Isolated Lactobacillus strains, Compositions Comprising The Same, And Uses

Abstract

The present disclosure provides isolated Lactobacillus strains, compositions comprising the same, and uses thereof, which can be used for promoting growth and development, enhancing immunity, or promoting intestinal health, etc.

Description

Technical Field

[0001] The present disclosure relates to the field of microorganisms, and more particularly to isolated Lactobacillusaceae strains, compositions comprising the same, and uses thereof. Background Art

[0002] Human growth and development refers to the maturation process from fertilized egg to adult. Growth refers to the growth and morphological changes of various organs and systems in the body, which is a quantitative change; development refers to the differentiation and perfection of cells, tissues and organs and the functional maturation, and the change of production quality. The two are closely related, and growth is the material basis of development. The state of development and maturity is reflected in the change of growth quantity. Studies have found that intestinal bacteria can affect the growth and development of the host, and some flora can mediate the growth and development process of the host by affecting growth hormone (GH) / insulin-like growth factor-1 (IGF-1).

[0003] Immunity refers to the body's ability to resist foreign invasion and maintain the stability of the internal environment. For children, the development and protection of the immune system is particularly important because it is directly related to the healthy growth of children. Infants or children with developmental delay have insufficient immunity and low resistance due to slow development of immune organs and gastrointestinal tract, and are prone to illness. Air, food, water and other environments are full of various microorganisms: bacteria, viruses, mycoplasma, chlamydia, fungi, etc. When children have insufficient immunity, they are easily infected by harmful bacteria, and then spread to guardians and classmates, and even cause serious influenza or diseases. Although the human body will produce corresponding antibodies to different pathogens to resist reinfection, antibodies are specific and time-limited. For example, streptococcal antibodies can only protect the body from reinvasion by streptococci in a short period of time, and cannot resist infection from other viruses. The bacteria or viruses that induce influenza mutate very quickly, and infants or children with low immunity find it difficult to resist the invasion of cold viruses. This is the real reason why he often catches colds.

[0004] Probiotics are living microorganisms that, when given in adequate amounts, can bring health benefits to the host. Studies have shown that probiotics can maintain the balance of intestinal flora through their own metabolism and inhibit the growth of harmful bacteria. Therefore, intestinal probiotics are closely related to human health. Probiotic strains have the effects of regulating intestinal health and enhancing immunity.

[0005] Lactobacillus crispatus and Lactobacillus paracasei are common probiotics of the genus Lactobacillus, and their safety has been widely verified. In terms of promoting animal growth and development, the main research of Lactobacillus crispatus focuses on promoting the production performance of poultry and pigs, including increasing the average daily weight gain, the average daily feed intake and the feed utilization rate, and improving the absorption and utilization rate of calcium and phosphorus in the feed. For example, CN114891657B discloses that Lactobacillus crispatus DC529 significantly improves the average daily weight gain and the average daily feed intake of broilers. CN108546663B discloses that Lactobacillus crispatus ZLC020 can increase the final weight of pigs by 5.52% (P<0.05), the average daily weight gain by 7.32%, and the feed-to-weight ratio by 5.68 (P<0.05) compared with the control group, indicating that the pig-derived Lactobacillus crispatus has a positive effect on improving the production performance of growing pigs. There are few reports that Lactobacillus crispatus can promote the growth and development of mice or infants.

[0006] For Lactobacillus paracasei, CN116875515A discloses that Lactobacillus paracasei GF027 promotes the growth of rat height, bone weight and bone density, and increases the insulin-like growth factor 1 in serum. The prior art mainly focuses on promoting the growth of bones and height for Lactobacillus paracasei to promote growth and development. However, children with developmental delay have low resistance and are prone to illness, which is related to the slow development of immune organs and gastrointestinal tract. There are few studies on the probiotics for promoting the growth and development of internal organs such as immune organs and digestive organs.

[0007] The number of microbial resources is extremely large, and screening out new species or strains that can promote the growth and development of internal organs such as immune organs and digestive organs, and further promote growth and development, enhance immunity, and / or improve intestinal health is a huge challenge, but it also represents a huge unmet need. Summary of the invention

[0008] The present disclosure has isolated a new strain of the family Lactobacillaceae. The new strain disclosed in the present disclosure can 1. promote the development of major immune organs such as the thymus and spleen of developmentally retarded subjects (including increasing the weight of immune organs such as the thymus and spleen, immune organ development index (immune organ weight / body weight), etc.); 2. promote the development and proliferation of immune cells such as T cells, and promote the immunity of developmentally retarded subjects; 3. increase the content of serum insulin-like growth factor 1 (IGF-1) in developmentally retarded subjects, promote weight and body length growth, promote bone and muscle growth, such as promoting the growth of tibia length, femur length, soleus and gastrocnemius weight; 4. increase the weight of the thymus, spleen, kidney, and intestinal length (large intestine length and / or small intestine length) of developmentally retarded subjects; increase the content of short-chain fatty acids in the intestine; improve intestinal flora; and promote intestinal development.

[0009] Therefore, the new strain disclosed in the present invention can promote the development of organs while promoting height and weight, especially promoting the development of immune organs, enhancing immunity, promoting intestinal development, improving intestinal health, etc. It can be used to improve malnutrition or developmental delay caused by poor absorption and digestion ability of children, and achieve the function of improving / treating / reversing developmental delay; achieving the purpose of promoting growth and development, enhancing immunity, or promoting intestinal health.

[0010] In a first aspect, the present disclosure provides an isolated Lactobacillusaceae strain having an average nucleotide identity (ANI) value of at least 95%, at least 95.5%, at least 96%, at least 96.5%, at least 97%, at least 97.1%, at least 97.2%, at least 97.3%, at least 97.4% or at least 97.5% with MNH06277 deposited with GDMCC No: 65552; ​​and / or having a 16S rRNA sequence that is at least 98.65% identical to the sequence shown in SEQ ID NO: 3; or The average nucleotide identity (ANI) value with MNH45330 with a deposit number of GDMCC No: 65556 is at least 95%, at least 95.5%, at least 96%, at least 96.5%, at least 97%, at least 97.1%, at least 97.2%, at least 97.3%, at least 97.4% or at least 97.5%; and / or, it has a 16S rRNA sequence that is at least 98.65% identical to the sequence shown in SEQ ID NO: 4.

[0011] In some embodiments, the Lactobacillusaceae strain has a 16S rRNA sequence that is at least 98.65%, at least 98.7%, at least 98.8%, at least 98.9%, at least 99.0%, at least 99.1%, at least 99.2%, at least 99.3%, at least 99.4%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% identical to the sequence shown in SEQ ID NO:3, or SEQ ID NO:4.

[0012] In some embodiments, the Lactobacillaceae strains are two new strains of Lactobacillaceae.

[0013] In some embodiments, the name of the Lactobacillaceae strain is: Lactobacillus paracasei ( Lacticaseibacillus paracasei )MNH06277, deposited in Guangdong Microbiological Culture Collection Center (GDMCC), with the deposit number GDMCC No: 65552, the deposit date is November 28, 2024, the address is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Institute of Microbiology, Guangdong Academy of Sciences, the deposit name is Lacticaseibacillus paracaseiMNH06277.

