CN115989836A - Nutritional composition containing lactoferrin and probiotics, food and application - Google Patents

Nutritional composition containing lactoferrin and probiotics, food and application Download PDF

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Publication number
CN115989836A
CN115989836A CN202211517832.1A CN202211517832A CN115989836A CN 115989836 A CN115989836 A CN 115989836A CN 202211517832 A CN202211517832 A CN 202211517832A CN 115989836 A CN115989836 A CN 115989836A
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bifidobacterium
cfu
lactoferrin
food
nutritional composition
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Inventor
崔东影
解庆刚
王象欣
石红丽
蒋士龙
张永久
冷友斌
陆思宇
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Heilongjiang Feihe Dairy Co Ltd
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Heilongjiang Feihe Dairy Co Ltd
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Abstract

The present invention relates to nutritional compositions. The nutritional composition comprises or consists of the following: lactoferrin and probiotics. The probiotic is preferably one or more selected from Bifidobacterium infantis, bifidobacterium longum, bifidobacterium bifidum, bifidobacterium adolescentis, bifidobacterium lactis, bifidobacterium breve, lactobacillus acidophilus and Lactobacillus rhamnosus; preferably bifidobacterium lactis, more preferably bifidobacterium lactis Bb12. The invention also relates to a food product comprising said nutritional composition. The invention also relates to the use of lactoferrin, probiotics or a nutritional composition or food product comprising both, to promote development of the immune system and improve non-therapeutic purposes in the intestinal flora of offspring via ingestion by humans or animal precursors.

Description

Nutritional composition containing lactoferrin and probiotics, food and application
Technical Field
The present invention relates generally to the field of food products. In particular, the present invention relates to a nutritional composition for promoting development of the immune system and/or improving the function of the immune system, and improving the intestinal flora of its progeny, a food comprising the nutritional composition, and the use of the nutritional composition or its nutritional ingredients, or a food comprising the nutritional composition. More particularly, the present invention relates to a nutritional composition comprising lactoferrin and probiotics, a food product comprising the nutritional composition, and the use of the lactoferrin or probiotics or the nutritional composition or the food product for the non-therapeutic purpose of promoting development of the immune system and improving the intestinal flora of offspring via maternal consumption.
Background
"1000 days early in life" refers to the period of time from the start of conception to 2 years of age. During this period, the cells are in a state of vigorous division, proliferation and differentiation, and tissues and organs begin to form. The nutritional exposure during this period affects the establishment of metabolic patterns in the body which persist throughout life, affecting the risk of developing metabolic diseases in childhood or even adulthood. Early 1000 days of life can be divided into 3 phases: fetal (intrauterine nutrition), breast milk or infant formula, supplementary food addition and early dietary period.
"fetal origin hypothesis" the fetal term is considered to be extremely sensitive to nutrient supply during fetal life, and fetal adaptation to a failure of the mother to ingest sufficient nutrients or uterine dysplasia may be performed by the fetus, including: reducing blood flow of viscera such as kidney, ensuring blood supply of heart and brain, reducing hormone secretion, and reducing sensitivity of organism to hormone. Such changes may result in permanent changes in the phenotype of the body through apparent modification of the gene, resulting in subsequent disease development, and may be inherited to the next generation. The epigenetic regulation mainly comprises covalent modification of histone, DNA methylation, genome imprinting, non-coding RNA and the like. Many nutrients such as folic acid, vitamin B12, contain methyl structures in their components, and participate in the methylation process of DNA, resulting in reduced agouti protein expression, and the body shows different phenotypes under the condition of unchanged genotypes. Thus, the risk of obesity in offspring can be effectively reduced after the parent ingests proper nutrients. All of the above indicate that while nutrients cannot alter the coding sequence of DNA, the gene phenotype can be altered by modification of DNA. Early life malnutrition affects the growth and development of fetuses and children, and has long-term effects on the health level and quality of life after adulthood. Some have reduced alpha-linolenic acid intake during pregnancy and lactation in female mice and explored the impact of early n-3PUFAs in life on adult metabolic syndrome and susceptibility to parkinson's disease.
Intestinal flora plays an important role in early life. It is generally accepted that infants, prior to birth, have their gastrointestinal tract sterile and begin to colonise with bacteria within 48 hours after birth. The initial bacteria in the gastrointestinal tract of neonates are from the birth canal of the mother, the environment and breast milk. Natural delivery is established earlier than infant gastrointestinal microecology by caesarean section. Breast-fed infants differ from artificial-fed infants in their original gastrointestinal microecology. The microecology of the term infants fed by breast milk is mainly bifidobacteria, and the intestinal flora diversity of the infants fed by human is increased, and the intestinal flora is accompanied by the remarkable increase of the number of the bacteroides.
Lactoferrin (LF) is an iron-binding glycoprotein with a variety of biological activities. At present, LF has become the main functional additive component of infant formulas and the impact in infant health has been studied extensively. Some clinical studies have shown that feeding breast milk can significantly reduce the incidence and mortality of necrotizing colitis in newborns by about 13% compared to premature infants fed infant formula, the reason being attributed to the beneficial effect of the bioactive ingredient represented by LF contained in breast milk for the prevention and treatment of IBD. LF can also regulate intestinal flora, inhibit growth of harmful bacteria, colonize intestinal tracts, and promote growth of probiotics. No studies have shown the effect of maternal lactoferrin supplementation on sub-immune or intestinal flora. At present, the lactoferrin is mainly aimed at immunity and intelligence, and the patent layout transferred by mother and infant still belongs to the blank stage.
The prior art has shown that even if a nutritional substance is considered beneficial for fetal/infant development and health, it is not necessarily able to promote development and health of offspring via the maternal-to-infant delivery pathway in the case of maternal consumption intake.
For example, boyle et al, "Lactobacillus GG treatment during pregnancy for the prevention of eczema: a randomized controlled trial", allergy 2011;66:509-516, it is generally believed that the probiotic Lactobacillus rhamnosus GG (LGG) is believed to treat eczema, but the results of the study indicate that prenatal treatment with Lactobacillus rhamnosus GG is insufficient to prevent eczema: LGG treatment of pregnant women from 36 weeks gestation to delivery did not reduce the risk of eczema in infants at high risk of developing allergic diseases. Prenatal LGG has no effect on neonatal immune responses and results in reduced levels of breast milk sCD14 and IgA.
Karen M Switkowski et al, "Higher Maternal Protein Intake during Pregnancy Is Associated with Lower Cord Blood Concentrations of Insulin-like Growth Factor (IGF) -II, IGF Binding Protein 3,and Insulin,but Not IGF-I, in a Cohort of Women with High Protein Intake", the Journal of Nutrition Nutritional Epidemiology,2017, 6/7; the results of the study indicate that, in a group of pregnant women with relatively high average protein intake, higher intake is associated with lower concentrations of somatotrophic hormone in cord blood, although prenatal exposure to dietary proteins is generally thought to modulate somatotrophic hormone and thus affect the early life growth patterns and risk of chronic diseases associated with later stages of life, as indicated in https:// doi. Org/10.3945/jn.117.250589.
While the use OF probiotics is generally believed to reduce the risk OF asthma in infants, as indicated by Xiaochen Wei et al in "Association between probiotic supplementation and asthma incidence in infants:aMeta-analysis OF randomized controlled trials", JOURNAL OF ASTHMA,2019,DOI:10.1080/02770903.2018.1561893, the results OF the study indicate that supplementing probiotics during pregnancy or early in life has no significant relevance to reducing the incidence OF asthma or wheezing in infants.
Thus, there is a need to find a method that can promote the development of the offspring's immune system and/or improve the function of the offspring's immune system, and improve the offspring's intestinal flora, via maternal consumption.
Disclosure of Invention
The present invention has been made in view of the above-described problems.
It is an object of the present invention to provide a nutritional composition for promoting development of the immune system and/or improving the immune system function, and improving the intestinal flora of its offspring. In particular, the nutritional composition is capable of promoting development of the immune system and improving intestinal flora of its offspring via ingestion by the parent (human or animal).
It is another object of the present invention to provide a food product comprising the nutritional composition.
