Methods for preventing, delaying, or ameliorating atopic diseases

Abstract

Compositions and methods are provided for preventing, delaying, or ameliorating atopic dermatitis in breastfed infants at high risk of developing atopic disease, the methods comprising administering a composition comprising administering Bifidobacterium to the breastfed infant.

Description

Technical Field

[0001] The present invention generally relates to methods for preventing, delaying, or improving atopic diseases such as atopic dermatitis, and particularly to methods comprising administering a composition comprising an effective amount of Bifidobacterium to infants being breastfed.

Background Art

[0002] Atopic diseases are a class of diseases in which the immune system generates immunoglobulins against common environmental allergens that are generally considered harmless. An example of an atopic disease is atopic dermatitis ("AD"). AD, also known as atopic eczema, is a chronic inflammatory allergic skin disorder that frequently occurs in childhood. AD is characterized by eczematous lesions that become scaly with strong itching, and in the chronic case, with cracks and lichenification, i.e., erythematous spots with exanthema, blister formation, and crust formation.

[0003] As the most common pediatric allergic disease, AD affects up to 30% of children. The incidence of AD peaks in infancy, but the disease generally persists and / or recurs in adulthood. There are numerous topical treatments (lotions, ointments, etc.), but none can completely cure eczema.

Summary of the Invention

Problems to be Solved by the Invention

[0004] Therefore, there is a continuing need for a treatment that is effective in preventing, delaying, and / or improving the onset of atopic dermatitis (as well as other atopic diseases) in infants.

Means for Solving the Problems

[0005] Therefore, one aspect of the present invention relates to a method for preventing, delaying, or improving atopic dermatitis in breastfed infants. In one or more embodiments, the method is used for B. longum, B. breve, B. bifidum, B. pseudocatenulatum, B. globosum, B. adolescentis, B. moukalabense, B. reuteri, B. pseudolongum, B. dentium, B. catenulatum, B. sp002742445, B. callitrichos, B. scardovii, B. tissieri, B. subtile, B. gallinarum, B. choerinum, B. angulatum, B. primatium, B. myosotis, B. mongoliense, B. merycicum, B. lemurum, B. stellenboschense, B. scaligerum, B. saguini, B. pullorum, B. fels The method involves administering to a breastfed infant a composition containing an effective amount of Bifidobacterium, selected from the group consisting of B. ineum, B. eulemuris, B. cuniculi, B. callitrichos_A, B. biavatii, B. anseris, B. vansinderenii, B. sp900551485, B. sp003952945, B. sp003952025, B. sp003952005, B. simiarum, B. pseudolongum_C, B. parmae, B. margollesii, B. kashiwanohense_A, B. italicum, B. imperatoris, B. cricetid, B. catulorum, B. callitrichidarum, B. animalis, B. aesculapii, and combinations thereof.

[0006] In some embodiments, Bifidobacterium is selected from the group consisting of B. longum, B. breve, B. kashiwanohense, and combinations thereof. In one or more embodiments, Bifidobacterium is a B. longum subspecies selected from the group consisting of longum, suis, infantis, and combinations thereof. In some embodiments, Bifidobacterium is B. infantis. In one or more embodiments, Bifidobacterium is mixed with breast milk before administering it to a breastfed infant. In some embodiments, Bifidobacterium is mixed with infant formula before administering it to a breastfed infant. In one or more embodiments, Bifidobacterium is mixed with about 3 to about 5 mL of breast milk, infant formula, or water before administering it to a breastfed infant. In some embodiments, breastfed infants are breastfed for more than 50% of their time. In one or more embodiments, breastfed infants are exclusively breastfed. In some embodiments, Bifidobacterium is in powder form mixed with lactose. In one or more embodiments, approximately 5 to 15 billion CFU of Bifidobacterium are administered to breastfed infants. In some embodiments, B. infantis is administered once daily. In one or more embodiments, B. infantis is first administered within the first two weeks of life. In some embodiments, B. infantis is administered before the first 12 weeks of life. In one or more embodiments, Bifidobacterium is first administered within the first two weeks of life and by the 12th week of life. In some embodiments, breastfed infants are at high risk of developing atopic diseases. In one or more embodiments, Bifidobacterium contains the EVC001 strain.

[0007] Another embodiment relates to a method for preventing, delaying, or improving atopic dermatitis in a breastfed infant having at least one first-degree relative with a history of atopic disease, the method comprising administering an effective amount of B. infantis mixed with breast milk to the breastfed infant once daily, wherein the breastfed infant is at least 90% breastfed, and the B. infantis is first administered within the first two weeks after birth.

[0008] In some embodiments, B. infantis contains the EVC001 strain. In one or more embodiments, B. infantis is mixed with about 3 to 5 mL of breast milk or infant formula before being administered to breastfed infants. In some embodiments, breastfed infants are exclusively breastfed. In one or more embodiments, B. infantis is in powder form mixed with lactose. In some embodiments, 8 billion CFU of B. infantis is administered to breastfed infants. In one or more embodiments, B. infantis is administered once daily until 12 weeks of age.

[0009] Another embodiment relates to a method for preventing, delaying, or improving atopic diseases selected from the group consisting of food allergies, allergic rhinitis, asthma, and combinations thereof in breastfed infants, wherein the method comprises administering a composition containing an effective amount of Bifidobacterium.

[0010] In some embodiments, Bifidobacterium is used to feed breastfed infants B. longum, B. breve, B. bifidum, B. pseudocatenulatum, B. globosum, B. adolescentis, B. moukalabense, B. reuteri, B. pseudolongum, B. dentium, B. catenulatum, B. sp002742445, B. callitrichos, B. scardovii, B. tissieri, B. subtile, B. gallinarum, B. choerinum, B. angulatum, B. primatium, B. myosotis, B. mongoliense, B. merycicum, B. lemurum, B. telllenboschense, B. scaligerum, B.saguini, B.pullorum, B.felsineum, B.eulemuris, B.cuniculi, B.callitrichos_A, B.biav atii, B.anseris, B.vansinderenii, B.sp900551485, B.sp003952945, B.sp003952025, B.sp003952005, B.si miarum, B. pseudolongum_C, B. parmae, B. margollesii, B. kashiwanohense_A, B. italicum, B. imperatoris, B. cricetid, B. catulorum, B. callitrichidarum, B. animalis, B. aesculapii, and combinations thereof. In one or more embodiments, Bifidobacterium is selected from the group consisting of B. longum, B. breve, B. kashiwanohense, and combinations thereof. In some embodiments, Bifidobacterium is a subspecies of B. longum selected from the group consisting of longum, suis, infantis, and combinations thereof. In one or more embodiments, Bifidobacterium is B. infantis.In some embodiments, Bifidobacterium is mixed with breast milk before being administered to breastfed infants. In one or more embodiments, Bifidobacterium is mixed with infant formula before being administered to breastfed infants. In some embodiments, Bifidobacterium is mixed with about 3 to about 5 mL of breast milk, infant formula, or water before being administered to breastfed infants. In one or more embodiments, breastfed infants are breastfed for more than 50% of their diet. In some embodiments, breastfed infants are exclusively breastfed. In one or more embodiments, Bifidobacterium is in powder form mixed with lactose. In some embodiments, about 5 to about 15 billion CFU of Bifidobacterium is administered to breastfed infants. In one or more embodiments, B. infantis is administered once daily. In some embodiments, B. infantis is first administered within the first two weeks of life. In one or more embodiments, B. infantis is administered before the first 12 weeks of life. In some embodiments, Bifidobacterium is first administered within the first two weeks of life and by the 12th week of life. In one or more embodiments, breastfed infants are at higher risk of developing atopic diseases. In some embodiments, Bifidobacterium includes the EVC001 strain.

[0011] Another embodiment relates to a method for improving infant colic, infant sleep, or infant physical measurement in breastfed infants, the method comprising administering a composition containing an effective amount of bifidobacterium.

