloating, and effects on gas. As part of the weekly questionnaire, participants were asked to score their experience of these symptoms on any scale from 1 to 5, with 1 being the least severe and 5 being the most severe.

Results The results show that supplementing the diet of overweight or obese individuals with the probiotic Pediococcus acidilactici NRRL B-50517 affects body fat storage and affects the concentration of inflammatory biomarkers associated with obesity pathology Show that for the first time. The clear difference between body fat percentage and between IL-6 and IL-23 levels observed between the placebo group and the 5051 treatment group at the end of the study was reduced without traditional dietary changes or exercise. Shows the effect of probiotics to help. The results are shown in FIGS. Error bars for all three graphs indicate that the body fat percentage, IL-6, and IL-23 values for the placebo and probiotic treatment groups are not within one standard deviation of each other. Indicating a significant difference between the two.

  P. The effect of acidilactici 5051 probiotics on body fat percentage is shown in FIG. Participants received a P. pylori in combination with peach fruit powder reaching a dose of 4 billion CFU Pediococcus probiotics / day. Either two gelatin capsules containing the compound of acidilactici NRRL B-50517 fermentation culture or twice a capsule containing only peach powder were administered. Body fat percentage was determined by a bioelectrical impedance analysis (BIA) test. Values are based on a double-blind field evaluation of Pediococcus probiotics for a total of 30 volunteers for 3 months of treatment. P = 0.0264, t = 2.4073, placebo group median = 0.280 ± 0.190 (SEM); NutriLots ™ median = −0.864 ± 0.418 (SEM). The significantly greater difference in body fat percentage observed in the probiotic treatment group (NutriLots) compared to the placebo group after 3 months of treatment Acidilactici NRRL B-50517 supplementation may result in accelerated weight loss without changing the diet or exercise pattern.

Effect of Pediococcus probiotic supplementation on the inflammatory biomarkers IL-6 and IL-23.
P. The effect of acidilactici 5051 probiotics on IL-23 is shown in FIG. Blood samples were collected from each participant, either placebo or probiotic, before the start of the study and after the end of the treatment period to determine changes in the presence of IL-6 and IL-23. A significant decrease in both IL-6 and IL-23 was observed in the Pediococcus probiotic treatment group. Values are based on a double-blind field evaluation of Pediococcus probiotics for a total of 30 volunteers for 3 months of treatment. P = 0.0295, t = 2.4239, median placebo = 3.058 ± 0.867 (SEM); NutriLots ™ median = −0.612 ± 0.221 (SEM). The lower ratio of IL-23 in the probiotic treatment group (NutriLots) suggests that 5051 has the ability to reduce obesity-related inflammation.

  P. The effect of acidilactici 5051 probiotics on IL-6 is shown in FIG. Values are based on a double-blind field evaluation of Pediococcus probiotics for a total of 30 volunteers for 3 months of treatment. P = 0.068, t = 3.0194, placebo median = 1.714 ± 0.377 (SEM); NutriLots ™ median = −0.648 ± 0.137 (SEM). The reduced ratio of IL-6 in the probiotic treatment group (NutriLots) indicates that 5051 has the ability to reduce obesity-related inflammation.

Discussion In this study, 12 weeks P.P. The acidilactici NRRL B-50517 probiotic treatment resulted in a significant reduction in body fat percentage, interleukins 6 and 23 when administered to various BMI participants (FIGS. 1, 2 and 3). Consistent results across the board are It shows that the power of the acidilactici action is not limited to individuals with obese body weight, but is the same for individuals with lower BMI. Most of the previous studies have shown the efficacy of LAB probiotic treatments for obese subjects only. When other probiotic strains were not effective in reducing the presence of obesity-related inflammation, 5051 reduced the levels of both interleukins 6 and 23 compared to the placebo group.

  The safety of this probiotic was confirmed in a separate published study conducted in parallel with this study. No significant differences in appetite, defecation, bloating, stool quality, vitality level, or scoring by gas participants were observed between the start and end of the study period in either the placebo or probiotic treatment groups .

  These discoveries have a profound impact on future treatment and prevention of metabolic diseases. Since most cases of chronic conditions such as cardiovascular disease (CVD) and type 2 diabetes develop in parallel with obesity, just improving the management of this disease has been found in several other works on public health. It has the potential to greatly reduce the incidence of obvious threats.

  Adipocytes have previously been thought to function only as containers that store excess heat in the form of triglycerides, but have been found to play a complex role in metabolism, immunity, and cancer (Calabro P, Yeh). ETH. 2007. Obesity, Inflammation, and Vassal Disease: the role of the adipose tissue as an endocrine organ. Subcellular Biochemistry. 42: 63-91). White adipocytes secrete proteins including cytokines and hormone-like factors such as adiponectin, leptin, and resistin. This phenomenon is of particular interest as these molecules are involved in vascular and metabolic complications (Calabro P, Yeh ETH. 2007. Obesity, Inflammation, and Vascular Disease: the role of the. adipose tissue as an endocrine organ. Subcellular Biochemistry. 42: 63-91). In the majority of obese patients, low-grade inflammation of white adipose tissue (WAT) resulting from chronic activation of innate immunity may have increased final expression of insulin resistance, impaired glucose tolerance, and diabetes propensity Bastard JP, Maachi M, Lagathu C, Kim MJ, Caron M, Vidal H, Capeau J, Feve B. 2006. Regent advancements in the relations in the rebet. 17 (1): 4-12). Obesity WAT macrophage infiltration serves as a source of pro-inflammatory cytokines and further contributes to the cause of insulin resistance. On the other hand, circulating levels of adiponectin, an insulin-sensing effector highly expressed in WAT, is lower in obese subjects than in normal weight subjects. WAT in these individuals overproduces and secretes increased levels of many inflammatory molecules including IL-6, another regulator of insulin sensitivity. For this reason, the pro-inflammatory causes of obesity and the systemic manifestation of insulin resistance are closely related and linked by the regulation of WAT.

  Regulation of calorimetric extraction from food materials can be considered as a possible mechanism of probiotic action on the results presented herein. The composition of the human intestinal microflora has been consistently implicated as a determinant of body weight as a result of its important role in nutrient acquisition and thermopower collection and regulation (Tennyson CA, Friedman G. 2008. Microecology, obesity, and probiotics.Curr.Opin.Endocr.Diab.Obes.15 (5): 422-7, DiBaise JK, Zhang H, Crowell MD, Krajmalnik-Brown R, Decker GA, Brit 200, Rit. possible relationship with obesity.May Clinic Proceedings.83 (4): 4 0-69.). There is a reason that the optimization of targeted microbial communities through the introduction of probiotics is considered to have potential as a novel therapeutic in the treatment of metabolic diseases. Conscious editing of the microflora can be key to adjusting the imbalance between caloric intake and consumption due to obesity.

  The absence of changes in the reported scores for appetite in both the placebo and probiotic treatment groups indicates that there is no change in possible dietary patterns or the satiety hormone leptin.

  In reducing serum fat-6 levels and at the same time reducing body fat percentage, It can be presumed that acidilactici 5051 can contribute to reducing the risk of type 2 diabetes because it can increase insulin sensitivity and reduce overall systemic inflammation. Since IL-6 is also associated with vascular injury in obese individuals, reduced serum levels of cytokines are likely to reduce the risk of CVD (Calabro P, Yeh ETH. 2007. Obesity, Inflammation, and Vascular Disease: the role of the adipose tissue as an endocrine organ. Subcellular Biochemistry. 42: 63-91). It can be hypothesized that the inflammatory activity in obese individuals that increases following WAT macrophage infiltration is reduced upon loss of body fat.