It is a further object of the present invention to provide the use of the nutritional composition or a defined nutritional ingredient thereof or a food product comprising the nutritional composition for non-therapeutic purposes (nutritional and/or health care) in promoting development of the immune system and improving intestinal flora of offspring via ingestion of the offspring via a parent (e.g. human or animal) diet.
The present inventors have found that by supplementing one or more of lactoferrin, probiotics (e.g. bifidobacterium infantis (bifidum), bifidobacterium longum (bifidum), bifidobacterium bifidum (bifidum), bifidobacterium adolescentis (bifidum), bifidobacterium lactis (bifidum lactis), bifidobacterium breve (bifidum breve), lactobacillus acidophilus (Lactobacillus acidophilus) and lactobacillus rhamnosus (Lactobacillus rhamnosus), preferably bifidobacterium lactis, more preferably bifidobacterium lactis Bb 12), or a combination of lactoferrin and probiotics during maternal pregnancy and/or lactation, it is possible to promote development of the immune system and improve the intestinal microbiota thereof (via maternal transfer).
In particular, when lactoferrin is used in combination with probiotics (e.g. one or more of bifidobacterium infantis, bifidobacterium longum, bifidobacterium bifidum, bifidobacterium adolescentis, bifidobacterium lactis, bifidobacterium breve, lactobacillus acidophilus and lactobacillus rhamnosus; preferably bifidobacterium lactis, more preferably bifidobacterium lactis Bb 12), there is a synergistic effect between the two, which can synergistically promote development of the immune system of the offspring via dietary intake by the parent (e.g. human (i.e. pregnant woman) or animal (i.e. pregnant female)) and can improve, e.g. synergistically improve, the intestinal flora of the offspring.
In particular, the invention is realized by:
1. a nutritional composition comprising or consisting of:
-lactoferrin; and
-probiotics.
2. The nutritional composition of item 1, wherein the probiotic is one or more selected from the group consisting of bifidobacterium infantis, bifidobacterium longum, bifidobacterium bifidum, bifidobacterium adolescentis, bifidobacterium lactis, bifidobacterium breve, lactobacillus acidophilus, and lactobacillus rhamnosus.
3. The nutritional composition of any one of items 1-2, wherein the probiotic is bifidobacterium lactis.
4. The nutritional composition of any one of items 1-3, wherein the probiotic is bifidobacterium lactis Bb12.
5. The nutritional composition of any one of items 1-4, wherein lactoferrin is provided in the form: lactoferrin (LF) enriched nutritional ingredients such as lactoferrin enriched whey protein powder and/or lactoferrin powder and the like, preferably lactoferrin powder.
6. The nutritional composition of any one of items 1-5, wherein:
the amount of probiotics was 5×10 relative to 1mg lactoferrin 2 ～5×10 10 cfu, preferably 5X 10 3 ～10 10 cfu, preferably 5X 10 4 ～5×10 9 cfu, preferably 5X 10 5 ～2×10 9 cfu, preferably 10 6 ～1.5×10 9 cfu。
7. A food product comprising the nutritional composition of any one of items 1-6.
8. The food product of item 7, wherein the food product is in powder or liquid form.
9. The food product of any one of items 7 to 8, which is an infant food product, a child food product, a teenager food product, or an adult food product; preferably the food is infant formula, infant complementary food, child formula, child snack, parturient formula, maternal formula, middle aged and elderly milk, or a nutritional or dietary supplement; preferably the food is maternal formula or maternal formula.
10. The food product of any one of items 7-9, wherein the nutritional composition is added in an amount such that, relative to the total mass of the food product:
the mass content of lactoferrin is at least 0.02mg/g, preferably at least 0.1mg/g, preferably at least 0.2mg/g, and preferably at most 200mg/g, preferably at most 100mg/g, preferably at most 20mg/g; and
the content of probiotics is at least 10 5 cfu/g, preferably at least 5X 10 5 cfu/g, preferably at least 10 6 cfu/g, and preferably at most 10 9 cfu/g, preferably at most 5X 10 8 cfu/g, preferably at most 10 8 cfu/g。
11. The food product of any one of items 7-10, wherein the nutritional composition is added in an amount such that, relative to the total mass of the food product:
The mass content of lactoferrin is at least 0.2mg/g and at most 20mg/g,
the amount of probiotics is at least 10 6 cfu/g and at most 10 8 cfu/g, and
the amount of probiotics was 5×10 relative to 1mg lactoferrin 4 ～5×10 9 cfu, preferably 5X 10 5 ～2×10 9 cfu, preferably 10 6 ～1.5×10 9 cfu。
12. Use of lactoferrin, probiotics or a nutritional composition as defined in any one of clauses 1-6 or a food product as defined in any one of clauses 7-11 for the non-therapeutic purpose of promoting development of the immune system of its offspring and improving the intestinal flora of its offspring via ingestion by a human or animal parent.
Drawings
FIG. 1 shows a typical experimental procedure diagram for a female mouse, wherein A is a vaginal smear before mating of the female mouse; b is a mating success picture; c is a photograph of a mouse in a 12-day state of birth.
FIG. 2 shows a representative diagram of macrophages phagocytosing chicken erythrocytes, wherein the lower arrow represents phagocytosing chicken erythrocytes by macrophages; the arrow above is the non-phagocytosed chicken erythrocytes.
FIG. 3 shows lactobacillus standard curve and sample amplification curve, wherein A is standard curve; b is a plasmid amplification curve; c is the sample amplification curve.
FIG. 4 shows a bifidobacterium standard curve and a sample amplification curve, wherein A is the standard curve; b is a plasmid amplification curve; c is the sample amplification curve.
Detailed Description
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs, if not otherwise defined, to which this invention belongs.
As used herein, the following terms have the following meanings.
The term "infant" refers to a person from 0 to 6 months of age.
The term "older infant" refers to a person of 6 to 12 months of age.
The term "young child" refers to a person of 12 to 36 months of age.
The term "infant" refers to a person 0-36 months of age.
The term "child" refers to a person of 3-6 years of age.
The term "juvenile" refers to a person of 7-17 years of age.
The term "adult" refers to a person over 18 years of age.
The term "young person" refers to a person of 18-40 years of age.
The term "adolescent" refers to a person 7-40 years of age.
The term "middle aged" refers to a person of 41-65 years of age.
The term "elderly" or "elderly" refers to people over 65 years of age.
The term "infant formula" as used herein encompasses infant formulas, older infant formulas, and toddler formulas. Typically, infant formulas are used as a breast milk substitute from the birth of the infant, and older infant formulas are used as breast milk substitutes from 6 to 12 months after the birth of the infant, and toddler formulas are used as breast milk substitutes from 12 to 36 months after the birth of the infant.
The term "infant formula" refers to a liquid or powder product produced by physical means only, using milk and milk protein products or soy and soy protein products as the main raw material, adding appropriate amounts of vitamins, minerals and/or other ingredients. Is suitable for normal infants, and the energy and nutrient components of the infant formula can meet the normal nutritional requirements of infants in 0-6 months.
The term "infant formula" refers to liquid or powder products made from milk and dairy products or soy and soy protein products as the main raw material, with the addition of appropriate amounts of vitamins, minerals and/or other ingredients, and by physical means only. Is suitable for older infants, and the energy and nutrient components of the older infants can meet the partial nutrient requirements of older infants in 6-12 months.
The term "infant formula" refers to a liquid or powder product which is produced by using milk and milk protein products or soybean and soybean protein products as main raw materials, adding proper amount of vitamins, minerals and/or other ingredients, and producing and processing by using only a physical method. Is suitable for infants, and the energy and nutrient components of the infant feed can meet the partial nutritional requirements of the infants in 12-36 months.
The term "breast milk" is understood to mean the mother's breast milk or colostrum.
The term "completely breast-fed infant or young child" has its ordinary meaning and refers to infants whose vast majority of nutrients and/or energy is derived from human breast milk.
The term "infant/follow-up/toddler fed mainly with infant formula" has its usual meaning, meaning that the source of nutrition for nutrients and/or energy is mainly derived from infants or toddlers physically produced and processed into infant formula, follow-up or growing-up milk. The term "primarily" refers to at least 50%, such as at least 75%, of those nutrients and/or energy.