[0012] In one or more embodiments, Bifidobacterium is used to feed breastfed infants B.longum, B.breve, B.bifidum, B.pseudocatenulatum, B.globosum, B.adolescentis, B.moukalabense, B.reuteri, B.pseudolongum, B.dentium, B.catenulatum, B.sp002742445, B.callitrichos, B.scardovii, B.tissieri, B.subtile, B.gallinarum, B.choerinum, B.angulatum, B.primatium, B.myosotis, B.mongoliense, B.merycicum, B.lemurum, B.stellenboschense, B. scaligerum, B.saguini, B.pullorum, B.felsineum, B.eulemuris, B.cuniculi, B.callitrichos_A, B.biav atii, B.anseris, B.vansinderenii, B.sp900551485, B.sp003952945, B.sp003952025, B.sp003952005, B.si miarum, B. pseudolongum_C, B. parmae, B. margollesii, B. kashiwanohense_A, B. italicum, B. imperatoris, B. cricetid, B. catulorum, B. callitrichidarum, B. animalis, B. aesculapii, and combinations thereof. In some embodiments, Bifidobacterium is selected from the group consisting of B. longum, B. breve, B. kashiwanohense, and combinations thereof. In one or more embodiments, Bifidobacterium is a subspecies of B. longum selected from the group consisting of longum, suis, infantis, and combinations thereof. In some embodiments, Bifidobacterium is B. infantis.In one or more embodiments, Bifidobacterium is mixed with breast milk before being administered to breastfed infants. In some embodiments, Bifidobacterium is mixed with infant formula before being administered to breastfed infants. In one or more embodiments, Bifidobacterium is mixed with about 3 to about 5 mL of breast milk, infant formula, or water before being administered to breastfed infants. In some embodiments, breastfed infants are breastfed for more than 50% of their diet. In one or more embodiments, breastfed infants are exclusively breastfed. In some embodiments, Bifidobacterium is in powder form mixed with lactose. In one or more embodiments, about 5 to about 15 billion CFU of Bifidobacterium are administered to breastfed infants. In some embodiments, B. infantis is administered once daily. In one or more embodiments, B. infantis is first administered within the first two weeks of life. In some embodiments, B. infantis is administered before the first 12 weeks of life. In one or more embodiments, Bifidobacterium is first administered within the first two weeks of life and by the 12th week of life. In some embodiments, breastfed infants are at higher risk of developing atopic diseases. In one or more embodiments, Bifidobacterium includes the EVC001 strain.

[0013] These and other features and advantages of the present invention will be readily apparent from the following "Modes for Carrying Out the Invention". [Modes for carrying out the invention]

[0014] Where used herein, the term “effective dose” means an amount sufficient to induce the desired effect. The term “safe dose” means an amount low enough to avoid serious side effects. The safe and / or effective dose of a compound, extract, or composition will vary depending, for example, the age, health condition, and environmental exposure of the end user, the duration and nature of the treatment, the specific extract, component, or composition used, the specific pharmaceutically acceptable carrier used, and similar factors.

[0015] As used herein, “essentially free” or “substantially free” of an ingredient means that the ingredient is present in less than 0.1 weight percent or less than 0.01 weight percent, or is not present at all.

[0016] One aspect of the present invention relates to a method for preventing, delaying, or improving atopic disease in breastfed infants at high risk of developing atopic disease, the method comprising administering a composition containing an effective amount of Bifidobacterium to the breastfed infant. Another aspect of the present invention relates to the use of Bifidobacterium for the prevention, delaying, or improving atopic disease in breastfed infants at high risk of developing atopic disease. Atopic disease may be selected from the group consisting of food allergies, allergic rhinitis, asthma, and combinations thereof. Atopic disease may be atopic dermatitis.

[0017] AD arises from a combination of impaired epidermal barrier function, T-cell activation, and enterotoxosis of skin commensal microorganisms, and can precede the development of other atopic diseases, such as food allergies, asthma, and allergic rhinitis, in the so-called "allergic march." Approximately one-third of children with AD develop food allergies due to transcutaneous allergic sensitization via eczematous skin. In fact, a distinct AD endotype associated with food allergies has been identified, characterized by altered terminal epidermal differentiation with changes in collagen expression, T-helper 2 (Th2) immunotranscripts, inadequate skin barrier function, and a predisposition to cutaneous Staphylococcus aureus colonization and infection.

[0018] Another aspect of the present invention relates to a method for improving infant colic, infant sleep, or anthropometric measurements in breastfed infants at high risk of developing atopic diseases, wherein the method comprises administering a composition containing an effective amount of Bifidobacterium. Another aspect of the present invention relates to the use of Bifidobacterium in improving infant colic, infant sleep, or anthropometric measurements in breastfed infants at high risk of developing atopic diseases.

[0019] Improvement or enhancement of any of the above conditions can be measured by methods known in the art. For example, improvement in the severity of atopic dermatitis can be measured using the Eczema Area and Severity Index (EASI), further defined below. Improvement in infant colic can be indicated by meeting fewer Rome IV criteria (defined below) or by a reduction in crying or fussing events. Improvement in infant sleep can be measured using the BISQ-R score (further discussed below) or by a reduction in one or more sleep pattern events (e.g., sleep latency, number and duration of nighttime awakenings, maximum length of sleep, total nighttime sleep). Improvement or enhancement may relate to conditions in infants not treated with Bifidobacterium.

[0020] As used herein, the term “high risk of developing atopic disease” refers to an infant who has one first-degree relative with a history of atopic disease (i.e., a biological parent or full sibling with AD, allergic rhinitis, or asthma reported by the mother and diagnosed by a physician). The infant may be born vaginally or by cesarean section.

[0021] Surprisingly, the incidence of Alzheimer's disease (AD), other atopic diseases, infant colic, infant sleep disorders, or infant physical measurements has been found to be affected even in breastfed infants. The greatest risk factor for AD is a family history of atopic disease; if one parent has atopic disease, the risk of the disease increases by 50%, and if one parent has AD, the risk increases by 300%. Genetic susceptibility to AD arises from multiple loci, including semi-dominant loss-of-function mutations in the filaggrin (FLG) gene, which impairs skin barrier function, and the Th2 cytokine cluster locus on chromosome 5q31.1. Juvenile determinants play a role in the development of atopic diseases, including AD. The gut microbiota significantly influences the development of childhood-onset AD and allergies due to its central role in the development and regulation of a healthy immune system against dysregulated responses associated with allergic diseases. Studies on the association between neonatal and infant gut microbiota and Alzheimer's disease (AD) have shown altered gut microbiota in the neonatal period, characterized by decreased diversity and increased concentrations of fecal Bacteroidaceae and Enterobacteriaceae. Conversely, lower relative levels of Bifidobacteriaceae and Lactobacillaceae are associated with the development of AD.

[0022] The results of some, though not all, studies conducted in this field contradict the finding that probiotics reduce the incidence and severity of Alzheimer's disease (AD). The pathophysiology of AD is complex and multifactorial, involving elements of impaired skin barrier function (sometimes mediated by loss-of-function mutations in the FLG gene), altered cell-mediated immune responses, IgE-mediated hypersensitivity, and environmental factors. The gut microbiota interacts with the skin as one of the major regulators in the gut-skin axis. For example, the gut microbiota can influence both innate and adaptive immunity through the production of secretory factors or metabolites that can enter circulation and thus induce systemic effects. This relationship plays a crucial role in maintaining skin homeostasis by supporting epithelial differentiation and immunomodulation. Conversely, the gut microbiota is involved in the pathophysiology of inflammatory skin disorders, including psoriasis and AD.

[0023] The variability in findings is partly due to heterogeneity in design, study population, and diagnostic criteria, as well as the use of different probiotics that, in all cases, have never been demonstrated to colonize the infant gut or affect infant immunotraining.

[0024] While we do not wish to be bound by any particular theory, it is thought that dominant colonization of the gut by B. infantis or similar Bifidobacterium in infancy may promote immunomodulation and reduce the risk of developing Alzheimer's disease (AD) in the first year of life in at-risk infants.

[0025] The intestine of an infant is sterile before birth, and the microbiota that develops afterwards is greatly influenced by how the infant is born and whether the infant is breastfed. In general, infants born via cesarean section (C-section) do not pass through the birth canal and are thought to develop a different gut microbiota profile than vaginally born infants. Instead, C-section infants are more likely to be exposed to the bacteria in their environment. Furthermore, breast milk itself has probiotic properties and contains oligosaccharides that help stimulate the growth of Bifidobacteriaceae and Lactobacillaceae. Therefore, breastfeeding an infant would be expected to help restore the infant's microbiota profile to an ideal profile, regardless of how the infant was born.