  The results presented here reflect what was observed in the analysis of post-surgical results in morbidly obese individuals. After bariatric surgery, patients have shown clinically significant reductions in IL-6, triglycerides, cholesterol, LDL, glucose, and insulin that correlate with BMI, which is related to the relationship between body weight and inflammatory profile The existence was demonstrated and the relationship between BMI and biochemical parameters in chronic metabolism and vascular pathology was further elucidated (Illan-Gomez F, Gonzalvez-Ortega M, Orea-Soller I, Alcaraz-Tafalla MS, Aragon- Alonso A, Pascal-Diaz M, Perez-Paredes M, Lozano-Almela ML. 2012. Obesity and information: change in C-reactive protein, tumor necrosis factor-alpha and interleukin-6 after vari- able surcharge. Obes. Surg. 22: 950-55).

  The significant decrease in IL-23 concentration observed in the probiotic treatment group is also a powerful indicator of reduced disease risk. IL-23 / IL-17 is strongly associated with the activation of signal pathways that lead to tumorigenesis and carcinogenic pathways. Diets associated with the development of obesity rather than the obesity state itself, as IL-23 / IL-17 axis stimulation was observed in obese women independent of increased abdominal fat, insulin resistance, leptin, or MIF levels And it is reasonable to assume that the behavioral pattern is the cause (Sumarac-Dumanovic M, Stevanovic D, Ljubic A, Jorga J, Simik M, Stamenkovic-Pejkovic D, Starvic V. Trajkovic. activity of interleukin-23 / interleukin-17 proxilamation axis in obese woman.Int.J.Obes 33: 151-56).

Conclusion In summary, the probiotic P.I. acidilactici NRRL B-50517 has a decreasing effect on body fat percentage, IL-6, and IL-23, suggesting its beneficial impact on weight management and metabolic diseases. In light of the evidence presented in this study, Pediococcus acidilactici NRRL B-50517 is found to be effective in reducing body fat and inflammation in individuals desiring to lose weight.

Example 4-Effect of Pediococcus Probiotics on Dogs and Cats with Pancreatitis A 14-year-old female contraceptive toy poodle with significant abdominal pain, vomiting, and loss of appetite may have pancreatitis Diagnosed. Ultrasound images showed high fat echoes from the stomach to the duodenum, and serum analysis showed 432 ug / L spec cPL (canine pancreatic specific lipase). The dog immediately switched to a low-fat diet and was treated with 100 mg KAMOSTAAL 100 twice a day, but the spec cPL remained higher than 400, and after about 2.5 months, diarrhea and vomiting And reached 610 ug / L. At that time, the dogs were treated with 200 mg Pediococcus probiotics twice daily, combined with a low fat diet and current treatment of 100 mg KAMOSTAAL 100 twice daily. Interestingly, not only did diarrhea and vomiting cease, but spec cPL returned to normal at 163 ug / L within about 6 weeks. For this reason, treatment with both Pediococcus probiotics and KAMOSTAAL100 was stopped. However, after about 3 months, the dog relapsed and the spec cPL increased to 276 ug / L. At this point, the dogs were treated with only 200 mg of Pediococcus probiotic twice daily. Treatment continued for about 7 months with good control of diarrhea, vomiting, and loss of appetite, and spec cPL was shown to be normal at 108 ug / L (Figure 4).

  Spec cPL (canine pancreatic specific lipase) and spec fPL (feline pancreatic specific lipase) are canine normal spec cPL and well-established assays for pancreatitis in dogs and cats. In healthy dogs and cats, the spec cPL is less than 200 ug / L and the spec fPL is 0.7-3.5 ug / L. Dogs and cats are considered to have pancreatitis if the spec cPL is greater than 400 ug / L and the cat spec fPL is greater than 5.4 ug / L. Based on this criterion, Pediococcus probiotics were applied to two dogs and one cat, both of which have a spec cPL> 600 ug / L and a cat has a spec fPL of 50 ug / L. And suffered from pancreatitis. All of these dogs and cats not only stopped vomiting and diarrhea, but spec cPL and spec fPL were controlled and returned to normal. In addition, two dogs who had elevated spec cPL (303 ug / L and 205 ug / L) and could have suffered from pancreatitis were also treated.

Example 5-Effect of Pediococcus probiotics on dogs suffering from cancer undergoing chemotherapy Treatment of 4 dogs with various cancers and undergoing chemotherapy by medication of Pediococcus probiotics did. After a short period of treatment with Pediococcus probiotics, these dogs experienced improvement in symptoms.
*: Chemotherapy and Pediococcus probiotic administration treatment were performed at Daktar Animal Hospital Central, Torri and Yaizu.

Example 6 Production of Pediococcus acdilactici NRRL B-50517 First, the strains required for the fermentation process were selected from bacteria frozen in a -70 ° C freezer. Grow cultures in sterile media bottles. Once grown, samples were taken to confirm cleanliness and basic phenotypic purity. If it is clean and the cells coincide with the initial Gram stain smear, permission to make the tank medium (fermentation broth) is granted. Prior to inoculation, the inoculum tank is CIP (Position Clean) with caustic and acidic solutions. The tank is sterilized before filling with broth fermentation ingredients to be released, mixed and sterilized. The tank medium is sterilized at 220 ° F. to 250 ° F. for 30 minutes to 1 and a half hours depending on the volume of the tank. The temperature is then lowered to 85-95 ° F and the tank is seeded with the inoculum. Once the inoculation tank has grown, cool the tank to 55-65 ° F. Once cooled, samples are taken for repeated purity checks. If release is approved, repeat steps for tank preparation and inoculation. The product will inoculate the tank with a grown and approved inoculum bottle. After all desired tanks have been grown, the purity check is repeated, after which the centrifuge is prepared and the cells are concentrated to a concentrated liquid. Place the concentrated liquid culture in a sterile storage tank. A sterile liquid cryoprotectant solution is added to the centrifuged culture.

  This is homogenized in a storage tank by means of a stirrer. Once homogenized, the culture is pumped into a sterile kettle (functional aliquot for product freezing or pelleting) and pelleted in a liquid nitrogen vat. When complete, the frozen pellet is lyophilized or freeze-dried. After drying, the freeze-dried pellets are milled to a fine powder. After taking the ground culture and homogenizing to ensure homogeneity, the culture is sampled for quality assurance by morphological, physiological, 16S rRNA DNA sequence, and high temperature stress assay. The product is removed from the blender used to homogenize the material, then placed in a bag and stored in a cool place at room temperature.

Example 7-Formulation and Testing of Pediococcus acdilactici NRRL B-50517 in Food As knowledge of the health benefits of probiotics spreads and the demand for foods infused with probiotics continues to increase, food companies are biotech companies Face a series of new challenges as we begin to work with. First, one or more probiotic strains need to be selected from a large number of available options. Ideally, the bacteria selected are: 1. Need to survive any manufacturing stress such as high heat treatment. 2. need to be compatible with the chemical and physical properties of the desired food matrix; 3. Need to maintain viability in food for the shelf life of the product after it has been incorporated In order to impart its health benefits to the host, it is necessary to resist destruction by the digestive mechanism. Many probiotic strains (such as Lactobacillus and Bifidobacterium) that are common in commercially available supplements are not suitable for industrial food production because they do not effectively meet these requirements. Due to the lack of critical high heat resistance necessary for survival in recently sterilized food, the use of these two lactic acid bacteria (LAB) is very limited in this context. The instability of these strains at room temperature can present further problems in transportation and storage to grocery stores and prospective consumers. As facultative anaerobic to obligate anaerobes, Lactobacillus and Bifidobacterium are particularly vulnerable to loss of viability upon any exposure to oxygen, further reducing their potential for incorporation into food. More versatile and reliable strains are required for effective probiotic infused food formulations.