In addition, in the context of the present invention, the term "comprising" or "comprises" does not exclude other possible elements. The compositions of the present invention (including embodiments described herein) may comprise, consist of, or consist essentially of the following elements: the essential elements of the invention described herein and any of the other or optional ingredients, components or limitations described herein or otherwise as desired.
The subject of the invention is suitable for normal humans and may be infants and/or older infants, and/or young children, and/or young adults, and/or middle-aged adults, and/or elderly adults.
All percentages are by mass unless otherwise indicated.
The invention will now be described in more detail. It should be noted that the various aspects, features, embodiments, examples, and advantages thereof described herein may be compatible and/or may be combined together.
The invention will now be described in more detail. It should be noted that the various aspects, features, embodiments, examples, and advantages thereof described herein may be compatible and/or may be combined together.
The present invention relates to a nutritional composition for promoting immune system development and/or improving immune system function, and improving intestinal flora, in particular a nutritional composition capable of promoting the development of the immune system and improving the intestinal flora of offspring via maternal consumption, a food product comprising the nutritional composition, the nutritional composition or a nutritional ingredient thereof or a food product comprising the nutritional composition for non-therapeutic purposes in promoting the development of the immune system of offspring and improving the intestinal flora of offspring via maternal consumption by a human or animal.
The present invention will be specifically described below.
Nutritional composition
In one aspect, the present invention provides a nutritional composition comprising:
-lactoferrin; and
-probiotics.
Lactoferrin is an iron-binding glycoprotein with a molecular weight of about 80kDa, belonging to the transferrin family. Lactoferrin is present in high levels in colostrum and milk and in lower levels in mucosal secretions such as tears, saliva, semen, nasal and bronchial secretions, bile and gastrointestinal fluids. In addition, lactoferrin is also a constituent of neutrophils. Lactoferrin is one of the main defensive molecules of the body, and in addition to helping transport and transfer iron ions, increasing the bioavailability of iron element, it has various biological immune activities including broad spectrum antibacterial, antiviral, antioxidant and immune-modulating activities.
Probiotics are a class of active microorganisms beneficial to a host by colonizing the human body and altering the flora composition of a part of the host. By regulating the immune function of host mucous membrane and system or regulating the balance of flora in intestinal tract, the effect of promoting nutrient absorption and maintaining intestinal health is achieved, so that single microorganism or mixed microorganism with definite composition beneficial to health is produced.
In one embodiment, the probiotic is one or more selected from the group consisting of bifidobacterium infantis, bifidobacterium longum, bifidobacterium bifidum, bifidobacterium adolescentis, bifidobacterium lactis, bifidobacterium breve, lactobacillus acidophilus and lactobacillus rhamnosus.
In one embodiment, the probiotic is bifidobacterium lactis.
In one embodiment, the probiotic or bifidobacterium lactis is bifidobacterium lactis Bb12.
Bifidobacterium lactis Bb12 (also sometimes referred to herein as bifidobacterium Bb12 or Bb12, for example) is belonging to the phylum actinomycota, bifidobacterium animalis, bifidobacterium lactis subspecies. It is a gram positive, polymorphous bacillus. The thallus form is in a bent rod shape or a branch rod shape, bb12 does not form spores, has no motility and is obligate anaerobic, the most suitable growth temperature is 36-38 ℃, and the most suitable growth ph value is 6.5-7.0.
The inventors have surprisingly found that when lactoferrin is used in combination with one or more of probiotics (e.g. bifidobacterium infantis, bifidobacterium longum, bifidobacterium bifidum, bifidobacterium adolescentis, bifidobacterium lactis, bifidobacterium breve, lactobacillus acidophilus and lactobacillus rhamnosus; preferably bifidobacterium lactis, more preferably bifidobacterium lactis Bb 12), there is a synergistic effect between the two components, which is capable of synergistically promoting development of the immune system of the offspring via dietary intake by the parent (e.g. human (i.e. pregnant woman) or animal (i.e. pregnant female)) and which may improve, e.g. synergistically improve, the intestinal microbiota of the offspring.
In one embodiment, the nutritional composition consists of:
-lactoferrin; and
-probiotics, preferably selected from one or more of bifidobacterium infantis, bifidobacterium longum, bifidobacterium bifidum, bifidobacterium adolescentis, bifidobacterium lactis, bifidobacterium breve, lactobacillus acidophilus and lactobacillus rhamnosus; preferably bifidobacterium lactis, more preferably bifidobacterium lactis Bb12.
In one embodiment, the probiotic is bifidobacterium lactis.
In one embodiment, the probiotic is bifidobacterium lactis Bb12.
In one embodiment, the lactoferrin may be derived from or provided in the form: lactoferrin (LF) enriched nutritional ingredients such as lactoferrin enriched whey protein powder and/or lactoferrin powder and the like, preferably lactoferrin powder.
In one embodiment, the amount of probiotics (e.g. one or more selected from the group consisting of bifidobacterium infantis, bifidobacterium longum, bifidobacterium bifidum, bifidobacterium adolescentis, bifidobacterium lactis, bifidobacterium breve, lactobacillus acidophilus and lactobacillus rhamnosus; e.g. bifidobacterium lactis, e.g. bifidobacterium lactis Bb 12) is 5 x 10 relative to 1mg of lactoferrin in the nutritional composition 2 ～5×10 10 cfu, preferably 5X 10 3 ～10 10 cfu, preferably 5X 10 4 ～5×10 9 cfu, preferably 5X 10 5 ～2×10 9 cfu, preferably 10 6 ～1.5×10 9 cfu may be, for example, (5.0, 5.1, 5.2, 5.3, 5.4, 5.5. 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, or 9.9). Times.10 2 Or (1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, or 9.9) ×10 3 Or (1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, or 9.9) ×10 4 Or (1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, or 9.9) ×10 5 Or (1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0),2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, or 9.9) ×10 6 Or (1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, or 9.9) ×10 7 Or (1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, or 9.9) ×10 8 Or (1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4) 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0. 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, or 9.9). Times.10 9 Or (1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or 5.0). Times.10 10 cfu, or a range defined by any two thereof.
The nutritional composition may synergistically promote development of the immune system of its offspring via ingestion by a parent, such as a human (i.e. pregnant woman) or an animal (i.e. pregnant female), and may improve, e.g. synergistically improve, the intestinal flora of its offspring. When the ratio of lactoferrin to probiotics (e.g. one or more selected from the group consisting of bifidobacterium infantis, bifidobacterium longum, bifidobacterium bifidum, bifidobacterium adolescentis, bifidobacterium lactis, bifidobacterium breve, lactobacillus acidophilus and lactobacillus rhamnosus; e.g. bifidobacterium lactis, e.g. bifidobacterium lactis Bb 12) in the nutritional composition is within the above-mentioned range, ingestion via a parent (e.g. a human (i.e. pregnant woman) or an animal (i.e. pregnant female)) is able to more significantly, in particular synergistically, promote development of the immune system of the offspring, and may improve, e.g. synergistically, the intestinal microbiota of the offspring.
Food products
In another aspect, the invention also relates to food products (including health products) comprising said nutritional composition.
The food product of the present invention may be in powder form or in liquid form.
The food of the present invention may be infant food, child food, juvenile food, or adult food; preferably the food is infant formula, infant complementary food, child formula, child snack, parturient formula, maternal formula, middle aged and elderly milk, or a nutritional or dietary supplement; preferably the food is maternal formula or maternal formula.
In one embodiment, the nutritional composition is added in an amount such that the mass content of lactoferrin relative to the total mass of the food product is at least 0.02mg/g, preferably at least 0.1mg/g, preferably at least 0.2mg/g, and preferably at most 200mg/g, preferably at most 100mg/g, preferably at most 20mg/g. For example, the total mass of the food product, the lactoferrin may be present in a mass content of 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200mg/g, or a range defined by any two thereof.