[0026] Bifidobacterium Bifidobacterium is a genus of Gram-positive anaerobic bacteria found in the gastrointestinal tract, vaginal tract, and oral tract of mammals, including humans. A suitable Bifidobacterium may have at least one human milk oligosaccharide (HMO) gene cluster. A Bifidobacterium may be similar to B. infantis. Bifidobacterium includes B.longum, B.breve, B.bifidum, B.pseudocatenulatum, B.globosum, B.adolescentis, B.moukalabense, B.reuteri, B.pseudolongum, B.dentium, B.catenulatum, B.sp002742445, B.call itrichos, B.scardovii, B.tissieri, B.subtile, B.gallinarum, B.choerinum, B.angulatum, B.primatium, B.myosotis, B.mongoliense, B.merycicum, B.lemurum, B.stellenboschense, B.scaligerum, B.sag You may choose from the group consisting of uini, B.pullorum, B.felsineum, B.eulemuris, B.cuniculi, B.callitrichos_A, B.biavatii, B.anseris, B.vansinderenii, B.sp900551485, B.sp003952945, B.sp003952025, B.sp003952005, B.simiarum, B.pseudolongum_C, B.parmae, B.margollesii, B.kashiwanohense_A, B.italicum, B.imperatoris, B.cricetid, B.catulorum, B.callitrichidarum, B.animalis, B.aesculapii, and combinations thereof. Bifidobacterium may be selected from the group consisting of B. longum, B. breve, B. kashiwanohense, and combinations thereof.The Bifidobacterium may be B. longum. It may also be a subspecies of B. longum selected from the group consisting of Bifidobacterium, longum, suis, and infantis.

[0027] As used herein, the terms "Bifidobacterium infantis" or "B. infantis" mean to refer to a subspecies of Bifidobacterium longum subspecies infantis. B. infantis may include strain EVC001. B. infantis can be isolated and cultured using methods known in the art.

[0028] B. infantis may be administered in combination with one or more other probiotics (i.e., other bacteria intended to have health benefits). The other probiotics may be strains selected from the genera Lactobacillus, Lacticaseibacillus, and Bifidobacterium. Examples of Bifidobacterium species include B. infantis, B. longum (subspecies other than B. infantis), B. Breve, B. catenulatum, B. adolescentis, B. animalis, B. gallicum, B. lactis, B. pseudocatenulatum, and other strains of B. bifidum. Examples of Lactobacillus strains include L. paracasei, L. acidophilus, L. johnsonii, L. delbrueckii, L. crispus, L. gasser, L. zeae, and combinations thereof. Examples of Lacticaseibacillus include L. casei, L. rhamnosus, and combinations thereof. Other probiotics include Lactiplantibacillus plantarum, Limosilactobacillus fermentum, and Ligilactobacillus salivarius. Alternatively, Bifidobacterium may be administered without any other probiotics; that is, Bifidobacterium may be formulated to essentially contain no other probiotics.

[0029] The gut microbiota profile of infants can be tested and monitored using methods known in the art to determine colonization by Bifidobacterium. Fecal samples can be used in such methods.

[0030] composition Bifidobacterium can be formulated into compositions that are easy to use and allow for consistent dosing. Fermentation products from Bifidobacterium production can be concentrated and freeze-dried to provide concentrated powders. The compositions may contain approximately 1 million, 500 million, 1 billion, 2 billion, 3 billion, 4 billion, 5 billion, 6 billion, 7 billion, 8 billion, 9 billion, 10 billion, or 12 billion colony-forming units (CFUs) per gram of dry weight.

[0031] Bifidobacterium may also be formulated with oligosaccharides. As used herein, the term “oligosaccharide” refers to a sugar polymer containing 2-20, 2-10, 3-20, or 3-10 monosaccharide units. Oligosaccharides may be found in the milk of mammals (e.g., humans or cattle). Oligosaccharides may be synthesized.

[0032] Compositions containing bifidobacterium may also contain auxiliary components. Such auxiliary components are commonly used in the art and may be selected from metabolites, fluids, or combinations thereof. Examples of fluids include starch, silicon dioxide, cellulose, sodium bicarbonate, and calcium silicate. The auxiliary components may also be milk proteins or components. The auxiliary components may include lactose. That is, in such examples, bifidobacterium is in powder form mixed with lactose.

[0033] The final form of the composition may be any known in the art. As described above, Bifidobacterium may be in a dry form as a powder (e.g., spray-dried or freeze-dried). The powder may be administered as packets, sachets, tablets, foods, capsules, lozenges, tablets, suspensions, dry forms, etc.

[0034] A Bifidobacterium product preferred according to one or more embodiments of the present invention is Evivo® probiotics, available from Evolve BioSystems (Davis, CA), which is packaged in a sachet containing 8 billion CFU of B. infantis (EVC001) co-formulated with lactose.

[0035] Administration of Bifidobacterium As used herein, the term “administer” means providing a given dose of Bifidobacterium to an infant as part of their diet (i.e., it is used as a dietary supplement). Bifidobacterium may be mixed with any medium that can be consumed by the infant, including breast milk, infant formula, water, or food, before administering Bifidobacterium to the infant. Bifidobacterium may be mixed with breast milk before administering Bifidobacterium to a breastfed infant. Alternatively, Bifidobacterium may be mixed with infant formula before administering Bifidobacterium to a breastfed infant. Bifidobacterium is mixed with sufficient infant formula or breast milk so that the infant can fully incorporate Bifidobacterium and the infant can still potentially and may ingest the full dose of Bifidobacterium. Therefore, Bifidobacterium may be mixed with about 3 to 5 mL of breast milk or infant formula before administering Bifidobacterium to a breastfed infant. The Bifidobacterium composition may be mixed by any suitable means, including simply stirring the composition with a medium (e.g., infant formula, breast milk, water) in a bowl (or any other suitable means for obtaining the mixture). The composition mixed with infant formula or breast milk may then be supplied to the infant by any suitable means. Preferred means of feeding to the infant include the use of a feeding syringe, spoon, or bottle. Bifidobacterium may also be administered to the infant before feeding when the infant is more likely to be hungry, as this is thought to increase the likelihood that the infant will consume the entire dose.

[0036] Dosage and frequency of administration can be selected as desired. For example, Bifidobacterium may be administered once daily. In such an example, the once-daily dose may contain approximately 5 to 15 billion or approximately 8 billion CFU. It is also conceivable to divide the desired total dose into smaller doses. Examples include several smaller doses throughout the day (e.g., 2, 3, 4, or 5 times per day).

[0037] The total daily dose may range from approximately 1 million, 500 million, 1 billion, 2 billion, 3 billion, 4 billion, 5 billion, 6 billion, 7 billion, 8 billion, 9 billion, 10 billion, or 12 billion to approximately 8 billion, 9 billion, 10 billion, 20 billion, 30 billion, 40 billion, 50 billion, 60 billion, 70 billion, 80 billion, 90 billion, 100 billion, 200 billion, 250 billion, or 500 billion colony-forming units (CFUs) of Bifidobacterium. The total daily dose may range from approximately 5 billion to approximately 15 billion CFUs, or approximately 8 billion CFUs. Such a total dose value may be given in a single dose.

[0038] Bifidobacterium may be administered starting at 1, 2, 3, 4, 5, or 6 days postnatally, or at 1 week postnatally, or within the first 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks postnatally, or at 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months postnatally. As used herein, the term “postnatal” means from birth. Once initiated, Bifidobacterium may continue to be administered until 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks postnatally, or until 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months postnatally. Bifidobacterium may be initially administered within the first two weeks postnatally. Bifidobacterium may be initially administered within the first two weeks postnatally and by 12 weeks postnatally.

[0039] An infant may be a breastfed infant. As used herein, the term “breastfed” means that the infant obtains at least a portion of its nutrition from human breast milk. The infant may be breastfed, or the breast milk may be expressed (e.g., pumped or hand-expressed) and given to the infant. A breastfed infant may be fed at least about 50, 60, 75, 80, 90%, or 95% breast milk. The remainder of the infant’s nutrition may come from infant formula or other foods. Alternatively, a breastfed infant may be exclusively breastfed. As used herein, the term “exclusively breastfed” means that the infant does not receive infant formula, except that a small amount of infant formula may be used solely for the purpose of administering it to the infant in combination with Bifidobacterium. Any caloric contributions from other sources during the first three months of life, including drugs, Bifidobacterium compositions, or any media used to deliver Bifidobacterium, are considered negligible.