[0014] In some embodiments, the name of the Lactobacillaceae strain is: Lactobacillus crispatus ( Lactobacillus crispatus )MNH45330, deposited in Guangdong Microbiological Culture Collection Center (GDMCC), with the deposit number GDMCC No: 65556, the deposit date is November 28, 2024, the address is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Institute of Microbiology, Guangdong Academy of Sciences, the deposit name is Lactobacillus crispatus MNH45330.

[0015] In a second aspect, the present disclosure provides a composition comprising the Lactobacillus strain of the first aspect, a culture thereof, and / or a metabolite thereof.

[0016] The present disclosure provides a composition for promoting growth and development, which comprises Lactobacillus paracasei ( Lacticaseibacillus paracasei ) MNH06277 and / or Lactobacillus crispatus ( Lactobacillus crispatus ) culture, live bacteria, freeze-dried bacteria or inactivated bacteria of MNH45330, wherein the Lactobacillus paracasei ( Lacticaseibacillus paracasei ) The deposit number of MNH06277 is GDMCC No: 65552; ​​the Lactobacillus crispatus ( Lactobacillus crispatus ) The deposit number of MNH45330 is GDMCC No: 65556.

[0017] In some embodiments, the Lactobacillus paracasei ( Lacticaseibacillus paracasei ) MNH06277 and / or Lactobacillus crispatus ( Lactobacillus crispatus ) The culture of MNH45330 includes any one of the following A) to D): A) the Lactobacillus paracasei ( Lacticaseibacillus paracasei ) MNH06277 and / or Lactobacillus crispatus ( Lactobacillus crispatus ) fermentation broth of MNH45330; B) Lactobacillus paracasei ( Lacticaseibacillus paracasei ) MNH06277 and / or Lactobacillus crispatus ( Lactobacillus crispatus ) fermentation supernatant of MNH45330; C) the Lactobacillus paracasei ( Lacticaseibacillus paracasei ) MNH06277 and / or Lactobacillus crispatus ( Lactobacillus crispatus ) Fermentation broth of MNH45330 inactivated; D) The concentrated or dried product of any one of A) to C) above.

[0018] In some embodiments, the culture of the Lactobacillusaceae strain includes a solid culture, a fermentation culture, a supernatant of a fermentation culture, or a dried product thereof.

[0019] In some embodiments, the fermentation culture or fermentation culture supernatant is a fermentation culture or fermentation culture supernatant obtained under anaerobic culture conditions using a liquid culture medium.

[0020] In some embodiments, the composition is provided in liquid form or in solid form.

[0021] In some embodiments, the composition comprises 1×10 4 Up to 1×10 12 cfu / mL or 1×10 4 Up to 1×10 12 cfu / mg of live bacteria of the Lactobacillusaceae strain.

[0022] In some embodiments, the composition comprises 1×10 5 Up to 1×10 11 cfu / mL or 1×10 5 Up to 1×10 11 cfu / mg of live bacteria of the Lactobacillusaceae strain.

[0023] In some embodiments, the composition comprises 1×10 6 Up to 1×10 10 cfu / mL or 1×10 6 Up to 1×10 10 cfu / mg of live bacteria of the Lactobacillusaceae strain.

[0024] In some embodiments, the composition comprises 1×10 7 Up to 1×10 9 cfu / mL or 1×10 7 Up to 1×10 9 cfu / mg of live bacteria of the Lactobacillusaceae strain.

[0025] In some embodiments, each gram of the composition comprises 1×10 3 Up to 1×10 17 colony forming units (CFU); for example, 1×10 4 Up to 1×10 12 pcs, 1×10 5 Up to 1×10 11 or 1×10 6 Up to 1×10 10 Colony forming units (CFU), for example, 1×10 3 , 2×103 、3×10 3 、4×10 3 、5×10 3 、6×10 3 、7×10 3 、8×10 3 、9×10 3 、1×10 4 、2×10 4 、3×10 4 、4×10 4 、5×10 4 、6×10 4 、7×10 4 、8×10 4 、9×10 4 、1×10 5 、2×10 5 、3×10 5 、4×10 5 、5×10 5 、6×10 5 、7×10 5 、8×10 5 、9×10 5 、1×10 6 、2×10 6 、3×10 6 、4×10 6 、5×10 6 、6×10 6 、7×10 6 、8×10 6 、9×10 6 、1×10 7 、2×10 7 、3×10 7 、4×10 7 、5×10 7 、6×10 7 、7×10 7 、8×10 7 、9×10 7 、1×10 8 、2×10 8 、3×10 8 、4×10 8 、5×10 8 、6×10 8 、7×10 8 、8×10 8 、9×10 8 、1×10 9 、2×10 9 、3×10 9, 4×10 9 , 5×10 9 , 6×10 9 ,7×10 9 , 8×10 9 ,9×10 9 , 1×10 10 , 2×10 10 , 3×10 10 , 4×10 10 , 5×10 10 , 6×10 10 ,7×10 10 , 8×10 10 ,9×10 10 , 1×10 11 , 2×10 11 , 3×10 11 , 4×10 11 , 5×10 11 , 6×10 11 ,7×10 11 , 8×10 11 ,9×10 11 , 1×10 12 , 2×10 12 , 3×10 12 , 4×10 12 , 5×10 12 , 6×10 12 ,7×10 12 , 8×10 12 ,9×10 12 , 1×10 13 , 2×10 13 , 3×10 13 , 4×10 13 , 5×10 13 , 6×10 13 ,7×10 13 , 8×10 13 ,9×10 13 or any value in between of colony forming units (CFU).

[0026] In some embodiments, the concentration of the Lactobacillusaceae strain as an active ingredient in the composition is 10 7 Up to 10 12 CFU / g.

[0027] In some embodiments, the Lactobacillus strain in the composition is a live bacterium, an attenuated bacterium, a freeze-dried bacterium, or an inactivated bacterium, for example, a heat-inactivated bacterium, preferably a pasteurized bacterium.

[0028] In some embodiments, the composition is in the form of a liquid, foam, cream, spray, powder (eg, lyophilized powder), or gel.

[0029] In some embodiments, the composition is in the form of a powder, microencapsulated powder, capsule, tablet, lozenge, granule, oral solution, suspension, emulsion, liquid preparation, sustained release preparation, nanoformulation, or microencapsulated capsule.

[0030] In some embodiments, the composition is in the form of an oral dosage or an injection.

[0031] In some embodiments, the composition further comprises one or more pharmaceutically and / or food-acceptable excipients.

[0032] The pharmaceutically and / or food acceptable excipients are well known to those skilled in the art.

[0033] In some embodiments, the auxiliary material may be at least one selected from a carrier, an excipient, a diluent, a lubricant, a wetting agent, an emulsifier, a suspension stabilizer, a preservative, a sweetener, and a flavor.