In one embodiment, the nutritional composition is added in an amount such that the content of probiotics (e.g. one or more selected from the group consisting of bifidobacterium infantis, bifidobacterium longum, bifidobacterium bifidum, bifidobacterium adolescentis, bifidobacterium lactis, bifidobacterium breve, lactobacillus acidophilus and lactobacillus rhamnosus; e.g. bifidobacterium lactis, e.g. bifidobacterium lactis Bb 12) is at least 10 relative to the total mass of the food product 5 cfu/g, preferably at least 5X 10 5 cfu/g, preferably at least 10 6 cfu/g, and preferably at most 10 9 cfu/g, preferably at most 5X 10 8 cfu/g, preferably at most 10 8 cfu/g. For example, the content of probiotics may be (1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.1, 3.7, 3.6, 3.8),5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, or 9.9) ×10 5 Or (1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, or 9.9) ×10 6 Or (1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, or 9.9) ×10 7 Or (1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, or 9.9) ×10 8 Or 1.0X10 9 cfu/g, or a range defined by any two thereof.
When the content of lactoferrin and probiotics (e.g. one or more selected from the group consisting of bifidobacterium infantis, bifidobacterium longum, bifidobacterium bifidum, bifidobacterium adolescentis, bifidobacterium lactis, bifidobacterium breve, lactobacillus acidophilus and lactobacillus rhamnosus; e.g. bifidobacterium lactis, e.g. bifidobacterium lactis Bb 12) in the foodstuff is within the above-mentioned ranges, the intake is significantly, in particular synergistically, promoted for the development of the immune system of the offspring thereof, and the intestinal flora of the offspring thereof, e.g. can be improved, e.g. synergistically, and the (other) aspects of nutrition required by e.g. the human or animal body, including the parent and offspring, can also be balanced at the same time, when consumed via the parent, i.e. pregnant woman or animal, i.e. pregnant female.
In a preferred embodiment, the nutritional composition is added in an amount such that the mass content of lactoferrin relative to the total mass of the food product is at least 0.2mg/g and at most 20mg/g, or any other value or range mentioned above within this range, and the amount of probiotics (e.g. one or more selected from bifidobacterium infantis, bifidobacterium longum, bifidobacterium bifidum, bifidobacterium adolescentis, bifidobacterium lactis, bifidobacterium breve, lactobacillus acidophilus and lactobacillus rhamnosus; e.g. bifidobacterium lactis, e.g. bifidobacterium lactis Bb 12) is at least 10 6 cfu/g and at most 10 8 cfu/g, or any other value or range mentioned above within this range, and the amount of probiotics relative to 1mg lactoferrin is 5 x 10 4 ～5×10 9 cfu, preferably 5X 10 5 ～2×10 9 cfu, preferably 10 6 ～1.5×10 9 cfu, or any other value or range mentioned above within this range. When the ratio of lactoferrin to probiotics (e.g. bifidobacteria Bb 12) and the amount are within the above-mentioned ranges or preferred ranges, the development of the offspring immune system can be more significantly, in particular synergistically, promoted when ingested via the mother (e.g. a human (i.e. pregnant woman) or an animal (i.e. pregnant female)), and the offspring intestinal flora can be improved, e.g. synergistically, and at the same time (other) aspects of nutrition, e.g. needed by the human or animal body (including the mother and offspring) can be balanced.
In addition to the components described above for the nutritional composition, the food product may also comprise other ingredients, such as proteins/amino acids, carbohydrates, fats, vitamins, minerals etc. which are often contained in a formula, e.g. an infant formula such as milk powder.
Use of the same
In a further aspect, the invention relates to the use of the above lactoferrin or probiotics (e.g. one or more selected from the group consisting of bifidobacterium infantis, bifidobacterium longum, bifidobacterium bifidum, bifidobacterium adolescentis, bifidobacterium lactis, bifidobacterium breve, lactobacillus acidophilus and lactobacillus rhamnosus; e.g. bifidobacterium lactis, e.g. bifidobacterium lactis Bb 12) or a nutritional composition or a food product as described above for non-therapeutic purposes (nutrition and/or health care) in promoting development of the immune system and improving the intestinal flora of offspring thereof via maternal consumption.
In one embodiment, the invention relates to the use of lactoferrin for non-therapeutic purposes (nutritional and/or health) in promoting development of the immune system and improving the intestinal flora of offspring via maternal consumption.
In one embodiment, the invention relates to the use of probiotics (e.g. one or more selected from the group consisting of bifidobacterium infantis, bifidobacterium longum, bifidobacterium bifidum, bifidobacterium adolescentis, bifidobacterium lactis, bifidobacterium breve, lactobacillus acidophilus and lactobacillus rhamnosus; e.g. bifidobacterium lactis, e.g. bifidobacterium lactis Bb 12) for promoting development of the immune system and improving non-therapeutic purposes (nutritional and/or health) in the intestinal flora of offspring thereof via maternal consumption.
In one embodiment, the present invention relates to the use of the above-described nutritional composition or food product comprising lactoferrin and a probiotic (e.g. one or more selected from the group consisting of bifidobacterium infantis, bifidobacterium longum, bifidobacterium bifidum, bifidobacterium adolescentis, bifidobacterium lactis, bifidobacterium breve, lactobacillus acidophilus and lactobacillus rhamnosus; e.g. bifidobacterium lactis, e.g. bifidobacterium lactis Bb 12) for promoting development of the immune system and improving non-therapeutic purposes (nutrition and/or health) in the intestinal flora of its offspring via maternal consumption.
Those skilled in the art will readily appreciate that the promotion is by maternal and fetal nutrient transfer (i.e., pregnancy) or maternal and infant (i.e., lactation).
Examples
The present invention will be described in further detail with reference to the following examples in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.
1. Experimental materials
1.1. Experimental main reagent
Figure BDA0003970836120000141
1.2. Main experimental instrument
Figure BDA0003970836120000151
2. Experimental method
To verify the effect of each experiment, the following animal (mouse) experiment was designed.
2.1. Experimental animal
Female 75 mice and 25 male mice of sexually mature ICR mice, with a mass of 20+ -2 g. Beijing Fukang biotechnology Co., ltd., license number: SCXK (jing) 2019-0008. Feeding conditions: indoor temperature 25+ -1deg.C, humidity 45+ -5%, standard 12h light/12 h dark rhythm. The feed, the drinking water and the like are uniformly matched, so that good ventilation of the indoor environment is kept, the indoor environment is cleaned every day, and good sanitary environment in the cage is maintained; animal welfare: in the whole treatment process of the experiment, reasonable means are carried out on the mice, which accords with the relevant regulations of the ethical committee of experimental animals, and the mice are specifically carried out according to IAC and other relevant standard operation regulations (SOP); the method for killing comprises the following steps: and (3) carrying out anesthesia on the mice after the experiment is finished and the mice are removed, carrying out abdominal artery bloodletting and sacrifice, then storing the cadavers in a special refrigerator container, and finally carrying out harmless unified treatment.
2.2. Experimental grouping and administration
SPF-class 75 female mice were fed at room temperature and were fed with water ad libitum. After 1 week of acclimatization, the female and male mice were then caged (2:1 or 3:1) in a given ratio and the female mice were checked for vagina in the morning (8:00) daily. The yellow-white embolus found in the vagina is the condensation of semen, namely the vaginal embolus, the vaginal embolus is the mated behavior, 8:00 in the morning of the vaginal embolus is found to be the 0 th day of pregnancy, and the gestation condition of the mated mice is observed in 0-6d, so that the pregnant mice are not removed and supplemented, and 55 pregnant mice are produced. FIG. 1 shows a typical experimental procedure wherein A is a pre-mating vaginal smear of a female mouse; b is a mating success picture; c is a photograph of a mouse in a 12-day state of birth.
Pregnant mice were randomly divided into 11 groups of 5 mice each. The test subjects were weighed and labeled in the blank control group and the test subjects 2 to 11 groups, respectively. Each mouse is respectively administrated with a test substance according to 100g/mL stomach irrigation (the test substance is prepared into corresponding dosage by purified water); the blank group was perfused with purified water until the offspring mice were naturally born.
Figure BDA0003970836120000161
Note that: the bifidobacterium is converted into a gastric lavage dose of 25mg/kg according to 0.75mg of mice with gastric lavage of 30 g.