[0040] While the foregoing description represents exemplary embodiments of the present invention, it will be understood that various additions, modifications, and substitutions can be made herein without departing from the spirit and scope of the invention. In particular, it will be apparent to those skilled in the art that the invention can be embodied in other specific forms, structures, arrangements, proportions, and using other elements, materials, and components without departing from its spirit or essential characteristics. Those skilled in the art will recognize that the invention can be used in various ways, with numerous modifications to the structure, arrangement, proportions, materials, and components used in practice of the invention, particularly adapted to specific environmental and operating requirements, without departing from the principles of the invention. Accordingly, the embodiments disclosed herein should be considered exemplary in all respects, not limiting, and the scope of the invention is indicated by the appended claims and is not limited to the foregoing description. In the claims, the term “includes / contains” will be understood not to exclude the existence of other elements or processes. Furthermore, singular references do not exclude plurals. Terms such as "a," "an," "first," and "second" do not exclude the possibility of plurality.

[0041] To provide a more precise explanation, some of the quantitative expressions presented herein are not modified with the term “about.” Whether the term “about” is explicitly used or not, all quantities given herein are meant to refer to actual given values, and also to approximate values ​​of such given values ​​that can be reasonably inferred on the basis of ordinary skill in the art, including approximations of such given values ​​under experimental and / or measurement conditions.

[0042] To provide more accurate information, in this specification, some quantitative expressions are referred to as a range from about X to about Y. Where a range is given, it is understood that the range includes the entire range from about X to about Y, or any quantity or range within that range, and is not limited to the upper and lower limits given.

[0043] All percentages, parts, and ratios are based on the total weight of the compositions of the present invention unless otherwise specified. All such weights relating to the listed components are based on the level of the specific component described and, unless otherwise specified, do not include carriers or by-products that may be present in commercially available substances.

[0044] Predictive Examples This clinical trial will investigate the clinical and immunological effects of B. infantis (EVC001 strain, Evolve BioSystems, Inc., Davis, CA) supplementation, initiated within 14 days of birth and continued for 12 weeks. A randomized, placebo-controlled experimental study is intended to reduce or eliminate known confounding effects from factors other than the study intervention.

[0045] Therefore, this type of study design is considered rigorous because it allows for testing of the causal relationship between the research intervention (in this case, the research supplement) and the research endpoint.

[0046] the purpose The first objective of this study is to evaluate the effect of B.infantis(EVC001) compared to placebo on the cumulative incidence of physician-diagnosed AD in the first year of life in healthy, breastfed infants at risk of developing AD. The second objective of this study is to evaluate the effect of B.infantis(EVC001) compared to placebo on the following in healthy, breastfed infants at risk of developing AD: • Percentage of infants experiencing adverse events (AEs) to assess safety and tolerability. • Cumulative incidence of AD at the time of addition • Time until the onset of AD • Establishment of B. infantis in the infant intestines • Severity of AD in individuals with AD

[0047] Further objectives of this research are as follows: 1) Evaluate the effects of B.infantis (EVC001) compared to placebo in healthy, breastfed infants at risk of developing Alzheimer's disease (AD) regarding the following: • Incidence of atopic diseases other than AD (food allergies, allergic rhinitis, asthma) • Allergic sensitization • Gut and skin microbiome • Skin immunology biomarker profiles • Incidence of infant colic • Infant sleep • Infant physical measurements • Relationship between the development of the maternal gut microbiota at baseline and the infant gut microbiota. 2) Characterize and compare the microbiome and immunological profiles of infants who do not develop Alzheimer's disease (AD) over a year and those who do, in the B. infantis (EVC001) supplementation group and the placebo supplementation group. 3) Determine the phenotype of immune responders in the B. infantis (EVC001) supplementation group and the placebo supplementation group. 4) Identify the target gene profile associated with benefiting from supplement intervention.

[0048] Overview This trial is a randomized, double-blind, placebo-controlled, two-arm, parallel-group (groups 1 and 2) trial. The study will enroll approximately 286 infants who have at least one first-degree relative with a history of atopic disease (i.e., a biological parent or full sibling with AD, allergic rhinitis, or asthma reported by the mother and diagnosed by a physician), are currently breastfed, and whose mothers intend to maintain exclusive breastfeeding for at least 12 weeks (approximately 3 months). Each infant will participate in the study under the supervision of their biological mother ("guardian").

[0049] Infant eligibility assessment will be conducted within the first 14 days of life. Eligible infants will be enrolled and randomized equally (1:1) to one of two groups: placebo (Group 1) or B. infantis (EVC001) (Group 2). Randomization will be stratified by the number of first-degree relatives (1 to 2 or more) of the infant with a history of the relevant atopic disease (as defined above). All baseline assessments will be conducted prior to the first dose of the assigned research supplement.

[0050] The administration of the assigned research supplement will begin on day 0 and continue for 12 weeks. Parents will be instructed to make their best efforts to maintain exclusive breastfeeding for at least this 12-week period and will be encouraged to continue breastfeeding for as long as possible during the first year of life. Following the 12-week supplementation period, infants will be followed up to 104 weeks (approximately 2 years) through scheduled and unscheduled visits.

[0051] Sub-studies will be planned to use a subset of the main study population (approximately 80-100 participants) to evaluate possible relationships between specific immune cells in peripheral circulation, gut microbiota, frequency, and function, circulating cytokine profiles, and the development of Alzheimer's disease (AD).

[0052] Selection and registration of target Eligibility criteria are designed to select subjects for whom the protocol procedures are deemed appropriate. Infant eligibility will be assessed within the first 14 days after birth. Initial eligibility verification may be performed by individuals who are not qualified physicians.

[0053] The inclusion criteria for infants include the following: 1) Male or female newborns 14 days old or younger at the time of study registration (day 0). 2) A healthy, full-term infant. 3) Having at least one first-degree relative (i.e., a biological parent or full sibling) who has an atopic disease (i.e., AD, allergic rhinitis, or asthma reported by the mother and diagnosed by a physician). 4) Established breastfeeding at the time of study registration (day 0), with the mother's intention to maintain exclusive breastfeeding for 12 weeks or more.

[0054] The exclusion criteria for infants are as follows: 1) Premature birth (gestational age less than 36 weeks [252 days]). 2) Admission to the neonatal unit for problems other than establishing normal feeding. 3) In the opinion of the PI or nominee, evidence of a baseline disease / condition (e.g., abnormal birth weight) or a significant risk of developing a disease / condition (based on an assessment of maternal / pregnancy information) that would pose a significant safety concern if the infant were enrolled in the study, or would otherwise exclude participation in the study. 4) A significant birth defect / complication (e.g., abdominal wall abnormality, congenital heart defect) that, in the opinion of the PI or the nominee, would raise safety concerns or otherwise disrupt the study. 5) Severe, widespread skin conditions (e.g., collodion). 6) In the opinion of the PI or the person named, any medical condition (in infants) or maternal medication / supplement use (e.g., daily or routine antibiotics or systemic antifungals) that could significantly alter the gut or skin microbiome. 7) The individual has consumed prebiotics or (a) a probiotic supplement / milk / formulated milk containing Bifidobacterium longum prior to registration (day 0). 8) The child consumed more than 100 mL of formula milk per day within 48 hours prior to registration (day 0). 9) A medical condition (infant) or surgery / injury / condition of the mother that would interfere with breastfeeding. 10) Known infant susceptibility or intolerance to soy or milk protein consumption. 11) Maternal infection due to human immunodeficiency virus, tuberculosis, hepatitis C, or hepatitis B. 12) In the opinion of the PI or the nominee, any parental condition that would prevent the parent and / or infant from complying with the research protocol requirements. 13) Twins or multiple births.

[0055] During the study, infant caregivers will be instructed to make their best efforts to ensure their infants are exclusively breastfed until at least 12 weeks of age and will be encouraged to continue breastfeeding for as long as possible during the first year of life. Caregivers will also administer the assigned research supplement to their infants once daily for 12 weeks, following the instructions and training provided. Caregivers will be instructed to avoid routine infant intake of probiotics during the first 12 weeks of the study (or, if longer, during the breastfeeding period) unless specifically prescribed by an HCP, for example, to prevent or treat antibiotic-related diarrhea or to treat gastroenteritis. Caregivers will also be instructed to ensure their infants do not ingest any prebiotics or any Bifidobacterium-containing probiotic supplements / milk / formula during the first 24 weeks of the trial.