[0034] In some embodiments, the composition comprises one or more of a buffer (e.g., sodium bicarbonate, infant formula or sterilized human milk or other agents that allow bacteria to survive and grow (e.g., survive in the acidic environment of the stomach and grow in the intestinal environment)), a lyoprotectant, a preservative, a stabilizer, a binder, a compacting agent, a lubricant, a dispersion enhancer, a disintegrant, an antioxidant, a flavoring agent, a sweetener, and a coloring agent.

[0035] In some embodiments, the composition further comprises one or more other active agents for promoting growth and development, enhancing immunity, or promoting intestinal health.

[0036] In some embodiments, the growth promotion includes: promoting at least one of the development of organs, height / body length, weight, bones, and muscles.

[0037] In some embodiments, the organ comprises at least one of heart, liver, spleen, kidney, and intestine.

[0038] In some embodiments, the bone comprises at least one of a femur and a tibia.

[0039] In some embodiments, the muscle comprises at least one of the soleus muscle and the gastrocnemius muscle.

[0040] In some embodiments, the other active agent may be one or more of a probiotic, a prebiotic, or a combination thereof; In some embodiments, the probiotic is selected from at least one of Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium animalis, Bifidobacterium breve, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus delbrueckii, Lactobacillus gasseri, Lactobacillus helveticus, Lactobacillus johnsonii, Lactobacillus krum, Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus fermentum, Lactobacillus reuteri, Lactobacillus plantarum, Lactobacillus salivarius, Lactobacillus curvatus, Lactobacillus sakei, Lactococcus lactis, and Lactococcus creamer.

[0041] In some embodiments, the prebiotic is selected from inulin, mulberry leaf extract, berberine, ganoderma lucidum, green coffee bean extract, oats, pectin, potato or its extract, citrus polyphenols, Ceylon cinnamon, chromium, ergothioneine, astaxanthin, quercetin, curcumin, proanthocyanidins, resistant dextrin, yeast β-glucan, ginseng or its extract, nutritional compounds, biotin, polydextrose, fructooligosaccharide (FOS), galacto-oligosaccharide (GOS), starch, cellulose, b-glucan, hemicellulose, lactulose, manno-oligosaccharide, manno-oligosaccharide (MOS), inulin rich in fructooligosaccharide, oligo-glucose, tagatose, trans-galacto-oligosaccharide, pectin, resistant starch, xylo-oligosaccharide (XOS) and any combination thereof.

[0042] In some embodiments, the composition can be formulated as a frozen composition, such as a frozen composition prepared by quick freezing and drying, or lyophilization, for storage and / or transportation.

[0043] In some embodiments, the composition is obtained by spray drying. In some embodiments, the composition is obtained by electrostatic spray drying.

[0044] In some embodiments, the strain in the composition is freeze-dried or spray-dried. In some embodiments, the strain in the composition is electrostatically spray-dried. In some embodiments, the strain in the composition is freeze-dried or spray-dried and is alive. In some embodiments, the strain in the composition is freeze-dried or spray-dried and can partially or completely colonize the intestine. In some embodiments, the strain is reconstituted before administration. In some cases, the reconstitution is carried out by using a diluent as described herein.

[0045] In some embodiments, the composition can be administered alone or in combination with a carrier such as a pharmaceutically acceptable carrier or a biocompatible scaffold.

[0046] In some embodiments, the composition is formulated for oral administration. In some embodiments, the composition is an enteric preparation. In some embodiments, the enteric preparation is a dosage form with an enteric coating. For example, the enteric preparation can be an enteric granule, an enteric tablet, or an enteric capsule. In some embodiments, the composition is a capsule. In some embodiments, the capsule is a hard capsule or a soft capsule; or the capsule is a sustained release capsule, a controlled release capsule, or an enteric capsule, or the capsule can be a microencapsulated capsule, or a microcapsule.

[0047] In some embodiments, the composition is a medicine, or a health product or a food.

[0048] In some embodiments, the composition is in an infant-suitable dosage form, a child-suitable dosage form, or an adult-suitable dosage form.

[0049] In some embodiments, the composition is in a parenteral dosage form.

[0050] In a third aspect, the present disclosure provides the use of the Lactobacillus strain described in the first aspect or the composition described in the second aspect in the preparation of medicines, health products or foods for promoting growth and development, enhancing immunity, or promoting intestinal health.

[0051] The present disclosure provides the use of the composition described in the second aspect in medicines, health products or foods for promoting growth and development, enhancing immunity, or promoting intestinal health.

[0052] The present disclosure provides the use of the composition described in the second aspect in the preparation of medicines, health products or foods for promoting growth and development, enhancing immunity, or promoting intestinal health.

[0053] In some embodiments, the growth promotion includes: promoting at least one of the development of organs, height / body length, weight, bones, and muscles.

[0054] In some embodiments, the organ comprises at least one of heart, liver, spleen, kidney, and intestine.

[0055] In some embodiments, the bone comprises at least one of a femur and a tibia.

[0056] In some embodiments, the muscle comprises at least one of the soleus muscle and the gastrocnemius muscle.

[0057] In some embodiments, the medicine, health product or food promotes growth and development by increasing hormone content and / or promoting digestion and absorption.

[0058] In some embodiments, the hormone comprises insulin-like growth factor 1 (IGF-1) and / or growth hormone (GH).

[0059] In some embodiments, the hormone is from a body fluid; further from blood; further from serum.

[0060] In some embodiments, the medicine, health product or food enhances immunity by promoting the development of the immune system.

[0061] In some embodiments, the promoting the development of the immune system comprises at least one of promoting the development of immune organs and promoting the generation of immune cells.

[0062] In some embodiments, the promoting the development of immune organs comprises promoting the development of the thymus, spleen, bone marrow, and / or lymph nodes; and further comprises promoting the development of the thymus and / or spleen.

[0063] In some embodiments, the promoting the generation of immune cells comprises promoting the generation of T cells, B cells, and / or NK cells (i.e., increasing the ratio of T cells, B cells, and / or NK cells; further comprising promoting the generation of T cells.

[0064] In some embodiments, the medicine, health product or food promotes intestinal health by increasing short-chain fatty acids and / or improving intestinal flora.

[0065] In some embodiments, the short-chain fatty acid comprises at least one of propionic acid, butyric acid, isovaleric acid, isobutyric acid, and 2-methylbutyric acid.

[0066] The composition comprises an isolated and purified consortium of viable microbial populations to increase body weight by at least 2%, 3%, 4%, 5%, 6%, or 7% in a subject compared to the subject's body weight prior to administration of the isolated and purified consortium of viable microbial species.