2.3. General observations
During the period of stomach irrigation, all the conditions of body weight, eating, drinking water, activity, defecation and urination and the like of the female mice are observed. After the female mice are delivered, the number of offspring mice in each nest, whether deformity exists or not and the development condition of offspring mice are recorded.
2.4. Immune index detection
The 11 groups of offspring mice are sacrificed 10-12 days after birth, spleen and blood samples are collected and used for immune index detection, and specific index detection is as follows:
2.4.1. determination of immune organ coefficients of mice
9 mice are taken from each group, thymus and spleen of each mouse are respectively taken, washed by normal saline, and then the mice are sucked dry by filter paper, weighed by an electronic balance and brought into a formula for calculation.
Organ coefficient calculation formula: organ coefficient (mg/g) =organ weight/corresponding mouse weight×100%
2.4.2. Detection of proliferation of spleen lymphocytes in mice
Taking 6 mice from each group, rapidly placing the weighed spleen in Hanks liquid containing 2mL to be cut into three sections, grinding the spleen to obtain cell suspension, filtering the cell suspension through a 200-mesh nylon net, transferring the cell suspension into a 1.5mL sterile centrifuge tube, centrifuging at 1500r/min at 4 ℃ for 5min, and discarding the supernatant; adding erythrocyte lysate, mixing, standing for 5min, centrifuging, and removing supernatant; washing with 1mL Hanks solution twice, centrifuging, and discarding the supernatant; it was made into a cell suspension with complete medium (45 mL 1640 medium+5 mL foetal calf serum) and its cell concentration was adjusted to 2.5X106 cells/mL with PBS. The cell suspension was added to a 96-well plate at 100. Mu.L/well, 2 multiplex wells. 1mg/mL of LPS medium was prepared, and 100. Mu.L of LPS-containing medium was added to one of the 8:30 wells on the morning, and no blank medium was added to the other well, as a control. The cells were incubated at 37℃in a 5% CO2 incubator for 72 hours. After the completion of the incubation, 10. Mu.L of CCK8 reagent was added to each well, and the OD value was measured by an ELISA reader at a wavelength of 490nm, and the result was expressed as OD value.
2.4.3. Determination of phagocytic function of mouse macrophages
Each group was prepared from 6 10-day mice, each of which was intraperitoneally injected with 0.8mL of a 5% peptone starch suspension. After 3 days, 0.8mL of 1% chicken erythrocyte suspension is injected into the abdominal cavity, and the abdomen of the mouse is massaged. After 30 minutes, the mice were sacrificed, and the liquid in the abdominal cavity was aspirated with a pipette, and smears were prepared. After the smear is completely dried, the smear is dyed for 5min by the Rayleigh dye liquor, and the smear is dried and sealed by neutral resin. The phagocytic percentage was calculated by counting the number of macrophages that engulf the chicken erythrocytes.
Percent phagocytosis (%) = number of macrophages phagocytosed with chicken erythrocytes/total number of macrophages 100%2.4.4. Determination of immunoglobulin (Ig G, ig a, ig M) content in serum
Taking 6 mice from each group, taking blood from broken ends, collecting blood samples, centrifuging 3500r/min for 10min and 6min respectively after water bath at 37 ℃ for 1h, sucking and preparing serum, and storing the serum in a refrigerator at-80 ℃ for unified measurement. 3 mouse sera were selected from each group and tested for Ig G, ig A, ig M content in the sera by enzyme-linked immunosorbent assay (ELISA). The specific operation method is as follows:
1) All reagents, samples were equilibrated to room temperature prior to testing. All required reagents and working concentration standards were prepared.
2) Soaking the ELISA plate: 300. Mu.L of 1 Xthe washing reagent was added thereto and the mixture was allowed to stand still for 30 seconds. Soaking is necessary to obtain the desired experimental results. After the wash liquid was discarded, the microplate was patted dry on absorbent paper. Immediately after the plate washing is completed, the microplate is used without drying the microplate.
4) Adding a standard substance: standard wells were added with 100 μl of 2-fold diluted standard. Blank wells were added with 100 μl of 1 x detection buffer (serum/plasma samples) or culture medium (cell culture supernatant samples).
5) Adding a sample: serum/plasma: sample wells were added with 90 μl of 1 x detection buffer and 10 μl of pre-diluted samples (10000-fold dilution of all samples). Cell culture supernatant: sample wells were added with 100 μl of cell culture supernatant. And (5) ensuring continuous sample adding in the steps 4 and 5 without interruption. The loading process was completed within 15 minutes.
6) Incubation: a sealing plate membrane seal plate is used. Shaking at 300 rpm, and incubating at room temperature for 2 hours.
7) Washing: the liquid was discarded, and 300. Mu.L of wash solution was added to wash the plate 6 times per well. And (5) washing the board each time, and drying the board by beating on absorbent paper. To obtain the desired experimental performance, the residual liquid must be thoroughly removed.
8) Adding a detection antibody: mu.L (1:100 dilution) of diluted detection antibody was added to each well.
9) Incubation: a new sealing plate membrane sealing plate is used. Shaking at 300 rpm, and incubating at room temperature for 1 hour.
10 Washing: and 7, repeating the step 7.
11 Color development of the substrate: mu.L of chromogenic substrate TMB was added to each well and incubated at room temperature for 5-30 min in the absence of light.
12 Adding a stop solution: 100. Mu.L of stop solution was added to each well. The color changes from blue to yellow. If the color is green or the color change is obviously uneven, the plate frame is gently tapped and fully mixed.
13 Detection reading): within 30 minutes, a dual wavelength detection was performed using an enzyme-labeled instrument, and OD values at a 450nm maximum absorption wavelength and a 570nm reference wavelength were determined. The OD value after calibration was measured at 450nm minus 570 nm.
And (3) detecting the absolute content of lactobacillus and bifidobacterium in the mouse excrement by RT-PCR.
2.5.1. Experimental reagent
2X SYBR Green pro TaqHS Premix (Ai Kerui, A0147); DNase, RNase-free Water (Thermo Fisher, R0581)
2.5.2. Experimental instrument
Centrifuge (thermosusher, legend Micro 21R), micro ultraviolet spectrophotometer (thermosusher, nanodrop 2000), fluorescent quantitative PCR instrument (ABI, 7900 HT).
2.5.3. Vector construction
Insertion of Gene information
Figure BDA0003970836120000181
Calculating the dosage
Vector usage (ng) =3×17.5 (PCR product length (bp)/2974)
Gene name Bifidobacterium
PCR product Length (bp) 511
Vector quantity (ng) 17.5
Carrier length (bp) 2974
Optimum usage (ng) 9.02
Inserted gene volume (μl) 0.66
Ligation reaction
Figure BDA0003970836120000191
Transformation, monoclonal selection and shaking
1) Adding part of the ligation product into 50-100 μl TOP10 competent cells (the competent cells should be taken out from a refrigerator at-70deg.C and placed on ice bath, adding ligation product when thawing, and adding ligation product in an amount not exceeding one tenth of the volume of competent cells), flicking, mixing, and ice-bathing for 30min;
2) Placing the centrifuge tube at a constant temperature of 42 ℃ for 90sec, immediately placing the centrifuge tube in an ice bath after the centrifuge tube is taken out, and placing for 2-3min without shaking the centrifuge tube during the process;
3) Adding 250-500 μl of LB (without antibiotics) culture medium preheated at 37deg.C into the centrifuge tube, and shake culturing at 150rpm and 37deg.C for 45min for the purpose of expressing related resistance marker genes on plasmid to resuscitate thallus;
4) The bacterial liquid in the centrifuge tube is evenly mixed, 100 mul of the bacterial liquid is absorbed and added on LB solid agar medium containing ampicillin, and the cells are gently and evenly spread by a sterile elbow glass rod. After the surface of the plate is dried, the plate is inverted and incubated at 37℃for 12-16h.
5) In an ultra clean bench, single macroscopic colonies were selected, picked into LB medium containing ampicillin, and cultured overnight (12-16 h, preferably with bacterial turbidity) at 37℃with shaking, 150rpm, one colony per tube.