[0056] Sample size, randomization / study supplement allocation, and blinding. Approximately 200-400 participants will be randomly assigned in a 1:1 ratio to receive either B.infantis (EVC001) or a placebo. Enrolled infants will be randomly assigned equally (1:1) to either the active supplement group or the placebo supplement group according to the randomization schedule. Infants will be stratified by the number of first-degree relatives with a history of atopic dermatitis (1 to 2 or more). The study will be double-blind, and as a result, parents and PIs / designated participants will not know which study supplement they will receive.

[0057] Identification and Use of Research Supplements As shown in Table 1 below, the following research supplements will be provided.

[0058] [Table 1]

[0059] The active supplement is Evivo® probiotic powder, a B. infantis supplement available from Evolve BioSystems, Inc., contained in a sachet. Each serving contains purified lactose and B. longum subspecies Infantis EVC001. Each sachet contains 625 mg of probiotic powder and 8 billion CFU of B. infantis (EVC001). The placebo sachet contains 625 mg of lactose.

[0060] Infants will receive the contents of a single sachet of either B.infantis (EVC001) or a matching placebo study supplement once daily for 12 weeks. At each supplementation, the contents of the sachet will be mixed with approximately 3-5 mL of expressed or pumped breast milk (or infant formula, if necessary) in the provided reservoir. Using the provided syringe, the mixture will be dispensed into the side of the infant's mouth to ensure the infant receives the full dose.

[0061] Test period, procedures, and evaluation schedule Infant eligibility assessment will be conducted within the first 14 days of the infant's life. Following enrollment and randomization (day 0), the research supplement will be administered daily from day 0 to week 12. Parents will be instructed to make every effort to maintain exclusive breastfeeding for at least 12 weeks. Following the 12-week supplementation period, subjects will be followed for a further 92 weeks to complete this 104-week (2-year) study.

[0062] Details of evaluation tools and additional research procedures Questionnaire regarding medical, family, medication, and supplement history: During screening, researchers will interview each infant's guardian to complete a medical, family, medication, and supplement history questionnaire to document the infant's medical and medication / supplement history, including pregnancy, childbirth, and breastfeeding / exposure from these. The questionnaire will also record information reported by the mother regarding a history of atopic diseases, e.g., current or past history of AD (including age of onset and details of diagnostic tests), allergic rhinitis / hay fever (including allergen identification), asthma, food reactions / allergies (including details of any other formal allergy tests, such as food and type of reaction), other skin conditions, or first-degree relatives of the infant with immune-mediated disorders.

[0063] Diagnosis of AD: Infants with suspected cases of atopic dermatitis identified by researchers will be evaluated by physicians trained for assessment and diagnosis. Briefly, a diagnosis of atopic dermatitis would be made if three of the following four criteria are met: 1) pruritus, 2) typical form and distribution (involvement of the face and extensors), 3) chronic or chronically recurrent dermatitis, and 4) personal or family history of atopic disease (Rajka G, Langeland T. Grading of the severity of atopic dermatitis. Acta Derm Venereol Suppl (Stockh) 1989;144:13-4.; Ganemo A, Svensson Å, Svedman C, Gronberg BM, Johansson AC, Wahlgren CF. Usefulness of Rajka & Langeland Eczema Severity Score in clinical practice. Acta Derm Venereol 2016;96:521-4.).

[0064] If an infant is diagnosed with Alzheimer's disease (AD), the severity will be assessed at the time of diagnosis and at 12, 52, and 104 weeks of gestation using the EASI and POEM tests described below.

[0065] Eczema Area and Severity Index (EASI) for children under 8 years old: The EASI is a tool used to measure the extent (area) and severity of atopic dermatitis (AD) (Hanifin JM, Thurston M, Omoto M, Cherill R, Tofte SJ, Graeber M. The eczema area and severity index (EASI): assessment of reliability in atopic dermatitis. EASI Evaluator Group. Exp Dermatol 2001;10:11-8). This device assesses four body regions: head and neck (including face, neck, and scalp), torso (including genital area), upper extremities (including hands), and lower extremities (including buttocks and feet), which are assigned proportional body surface area percentages of 20%, 30%, 20%, and 30%, respectively. Area scores are determined for each of these four body regions based on the percentage of skin affected by AD (defined by the four key signs listed below) within that region (0=none, 1=1-9%, 2=10-29%, 3=30-49%, 4=50-69%, 5=70-89%, 6=90-100%). Each of the four body regions is also assessed for the severity of the four key signs of AD: erythema, induration / papulogenesis / edema, epidermal exfoliation, and lichenification, using a scale of 0-3, where 0=none, 1=mild, 2=moderate, and 3=severe (note: half points are acceptable). The total score for each body region is determined by multiplying the area score by the sum of the severity scores for the four key signs, and then multiplying the result by a fixed body surface area assigned to that region. The total EASI score is the sum of the body region scores and ranges from 0 to 72. A designated, trained physician will use the EASI to assess the severity of AD at the time of diagnosis, and (for subjects with AD only) at weeks 12, 52, and 104.

[0066] Patient-Oriented Eczema Measure (POEM): POEM is a simple, effective, easily interpretable, and reproducible tool for assessing Alzheimer's disease (AD) and monitoring aspects of the disease that are important to the patient (Charman CR, Venn AJ, Williams HC. The Patient-Oriented Eczema Measure: Development and initial validation of a new tool for measuring atopic eczema severity from the patients' perspective. Arch Dermatol 2004;140:1513-9.; Charman CR, Venn AJ, Ravenscroft JC, Williams HC. Translating Patient-Oriented Eczema Measure (POEM) scores into clinical practice by suggesting severity strata derived using anchor-based methods. Br J Dermatol 2013;169:1326-32.). Researchers will interview parents at the time of AD diagnosis, and (only for subjects with AD) at weeks 12, 52, and 104, to assess the frequency of seven symptoms (itchy skin, sleep disturbances, bleeding skin, exudation / capillary bleeding, peeling skin, cracking skin, and dry / rough skin) during the previous week (days 0, 1-2, 3-4, and 5-6) using a 5-point scale. The maximum total POEM score is 28.

[0067] Infant colic Researchers will interview parents to document signs and symptoms of infant colic at baseline (day 0) and at weeks 6, 12, and 24, based on the Rome IV criteria (Benninga M, Nurko S, Faure C, Hyman P, St. James-Roberts I, Schechter N. Childhood functional gastrointestinal disorders: neonate / toddler. Gastroenterology 2016;150:1443-55). The occurrence of infant colic, as defined by the Rome IV criteria, will be determined using these parent interviews in combination with daily parental entries in a diary. Parents will use the diary to document the occurrence and duration of other related symptoms, such as crying, fussiness, and bowel movements, daily during baseline and between weeks 5–7, 11–13, and 23–24. "Fretting" refers to intermittent vocalizations accompanied by distress, and is defined as "not crying, but also not being aroused or satisfied [behavior]" (Benninga, 2016; Zeevenhooven J, Koppen IJ, Benninga MA. The new Rome IV criteria for functional gastrointestinal disorders in infants and toddlers. Pediatr Gastroenterol Hepatol Nutr 2017;20:1-13). As defined by the Rome IV criteria, the occurrence of infant colic is determined using parental journal entries in combination with parental interviews conducted at the time of visit.

[0068] Infant physical measurements Infants will have their length / height (using an infant length board), weight, and head circumference measured during all research visits. The Body Mass Index (BMI) will be calculated based on the weight and length / height measurements according to the following formula: BMI (kg / m 2 ) = weight (kg) / ([length or height in cm / 100] 2 ).