[0067] As used herein, a microbial population generally refers to a microbial population consisting essentially of a single strain, species or genus, which may be the case when a population is cultured from a subpopulation of such strain, species or genus that is separated and purified. Thus, for a given microbial population, if cultured from an isolated microbial species or strain, such a population will be referred to herein as purified or substantially pure. The resulting population may typically be at least 80% pure for the microbial species or strain, at least 90% pure, at least 95% pure, at least 98% pure, at least 99% pure, at least 99.5% pure, or at least 99.9% pure relative to other microbial species or strains within the particular population. In contrast, the level of non-desirable strains in any particular desired microbial population will be less than 20%, less than 10%, less than 5%, less than 2%, less than 1%, less than 0.5% or less than 0.1%. For example, the level of impurities in a purified population population, such as other undesirable microbial strains or species, may be proportional to or less than the above levels for each desired population. Less than 2%, less than 1%, less than 0.5% or less than 0.1%. In the case where the composition comprises a consortium of multiple microbial populations, each population can have the above-mentioned purities, either before it is incorporated into the composition, or when the consortium is measured in an aggregate. For example, the level of impurities in the purified population consortium, such as other undesirable microbial strains or species, can be proportional to the above-mentioned levels for each desired population or below the above-mentioned levels. Less than 2%, less than 1%, less than 0.5% or less than 0.1%. In the case where the composition comprises a consortium of multiple microbial populations, each population can have the above-mentioned purities, either before it is incorporated into the composition, or when the consortium is measured in an aggregate. For example, the level of impurities in the purified population consortium, such as other undesirable microbial strains or species, can be proportional to the above-mentioned levels for each desired population or below the above-mentioned levels.

[0068] The compositions of the present disclosure may also include cellular components, metabolites, secreted molecules and compounds metabolized by the Lactobacillus strains, and the like. Cellular components or cell fractions, metabolites or secreted compounds may be extracted from the culture of the Lactobacillus strains, for example, by recovering the supernatant of the culture of the Lactobacillus strains or by extracting the cell components or cell fractions, metabolites or secreted compounds from the culture of the Lactobacillus strains; may correspond to components in isolated form from the Lactobacillus strains, or any mixture of one or more components from the Lactobacillus strains.

[0069] In some embodiments, the Lactobacillus family strains or compositions described in the present disclosure can be used to promote growth and development, enhance immunity, or promote intestinal health.

[0070] In some embodiments, the growth promotion includes: promoting at least one of the development of organs, height / body length, weight, bones, and muscles.

[0071] In some embodiments, the organ comprises at least one of a thymus, a heart, a liver, a spleen, a kidney, and an intestinal tract.

[0072] In some embodiments, the bone comprises at least one of a femur and a tibia.

[0073] In some embodiments, the muscle comprises at least one of the soleus muscle and the gastrocnemius muscle.

[0074] In some embodiments, the Lactobacillusaceae strain or composition promotes growth and development by increasing hormone content and / or promoting digestion and absorption.

[0075] In some embodiments, the hormone comprises insulin-like growth factor 1 (IGF-1) and / or growth hormone (GH).

[0076] In some embodiments, the hormone is from a body fluid; further from blood; further from serum.

[0077] In some embodiments, the Lactobacillusaceae strain or composition enhances immunity by promoting immune system development.

[0078] In some embodiments, the promoting the development of the immune system comprises at least one of promoting the development of immune organs and promoting the generation of immune cells.

[0079] In some embodiments, the promoting the development of immune organs comprises promoting the development of the thymus, spleen, bone marrow, and / or lymph nodes; and further comprises promoting the development of the thymus and / or spleen.

[0080] In some embodiments, the promoting the generation of immune cells comprises promoting the generation of T cells, B cells, and / or NK cells (i.e., increasing the proportion of T cells, B cells, and / or NK cells); and further comprises promoting the generation of T cells.

[0081] In some embodiments, the Lactobacillus family strain or composition promotes intestinal health by increasing short-chain fatty acids and / or improving intestinal flora.

[0082] In some embodiments, the short-chain fatty acid comprises at least one of propionic acid, butyric acid, isovaleric acid, isobutyric acid, and 2-methylbutyric acid.

[0083] In some embodiments, the Lactobacillus family strain or composition of the present disclosure can increase heart, liver, spleen, and / or kidney weight, and / or intestinal length, thereby promoting organ development.

[0084] In some embodiments, the Lactobacillus strain or composition of the present disclosure can increase body weight, body length, tibia length, femur length, soleus muscle and / or gastrocnemius muscle weight, thereby promoting height (body length), weight, bone, and muscle development.

[0085] In some embodiments, the Lactobacillus strain or composition of the present disclosure can increase the content of serum insulin-like growth factor 1 (IGF-1) and / or growth hormone (GH), thereby promoting growth and development.

[0086] In some embodiments, the Lactobacillus family strains of the present disclosure or compositions comprising the same can promote digestion and absorption, thereby promoting growth and development without increasing cumulative food intake.

[0087] In some embodiments, the Lactobacillus strain or composition disclosed herein can promote the development of major immune organs such as the thymus and spleen, and / or increase the proportion of T cells, thereby promoting the development of the immune system and enhancing immunity.

[0088] In some embodiments, the Lactobacillus strain or composition of the present disclosure can increase the content of short-chain fatty acids and / or improve intestinal flora, thereby promoting intestinal health.

[0089] In some embodiments, the drug or health product or food has at least one effect selected from the following: increase the weight of the heart, liver, spleen, and / or kidney, and / or intestinal length (large intestine length and / or small intestine length); do not increase food intake; increase body weight, body length, tibia length, femur length, soleus muscle and / or gastrocnemius muscle weight; promote weight growth, body length growth, bone and muscle growth (wherein, promoting bone growth includes growth in bone length and density, and promoting muscle growth includes promoting specific muscle development and maintaining healthy and benign growth characteristics, such as: low body fat rate (calf muscle / body weight) and / or high muscle weight (calf muscle / calf weight)); Increase the content of serum insulin-like growth factor 1 (IGF-1) and / or growth hormone (GH); promote the development of major immune organs such as the thymus and spleen (including increasing the weight of immune organs such as the thymus and spleen, the immune organ development index (immune organ weight / body weight), etc.), increase the proportion of T cells; increase the content of short-chain fatty acids; improve intestinal flora; promote intestinal development; improve lesions or inflammation caused by developmental delay to achieve the function of improving / treating / reversing developmental delay; promote organ development; promote digestion and absorption; promote height (body length), weight, bone, and muscle development; promote the development of the immune system and enhance immunity; promote intestinal health; can be used to promote growth and development, enhance immunity, or promote intestinal health.

[0090] A method for promoting growth and development, enhancing immunity, or promoting intestinal health, comprising administering an effective amount of the Lactobacillus strain described in the first aspect of the present disclosure or the composition described in the second aspect to a subject in need.

[0091] In some embodiments, the growth promotion includes: promoting at least one of the development of organs, height / body length, weight, bones, and muscles.

[0092] In some embodiments, the organ comprises at least one of a thymus, a heart, a liver, a spleen, a kidney, and an intestinal tract.

[0093] In some embodiments, the bone comprises at least one of a femur and a tibia.

[0094] In some embodiments, the muscle comprises at least one of the soleus muscle and the gastrocnemius muscle.

[0095] In some embodiments, the Lactobacillusaceae strain or composition promotes growth and development by increasing hormone content and / or promoting digestion and absorption.

[0096] In some embodiments, the hormone comprises insulin-like growth factor 1 (IGF-1) and / or growth hormone (GH).