Plasmid extraction: plasmids were extracted using the TIANprep Rapid Mini Plasmid Kit kit.
2.5.4. RT-PCR detection of bifidobacteria and lactobacillus
(1) Standard curve preparation: the cloned plasmid was subjected to 10-fold gradient dilution, 45. Mu.L of the dilution+5. Mu.L of plasmid, and 4-7 spots were generally selected and a suitable standard was selected for preparing a standard curve by preliminary experiments. Clone plasmid copy number calculation formula (copies/. Mu.L) = [ plasmid concentration (ng/ul). Times.10-9X 6.02X 1023]/[ clone product base number X660 (g/mol) ]
Figure BDA0003970836120000201
(2) Extraction of fecal DNA samples: 6 parts of 4-5 intestinal faeces of the mixed mice are collected in each group, placed in a sterile EP tube and weighed. All bacterial genomic DNA in the feces was extracted with a stool DNA miniprep kit and stored at-80 ℃. Carrying out real-time fluorescence quantitative PCR reaction on the DNA extract of the mouse fecal sample according to a standard reaction system and reaction conditions.
(3) Designing the gene sequence of lactobacillus and bifidobacterium 16S rDNA: the lactobacillus and bifidobacterium specific PCR amplification primer pairs were queried according to the literature.
TABLE 2RT-PCR amplification primer sequences
Figure BDA0003970836120000202
(4) Quantitative PCR assay: the bifidobacterium and lactobacillus freeze-dried bacterial powder is also extracted by using a small amount of purification kit of the Takara bacterial genome DNA. 10. Mu.L of a standard PCR reaction system, 2X SYBR Green pro TaqHS Premix 5.0. Mu.L of a reagent, 0.4. Mu.L of upper and lower primers, 0.2. Mu.L of a ROX reagent, 1.0. Mu.L of DNA Template and 3.0. Mu.L of DNase were sequentially added dropwise according to a PCR kit, and reaction detection was performed. The corresponding reaction conditions: signals were collected by pre-denaturation at 95℃for 60s,40 cycles (denaturation at 95℃for 10s, annealing at 60℃for 30s, extension at 72℃for 30 s) and extension at 72℃for 10 min.
(5) And (3) calculating: the Ct values obtained for each sample were carried into a standard curve, and the copy numbers of the genes of Lactobacillus and Bifidobacterium were calculated per microgram of DNA.
2.6. Statistical treatment
Mean ± standard deviation of experimental results
Figure BDA0003970836120000203
Representing, and performing differential analysis on experimental data by adopting SPSS 15.0 statistical software, wherein single-factor analysis of variance (ANOVA) variance homogeneity is tested by adopting LSD; variance is measured using a non-parametric rank sum test or a two-sample independent T test. P (P)<0.05 indicates a statistical significance.
3. Experimental results
3.1. General Condition observations
The female mice drink water normally during the whole pregnancy, the weight of the female mice increases slowly in the early period and increases rapidly in the later period. The mice have no deformity after birth and good growth state.
3.2. Effects on immune organ coefficients in mice
3.2.1. Effect on spleen index in mice
The effect of different doses of lactoferrin and/or Bb12 on the immune organ index of mice was studied. The results are shown in Table 3-1.
Table 3-1 spleen and thymus index of mice in each group
Figure BDA0003970836120000211
Figure BDA0003970836120000221
Note that: p <0.05, < P <0.01, < P <0.001 compared to the placebo group; t test was used.
The development of immune organs directly reflects the immune condition of an organism, spleen and thymus are taken as important immune centers of the organism, are main sites of differentiation, development and maturation of T cells and B cells and are also the centers of cellular immunity and humoral immunity of the organism, so that spleen and thymus indexes can reflect the immune capacity of the organism. Each group of experiments performed in good condition.
From Table 3-1, it is clear that ICR pregnant mice have spleen and thymus indexes increased under different doses of lactoferrin, compared with the blank control group, so that the immune organ growth of mice is promoted to a certain extent by the intervention of lactoferrin in the matrix, the immune organ indexes of the mice can be improved, and the development of the immune system is promoted.
Under the intervention of Bb12 in different doses, spleen and thymus indexes of mice in each group are increased compared with that of mice in a blank control group, which indicates that the intervention of Bb12 in a parent body has a certain promoting effect on the immune organ growth of mice, can improve the immune organ indexes of the mice and promote the development of immune systems.
The inventors have further found that when lactoferrin and Bb12 are used in combination, the IgG content can be significantly increased, and that there can be a synergistic effect between the two.
In particular, with respect to the components used and the amounts used, group 10 corresponds to the combination of groups 3 and 5, and groups 10, 3, 5 increase the spleen index by 1.7, 1.09, 0.57, respectively, relative to the blank, with the increase in spleen index (1.7) in group 10 being greater than the sum of the respective increases (1.66) of groups 3 and 5.
3.3. Effect on serum immunoglobulins in mice
Immunoglobulin is a protein with antibody activity, which exists in blood or body fluid, can kill pathogenic microorganisms under the synergistic effect of complement, and is an important part of body humoral immunity. IgG is the highest content immunoglobulin in serum, is the only immunoglobulin capable of passing through placenta, is the main immunoglobulin mediating the humoral immunity, and is also the main immunoglobulin for serological diagnostic detection. IgA content was next to IgG. IgM is the earliest produced immunoglobulin of the organism, and the immune level of the organism can be reflected by the content of the immunoglobulin in serum.
3.3.1. Effect on mouse serum immunoglobulin IgG
Table 3-2 changes in serum immunoglobulin IgG levels in groups of mice
Figure BDA0003970836120000231
Note that: p <0.05, < P <0.01, < P <0.001 compared to the placebo group; t-test is adopted between groups
Under the intervention of different doses of lactoferrin, the ICR pregnant mice have higher IgG of each group of mice compared with a blank control group, which indicates that the immune globulin of the mice is increased to a certain extent by the intervention of the lactoferrin in the matrix, and the humoral immunity of the mice can be improved.
Under the intervention of Bb12 in different doses, the ICR pregnant mice have higher IgG of each group of mice compared with a blank control group, which shows that the intervention of Bb12 in the matrix has a certain promotion effect on the growth of immunoglobulin of the mice, and can improve the humoral immunity of the mice.
The inventors have further found that when lactoferrin and Bb12 are used in combination, the IgG content can be significantly increased, and that there can be a synergistic effect between the two.
In particular, group 8 corresponds to the combination of groups 4 and 5 in terms of the components used and the amounts used, and groups 8, 4, 5 increase the IgG content by 7.23, 5.41, 1.33, respectively, relative to the blank, with the IgG increase (7.23) in group 8 being greater than the sum of the respective increases (6.74) of groups 4 and 5. 3.3.2. Effect on murine serum immunoglobulin IgA
Tables 3-3 variation in serum immunoglobulin IgA content of mice of each group
Figure BDA0003970836120000241
Note that: p <0.05, < P <0.01, < P <0.001 compared to the placebo group; t-test is adopted between groups
Under the intervention of different doses of lactoferrin, the ICR pregnant mice have higher IgA of each group of mice compared with a blank control group, which shows that the immune globulin of the mice is increased to a certain extent by the intervention of the lactoferrin in the matrix, and the humoral immunity of the mice can be improved.
Under the intervention of Bb12 in different doses, the ICR pregnant mice have higher IgA than that of blank control groups, which shows that the intervention of Bb12 in the matrix has a certain promotion effect on the growth of immunoglobulin of the mice, and can improve the humoral immunity of the mice.
3.3.3 Effect on mouse serum immunoglobulin IgM
Serum immunoglobulin IgM content of mice of each group of tables 3-4
Figure BDA0003970836120000242
/>
Figure BDA0003970836120000251
Note that: p <0.05, < P <0.01, < P <0.001 compared to the placebo group; t-test is adopted between groups
Compared with a blank control group, ICR pregnant mice have elevated IgM under the intervention of different doses of lactoferrin, which indicates that the immune globulin of the mice is increased to a certain extent by the intervention of the lactoferrin in the matrix, and the humoral immunity of the mice can be improved.