[0069] Brief Infant Sleep Questionnaire - Revised (BISQ-R) At weeks 12, 24, 52, and 76, parents will use the BISQ-R to document their infant's sleep habits and patterns over the past two weeks. The BISQ-R is an age-based standard reference scoring system that provides a comprehensive assessment of infant and toddler sleep patterns (5 items related to sleep latency, number and duration of nighttime awakenings, maximum length of sleep, and total nighttime sleep), as well as parental perceptions that may influence sleep outcomes (3 items related to difficulty falling asleep, nighttime sleep, and overall childhood sleep problems), and parental behaviors (11 items related to bedtime routine consistency, bedtime, parental behavior at bedtime and subsequent nighttime awakenings, and sleep position at bedtime and subsequent nighttime awakenings) (Mindell JA, Gould RA, Tikotzy L, Leichman ES, Walters RM. Norm-referenced scoring system for the Brief Infant Sleep Questionnaire-Revised (BISQ-R). Sleep Med 2019;63:106-14.). A score ranging from 0 to 100 is derived for each subscale, with higher scores indicating better sleep quality, a more positive perception of sleep quality, and parental habits that promote healthy sleep behaviors and independent infant sleep, respectively. The total score is calculated as the average of the subscale scores for infant sleep, parental perception, and parental behavior.

[0070] Checklist for introducing solid foods Parents should document when their infant first consumes a new solid food, including the type and amount of the solid food, if applicable.

[0071] Records of infant feeding and use of research supplements Parents will keep a log detailing the mother's breastfeeding, the use of infant formula (if applicable), and the administration of research supplements up to week 12, and optionally daily thereafter. Parents will also log any forgotten, incomplete, or extra doses of research supplements, and any missed doses will not be supplemented on subsequent days.

[0072] blood sample 2.0 mL of venous whole blood samples will be collected from infants at 24, 52, and 104 weeks of gestation. The blood will be processed and stored according to the instructions provided in the Laboratory Reference Manual for future analysis. To assess the development of allergic sensitization, allergen-specific serum IgE testing will be performed using venous whole blood samples. In addition, blood samples may also be used to investigate the phenotype of the infant's immune responder by comparing the serial RNA expression profiles of the subject in the B. infantis (EVC001) supplementation group and the placebo supplementation group. Blood samples may also be used for exome sequencing to determine the presence of mutations in different genes, including the FLG gene, that may be associated with atopic diseases (e.g., AD, asthma, and allergic sensitization) and ichthyosis vulgaris.

[0073] Fecal sample If the mother consents to collection (optional), maternal fecal samples will be collected between 2 and 6 weeks postpartum for future microbiome analysis. The caregiver will collect infant fecal samples within 5 days (preferably within 3 days) prior to baseline (day 0), and at visits at 6, 12, 24, 52, and 104 weeks. The baseline sample should be a non-meconium sample, and if this is not possible before the start of supplementation, the deviation should be recorded, and a non-meconium sample should be collected as soon as possible (preferably within 24 hours) after the start of supplementation.

[0074] All fecal samples will be collected using the provided supplies and frozen in the provided collection tubes and medical waste bags until collected by researchers during actual hospital visits. Soft-side coolers fitted with ice packs will be used to transport the frozen fecal samples to the research facility, where they will be stored at -80°C for future analysis.

[0075] Fecal samples collected at baseline (day 0) and at weeks 6, 12, 24, 52, and 104 will be analyzed for the overall bacterial profile of the intestine, including the presence of B. infantis, to determine the degree of colonization (percentage of B. infantis colonization within each infant) and incidence (percentage of infants with intestinal B. infantis colonization). An infant's intestine will be considered colonized if the fecal concentration of B. infantis, as determined by shotgun sequencing, is 50% or greater of the total bacteria.

[0076] Skin swab sample Parents will be instructed to avoid applying any topical treatments within three hours prior to the research visit to avoid interfering with the collection of skin samples. All collected skin samples, as detailed below, will be frozen and stored at -80°C in the research facility for future analysis, as detailed in the Laboratory Reference Manual.

[0077] Skin Biomarkers: To evaluate the effects of B. infantis (EVC001) supplementation on immunomodulation in the skin and to learn about biomarkers associated with the development of Alzheimer's disease (AD), researchers will collect two skin samples using FibroTX skin sample collection swabs at baseline (day 0), week 12, and week 52. If AD is diagnosed, two additional samples will be collected at these time points from the lesion site and adjacent clearly non-lesional sites. FibroTX skin sample collection swabs are a highly sensitive, multi-sample research tool for non-invasive biomarker measurement directly from the skin.

[0078] Skin Microbiota: To evaluate the systemic effects of research supplements on the skin microbiota, researchers will collect two skin samples at baseline (day 0), week 6, week 12, and week 52 using pre-moistened swabs. One sample will be collected from the cubital fossa (elbow crease) and the other from a location determined by the researcher or sponsor. If AD is confirmed, two additional samples will be obtained at these time points from the affected skin and adjacent clean, non-affected skin.

[0079] Research visits and evaluations: Table 2 below outlines the research procedure and evaluation schedule.

[0080] [Table 2]

[0081] [Table 3]

[0082] Research endpoints and data analysis Primary endpoint: Cumulative incidence of Alzheimer's disease (AD) up to week 52. The cumulative incidence of AD up to week 52 will be compared between the supplementation groups.

[0083] Secondary endpoints: The distribution of the time to the onset of Alzheimer's disease up to week 104 will be compared between the supplementation groups. • Cumulative incidence of AD up to week 24 and week 104. • Percentage of infants with intestinal colonization of B. infantis at 12 weeks: Fecal samples from 12 weeks will be analyzed for B. infantis colonization and overall bacterial load to determine the percentage of infants with intestinal colonization of B. infantis. The infant's intestines will be considered colonized as defined above. • AD severity based on EASI scores at the time of onset, and at weeks 12, 52, and 104. • AD severity based on POEM scores at the time of onset, as well as at weeks 12, 52, and 104.

[0084] Additional endpoints: • Cumulative incidence of atopic disorders other than AD up to weeks 24, 52, and 104: Researchers will document the onset of allergic rhinitis, asthma, and food allergies (confirmed by a specialist / pediatric allergist) throughout the study and determine the cumulative incidence of these disorders at weeks 24, 52, and 104. • Cumulative incidence of allergic sensitization up to weeks 24, 52, and 104: Researchers will document the occurrence of allergic sensitization (confirmed by specific serum IgE testing) to dietary allergens or inhaled allergens, and determine the cumulative incidence at weeks 24, 52, and 104. • Changes from baseline in gut and skin microbiome up to week 104: This will determine the changes from baseline in the fecal (weeks 6, 12, 24, 52, and 104) and skin (weeks 6, 12, and 52) microbiomes. • Percentage of infants with intestinal colonization of B. infantis at 24 weeks: Fecal samples collected in connection with the 24-week visit will be analyzed for intestinal colonization of B. infantis as detailed above. • Changes from baseline in skin immunological biomarker profiles at weeks 12 and 52: This will determine the changes from baseline in skin biomarkers at weeks 12 and 52. • Cumulative incidence of infant colic up to weeks 6, 12, and 24: Diagnosis of infant colic will be based on Rome IV criteria, including data generated from in-clinic interviews and parental diaries. • BISQ-R scores at weeks 12, 24, 52, and 76: This will determine the BISQ-R subscale scores for infant sleep, parental perception, and parental behavior, as well as the total BISQ-R score, at weeks 12, 24, 52, and 76. • Changes from baseline in infant physical measurements at weeks 6, 12, 24, 52, 76, and 104: Changes from baseline in the infant's length / height, weight, head circumference, and BMI at weeks 6, 12, 24, 52, 76, and 104 will be determined. • Influence of maternal gut microbiota on the development of infant gut microbiota from baseline to 2 years of age: The microbiota of maternal fecal samples collected 2–6 weeks postpartum will be evaluated for correlation with the development of the infant gut microbiota, including the colonization of B. infantis in the infant's gut, at baseline (day 0) and at weeks 6, 12, 24, 52, and 104. • Microbiome and immunological profiles in infants with and without AD: Outcome measures related to the microbiome and immunological profiles will be characterized and compared between infants who do not develop AD over a year and those who do, in the B.infantis (EVC001) supplementation group and the placebo supplementation group. • Continuous RNA expression analysis at weeks 24, 52, and 104: Outcome measures related to continuous RNA expression profiles will be characterized and compared between subjects in the B. infantis (EVC001) supplementation group and the placebo supplementation group at weeks 24, 52, and 104 to investigate the phenotype of the infant's immune response. • Genetic analysis of the subjects: Outcome measures related to the subjects' genetic profiles will be characterized and compared between infants who do not develop Alzheimer's disease (AD) over a year and those who do in the B.infantis (EVC001) supplementation group, in order to identify subjects who may benefit from the intervention.