[0097] In some embodiments, the hormone is from a body fluid; further from blood; further from serum.

[0098] In some embodiments, the Lactobacillusaceae strain or composition enhances immunity by promoting immune system development.

[0099] In some embodiments, the promoting the development of the immune system comprises at least one of promoting the development of immune organs and promoting the generation of immune cells.

[0100] In some embodiments, the promoting the development of immune organs comprises promoting the development of the thymus, spleen, bone marrow, and / or lymph nodes; and further comprises promoting the development of the thymus and / or spleen.

[0101] In some embodiments, the promoting the generation of immune cells comprises promoting the generation of T cells, B cells, and / or NK cells (i.e., increasing the proportion of T cells, B cells, and / or NK cells); and further comprises promoting the generation of T cells.

[0102] In some embodiments, the Lactobacillus family strain or composition promotes intestinal health by increasing short-chain fatty acids and / or improving intestinal flora.

[0103] In some embodiments, the short-chain fatty acid comprises at least one of propionic acid, butyric acid, isovaleric acid, isobutyric acid, and 2-methylbutyric acid. BRIEF DESCRIPTION OF THE DRAWINGS

[0104] The embodiments will be described below in conjunction with the accompanying drawings so that the above and other aspects and advantages of the present invention will become apparent and easily understood.

[0105] Figure 1 : shows the colony morphology photograph of strain MNH06277.

[0106] Figure 2 : Gram staining photographs of strain MNH06277 are shown.

[0107] Figure 3 : shows the electron micrograph of strain MNH06277.

[0108] Figure 4 : shows the colony morphology photograph of strain MNH45330.

[0109] Figure 5 : Gram staining photographs of strain MNH45330 are shown.

[0110] Figure 6 : shows the electron micrograph of strain MNH45330.

[0111] Figure 7 : Shows the results of tolerance of strain MNH06277 to different pH.

[0112] Figure 8 : Shows the tolerance results of strain MNH06277 to different concentrations of NaCl.

[0113] Fig. 9 : Shows the tolerance results of strain MNH06277 to different concentrations of bile salts.

[0114] Fig.10 : Shows the results of the tolerance of strain MNH45330 to different pH.

[0115] Fig.11 : Shows the tolerance results of strain MNH45330 to different concentrations of NaCl.

[0116] Fig.12 : Shows the tolerance results of strain MNH45330 to different concentrations of bile salts.

[0117] Fig.13 : The phylogenetic tree of strain MNH06277 is shown.

[0118] Fig.14 : The phylogenetic tree of strain MNH45330 is shown.

[0119] Fig.15:Shows that strains MNH06277 and MNH45330 can promote organ development (spleen weight). Data are presented as mean ± standard deviation (Mean ± SD). Statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the LFD-Control group; **, p < 0.01 compared with the LFD-Control group.

[0120] Fig.16 :Shows that strains MNH06277 and MNH45330 can promote organ development (kidney weight). Data are presented as mean ± SD. Statistical analysis was performed using Student's t test; **, p < 0.01 compared with the LFD-Control group; ****, p < 0.0001 compared with the LFD-Control group.

[0121] Fig.17 :Shows that strains MNH06277 and MNH45330 can promote organ development (intestinal length). Data are presented as mean ± standard deviation (Mean ± SD). Statistical analysis was performed using the Student's t test; **, p < 0.01 compared with the LFD-Control group.

[0122] Fig.18 :Shows that strains MNH06277 and MNH45330 promote body weight development (weight gain). Data are shown as mean ± standard deviation (Mean ± SD). Statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the LFD-Control group; ****, p < 0.0001 compared with the LFD-Control group.

[0123] Fig.19 :Shows that strains MNH06277 and MNH45330 promote the development of height (body length) (body length increment). Data are shown as mean ± standard deviation (Mean ± SD). Statistical analysis was performed using the Student's t test; **, p < 0.01 compared with the LFD-Control group; ****, p < 0.0001 compared with the LFD-Control group.

[0124] Fig. 20:Shows that strains MNH06277 and MNH45330 promote bone development (femur length). Data are shown as mean ± standard deviation (Mean ± SD). Statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the LFD-Control group; **, p < 0.01 compared with the LFD-Control group; ***, p < 0.001 compared with the LFD-Control group.

[0125] Fig.21 :Shows that strains MNH06277 and MNH45330 promote bone development (tibia length). Data are shown as mean ± standard deviation (Mean ± SD). Statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the LFD-Control group; **, p < 0.01 compared with the LFD-Control group; ***, p < 0.001 compared with the LFD-Control group.

[0126] Fig. 22 :Shows the muscle development (soleus muscle weight) of strains MNH06277 and MNH45330. Data are presented as mean ± standard deviation (Mean ± SD). Statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the LFD-Control group.

[0127] Fig.23 :Shows that strains MNH06277 and MNH45330 promote muscle development (gastrocnemius muscle weight). Data are shown as mean ± standard deviation (Mean ± SD). Statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the LFD-Control group; **, p < 0.01 compared with the LFD-Control group.

[0128] Fig.24 :Shows that strain MNH06277 increases serum insulin-like growth factor 1 (IGF-1) content. Data are shown as mean ± standard deviation (Mean ± SD). Statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the LFD-Control group; ****, p < 0.0001 compared with the LFD-Control group.

[0129] Fig.25: It is shown that strains MNH06277 and MNH45330 did not increase cumulative food intake (cumulative food intake line graph). Data are presented as mean ± standard deviation (Mean ± SD). Statistical analysis was performed using the Student's ttest analysis method.

[0130] Fig.26 : It is shown that strains MNH06277 and MNH45330 did not increase cumulative food intake (cumulative food intake bar graph). Data are presented as mean ± SD. Statistical analysis was performed using the Student's ttest.

[0131] Fig. 27 :Shows that strains MNH06277 and MNH45330 promote immune system development and enhance immunity (thymus weight). Data are presented as mean ± standard deviation (Mean ± SD). Statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the LFD-Control group; **, p < 0.01 compared with the LFD-Control group.

[0132] Fig.28 :Shows that strains MNH06277 and MNH45330 promote immune system development and enhance immunity (spleen weight). Data are presented as mean ± standard deviation (Mean ± SD). Statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the LFD-Control group; **, p < 0.01 compared with the LFD-Control group.

[0133] Fig.29 :Shows that strain MNH45330 promotes immune system development and enhances immunity (T cell percentage). Data are presented as mean ± standard deviation (Mean ± SD). Statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the LFD-Control group.

[0134] Fig.30 :Shows that strain MNH45330 promotes the production of short-chain fatty acids (propionic acid content) in cecal contents. Data are presented as mean ± standard deviation (Mean ± SD). Statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the LFD-Control group.

[0135] Fig.31:Shows that strain MNH45330 promotes the production of short-chain fatty acids (isobutyric acid content) in cecal contents. Data are shown as mean ± standard deviation (Mean ± SD). Statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the LFD-Control group, **, p < 0.01 compared with the LFD-Control group.