  Pediococcus acidilactici NRRL B-50517 is a uniquely formulated powder composed of bacterial strains that can withstand a wide range of temperature, osmotic pressure, and oxygen exposure. Probiotics, originally durable microorganisms isolated from plant materials, have demonstrated the ability to survive in a wide range of foods under a variety of environmental conditions and heat treatment procedures.

＊：０．２ｇの１０億（１Ｂ）ＣＦＵ／ｇのＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７を２０ｍＬの各スクロース溶液に添加し、室温で貯蔵した。生存能力試験を、０．１％の生理食塩水中のスクロース＋Ｐ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７溶液を連続希釈し、ＭＲＳ上にプレーティングし、一晩のインキュベーション後にプレートを数え上げることによって実施した。＊＊プレートは３日目に汚染され、安定性についての試験の継続は妨げられた。 P. in a sucrose solution with a concentration range of 10-50%. The survival of acidilactici NRRL B-50517 indicates the resistance of probiotics to osmotic pressure (Table 3). Weaker bacteria are P. aeruginosa even when they are more likely to lose viability in solutions with relatively high osmotic pressure. acidilactici NRRL B-50517 retains a remarkably stable cell number even at the highest concentrations tested. Similar results were obtained in lactose solutions in the same concentration range over 9 days (Table 4). In a solution of sterile water, 0.1-20% NaCl, and a combination of NaCl and sucrose, P.I. Acidilactici NRRL B-50517 maintained significant cell viability for up to 1 week in all assayed samples and showed the ability of probiotics to adapt to myriad chemical environments (Table 5).

*: 0.2 g of 1 billion (1B) CFU / g P.I. Acidilactici NRRL B-50517 was added to 20 mL of each sucrose solution and stored at room temperature. Viability testing was performed using sucrose + P.I. in 0.1% saline. The acidilactici NRRL B-50517 solution was serially diluted, plated on MRS, and enumerated after overnight incubation. ** The plate was contaminated on the third day, preventing continued testing for stability.

＊：０．２ｇの１Ｂ／ｇのＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７を２０ｍＬの各ラクトース溶液に添加し、室温で貯蔵した。生存能力試験を、０．１％の生理食塩水中のスクロース＋Ｐ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７溶液を連続希釈し、ＭＲＳ上にプレーティングし、一晩のインキュベーション後にプレートを数え上げることによって実施した。 Conclusion: acidilactici NRRL B-50517 maintains viability in sucrose solutions in the concentration range of 10-50%. 3 shows the resistance of acidilactici NRRL B-50517 to high osmotic pressure environment.

*: 0.2 g of 1 B / g P.I. Acidilactici NRRL B-50517 was added to 20 mL of each lactose solution and stored at room temperature. Viability testing was performed using sucrose + P.I. in 0.1% saline. The acidilactici NRRL B-50517 solution was serially diluted, plated on MRS, and enumerated after overnight incubation.

＊：０．２ｇの１Ｂ／ｇのＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７を、２０ｍＬの異なる濃度のＮａＣｌまたはスクロース＋ＮａＣｌ溶液に添加し、室温で貯蔵した。生存能力試験を、０．１％の生理食塩水中のＮａＣｌ＋Ｐ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７溶液を連続希釈し、ＭＲＳ上にプレーティングし、一晩のインキュベーション後にプレートを数え上げることによって実施した。 Conclusion: P. acidilactici NRRL B-50517 maintained a very stable viable count in 10-50% lactose solution.

*: 0.2 g of 1 B / g P.I. Acidilactici NRRL B-50517 was added to 20 mL of different concentrations of NaCl or sucrose + NaCl solution and stored at room temperature. Viability testing was performed with NaCl + P.I. in 0.1% saline. The acidilactici NRRL B-50517 solution was serially diluted, plated on MRS, and enumerated after overnight incubation.

＊：Ｐ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７及びピーナッツバターの試料を、６ｇの１００Ｂ／ｇのＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７粉末を２０ｇのピーナッツバターと混合した後、室温（２３℃）または３７℃のいずれかで貯蔵することによって調製した。安定性試験を、０．１ｇの混合物を５ｍＬの０．１％の生理食塩水に添加し、ＭＲＳ上でのプレーティングのために溶液を連続希釈し、４５℃で一晩インキュベートした後でプレートを数え上げることによって実施した。生存率を、Ｐ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７＋生理食塩水対照（０．２ｇの１００Ｂ／ｇのＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７を室温で１０ｍＬの０．１％の生理食塩水に添加したもの）の比率として計算した。 When assayed on a large scale for viability, 5051® maintains viable cell count (CFU / g) for up to 113 days after incorporation into peanut butter when stored at room temperature, and high storage stability (Table 6). The probiotics showed similar results when heated to 85 ° C. in peanut butter and then stored at room temperature. Stability was similarly constant when probiotics were incorporated into pudding heated to 85 ° C., and the bacterial count remained within log 1 for 29 days stored in the refrigerator (Table 7).

*: P.I. A sample of acidilactici NRRL B-50517 and peanut butter was added to 6 g of 100 B / g P. a. Acidilactici NRRL B-50517 powder was mixed with 20 g peanut butter and then stored at either room temperature (23 ° C.) or 37 ° C. The stability test was performed by adding 0.1 g of the mixture to 5 mL of 0.1% saline and serially diluting the solution for plating on MRS and incubating overnight at 45 ° C. It was carried out by counting. Survival rate was determined by P.P. It was calculated as the ratio of acidilactici NRRL B-50517 + saline control (0.2 g 100 B / g P. acidilactici NRRL B-50517 added to 10 mL 0.1% saline at room temperature).

＊：Ｐ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７及びピーナッツバターの試料を、１．２ｇの１００Ｂ／ｇのＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７粉末を３．８ｇのピーナッツバター中に混合することによって調製した。空の試験管を８５℃に加熱した後、０．５ｇのピーナッツバターとＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７との混合物を添加し、５分間ホットプレート上に置いた。１０分間の冷却期間の後、１０ｍＬの０．１％の生理食塩水を各試験管に添加した。生存能力試験を、生理食塩水中に連続希釈し、ＭＲＳ上にプレーティングし、４５℃で一晩インキュベートした後でプレートを数え上げることによって実施した。生存率を、Ｐ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７＋生理食塩水対照（０．２ｇの１００Ｂ／ｇのＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７を室温で１０ｍＬの０．１％の生理食塩水に添加したもの）の比率として計算した。 Conclusion: acidilactici NRRL B-50517 showed high bacterial counts (CFU / g) in peanut butter at room temperature over 113 days, indicating that products containing both ingredients can maintain high storage stability. Even when stored at 37 ° C., peanut butter and P.P. The mixture with acidilactici NRRL B-50517 showed similarly high viability over 22 days and declined in the viability test on days 22-113.

*: P.I. A sample of acidilactici NRRL B-50517 and peanut butter was added to 1.2 g of 100 B / g P. a. Acidilactici NRRL B-50517 powder was prepared by mixing in 3.8 g peanut butter. After heating the empty test tube to 85 ° C., 0.5 g peanut butter and P.I. A mixture with acidilactici NRRL B-50517 was added and placed on a hot plate for 5 minutes. After a 10 minute cooling period, 10 mL of 0.1% saline was added to each tube. Viability testing was performed by serial dilutions in saline, plating on MRS, and enumerating plates after incubation overnight at 45 ° C. Survival rate was determined by P.P. It was calculated as the ratio of acidilactici NRRL B-50517 + saline control (0.2 g 100 B / g P. acidilactici NRRL B-50517 added to 10 mL 0.1% saline at room temperature).