[0085] success criteria The success of the study will primarily be determined by the statistically significant effect of supplementation compared to placebo on the cumulative incidence of Alzheimer's disease (AD) up to week 52.

[0086] Summary of the Invention The above example constitutes a method comprising administering a composition containing Bifidobacterium (B. infantis) to breastfed infants at high risk of developing atopic diseases. Atopic dermatitis, food allergies, allergic rhinitis, and asthma will be monitored and evaluated for signs of prevention, delay, and / or improvement in breastfed infants. Infant colic, infant sleep, and infant physical measurements will also be monitored and evaluated.

[0087] [Implementation Method] (1) A method for preventing, delaying, or improving atopic dermatitis in breastfed infants, wherein the method is aBlongum, B.breve, B.bifidum, B.pseudocatenulatum, B.globosum, B.adolescentis, B.moukalabense, B.reuteri, B.pseudolongum, B.dentium, B.catenulatum, B.sp002742445, B.callitrichos, B.scardovii, B.tis sieri, B. subtile, B. gallinarum, B. choerinum, B. angulatum, B. primatium, B. myosotis, B. mongoliense, B. merycicum, B. lemurum, B. stellenboschense, B. scaligerum, B. saguini, B. pullorum, B. felsineum, B. eule A method comprising administering to a breastfed infant a composition containing an effective amount of Bifidobacterium selected from the group consisting of muris, B. cuniculi, B. callitrichos_A, B. biavatii, B. anseris, B. vansinderenii, B. sp900551485, B. sp003952945, B. sp003952025, B. sp003952005, B. simiarum, B. pseudolongum_C, B. parmae, B. margollesii, B. kashiwanohense_A, B. italicum, B. imperatoris, B. cricetid, B. catulorum, B. callitrichidarum, B. animalis, B. aesculapii, and combinations thereof. (2) The method according to Embodiment 1, wherein the Bifidobacterium is selected from the group consisting of B. longum, B. breve, B. kashiwanohense, and combinations thereof. (3) The method according to Embodiment 1 or 2, wherein the Bifidobacterium is a B. longum subspecies selected from the group consisting of longum, suis, infantis, and combinations thereof. (4) The method according to any one of embodiments 1 to 3, wherein the Bifidobacterium is B. infantis. (5) The method according to any one of embodiments 1 to 4, wherein the Bifidobacterium is mixed into breast milk before administering the Bifidobacterium to an infant being breastfed.

[0088] (6) The method according to any one of embodiments 1 to 5, wherein the Bifidobacterium is mixed into infant formula before administering the Bifidobacterium to the breastfed infant. (7) The method according to any one of embodiments 1 to 6, wherein the Bifidobacterium is mixed with about 3 to about 5 mL of breast milk, infant formula, or water before administering the Bifidobacterium to the breastfed infant. (8) The method according to any one of embodiments 1 to 7, wherein more than 50% of the breastfed infants are breastfed. (9) The method according to any one of embodiments 1 to 8, wherein the breastfed infant is breastfed exclusively. (10) The method according to any one of embodiments 1 to 9, wherein the Bifidobacterium is in the form of a powder mixed with lactose.

[0089] (11) The method according to any one of embodiments 1 to 10, wherein approximately 5 billion to approximately 15 billion CFU of Bifidobacterium is administered to the breastfed infant. (12) The method according to any one of embodiments 1 to 11, wherein B. infantis is administered once daily. (13) The method according to any one of embodiments 1 to 12, wherein B. infantis is first administered within the first two weeks after birth. (14) The method according to any one of embodiments 1 to 13, wherein B. infantis is administered 12 weeks before the first week of life. (15) The method according to any one of embodiments 1 to 14, wherein the Bifidobacterium is first administered within the first two weeks after birth and by the 12th week after birth.

[0090] (16) The method according to any one of embodiments 1 to 15, wherein the breastfed infant is at high risk of developing an atopic disease. (17) The method according to any one of embodiments 1 to 16, wherein the Bifidobacterium includes strain EVC001. (18) A method for preventing, delaying, or improving atopic dermatitis in a breastfed infant having at least one first-degree relative with a history of atopic disease, wherein the method is a. Administering an effective amount of B. infantis mixed with breast milk once daily to an infant being breastfed, A method wherein the breastfed infant is breastfed for at least 90% of the time, and B. infantis is first administered within the first two weeks after birth. (19) The method according to Embodiment 18, wherein B. infantis includes strain EVC001. (20) The method according to Embodiment 18 or 19, wherein B. infantis is mixed with about 3 to about 5 mL of breast milk or infant formula before administering B. infantis to the breastfed infant.

[0091] (21) The method according to any one of embodiments 18 to 20, wherein the breastfed infant is breastfed exclusively. (22) The method according to any one of embodiments 18 to 21, wherein B. infantis is in the form of a powder mixed with lactose. (23) The method according to any one of embodiments 18 to 22, wherein 8 billion CFUs of B. infantis are administered to the breastfed infant. (24) The method according to any one of embodiments 18 to 23, wherein B. infantis is administered once daily until the 12th week of life. (25) A method for preventing, delaying, or improving an atopic disease selected from the group consisting of food allergies, allergic rhinitis, asthma, and combinations thereof in a breastfed infant, wherein the method comprises administering to the infant a composition containing an effective amount of Bifidobacterium.

[0092] (26) The Bifidobacterium may contain B.longum, B.breve, B.bifidum, B.pseudocatenulatum, B.globosum, B.adolescentis, B.moukalabense, B.reuteri, B.pseudolongum, B.dentium, B.catenulatum, B.sp00274 to the breastfed infant. 2445, B.callitrichos, B.scardovii, B.tissieri, B.subtile, B.gallinarum, B.choerinum, B.angulatum, B.primatium, B.myosotis, B.mongoliense, B.merycicum, B.lemurum, B.stellenboschense, B.scaligerum, B The method according to Embodiment 25, selected from the group consisting of .saguini, B.pullorum, B.felsineum, B.eulemuris, B.cuniculi, B.callitrichos_A, B.biavatii, B.anseris, B.vansinderenii, B.sp900551485, B.sp003952945, B.sp003952025, B.sp003952005, B.simiarum, B.pseudolongum_C, B.parmae, B.margollesii, B.kashiwanohense_A, B.italicum, B.imperatoris, B.cricetid, B.catulorum, B.callitrichidarum, B.animalis, B.aesculapii, and combinations thereof. (27) The method according to embodiment 25 or 26, wherein the Bifidobacterium is selected from the group consisting of B. longum, B. breve, B. kashiwanohense, and combinations thereof. (28) The method according to any one of embodiments 25 to 27, wherein the Bifidobacterium is a B. longum subspecies selected from the group consisting of longum, suis, infantis and combinations thereof. (29) The method according to any one of embodiments 25 to 28, wherein the Bifidobacterium is B. infantis. (30) The method according to any one of embodiments 25 to 29, wherein the Bifidobacterium is mixed with breast milk before administering the Bifidobacterium to an infant being breastfed.

[0093] (31) The method according to any one of embodiments 25 to 30, wherein the Bifidobacterium is mixed into infant formula before administering the Bifidobacterium to the breastfed infant. (32) The method according to any one of embodiments 25 to 31, wherein the Bifidobacterium is mixed with about 3 to about 5 mL of breast milk, infant formula, or water before administering the Bifidobacterium to the breastfed infant. (33) The method according to any one of embodiments 25 to 32, wherein more than 50% of the breastfed infants are breastfed. (34) The method according to any one of embodiments 25 to 33, wherein the breastfed infant is breastfed exclusively. (35) The method according to any one of embodiments 25 to 34, wherein the Bifidobacterium is in the form of a powder mixed with lactose.

[0094] (36) The method according to any one of embodiments 25 to 35, wherein approximately 5 billion to approximately 15 billion CFU of Bifidobacterium is administered to the breastfed infant. (37) The method according to any one of embodiments 25 to 36, wherein B. infantis is administered once daily. (38) The method according to any of embodiments 25 to 37, wherein B. infantis is first administered within the first two weeks after birth. (39) The method according to any of embodiments 25 to 38, wherein B. infantis is administered before the first 12 weeks of life. (40) The method according to any one of embodiments 25 to 39, wherein the Bifidobacterium is first administered within the first two weeks after birth and by the 12th week after birth.