[0136] Fig.32 :Shows that strain MNH45330 promotes the production of short-chain fatty acids (2-methylbutyrate content) in cecal contents. Data are shown as mean ± standard deviation (Mean ± SD). Statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the LFD-Control group, **, p < 0.01 compared with the LFD-Control group.

[0137] Fig.33 :Shows that strain MNH45330 promotes the production of short-chain fatty acids (isovaleric acid content) in cecal contents. Data are shown as mean ± standard deviation (Mean ± SD). Statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the LFD-Control group, **, p < 0.01 compared with the LFD-Control group.

[0138] Fig.34 :Shows that strain MNH06277 promotes the production of short-chain fatty acids (butyrate ratio) in cecal contents. Data are presented as mean ± standard deviation (Mean ± SD). Statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the LFD-Control group.

[0139] Deposit of strains

[0140] The strain Lactobacillus paracasei ( Lacticaseibacillus paracasei )MNH06277, deposited in Guangdong Microbiological Culture Collection Center (GDMCC), with the deposit number GDMCC No: 65552, the deposit date is November 28, 2024, the address is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Institute of Microbiology, Guangdong Academy of Sciences, the deposit name is Lacticaseibacillus paracasei MNH06277, taxonomic name Lacticaseibacillus paracasei .

[0141] Lactobacillus crispatus strain ( Lactobacillus crispatus)MNH45330, deposited in Guangdong Microbiological Culture Collection Center (GDMCC), with the deposit number GDMCC No: 65556, the deposit date is November 28, 2024, the address is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Institute of Microbiology, Guangdong Academy of Sciences, the deposit name is Lactobacillus crispatus MNH45330, taxonomic name Lactobacillus crispatus . DETAILED DESCRIPTION

[0142] The present invention separates two new strains of the Lactobacillaceae family, with the deposit numbers GDMCC No: 65552 and GDMCC No: 65556, respectively, and identifies them using traditional classification methods and molecular biology methods. The identification results show that the strain with the deposit number GDMCC No: 65552 belongs to a new strain of the species Lactobacillus paracasei, and the strain with the deposit number GDMCC No: 65556 belongs to a new strain of the species Lactobacillus crispatus. Further, the present invention studies the biochemical properties and therapeutic uses of the two strains.

[0143] It is known in the art that bacterial species can be classified and identified by traditional classification methods and molecular biological methods. Traditional classification methods include, but are not limited to, cell morphology observation, Gram staining, flagella staining, various metabolic experiments, etc. Molecular biological methods include, but are not limited to, ribosomal RNA sequence determination methods, determination methods based on whole genome sequencing, etc.

[0144] As used herein, the term "prebiotic" may be a general term referring to chemicals and / or ingredients that can affect the growth and / or activity of microorganisms in a host (e.g., can allow for specific changes in the composition and / or activity of a microbiome).

[0145] The terms "subject," "subject," "individual," "host," and "patient" are used interchangeably herein to refer to any animal subject, including: humans, mammals, laboratory animals, livestock, and domestic pets.

[0146] Compositions or preparations disclosed herein can be used as pharmaceutical preparations, therapeutic compositions, dietary supplements, nutritional supplements, medical probiotics or medical foods. In some cases, the composition is applied in the form of a pharmaceutical preparation. In some cases, the composition is applied in the form of a nutritional supplement. In some cases, the composition is applied in the form of a dietary supplement. In some cases, the composition is applied in the form of a medical food. In some cases, the composition is applied in the form of a medical probiotic. In some cases, the composition (e.g., dietary supplements, nutritional supplements, medical probiotics or medical foods) can be orally administered, for example as a capsule, pill or tablet.

[0147] 16S rRNA is a ribosomal RNA of prokaryotes. The 16S rRNA gene consists of a variable region and a conserved region. The conserved region is common to all bacteria, while the variable region has different degrees of difference between different bacteria. By comparing the 16S rRNA gene sequences of bacteria, according to the base number of sequence differences and their evolutionary distance, an evolutionary tree can be drawn. When the identity between the 16S rRNA gene sequences of two strains is less than 98.65%, they can be judged to belong to different species (see Kim, M., Oh, H.-S., Park, S.-C., & Chun, J. (2014). Towards a taxonomic coherence between average nucleotide identity and 16S rRNA gene sequence similarity for species demarcation of prokaryotes. International Journal of Systematic and Evolutionary Microbiology, 64(Pt 2), 346–351, and Liu, C., Du, M.-X., Abuduaini,R., Yu, H.-Y., Li, D.-H., Wang, Y.-J., Liu, S.-J. (2021). Enlightening the taxonomy darkness of human gut microbiomes with a cultured biobank. Microbiome, 9(1), P23).

[0148] The "identity" between two nucleic acid molecule sequences can be determined using known computer algorithms, such as the "FASTA" program, the GCG program package, BLASTN or FASTA. Commercial or publicly available programs may also be, for example, the DNAStar "MegAlign" program.

[0149] The second generation sequencing technology can also be used to identify bacterial species based on whole genome sequencing, making the identification results more accurate. The average nucleotide identity (ANI) of bacterial genomes refers to the similarity of homologous genes between two bacterial genomes. The ANI value can be calculated by methods such as BLAST. In the field of bacterial taxonomy, it is generally believed that the ANI value needs to reach more than 95% to be identified as belonging to the same species (Jain C, Rodriguez-R LM, Phillippy AM, et al. High throughput ANI analysis of 90K prokaryotic genomes reveals clear species boundaries[J]. Nature Communications, 2018, 9 (1): 5114.).

[0150] Various existing mature ANI value calculation tools can be used, such as local calculation software Jspecies ( / jspecies) and Gegenees ( / documentation.html), online calculation tools ANI caculator (http: / enveomics.gatech.edu / ), EzGenome ( / ezgenome / ani) and ANItools.

[0151] Utilize the above method, those skilled in the art can judge whether a strain separation belongs to the Lactobacillus paracasei kind or the Lactobacillus crispatus kind identified by the inventor. For example, when the Lactobacillus paracasei (GDMCC No: 65552) is separated from the present invention, the present invention can be used to separate the Lactobacillus paracasei kind. Lacticaseibacillus paracasei) has an average nucleotide identity ANI value of at least 95%, for example at least 95.1%, 95.2%, 95.3%, 95.4%, 95.5%, 95.6%, 95.7%, 95.8%, 95.9%, 96%, 96.1%, 96.2%, 96.3%, 96.4%, 96.5%, 96.6%, 96.7%, 96.8%, 96.9%, 97%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.9%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 98. 6%, 97.7%, 97.8%, 97.9%, 98%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or 100%, it can be determined that they belong to the same species (i.e., belong to the species Lactobacillus paracasei); or when they are identical to Lactobacillus crispatus with a deposit number of GDMCC No: 65556 ( Lactobacillus crispatus ) has an average nucleotide identity ANI value of at least 95%, for example at least 95.1%, 95.2%, 95.3%, 95.4%, 95.5%, 95.6%, 95.7%, 95.8%, 95.9%, 96%, 96.1%, 96.2%, 96.3%, 96.4%, 96.5%, 96.6%, 96.7%, 96.8%, 96.9%, 97%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98 ... .5%, 97.6%, 97.7%, 97.8%, 97.9%, 98%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or 100%, it can be determined that they belong to the same species (i.e., the Lactobacillus crispatus species).