Conclusion: P.P. acidilactici NRRL B-50517 maintains high viability in nut butter and The compatibility of acidilactici NRRL B-50517 with commercially manufactured nut products was supported.
PB1 component: roasted peanut, sugar, hydrogenated vegetable oil (cotton seed oil, soybean oil, and rapeseed oil) for prevention of separation, salt.

  Hazelnut spread ingredients: sugar, vegetable oil (palm oil and rapeseed oil), hazelnut, cocoa powder, skim milk, whey, lactose, sunflower lecithin (emulsifier), natural vanilla flavor.

＊：９００ｕＬの油をホットプレート上で８５℃に加熱した後、０．１ｇの１Ｂ／ｇのＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７を添加し、その後、一定期間そのままにした。次に、試験管を除去し、最低１０分間冷却させた後、０．１％の生理食塩水中に連続希釈し、ＭＲＳ上でプレーティングした。プレートを４５℃で一晩インキュベートし、翌日数え上げた。生存率を、生理食塩水＋Ｐ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７対照（０．２ｇの１Ｂ／ｇのＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７を室温で２０ｍＬの０．１％の生理食塩水に添加したもの）の比率として計算した。 P. in 5 highly heat-treated oils (corn oil, EVOO, LTOO, peanut oil, and vegetable oil). Testing acidilactici NRRL B-50517 gave results similar to those observed in peanut butter. Two of the oils, EVOO and corn oil (Table 8) showed excellent survival even after continuous exposure to 85 ° C. for 30 minutes (Table 9 and Table 10). The apparent durability of probiotics in oils particularly facilitates their use in traditional food preparation techniques with heat.

*: After heating 900 uL of oil on a hot plate to 85 ° C., 0.1 g of 1 B / g P.I. Acidilactici NRRL B-50517 was added and then left for a period of time. The test tube was then removed and allowed to cool for a minimum of 10 minutes before being serially diluted in 0.1% saline and plated on MRS. Plates were incubated overnight at 45 ° C. and counted the next day. Survival rate was determined by saline + P.V. Acidilactici NRRL B-50517 control (0.2 g of 1B / g P. acidilactici NRRL B-50517 added to 20 mL of 0.1% saline at room temperature) was calculated.

＊：９００ｕＬの油をホットプレート上で８５℃に加熱した後、０．１ｇのＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７を添加し、その後、一定期間そのままにした。試験管を熱源から除去し、最低１０分間冷却させた後、連続希釈し、ＭＲＳ上でプレーティングした。プレートを４５℃で一晩インキュベートし、翌日数え上げた。熱処理の各期間での生存率を、室温のＥＶＯＯにおける生存の比率として計算した。 Conclusion: acidilactici NRRL B-50517 maintains a high viable count (CFU / g) in various commercial oils after high temperature treatment at room temperature.

*: After heating 900 uL of oil on a hot plate to 85 ° C., 0.1 g of P.I. Acidilactici NRRL B-50517 was added and then left for a period of time. The test tube was removed from the heat source and allowed to cool for a minimum of 10 minutes before serial dilution and plating on MRS. Plates were incubated overnight at 45 ° C. and counted the next day. The survival rate at each period of heat treatment was calculated as the ratio of survival in EVOO at room temperature.

＊：９００ｕＬの油をホットプレート上で８５℃に加熱した後、０．１ｇのＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７を添加し、その後、一定期間そのままにした。試験管を熱源から除去し、最低１０分間冷却させた後、連続希釈し、ＭＲＳ上でプレーティングした。プレートを３７℃で一晩インキュベートし、翌日数え上げた。熱処理の各期間での生存率を、室温のトウモロコシ油における生存の比率として計算した。 Conclusion: Acidilactici NRRL B-50517 survives high heat treatment in EVOO and produces a high viable count even after 30 minutes at 85 ° C. The probiotics are likely to be compatible with a variety of food preparation techniques, including techniques involving heating.

*: After heating 900 uL of oil on a hot plate to 85 ° C., 0.1 g of P.I. Acidilactici NRRL B-50517 was added and then left for a period of time. The test tube was removed from the heat source and allowed to cool for a minimum of 10 minutes before serial dilution and plating on MRS. Plates were incubated overnight at 37 ° C. and enumerated the next day. Survival at each period of heat treatment was calculated as the percentage of survival in corn oil at room temperature.

  Conclusion: Acidilactici NRRL B-50517 survives high heat treatment in corn oil and produces a high viable count even after 30 minutes at 85 ° C. The probiotics are likely to be compatible with various food preparation techniques, including those involving heating.

＊：１００ｍＬカップのＳｈｉｎｙ Ｓｐｏｏｎ Ｐｕｄｄｉｎｇを、２つの５０ｍＬの試験管に空け、２０分間８５℃に加熱した。元の容器を石鹸及び水で洗浄し、乾燥させ、２ｇの１０Ｂ／ｇのＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７を充填した。次に、加熱したプリンを元の容器に戻し、２０分間冷却した後、一晩冷蔵庫で貯蔵した。翌日、プリンを混合し、まず２ｇのプリンを５ｍＬの生理食塩水中に希釈し、続いて、ＭＲＳ上でのプレーティングのために連続希釈することによって、生存能力についてアッセイした。プレートを一晩インキュベートし、翌日数え上げた。 P. acidilactici NRRL B-50517 mimics the sterilization procedures used in the food industry to treat different foods that have been heat treated at 85 ° C. It can survive after dispensing into different containers with acidilactici NRRL B-50517 (Tables 11 and 12) and retains viability in products with diverse physiochemical properties for weeks or months. This provides a novel approach for introducing raw probiotics into food.

*: A 100 mL cup of Shiny Spoon Pudding was emptied into two 50 mL test tubes and heated to 85 ° C. for 20 minutes. The original container was washed with soap and water, dried and 2 g of 10 B / g P.I. Acidilactici NRRL B-50517 was charged. Next, the heated pudding was returned to the original container, cooled for 20 minutes, and stored in the refrigerator overnight. The next day, the purines were mixed and assayed for viability by first diluting 2 g purines into 5 mL saline followed by serial dilutions for plating on MRS. Plates were incubated overnight and enumerated the next day.

＊：試料を次のように調製した。
１．１０ｍＬのケチャップを２０分間８５℃で加熱し、続いて０．４ｇの１Ｂ／ｇのＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７及び５ｍＬの滅菌水と混合し、２０分間冷却させた後、生存能力について試験した。
２．１００ｇのフルーツカップ混合物を４５分間８５℃で加熱した後、元の容器中、１ｇの１Ｂ／ｇのＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７上に注ぎ戻し、生存能力について試験した。
３．５ｍＬのＥＶＯＯ及びＧｒｅａｔ Ｖａｌｕｅの油の試験管を２０分間８５℃で加熱した後、０．２ｇの１Ｂ／ｇのＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７に添加し、生存能力について試験した。
４．５ｍＬの２．５％のラクトースを含む試験管を２０分間８５℃で加熱した後、０．２ｇの１Ｂ／ｇのＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７を含む１５ｍＬの試験管に添加した。試験管を２０分間冷却させた後、生存能力について試験した。
５．２８３ｇのシロップ中のイチゴの混合物をビーカーに注ぎ入れ、３０分間８５℃で加熱した後、元の容器中、２ｇの１Ｂ／ｇのＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７の上に注ぎ戻した。２０分間冷却した後、混合物を生存能力について試験した。
６．５ｍＬのオレンジジュースを２０分間８５℃で加熱した後、０．１ｇの１Ｂ／ｇのＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７に添加し、２０分間冷却し、生存能力について試験した。 Conclusion: After incorporation into either vanilla pudding or chocolate pudding heated to 85 ° C. under conditions similar to sterilization, Acidilactici NRRL B-50517 maintains a very stable cell count for about a month when stored at refrigerator temperature.