Compared with a blank control group, ICR pregnant mice have higher IgM in each group under the intervention of Bb12 at different doses, which shows that the intervention of Bb12 in the matrix has a certain promotion effect on the growth of immunoglobulin of the mice, and can improve the humoral immunity of the mice.
The inventors have further found that when lactoferrin and Bb12 are used in combination, the IgM content can be significantly increased, and that there can be a synergistic effect between the two.
In particular, with respect to the components used and the amounts used,
group 7 corresponds to the combination of groups 4 and 6, groups 7, 4, 6 increasing the IgM content by 0.15, 0.12, 0.01, respectively, relative to the blank, the IgG increase (0.15) in group 7 being greater than the sum (0.13) of the respective increases of groups 4 and 6.
3.4 Effect on proliferation of spleen lymphocytes in mice
Tables 3-5 changes in the percent proliferation of spleen lymphocytes in groups of mice
Figure BDA0003970836120000252
/>
Figure BDA0003970836120000261
Note that: p <0.05, < P <0.01 compared to the placebo group; t-test is adopted between groups
Spleen is the place of immune response of T, B lymphocytes, and the proliferation capacity of the lymphocytes reflects the strength of the body's immunity. ConA has the capacity of non-specifically inducing T cell activation, mouse T lymphocytes without ConA stimulation are in G0 phase, conA can enable the mouse T lymphocytes to enter G1 phase (DNA synthesis prophase) from G0 phase, and simultaneously express IL-2 receptor, so that T cells are activated and proliferated, spleen lymphocyte transformation experiments are currently becoming a common method for researching the effect of immunomodulators on lymphocytes, and are also basic indexes for reflecting the immune status of organism cells, and the proliferation capacity of the lymphocytes stimulated by antigens or mitogens reflects the height of lymphocyte functions.
From the table, it can be seen that ICR pregnant mice continuously proliferate spleen lymphocytes of each group of mice under the intervention of different doses of lactoferrin, compared with a blank control group, which indicates that the interference of the lactoferrin on the parent body has a certain promotion effect on the proliferation of the spleen lymphocytes of the mice, and can improve the cellular immunity of the mice.
Under the intervention of Bb12 in different doses, the ICR pregnant mice are compared with the blank control group, and spleen lymphocytes of the mice in each group are continuously proliferated, which indicates that the intervention of Bb12 in the parent body has a certain promoting effect on the proliferation of the spleen lymphocytes of the mice, and can improve the cellular immunity of the mice.
The inventors have further found that when lactoferrin and Bb12 are used in combination, there can be a synergistic effect between the two that can significantly increase spleen lymphocyte proliferation.
In particular, with respect to the components used and the amounts used,
group 7 corresponds to the combination of groups 4 and 6, groups 7, 4, 6 respectively multiplying the splenic lymphocytes by 22.81%, 16.86%, 1.21% relative to the blank, the increase in proliferation of the splenic lymphocytes in group 7 (22.81%) being greater than the sum of the respective increases of groups 4 and 6 (18.07%).
Group 8 corresponds to the combination of groups 4 and 5, groups 8, 4, 5, respectively, allow proliferation of spleen lymphocytes by 18.75%, 16.86%, 0.06%, the increase in proliferation of spleen lymphocytes in group 8 (18.75%) being greater than the sum of the respective increases of groups 4 and 5 (16.92%) with respect to the blank group.
Group 9 corresponds to the combination of groups 3 and 6, groups 9, 3, 6 respectively allow proliferation of spleen lymphocytes by 20.86%, 10.93%, 1.21% relative to the blank, the increase in proliferation of spleen lymphocytes in group 9 (20.86%) being greater than the sum of the respective increases of groups 3 and 6 (12.14%).
Group 10 corresponds to the combination of groups 3 and 5, groups 10, 3, 5, respectively, having a proliferation of spleen lymphocytes of 19.25%, 10.93%, 0.06% relative to the blank, the increase in proliferation of spleen lymphocytes in group 10 (19.25%) being greater than the sum of the respective increases of groups 3 and 5 (10.99%).
Group 11 corresponds to the combination of groups 2 and 6, groups 11, 2 and 6, respectively, allow proliferation of spleen lymphocytes by 22.14%, 6.35% and 1.21% with respect to the blank group, the increase in proliferation of spleen lymphocytes in group 11 (22.14%) being greater than the sum of the respective increases of groups 2 and 6 (7.56%).
3.5 Effect on phagocytic function of mouse macrophages
Tables 3-6 change in phagocytic function of mouse macrophages
Figure BDA0003970836120000271
Note that: p <0.05, < P <0.01, < P <0.001 compared to the placebo group; t-test is adopted between groups
In the aspect of nonspecific immunity, mononuclear macrophages are important executors of nonspecific immunity, are beneficial to phagocytosis of pathogenic microorganisms, clearance of self-aging injury cells, and are beneficial to uptake, processing, treatment, presentation and immune response of antigens.
FIG. 2 shows a representative graph of phagocytic chicken erythrocytes by macrophages. Wherein the lower arrow represents phagocytic chicken erythrocytes by macrophages; the arrow above is the non-phagocytosed chicken erythrocytes.
ICR pregnant mice have continuously enhanced phagocytic capacity of macrophages of the mice in each group compared with a blank control group under the intervention of different doses of lactoferrin, which indicates that the ICR pregnant mice have certain promotion effect on the phagocytic capacity of the macrophages of the mice by the intervention of the lactoferrin in the matrix.
Compared with a blank control group, ICR pregnant mice are continuously enhanced in phagocytic capacity under the intervention of Bb12 at different doses, which shows that the interference of Bb12 on the parent has a certain promotion effect on the phagocytic capacity of macrophages of mice, and the nonspecific immunity of the mice can be improved.
3.6 Effect on the Lactobacillus and Bifidobacterium content of the mouse feces
3.6.1 Effect on Lactobacillus plantarum in murine faeces
FIG. 3 shows lactobacillus standard curve and sample amplification curve, wherein A is standard curve; b is a plasmid amplification curve; c is the sample amplification curve. Drawing a standard curve with copy number (Copies) as an abscissa and CT value as an ordinate, wherein the bifidobacterium linear standard curve is y= -3.3537462log x+34.52568, R 2 ＝0.9989。
Tables 3 to 7 content of Lactobacillus in faeces of mice
Figure BDA0003970836120000281
Figure BDA0003970836120000291
And (3) injection: p <0.05, < P <0.01, < P <0.001 compared to the placebo group; t-test is adopted between groups
Under the intervention of different doses of lactoferrin, ICR pregnant mice can increase the abundance of lactobacillus compared with a blank control group, which indicates that the ICR pregnant mice have a certain improvement effect on intestinal flora of mice through the intervention of lactoferrin in the matrix.
Under the intervention of Bb12 in different doses, ICR pregnant mice can increase the abundance of lactobacillus in each dose compared with a blank control group, which indicates that the intestinal flora of the mice is improved to a certain extent by the intervention of Bb12 in the parent.
The inventors have further found that when lactoferrin and Bb12 are used in combination, the abundance of lactobacillus can be significantly increased, and there can be a synergistic effect between the two.
In particular, with respect to the components used and the amounts used, group 10 corresponds to the combination of groups 3 and 5, and groups 10, 3, 5 increase the abundance of lactobacillus by 4030.99, 1311.3, 1211.7, respectively, relative to the blank, with the increase in abundance of lactobacillus in group 10 (4030.99) being greater than the sum of the respective increases of groups 3 and 5 (2523).
3.6.2 Effect on Bifidobacterium on mouse faeces
FIG. 4 bifidobacterium standard curve and sample amplification curve, wherein A is the standard curve; b is a plasmid amplification curve; c is the sample amplification curve. Drawing a standard curve by taking a baby tree as an abscissa and a CT value as an ordinate, wherein the bifidobacterium linear standard curve is y= -3.5246546logx+39.383186, R 2 ＝0.9981。
Tables 3-8 changes in the Bifidobacterium content of the mouse feces of each group
Figure BDA0003970836120000292
Figure BDA0003970836120000301
Note that: p <0.05, < P <0.01, < P <0.001 compared to the placebo group; t-test is adopted between groups
Under the intervention of different doses of lactoferrin, ICR pregnant mice can increase the abundance of bifidobacteria at medium and high doses compared with a blank control group, which shows that the ICR pregnant mice have a certain improvement effect on intestinal flora of mice through the intervention of lactoferrin in a parent body.