[0152] For another example, when its 16S rRNA sequence has at least 98.65% identity with the sequence shown in SEQ ID NO: 3, for example, at least 98.7%, 98.8%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or 100%, it can be determined that they belong to the same species (i.e., they belong to the species Lactobacillus paracasei); when its 16S rRNA sequence has at least 98.65% identity with the sequence shown in SEQ ID NO: 4, for example, at least 98.7%, 98.8%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or 100%, it can be determined that they belong to the same species (i.e., they belong to the species Lactobacillus crispatus).

[0153] A "strain" refers to a member of a bacterial species that has a genetic characteristic that allows it to be distinguished from closely related members of the same bacterial species. The genetic characteristic can be the total or partial absence of at least one gene, the total or partial absence of at least one regulatory region (e.g., promoter, terminator, riboswitch, ribosomal binding site), the absence ("cure" of at least one native plasmid), the presence of at least one recombinant gene, the presence of at least one mutated gene, the presence of at least one exogenous gene (a gene from another species), the presence of at least one mutated regulatory region (e.g., promoter, terminator, riboswitch, ribosomal binding site), the presence of at least one non-native plasmid, the presence of at least one antibiotic resistance cassette, or a combination thereof. Genetic characteristics between different strains can be identified by PCR amplification, optionally followed by DNA sequencing of the genomic region of interest or the entire genome. In the case of a strain that gains or loses antibiotic resistance (compared to another strain of the same species) or gains or loses a biosynthetic ability (e.g., an auxotrophic strain), the strains can be distinguished by selection or counter-selection using antibiotics or nutrients / metabolites.

[0154] "Supernatant" or "supernatant" within the meaning herein refers to the culture supernatant of the bacterial strain according to the present disclosure, optionally comprising compounds and / or cell debris of said strain, and / or metabolites and / or molecules secreted by said strain.

[0155] The composition can be prepared using the Lactobacillus strains described herein, for example, by using pharmaceutically acceptable excipients. The pharmaceutical composition comprises a pharmaceutically effective amount of the Lactobacillus strain, for example, Lactobacillus paracasei (GDMCC No: 65552). Lacticaseibacillus paracasei ), or Lactobacillus crispatus with a deposit number of GDMCC No: 65556 ( Lactobacillus crispatus). Similarly, the Lactobacillus paracasei with a deposit number of GDMCC No: 65552 ( Lacticaseibacillus paracasei ), or Lactobacillus crispatus with a deposit number of GDMCC No: 65556 ( Lactobacillus crispatus ) can also be prepared into a pharmaceutical composition, for example, by using a pharmaceutically acceptable excipient, which comprises a pharmaceutically effective amount of the Lactobacillus family strain.

[0156] Suitable pharmaceutically acceptable excipients that may be used include, for example, carriers, excipients, diluents, lubricants, wetting agents, emulsifiers, suspension stabilizers, preservatives, sweeteners and flavors.

[0157] The composition herein can be formulated into any form suitable for enhancing the abundance of the Lactobacillusaceae strain in the subject. The composition can be administered orally (e.g., by oral gavage), intramuscular injection, inhalation, intracranial, intralymphatic, intraocular, intraperitoneal, intrapleural, intrathecal, intratracheal, intrauterine, intravascular, intravenous, intravesical, intranasal, intragastrointestinal, bile infusion, cardiac infusion, anterior anus, rectum, spinal subcutaneous, sublingual, topical, intravaginal, transdermal, ureteral or urethral.

[0158] Examples of dosage forms suitable for the composition herein include, but are not limited to, tablets, aerosols, chewable sticks, capsules, capsules containing coated granules, capsules containing sustained-release granules, capsules containing sustained-release granules, and concentrates.

[0159] In some embodiments, the composition is a sugar-coated tablet, a gel capsule, a gel, an emulsion, a tablet, a sheet capsule, a hydrogel, a nanofiber gel, an electrospun fiber, a food bar, a candy, a fermented milk, a fermented cheese, a chewing gum, a powder or a toothpaste, etc.

[0160] In some embodiments, administration can also be by inclusion in a subject's diet, such as inclusion in a functional food for humans or companion animals.

[0161] The compositions provided herein may include a pharmaceutically acceptable excipient, diluent or carrier. The pharmaceutically acceptable excipient, diluent or carrier is well known in the art.

[0162] In some embodiments, the lactobacillus strain in the composition of the present disclosure is freeze-dried. In some embodiments, the lactobacillus strain in the composition of the present disclosure is spray-dried. In some embodiments, the lactobacillus strain in the composition of the present disclosure is freeze-dried or spray-dried, and is alive. In some embodiments, the lactobacillus strain in the composition of the present disclosure is freeze-dried or spray-dried, and can be partially or completely colonized in the intestine. In some embodiments, the freeze-dried lactobacillus strain is reconstructed before administration. In some embodiments, the reconstruction is carried out using a diluent as described herein.

[0163] In some embodiments, the compositions of the present disclosure are administered orally. Oral administration may involve swallowing, thereby allowing the composition to enter the gastrointestinal tract, and / or administration through the mouth, tongue, or sublingually.

[0164] In some embodiments, the composition is prepared by freeze-drying or spray-drying.

[0165] The composition disclosed herein includes medicines, health products or foods.

[0166] The subject of the present disclosure may be a human or an animal, and the animal includes but is not limited to cattle, sheep, cats, dogs, horses, rabbits, monkeys, mice, rats, alpacas, camels, etc.

[0167] In 2020, ZHENG et al. conducted a study on Lactobacillus spp. Lactobacillus ) were used to study the core genes of 261 species, compare the average nucleotide identity (ANI) between species, analyze the characteristic genes, and analyze the physiological standards and ecology of the species. It was proposed to redivide the genus Lactobacillus into 25 genera, including Lactobacillus and 23 new genera. It was also suggested that the family Leuconostocaceae ( Leuconostocaceae ) is assigned to the Lactobacillaceae family ( Lactobacillaceae ). According to this study, the Lactobacillus casei Lactobacillus casei , Lactobacillus paracasei subsp. paracasei , Lactobacillus paracasei subsp. tolerans , Lactobacillus rhamnosus , Lactobacillus chiayiensis Renamed to Lacticaseibacillus casei , Lacticaseibacillus paracasei subsp. paracasei , Lacticaseibacillus paracasei subsp. tolerans , Lacticaseibacillus rhamnosus , Lacticaseibacillus chiayiensis Among them, the Lactobacillus genus contains many important probiotic resources, and changes in its classification may affect the update of microbial-related industry regulations and the application of bacterial species in various countries.