*: The sample was prepared as follows.
1. 10 mL of ketchup was heated at 85 ° C. for 20 minutes, followed by 0.4 g of 1 B / g P.I. Acidilactici NRRL B-50517 and 5 mL of sterile water were mixed and allowed to cool for 20 minutes before testing for viability.
2. After heating 100 g of fruit cup mixture for 45 minutes at 85 ° C., 1 g of 1 B / g P.I. Pour back onto acidilactici NRRL B-50517 and test for viability.
After heating a 3.5 mL EVOO and Great Value oil test tube at 85 ° C. for 20 minutes, 0.2 g of 1 B / g P.A. Acidilactici NRRL B-50517 was added and tested for viability.
A test tube containing 4.5 mL of 2.5% lactose was heated at 85 ° C. for 20 minutes, and then 0.2 g of 1 B / g P.A. Add to a 15 mL tube containing acidilactici NRRL B-50517. The tubes were allowed to cool for 20 minutes before being tested for viability.
5. Pour a mixture of strawberries in 283 g syrup into a beaker and heat at 85 ° C. for 30 minutes, then 2 g of 1 B / g P.A. in the original container. Pour back onto acidilactici NRRL B-50517. After cooling for 20 minutes, the mixture was tested for viability.
After heating 6.5 mL of orange juice for 20 minutes at 85 ° C., 0.1 g of 1 B / g P.P. Added to acidilactici NRRL B-50517, cooled for 20 minutes and tested for viability.

  ** All viability studies are The acidilactici NRRL B-50517 + heat treated food mixture was serially diluted in 0.1% saline, plated on MRS and incubated overnight before enumerating the plates. Survival rate was determined by saline + P.V. Acidilactici NRRL B-50517 control (0.2 g of 1B / g P. acidilactici NRRL B-50517 added to 20 mL of 0.1% saline at room temperature) was calculated.

  Conclusion: Acidilactici NRRL B-50517 maintains viability in various liquid and solid matrices after high heat treatment and is highly adapted for incorporation into many different foods after sterilization or other similar high heat sterilization procedures Showing gender.

Example 8-Increasing probiotic viability under heat treatment by applying an amphiphilic product as a carrier Amphiphilic molecules are molecules that have both polar and nonpolar moieties in their structure. is there. Chemical compounds characterized by these molecules are essential for acceptance of biological and industrial processes.

  In this test, amphiphilic products such as lecithin, peanut butter, almond butter, soy butter, and cookie butter are applied as alternative carriers for Pediococcus acidilactici NRRL B-50517 fermentation culture. Excessive amphiphilic products can be found in P.I. Acidilactici NRRL B-50517 fermentation culture can be protected from severe dry heat or wet heat treatment. After mixing the amphiphilic product dry powder and bacterial freeze-dried fermentation culture with oil, the inherent properties of the amphiphilic product form a lipid paste, Acts as a carrier for acidilactici fermentation culture and can withstand dry heat treatment or hot water treatment similar to sterilization for food processing.

  There are several approaches for including probiotics in a sterilized food / sample product. A desirable approach is to include a probiotic together in the food / sample ingredients followed by a sterilization process that may include hydrothermal treatment. Alternatively, food / sample ingredients can be dispensed into the probiotic composition. This is practiced by removing the hydrothermally treated juice / liquid after the juice / liquid reaches the specified sterilization temperature and immediately combining it with a mixture of probiotics and amphiphilic products. Can do. For this reason, the following experiments show that hot water / liquid reaches the desired temperature, and immediately after that, hydrothermal heat treatment is applied to the combined probiotic and amphiphilic product for 5 minutes and 10 minutes, after which The assay was performed to determine the number of probiotics alive after these heat treatments. In these efforts, it can be shown that amphiphilic products combined with probiotics can undergo a heat treatment such as a sterilization process.

  Lecithin is defined as phosphatidylcholine and represents a natural mixture of plant and / or animal derived neutral and polar fats. This is an excellent emulsifier with low water solubility. In aqueous solution, the phospholipid can form either liposomes, bilayer sheets, micelles, or lamellar structures, depending on hydration properties and temperature. This produces a type of surfactant that is usually classified as amphiphilic.

＊：マルトデキストリンを担体として有する０．２ｇのＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７フリーズドライ発酵培養菌（１０億ｃｆｕ／ｇ）をこれらの試験に使用した。全ての加熱した試料。温度をデジタルプローブで確認した。試料を６５℃で３０分間予熱した後、適当な時間にわたって８５℃の乾熱に加えた。粉末を熱から取り出し、室温で放置して１０分間冷却した。乾燥粉末を、５ｍｌの生理食塩水に添加し、生理食塩水中で５分間混合し固めた後、３７℃のインキュベーターで一晩、ＭＲＳプレート上にプレーティングし、生存能力について採点した。 In this test, amphiphilic products such as lecithin, peanut butter, almond butter, soy butter, and cookie butter are applied as alternative carriers for Pediococcus acidilactici NRRL B-50517 fermentation culture. Excessive amphiphilic products can be found in P.I. Acidilactici NRRL B-50517 fermentation culture can be protected from severe dry heat or wet heat treatment. After mixing the amphiphilic product dry powder and the bacterial freeze-dried fermentation culture with oil, the inherent properties of the amphiphilic product can form a lipid paste. It carries acidilactici fermentation culture and makes it resistant by heat such as hot water treatment.

*: 0.2 g of P.I. having maltodextrin as a carrier. Acidilactici NRRL B-50517 freeze-dried fermentation culture (1 billion cfu / g) was used for these studies. All heated samples. The temperature was confirmed with a digital probe. Samples were preheated at 65 ° C. for 30 minutes and then added to 85 ° C. dry heat for an appropriate time. The powder was removed from the heat and allowed to cool at room temperature for 10 minutes. The dry powder was added to 5 ml saline, mixed and hardened in saline for 5 minutes, then plated on MRS plates overnight in a 37 ° C. incubator and scored for viability.

  1.75 g sunflower lecithin and 1.75 g P.I. Acidilactici NRRL B-50517 (1 billion cfu / g) of 0.2 g (500 million cfu / g) of powder was transferred onto weight paper. The sample was heated at 65 ° C. for 30 minutes and then cooled for 10 minutes. The other 65 ° C. treated samples were transferred to 85 ° C. for 10 and 30 minutes and allowed to cool for 10 minutes. The temperature was confirmed with a digital probe. After dissolving in 5 ml saline, the desired dilution was performed and 100 ul was plated on MRS agar plates and incubated overnight at 37 ° C. for viability calculations.

＊：０．５ｇの試料［１．４ｇのＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７フリーズドライ発酵培養菌（１００億ｃｆｕ／ｇ）＋８ｇのヒマワリレシチン＋９ｍＬのオリーブ油］を、１ｍｌの蒸留水を含むエッペンドルフ型試験管に移し、異なる時間８０℃または８２℃の水槽でインキュベートし、試験管中の蒸留水が８０℃または８２℃に達した直後、８０℃または８２℃で５分または１０分間、インキュベートを継続し、水槽から試験管を取り出した後、１０分間冷却させ、望ましい希釈を行い、１００ｕｌをＭＲＳ寒天プレート上にプレーティングし、生存能力算定のために３７℃で一晩インキュベートした。温度をデジタルプローブで確認した。 result:
Lecithin was added to P.I. By applying it as a carrier for acidilactici freeze-dried fermentation cultures, the survival of bacteria under different heat treatments has been demonstrated by heat treated P. aeruginosa. Compared with acidilactici freeze-dried culture alone, it was improved 5-10 times.