[0095] (41) The method according to any one of embodiments 25 to 40, wherein the infant being breastfed is at high risk of developing the atopic disease. (42) The method according to any one of embodiments 25 to 41, wherein the Bifidobacterium comprises the EVC001 strain. (43) A method for improving infant colic, infant sleep, or infant physical measurement in a breastfed infant, wherein the method comprises administering to the infant a composition containing an effective amount of bifidobacterium. (44) The Bifidobacterium may contain B.longum, B.breve, B.bifidum, B.pseudocatenulatum, B.globosum, B.adolescentis, B.moukalabense, B.reuteri, B.pseudolongum, B.dentium, B.catenulatum, B.sp00274 to the breastfed infant. 2445, B.callitrichos, B.scardovii, B.tissieri, B.subtile, B.gallinarum, B.choerinum, B.angulatum, B.primatium, B.myosotis, B.mongoliense, B.merycicum, B.lemurum, B.stellenboschense, B.scaligerum, B The method according to Embodiment 43, selected from the group consisting of .saguini, B.pullorum, B.felsineum, B.eulemuris, B.cuniculi, B.callitrichos_A, B.biavatii, B.anseris, B.vansinderenii, B.sp900551485, B.sp003952945, B.sp003952025, B.sp003952005, B.simiarum, B.pseudolongum_C, B.parmae, B.margollesii, B.kashiwanohense_A, B.italicum, B.imperatoris, B.cricetid, B.catulorum, B.callitrichidarum, B.animalis, B.aesculapii, and combinations thereof. (45) The method according to embodiment 43 or 44, wherein the Bifidobacterium is selected from the group consisting of B. longum, B. breve, B. kashiwanohense, and combinations thereof.

[0096] (46) The method according to embodiment 45, wherein the Bifidobacterium is a B. longum subspecies selected from the group consisting of longum, suis, infantis, and combinations thereof. (47) The method according to any one of embodiments 43 to 46, wherein the Bifidobacterium is B. infantis. (48) The method according to any one of embodiments 43 to 47, wherein the Bifidobacterium is mixed with breast milk before administering the Bifidobacterium to an infant being breastfed. (49) The method according to any one of embodiments 43 to 48, wherein the Bifidobacterium is mixed into infant formula before administering the Bifidobacterium to the breastfed infant. (50) The method according to any one of embodiments 43 to 49, wherein the Bifidobacterium is mixed with about 3 to about 5 mL of breast milk, infant formula, or water before administering the Bifidobacterium to the breastfed infant.

[0097] (51) The method according to any one of embodiments 43 to 50, wherein more than 50% of the breastfed infants are breastfed. (52) The method according to any one of embodiments 43 to 51, wherein the breastfed infant is breastfed exclusively. (53) The method according to any one of embodiments 43 to 52, wherein the Bifidobacterium is in the form of a powder mixed with lactose. (54) The method according to any one of embodiments 43 to 53, wherein approximately 5 billion to approximately 15 billion CFU of Bifidobacterium is administered to the breastfed infant. (55) The method according to any one of embodiments 43 to 54, wherein B. infantis is administered once daily.

[0098] (56) The method according to any one of embodiments 43 to 55, wherein B. infantis is first administered within the first two weeks after birth. (57) The method according to any one of embodiments 43 to 56, wherein B. infantis is administered before the first 12 weeks of life. (58) The method according to any one of embodiments 43 to 57, wherein the Bifidobacterium is first administered within the first two weeks after birth and by the 12th week after birth. (59) The method according to any one of embodiments 43 to 58, wherein the infant being breastfed is at high risk of developing the atopic disease. (60) The method according to any one of embodiments 43 to 59, wherein the Bifidobacterium includes strain EVC001.

Claims

1. 1. A composition for use in a method of preventing, delaying, or ameliorating atopic dermatitis in a breast-fed infant having at least one first degree relative with a history of atopic disease, said composition comprising an effective amount of B. infantis mixed with breast milk, said method comprising: a. administering the composition to the breast-fed infant once daily; The method of claim 1, wherein the breast-fed infant is at least 90% breast-fed, and wherein the B. infantis is first administered within the first two weeks of life.

2. 2. The composition of claim 1, wherein the B. infantis comprises strain EVC001.

3. The composition of claim 1, wherein the B. infantis is mixed with about 3 to about 5 mL of breast milk or infant formula prior to administering the B. infantis to the breast-fed infant.

4. The composition of any one of claims 1 to 3, wherein the breast-fed infant is exclusively breast-fed.

5. The composition of any one of claims 1 to 4, wherein the B. infantis is in powder form mixed with lactose.

6. The composition of claim 1, wherein the method comprises administering 8 billion CFU of B. infantis to the breastfed infant.

7. The method of claim 1, wherein the B. infantis is administered once daily until the animal is 12 weeks of age.

8. A composition for use in a method for preventing, delaying, or ameliorating an atopic disease selected from the group consisting of food allergy, allergic rhinitis, asthma, and combinations thereof in a breastfed infant, said composition comprising an effective amount of Bifidobacterium, said method comprising administering said composition to said breastfed infant.

9. The Bifidobacterium is selected from the group consisting of B. longum, B. breve, B. bifidum, B. pseudocatenulatum, B. globosum, B. adolescentis, B. moukalabense, B. reuteri, B. pseudolongum, B. dentium, B. catenulatum, B. sp002742445, B. callitrichos, B. scardovii, B. tissieri, B. subtile, B. gallinarum, B. choerinum, B. cholera, B. cholera, B. cholera, B. cholera and B. cholera to the breast-fed infant. angulatum, B. primatium, B. myosotis, B. mongoliense, B. marycicum, B. lemurum, B. stellenboschense, B. scaligerum, B. saguini, B. pullorum, B. felsineum, B. eulemuris, B. cuniculi, B. callitrichos_A, B. biavatii, B. anseris, B. vansinderenii, B. sp900551485, B. sp003952945, B. sp003952025, B.

9. The composition of claim 8, wherein the B. simiarum, B. pseudolongum_C, B. parmae, B. margollesii, B. kashiwanohense_A, B. italicum, B. imperatoris, B. cricetid, B. catulorum, B. callitrichidarum, B. animalis, B. aesculapii, and combinations thereof.

10. 10. The composition of claim 8 or 9, wherein the Bifidobacterium is selected from the group consisting of B. longum, B. breve, B. kashiwanohense, and combinations thereof.

11. 11. The composition of any one of claims 8 to 10, wherein the Bifidobacterium is a B. longum subspecies selected from the group consisting of B. longum, suis, infantis, and combinations thereof.

12. The composition of any one of claims 8 to 11, wherein the Bifidobacterium is B. infantis.

13. A composition according to any one of claims 8 to 12, wherein in the method the Bifidobacterium is mixed into breast milk prior to administration of the Bifidobacterium to the breast-fed infant.

14. The composition of any one of claims 8 to 13, wherein the method comprises mixing the Bifidobacterium with an infant formula prior to administering the Bifidobacterium to the breastfed infant.

15. The composition of any one of claims 8 to 14, wherein the method comprises mixing the Bifidobacterium with about 3 to about 5 mL of breast milk, infant formula, or water prior to administering the Bifidobacterium to the breast-fed infant.

16. The composition of any one of claims 8 to 15, wherein the breast-fed infant is more than 50% breast-fed.

17. The composition of any one of claims 8 to 16, wherein the breast-fed infant is exclusively breast-fed.

18. 18. The composition of any one of claims 8 to 17, wherein the Bifidobacterium is in powder form mixed with lactose.

19. The composition of any one of claims 8 to 18, wherein said method comprises administering from about 50 to about 15 billion CFU of said Bifidobacterium to said breastfed infant.

20. The method of claim 8, wherein the B. infantis is administered once daily.

21. The composition of any one of claims 8 to 20, wherein the method comprises administering the B. infantis for the first time within the first two weeks of life.

22. The method of claim 8, wherein the B. infantis is administered prior to the first 12 weeks of life.

23. The composition of any one of claims 8 to 22, wherein the method comprises administering the Bifidobacterium initially within the first two weeks of life and no later than the 12th week of life.

24. The composition of any one of claims 8 to 23, wherein the breastfed infant is at high risk of developing the atopic disease.

25. The composition of any one of claims 8 to 24, wherein the Bifidobacterium comprises the EVC001 strain.