Under the intervention of Bb12 in different doses, ICR pregnant mice can increase the abundance of bifidobacteria in each dose compared with a blank control group, which shows that the ICR pregnant mice have certain improvement effect on intestinal flora of mice through the intervention of Bb12 in the parent body
The invention has the beneficial effects that: the invention provides researches on the transfer function of lactoferrin, probiotics (one or more selected from bifidobacterium infantis, bifidobacterium longum, bifidobacterium bifidum, bifidobacterium adolescentis, bifidobacterium lactis, bifidobacterium breve, lactobacillus acidophilus and lactobacillus rhamnosus, preferably bifidobacterium lactis, more preferably bifidobacterium lactis Bb 12) and the combination of the lactoferrin and the probiotics (such as bifidobacterium lactis Bb 12) in the mother and infant, and provides a new idea for the development of future functional foods. Lactoferrin and probiotics (such as bifidobacterium lactis Bb 12) have wide prospects in improving immune development of organisms.
What has been described above is merely an exemplary embodiment of the present invention. It should be noted herein that modifications to the invention can be made by those skilled in the art without departing from the inventive concept, and are intended to be within the scope of the invention.

Claims (10)

1. A nutritional composition comprising or consisting of:
-lactoferrin; and
-probiotics.
2. The nutritional composition of claim 1, wherein the probiotic is one or more selected from the group consisting of bifidobacterium infantis, bifidobacterium longum, bifidobacterium bifidum, bifidobacterium adolescentis, bifidobacterium lactis, bifidobacterium breve, lactobacillus acidophilus, and lactobacillus rhamnosus; preferably bifidobacterium lactis, more preferably bifidobacterium lactis Bb12.
3. The nutritional composition of claim 1, wherein lactoferrin is provided in the form: lactoferrin (LF) enriched nutritional ingredients such as lactoferrin enriched whey protein powder and/or lactoferrin powder and the like, preferably lactoferrin powder.
4. A nutritional composition according to any one of claims 1-3, wherein:
the amount of probiotics was 5×10 relative to 1mg lactoferrin 2 ～5×10 10 cfu, preferably 5X 10 3 ～10 10 cfu, preferably 5X 10 4 ～5×10 9 cfu, preferably 5X 10 5 ～2×10 9 cfu, preferably 10 6 ～1.5×10 9 cfu。
5. A food product comprising the nutritional composition of any one of claims 1-4.
6. The food product of claim 5, wherein the food product is in powder or liquid form.
7. The food according to claim 5 or 6, which is an infant food, a child food, a teenager food, or an adult food;
preferably the food is infant formula, infant complementary food, child formula, child snack, parturient formula, maternal formula, middle aged and elderly milk, or a nutritional or dietary supplement;
preferably the food is maternal formula or maternal formula.
8. The food product of any one of claims 5-7, wherein the nutritional composition is added in an amount such that, relative to the total mass of the food product:
the lactoferrin content is at least 0.02mg/g, preferably at least 0.1mg/g, preferably at least 0.2mg/g, and preferably at most 200mg/g, preferably at most 100mg/g, preferably at most 20mg/g; and
the content of probiotics is at least 10 5 cfu/g, preferably at least 5X 10 5 cfu/g, preferably at least 10 6 cfu/g, and preferably at most 10 9 cfu/g, preferably at most 5X 10 8 cfu/g, preferably at most 10 8 cfu/g。
9. The food product of any one of claims 5-8, wherein the nutritional composition is added in an amount such that, relative to the total mass of the food product:
The mass content of lactoferrin is at least 0.2mg/g and at most 20mg/g,
the amount of probiotics is at least 10 6 cfu/g and at most 10 8 cfu/g, and
the amount of probiotics was 5×10 relative to 1mg lactoferrin 4 ～5×10 9 cfu, preferably 5X 10 5 ～2×10 9 cfu, preferably 10 6 ～1.5×10 9 cfu。
10. Use of the nutritional composition according to any one of claims 1-4 or the food according to any one of claims 5-9 for non-therapeutic purposes in promoting development of the immune system and improving the intestinal flora of offspring via ingestion by humans or animal precursors.
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CN117018169A (en) * 2023-10-07 2023-11-10 广州菲勒生物科技有限公司 Nutritional composition preparation for preventing respiratory tract virus infection

Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102090558A (en) * 2010-12-09 2011-06-15 上海交大昂立股份有限公司 Composition containing probiotics, DHA (docosahexaenoic acid) and lactoferrin and applications thereof
CN102665428A (en) * 2009-12-08 2012-09-12 雀巢产品技术援助有限公司 Infant formula with probiotics and milk fat globule membrane components
CN102724886A (en) * 2009-10-29 2012-10-10 雀巢产品技术援助有限公司 Nutritional compositions comprising lactoferrin and probiotics and kits of parts thereof
CN106820156A (en) * 2017-02-17 2017-06-13 福建康是美生物科技有限公司 A kind of probiotic composition of regulating intestinal canal flora health
CN108029768A (en) * 2017-11-29 2018-05-15 上海晨冠乳业有限公司 A kind of baby formula milk powder containing lactoferrin and probiotics and preparation method thereof
CN110122589A (en) * 2019-05-29 2019-08-16 山东翌丰经贸有限公司 A kind of modulation milk powder and preparation method thereof containing lactoferrin and probiotics
CN111528486A (en) * 2020-05-19 2020-08-14 仙乐健康科技股份有限公司 Composition containing lactoferrin
CN112375727A (en) * 2020-09-17 2021-02-19 合生元（广州）健康产品有限公司 Application of lactoferrin in promoting proliferation of bifidobacteria and lactobacilli
CN113491337A (en) * 2021-06-15 2021-10-12 合生元（广州）健康产品有限公司 Composition for regulating infant intestinal tract short-chain fatty acid metabolism and application thereof
CN114532406A (en) * 2020-11-27 2022-05-27 内蒙古伊利实业集团股份有限公司 Formula milk powder for improving human immunity and preparation method and application thereof

Patent Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102724886A (en) * 2009-10-29 2012-10-10 雀巢产品技术援助有限公司 Nutritional compositions comprising lactoferrin and probiotics and kits of parts thereof
CN102665428A (en) * 2009-12-08 2012-09-12 雀巢产品技术援助有限公司 Infant formula with probiotics and milk fat globule membrane components
CN102090558A (en) * 2010-12-09 2011-06-15 上海交大昂立股份有限公司 Composition containing probiotics, DHA (docosahexaenoic acid) and lactoferrin and applications thereof
CN106820156A (en) * 2017-02-17 2017-06-13 福建康是美生物科技有限公司 A kind of probiotic composition of regulating intestinal canal flora health
CN108029768A (en) * 2017-11-29 2018-05-15 上海晨冠乳业有限公司 A kind of baby formula milk powder containing lactoferrin and probiotics and preparation method thereof
CN110122589A (en) * 2019-05-29 2019-08-16 山东翌丰经贸有限公司 A kind of modulation milk powder and preparation method thereof containing lactoferrin and probiotics
CN111528486A (en) * 2020-05-19 2020-08-14 仙乐健康科技股份有限公司 Composition containing lactoferrin
CN112375727A (en) * 2020-09-17 2021-02-19 合生元（广州）健康产品有限公司 Application of lactoferrin in promoting proliferation of bifidobacteria and lactobacilli
CN114532406A (en) * 2020-11-27 2022-05-27 内蒙古伊利实业集团股份有限公司 Formula milk powder for improving human immunity and preparation method and application thereof
CN113491337A (en) * 2021-06-15 2021-10-12 合生元（广州）健康产品有限公司 Composition for regulating infant intestinal tract short-chain fatty acid metabolism and application thereof

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117018169A (en) * 2023-10-07 2023-11-10 广州菲勒生物科技有限公司 Nutritional composition preparation for preventing respiratory tract virus infection

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