*: 0.5 g sample [1.4 g P.I. acidilactici NRRL B-50517 freeze-dried fermentation culture (10 billion cfu / g) +8 g sunflower lecithin + 9 mL olive oil] was transferred to an eppendorf tube containing 1 ml distilled water for different times in an 80 ° C. or 82 ° C. water bath. Incubate and immediately after the distilled water in the test tube reaches 80 ° C. or 82 ° C., continue the incubation at 80 ° C. or 82 ° C. for 5 or 10 minutes, remove the test tube from the water bath, and allow it to cool for 10 minutes. The desired dilutions were made, 100 ul was plated on MRS agar plates and incubated overnight at 37 ° C. for viability calculations. The temperature was confirmed with a digital probe.

＊：各試料は、７ｇのヒマワリレシチン粉末中に混合した１．４ｇの１０Ｂ／ｇの５０５１を有した（１．６７Ｂ ｃｆｕ／ｇ）。４つの試料を作製した。Ａ及びＢは室温、異なるＡ及びＢは３７℃のインキュベーションに配置した。乾燥材の小包に加えた。５ｍＬの生理食塩水中の０．２ｇの粉末を使用する希釈によって試験した。３０秒間のボルテックスによって均一に混合し、生理食塩水による連続希釈を行い、１００ｕｌの望ましい希釈物を一晩３７℃でＭＲＳプレート上にプレーティングし、個々のコロニーを計算した Results: P.P. A mixture of acidilactici NRRL B-50517 and sunflower lecithin was resuspended with olive oil as a paste and soaked for heat treatment at 80 ° C or 82 ° C. Once the heated bacterial paste was removed from the hot water temperature when it reached 80 ° C., the treated bacterial paste product was moved to room temperature for the cooling process and more than 37% viable bacteria were detected. Approximately 8.0% viable bacteria were detected with similar heat treatment up to 82 ° C. These heat treatment processes apply probiotics after the product has undergone a sterilization process and combine these hot products with amphiphilic products in a container. Dispensing directly into acidilactici fermentation culture.

*: Each sample had 1.4 g of 10 B / g 5051 mixed in 7 g sunflower lecithin powder (1.67 B cfu / g). Four samples were made. A and B were placed at room temperature and different A and B were placed at 37 ° C. Added to desiccant package. Tested by dilution using 0.2 g powder in 5 mL saline. Mix evenly by vortexing for 30 seconds, perform serial dilution with saline, plate 100 ul of desired dilution on MRS plates overnight at 37 ° C., and calculate individual colonies

＊：０．７グラムのＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＮＲＬ Ｂ−５０５１７、１００億ｃｆｕ／ｇ、３．５グラムのヒマワリレシチン、及び４．５ｍｌのオリーブ油を含む、０．５グラムのペーストを、１ｍｌの醤油を含むエッペンドルフ試験管に移し、８０℃の水槽でインキュベートした。醤油の温度が８０℃に達したら、１）直後に、２）８０℃で５分間インキュベートした後に、３）８０℃で１０分間インキュベートした後に、試験管を水槽から取り出し、３０秒間のボルテックスによって均一に混合し、生理食塩水による連続希釈を行い、１００ｕｌの望ましい希釈物を一晩３７℃でＭＲＳプレート上にプレーティングし、個々のコロニーを計算した。温度をデジタルプローブで確認した。

＊：０．５ｇのペースト（０．７ｇのＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ ｂ−５０５１７１０Ｂ／ｇと４．５ｍＬのオリーブ油との混合物と混合した３．５ｇのレシチン）を１ｍＬのジュース液中に移し、指定の時間、８０℃の水槽中で熱処理した。温度プローブを試験管中で使用して、内部温度が８０℃に達したときを決定した。ジュースの温度が８０℃に達したら、１）直後に、２）８０℃で５分間インキュベートした後に、３）８０℃で１０分間インキュベートした後に、試験管を水槽から取り出し、３０秒間のボルテックスによって均一に混合し、生理食塩水による連続希釈を行い、１００ｕｌの望ましい希釈物を一晩３７℃でＭＲＳプレート上にプレーティングし、個々のコロニーを計算した。温度をデジタルプローブで確認した。

＊：２８ｇのオート麦を加圧滅菌したガラスボウル中に秤量した。１．４ｇの１０Ｂ／ｇの５０５１を７ｇのＮＯＷヒマワリレシチン及び９ｍＬのオリーブ油と混合した。１．１ｇの１０Ｂ／ｇの５０５１を１０ｇのＳｋｉｐｐｙピーナッツバターと混合した。１ｇのレシチン（０．８Ｂ ｃｆｕ）、ＰＢ（１Ｂ ｃｆｕ）、または単独の１Ｂ／ｇの５０５１のいずれかを、オートミールの各ボウル（各３つ）に添加した。電子レンジで沸騰させた１１８ｍＬの水を、８２℃、８８℃、及び９３℃で各ボウルに添加した。温度をデジタルプローブで確認した。１ｇのオートミール混合物を、５ｍＬの生理食塩水中で希釈し、希釈し、ＭＲＳ上にプレーティングした。 Results: Lecithin and P.I. The mixture with acidilactici freeze-dried fermentation cultures was stable for more than 90 days at either room temperature or 37 ° C. P. The viability of acidilactici remains a similar viable amount through storage at both room temperature and 37 ° C.

*: 0.7 grams of P.I. Transfer 0.5 grams of paste containing acidilactici NNRL B-50517, 10 billion cfu / g, 3.5 grams of sunflower lecithin, and 4.5 ml of olive oil to an Eppendorf tube containing 1 ml of soy sauce, 80 ° C. Incubated in a water bath. When the temperature of the soy sauce reaches 80 ° C, immediately after 1), 2) after incubating at 80 ° C for 5 minutes, and after 3) incubating at 80 ° C for 10 minutes, the test tube is taken out of the water bath and uniformly by vortexing for 30 seconds And serial dilutions with saline, 100 ul of the desired dilution was plated on MRS plates overnight at 37 ° C. and individual colonies were calculated. The temperature was confirmed with a digital probe.

*: Transfer 0.5 g of paste (0.7 g of P. acidilactici NRRL b-5051710 B / g and 3.5 g of lecithin mixed with 4.5 mL of olive oil) into 1 mL of juice. Heat treatment was performed in a water bath at 80 ° C. for a time. A temperature probe was used in the test tube to determine when the internal temperature reached 80 ° C. When the juice temperature reaches 80 ° C, 1) immediately, 2) after 5 minutes incubation at 80 ° C, 3) after 10 minutes incubation at 80 ° C, the test tube is removed from the water bath and homogenized by vortexing for 30 seconds And serial dilutions with saline, 100 ul of the desired dilution was plated on MRS plates overnight at 37 ° C. and individual colonies were calculated. The temperature was confirmed with a digital probe.

*: 28 g of oats were weighed into a pressure sterilized glass bowl. 1.4 g 10B / g 5051 was mixed with 7 g NOW sunflower lecithin and 9 mL olive oil. 1.1 g of 10B / g 5051 was mixed with 10 g of Skippy peanut butter. Either 1 g of lecithin (0.8B cfu), PB (1B cfu), or single 1B / g 5051 was added to each bowl of oatmeal (3 each). 118 mL of water boiled in the microwave was added to each bowl at 82 ° C, 88 ° C, and 93 ° C. The temperature was confirmed with a digital probe. 1 g of oatmeal mixture was diluted in 5 mL of saline, diluted and plated on MRS.

Example 9-Effect of different amphiphilic products as a probiotic carrier on heat treatment In this example, the effect of different amphiphilic products was tested on their ability to enhance the heat resistance of Pediococcus acidilactici.

Peanut butter and P.I. A mixture of 0.5 g (1 billion cfu / g) with acidilactici NRRL B-50517 is placed in an Eppendorf tube and placed at 95 ° C. for 1, 2, 2.5, 5 and 10 minutes. Or placed at 85 ° C. for 1 minute, 2.5 minutes, 5 minutes, 10 minutes, and 20 minutes. The temperature was confirmed with a digital probe. 500 ul of saline was added, vortexed and diluted, and 100 ul of the desired dilution was plated on MRS plates overnight at 37 ° C. for viability calculations. Viability was obtained by dividing the number of viable bacteria upon heat treatment by the number of viable bacteria at room temperature (no heat treatment).

＊：空の試験管をホットプレート上で８５℃に予熱した。温度をデジタルプローブで確認した。空の試験管が温まったら、０．５ｍＬ（約０．５ｇのピーナッツバターとＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７との混合物（１０億ｃｆｕ／ｇ）を添加した。混合物を８５℃のホットプレート上に５分間置いたままにした。加熱後、試験管を約１０分間室温に置いて冷却させた。冷却後、混合物に１０ｍＬの生理食塩水を添加した。希釈を行い、１００ｕｌの望ましい希釈物を、生存能力算定のために３７℃で一晩、ＭＲＳプレート上にプレーティングした。生存率を、熱処理時の生菌の数を室温での（熱処理なし）生菌の数で割ることによって得た。

＊： Ｐ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７発酵培養菌とレシチンとの０．５ｇの混合物［３．５ｇのレシチンを０．７ｇのＰ．ａｃｉｄｉｌａｃｔｉｃｉ ＮＲＲＬ Ｂ−５０５１７（１００億ｃｆｕ／ｇ）及び４．５ｍＬのオリーブ油と混合することによって調製］。０．５ｇの混合物を１ｍＬの液体に秤量し、指定の時間、８０℃の水槽において熱処理した。温度プローブを試験管中で使用して、内部温度が８０℃に達したときを決定した。直後：温度到達直後に水槽から除去した。加熱後、試験管を約１０分間室温に置いて冷却させた。冷却後、混合物に１０ｍＬの生理食塩水を添加した。希釈を行い、１００ｕｌの望ましい希釈物を、生存能力算定のために３７℃で一晩、ＭＲＳプレート上にプレーティングした。生存率を、熱処理時の生菌の数を室温での（熱処理なし）生菌の数で割ることによって得た。 P. A mixture of 0.5 g (1 billion cfu / g) of acidilactici NRRL B-50517 fermentation culture and peanut butter was placed in a 1.5 ml test tube containing 1 ml of distilled water. Place a 1.5 ml test tube on the heat plate. After 5 minutes or until the temperature reaches 80 ° C., take 1 minute, 2.5 minutes, and 5 minutes. The temperature was confirmed with a digital probe. After heat treatment, the sample was vortexed and diluted and 100 ul of the desired dilution was plated on MRS plates overnight at 37 ° C. for viability calculations. Viability was obtained by dividing the number of viable bacteria upon heat treatment by the number of viable bacteria at room temperature (no heat treatment).

*: An empty test tube was preheated to 85 ° C. on a hot plate. The temperature was confirmed with a digital probe. Once the empty test tube was warmed, 0.5 mL (a mixture of about 0.5 g peanut butter and P. acidilactici NRRL B-50517 (1 billion cfu / g) was added. The mixture was placed on an 85 ° C. hot plate. After heating, the tube was allowed to cool for about 10 minutes at room temperature, after which it was cooled and 10 mL of saline was added to the mixture. Viability was plated on MRS plates overnight at 37 ° C. Viability was obtained by dividing the number of viable bacteria at heat treatment by the number of viable bacteria at room temperature (no heat treatment).

*: P.I. acidilactici NRRL B-50517 0.5 g mixture of fermented culture and lecithin [3.5 g lecithin 0.7 g P.P. prepared by mixing with acidilactici NRRL B-50517 (10 billion cfu / g) and 4.5 mL olive oil]. 0.5 g of the mixture was weighed into 1 mL of liquid and heat treated in a water bath at 80 ° C. for the specified time. A temperature probe was used in the test tube to determine when the internal temperature reached 80 ° C. Immediately after removing from the water tank immediately after reaching the temperature. After heating, the test tube was allowed to cool for about 10 minutes at room temperature. After cooling, 10 mL of physiological saline was added to the mixture. Dilutions were made and 100 ul of the desired dilution was plated on MRS plates overnight at 37 ° C. for viability calculations. Viability was obtained by dividing the number of viable bacteria upon heat treatment by the number of viable bacteria at room temperature (no heat treatment).

  While the presently preferred embodiment of the invention has been shown and described, those skilled in the art will appreciate that other and further embodiments depart from the spirit and scope of the invention as described in this application. It will be understood that this application includes all such modifications that may be made without departing from the intended scope of the claims set forth herein. All patents and publications mentioned and / or cited herein are to the same extent as if each individual publication was specifically and individually indicated as incorporated by reference in its entirety. Incorporated by reference.

Claims (81)

  A composition comprising probiotics, wherein the composition is in a mixture with an effective amount of an amphiphile that enhances the viability of the probiotic in the composition when subjected to heat. ,Composition.   The composition of claim 1, wherein the probiotic is mixed with the amphiphile as a lyophilized fermentation medium.   The composition according to claim 1, wherein the amphiphile is mixed with the probiotic as a dry powder.   The composition according to claim 1, further comprising an oil mixed with the amphiphile and probiotic.   The composition of claim 4, wherein the oil comprises peanut oil, corn oil, vegetable oil, soybean oil, or olive oil.   7. The ratio (w / w) of the amphiphile to the probiotic to the oil ranges from about 1: 10: 0.1 to about 10: 1: 25. A composition according to 1.   7. A composition according to any of claims 1 to 6, wherein the composition forms a lipid paste and the amphiphile serves as a carrier for the probiotic.   The composition according to any one of claims 1 to 7, wherein the probiotic is Pediococcus acidilactici.   The composition according to any of claims 1 to 8, wherein the probiotic is Pediococcus acidilactici NRRL B-50517.   The composition according to any of claims 1 to 9, wherein the amphiphile comprises lecithin, peanut butter, almond butter, soy butter, or cookie butter.   The composition according to claim 1, wherein the heat is dry heat.   12. A composition according to claim 11, wherein the dry heat is at least 50 <0> C.   The composition in any one of Claims 11-12 whose said dry heat is the range of 50 to 100 degreeC.   14. A composition according to any preceding claim, wherein the composition is subjected to heat immediately and removed from the heat immediately thereafter.   15. A composition according to any preceding claim, wherein the composition is subjected to heat for at least about 10 minutes.   16. The composition according to any one of claims 1 to 15, wherein the composition is subjected to a second temperature of dry heat after the composition is subjected to a first temperature of dry heat.   The composition of claim 16, wherein the first temperature is about 65 ° C. and the second temperature is about 85 ° C.   The composition is subjected to heat at the first temperature for at least about 1 second to at least overnight, and is subjected to heat at the second temperature for at least about 1 second to at least about 30 minutes. Item 18. The composition according to any one of Items 16 to 17.   18. A composition according to any of claims 16 to 17, wherein the composition is subjected to heat at the first temperature for about 30 minutes and is subjected to heat at the second temperature for about 10 minutes.   18. The composition of any of claims 16-17, wherein the composition is subjected to heat at the first temperature for about 30 minutes and is subjected to heat at the second temperature for about 30 minutes.   The composition according to claim 1, wherein the heat is liquid heat.   The composition of claim 21, wherein the liquid heat is at least about 65 ° C.   The composition of claim 21, wherein the liquid heat is at least about 80 ° C.   The composition of claim 21, wherein the liquid heat is at least about 82 ° C.   25. A composition according to any of claims 21 to 24, wherein the composition is subjected to the liquid heat for at least about 1 second.   25. A composition according to any of claims 21 to 24, wherein the composition is subjected to the heat for at least about 5 minutes.   25. A composition according to any of claims 21 to 24, wherein the composition is subjected to the heat for at least about 10 minutes.   28. The composition of any of claims 1-27, wherein the viability is enhanced at least about 2-fold compared to a composition lacking the effective amount of amphiphile.   28. The composition of any of claims 1-27, wherein the viability is enhanced at least about 10 times compared to a composition lacking the effective amount of amphiphile.   30. The composition according to any one of claims 1 to 29, wherein the amphiphile is sunflower lecithin.   31. The composition of any of claims 1-30, wherein the ratio (w / w) of the amphiphile to the probiotic ranges from about 1:10 to about 25: 1.   32. The composition of any of claims 1-31, wherein the ratio (w / w) of the amphiphile to the probiotic ranges from about 1: 1 to about 10: 1.   35. The composition according to any of claims 1-32, wherein the ratio (w / w) of the amphiphile to the probiotic is about 5: 1.   35. The composition according to any of claims 1-32, wherein the ratio (w / w) of the amphiphile to the probiotic is about 10: 1.   The container suitable for accommodating the foodstuff containing the composition in any one of Claims 1-34.   36. The container of claim 35, wherein the container is made from a material comprising glass, plastic, paper / carton, aluminum, or a dried plant fruit shell.   37. A container according to any of claims 35 to 36, wherein the composition is coated on the surface or part of the container.   The container according to any one of claims 35 to 37, further comprising a food.   The food comprises oil, vinegar, yogurt, fruit products, applesauce, dairy products, beverages, candy, snacks, juices, dessert foods such as cookies, cakes, JELLO, fruit bars, fruit custards, or tiramisu, 39. A container according to claim 38.   A container suitable for containing food, wherein a part or surface of the container is coated with an effective amount of Pediococcus acidilactici.   41. The container of claim 40, wherein the Pediococcus acidilactici is NRRL B-50517 strain.   42. A container according to any of claims 40 or 41, wherein the container is made from a material comprising glass, plastic, paper / carton, and aluminum.   44. A container according to any of claims 40 to 43, wherein the Pediococcus acidilactici is coated on the inner surface of the container.   The container according to any one of claims 40 to 43, further comprising a food.   45. The container of claim 44, wherein the food product comprises oil, vinegar, yogurt, fruit product, apple sauce, dairy product, beverage, candy, snack confectionery, and juice. A method for enhancing the viability of probiotics in a composition subjected to heat comprising:
i) adding an effective amount of an amphiphile to the composition comprising probiotics;
ii) subjecting the composition to heat,
Thereby, the viability of the probiotic in the composition subjected to the heat is enhanced.   47. The method of claim 46, wherein the amphiphile is mixed with the probiotic lyophilized fermentation medium.   48. A method according to any of claims 46 to 47, wherein the amphiphile is in the form of a dry powder.   49. A method according to any of claims 46 to 48, wherein the amphiphile is mixed with the probiotic and oil.   50. The method of claim 49, wherein the oil comprises peanut oil, corn oil, vegetable oil, soybean oil, or olive oil.   51. The ratio of the amphiphile to the probiotic to the oil (w / w) ranges from about 1: 10: 0.1 to about 10: 1: 25. The method described in 1.   51. A method according to any of claims 46 to 50, wherein the amphiphile forms a lipid paste and acts as a carrier.   53. A method according to any of claims 46 to 52, wherein the probiotic is Pediococcus acidilactici.   53. A method according to any of claims 46 to 52, wherein the probiotic is Pediococcus acidilactici NRRL B-50517.   55. A method according to any of claims 46 to 54, wherein the amphiphile comprises lecithin, peanut butter, almond butter, soy butter or cookie butter.   56. A method according to any of claims 46 to 55, wherein the heat is dry heat.   57. The method of claim 56, wherein the dry heat is at least 50 ° C.   57. A method according to any of claims 55 to 56, wherein the dry heat is in the range of 50 to 100 <0> C.   59. A method according to any of claims 46 to 58, wherein the composition is immediately subjected to heat and immediately removed from the heat.   60. The method of any of claims 46-59, wherein the composition is subjected to heat for at least about 10 minutes.   61. The method of any of claims 46-60, wherein the composition is subjected to a second temperature of dry heat after the composition is subjected to a first temperature of dry heat.   62. The method of claim 61, wherein the first temperature is about 65 degrees Celsius and the second temperature is about 85 degrees Celsius.   64. The composition is subjected to heat at the first temperature for at least about 1 to at least about 30 minutes and is subjected to heat at the second temperature for at least about 1 to at least about 30 minutes. 63. The method according to any of 62.   64. The method of any of claims 61-63, wherein the composition is subjected to heat at the first temperature for about 30 minutes and subjected to heat at the second temperature for about 10 minutes.   64. The method of any of claims 61-63, wherein the composition is subjected to heat at the first temperature for about 30 minutes and subjected to heat at the second temperature for about 30 minutes.   56. A method according to any of claims 46 to 55, wherein the heat is liquid heat.   68. The method of claim 66, wherein the liquid heat is at least about 50 ° C.   68. The method of claim 66, wherein the liquid heat is at least about 80 degrees Celsius.   68. The method of claim 66, wherein the liquid heat is at least about 82 degrees Celsius.   70. A method according to any of claims 66 to 69, wherein the composition is immediately subjected to the heat and immediately removed from the heat.   71. The method of any of claims 66-70, wherein the composition is subjected to the heat for at least about 5 minutes.   71. The method of any of claims 66-70, wherein the composition is subjected to the heat for at least about 10 minutes.   73. The method of any of claims 46-72, wherein the viability is enhanced at least about 2-fold compared to a composition lacking the effective amount of amphiphile.   73. The method of any of claims 46-72, wherein the viability is enhanced at least about 10 times compared to a composition lacking the effective amount of amphiphile.   75. A method according to any of claims 46 to 74, wherein the amphiphile is sunflower lecithin.   76. The method of any of claims 46-75, wherein the ratio (w / w) of the amphiphile to the probiotic ranges from about 1:10 to about 25: 1.   77. A method according to any of claims 46 to 76, wherein the ratio (w / w) of the amphiphile to the probiotic ranges from about 1: 1 to about 10: 1.   78. A method according to any of claims 46 to 77, wherein the ratio (w / w) of the amphiphile to the probiotic is about 5: 1.   78. A method according to any of claims 46 to 77, wherein the ratio (w / w) of the amphiphile to the probiotic is about 10: 1.   80. A method according to any of claims 46 to 79, wherein the composition is contacted with food and subjected to heat with the food. 47. Subjecting the composition to the heat step of part ii) comprises contacting the composition with food, wherein the food is already subjected to heat prior to contacting the composition. A method according to -79.

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