[0168] The scheme of the present invention will be explained below in conjunction with the embodiments. It will be appreciated by those skilled in the art that the following embodiments are only used to illustrate the present invention and should not be considered as limiting the scope of the invention. Where specific techniques or conditions are not indicated in the embodiments, the techniques or conditions described in the literature of the art or in accordance with the product or instrument instructions are used. All reagents or instruments are indicated by the manufacturer and can be purchased commercially.

[0169] Example

[0170] The anaerobic blood agar plate involved in the embodiment was purchased from Huankai Microorganisms, with a formula of 10 g / L casein pancreatic digest, 3 g / L heart pancreatic digest, 1 g / L corn starch, 5 g / L meat gastric digest, 5 g / L yeast extract powder, 5 g / L sodium chloride, 15 g / L agar, 50-100 mL / L sterile defibrinated sheep blood, pH 7.3±0.2.

[0171] The PYG liquid culture medium involved in the embodiment, each 1 L PYG liquid culture medium contains: Trypticase peptone (trypticase peptone) 5.0 g, Peptone (peptone) 5.0 g, Yeast extract (yeast extract powder) 10.0 g, Beef extract (beef extract powder) 5.0 g, Glucose (glucose) 5.0 g, K 2 HPO 4 2.0g, Tween 80 1.0ml, Resazurin 1.0mg, 40ml Salt solution 40ml (its components are as follows: CaCl 2 ·2H 2 O0.01g, MgSO 4 7H 2 O 0.02g, K 2 HPO 4 0.04 g, NaHCO 3 0.4g, NaCl 0.08g, balance water), balance water, pH 7.2±0.2 (25℃).

[0172] The MRS plate medium involved in the embodiment, each 1 L MRS plate medium contains: peptone 10.0 g, beef extract powder 10.0 g, yeast extract powder 5.0 g, glucose 20.0 g, Tween-80 1.0 mL, dipotassium hydrogen phosphate 2.0 g, sodium acetate 5.0 g, triammonium citrate 2.0 g, magnesium sulfate (MgSO 4 7H 2 O) 0.1 g, manganese sulfate (MnSO 4 ·4H 2O) 0.05 g, agar 15.0 g, the balance water.

[0173] The MRS liquid culture medium involved in the embodiment, each 1 L MRS liquid culture medium contains: peptone 10.0 g, beef extract powder 10.0 g, yeast extract powder 5.0 g, glucose 20.0 g, Tween-80 1.0 mL, dipotassium hydrogen phosphate 2.0 g, sodium acetate 5.0 g, triammonium citrate 2.0 g, magnesium sulfate (MgSO 4 7H 2 O) 0.1 g, manganese sulfate (MnSO 4 ·4H 2 O) 0.05 g, the balance being water.

[0174] The above culture medium can be prepared by conventional preparation methods and sterilization methods.

[0175] Example 1. Isolation and identification of strains

[0176] 1.1 Isolation and purification of strains MNH06277 and MNH45330 Two intestinal strains of Lactobacillus family (Lactobacillus paracasei Lacticaseibacillus paracasei MNH06277 and Lactobacillus crispatus Lactobacillus crispatus MNH45330) were isolated from fecal samples of healthy volunteers. The isolation method adopted the conventional strain isolation method, using gradient dilution, and then picking a single colony for isolation and culture, and then purifying the strain and anaerobic culture at 37°C. The pure culture strain was prepared into 20% glycerol / water-bacterial solution and stored at -80°C.

[0177] Specifically, the separation and purification methods of the strains are as follows: The donor takes 2-5 g of fresh feces and puts it into a sample collection and storage tube. After shaking and homogenization, the processed feces sample is placed in an ice box and sent to the laboratory for strain isolation within 24 hours.

[0178] Dispense physiological saline in a biosafety cabinet, 9 mL / tube; prepare anaerobic blood agar plates for strain isolation culture medium, and transfer them to the anaerobic workstation 24 hours in advance, marking the sample information, culture medium type, isolation date, etc.

[0179] Take a fresh stool sample and place it in an anaerobic workstation and shake it on a vortex shaker for 1 min to mix it. Pipette 1 mL of the sample into 9 mL of normal saline and mix it for 10 min. -1 dilution, and then serially diluted to 10 -6 Dilution solution for later use.

[0180] Take 10 -6Drop the diluted solution onto the anaerobic blood agar plate at a drop size of 100 μL / plate. Spread evenly. After the surface of the plate is dry, invert the plate and culture at 37°C for 3-5 days.

[0181] Observe the growth of the strain on the isolation medium (anaerobic blood agar plate) and pick a single colony with a sterilized toothpick for strain purification. The purified strain is placed at 37°C for anaerobically cultured. The pure culture strain is prepared into a 20% glycerol / water-bacteria solution and stored at -80°C.

[0182] 1.2 Morphological characteristics of strains MNH06277 and MNH45330 1.2.1 Morphological characteristics of strain MNH06277 The strain MNH06277 was inoculated on an MRS plate medium and cultured anaerobically at 37°C for 48 hours. Visible colonies were formed on the MRS plate medium. The colonies were round, with regular and smooth edges, about 1-2 mm in diameter, white, and opaque. The strain was Gram-positive. Microscopic morphological observation revealed that it had no flagella, was non-motile, rod-shaped, and about 0.5-1µm×1.5-3µm in size. The colony morphology of strain MNH 06277 cultured on an MRS plate for 48 hours is shown in Figure 1 The Gram staining photos of strain MNH06277 are shown in Figure 2 See the electron microscope photos for Figure 3 .

[0183] 1.2.2 Morphological characteristics of strain MNH45330 The strain MNH45330 was inoculated on an MRS plate medium and cultured anaerobically at 37°C for 48 hours. Visible colonies were formed on the MRS plate medium. The colonies were round, with regular and smooth edges, about 0.5-2 mm in diameter, white, and opaque. The strain was Gram-positive. Microscopic morphological observation revealed that it had no flagella, was non-motile, rod-shaped, and about 0.5-1 µm × 1.5-4 µm in size. The colony morphology of the strain MNH 45330 cultured on an MRS plate for 48 hours is shown in Figure 4 The Gram staining photos of strain MNH45330 are shown in Figure 5 See the electron microscope photos for Figure 6 .

[0184] 1.3 Physiological and biochemical characteristics of strains MNH06277 and MNH45330 1.3.1 Physiological and biochemical characteristics of strain MNH06277 The strain MNH06277 grows both aerobically and anaerobically. It can grow in the pH range of 4.0 to 10.0, with the optimal growth pH being 6.0 (see the results of the strain's tolerance to different pH values ​​for details). Figure 7); it can still grow on medium with NaCl content exceeding 6% (the results of strain tolerance to different concentrations of NaCl are shown in Figure 8 ); strain MNH06277 could not grow in medium with bile salt concentrations above 0.1% (the results of strain tolerance to different concentrations of bile salts are shown in Figure 9 ).

[0185] 1.3.2 Physiological and biochemical characteristics of strain MNH45330 The strain MNH45330 can grow in the pH range of 4.0 to 10.0, with the optimal growth pH being 8.0 (see the results of the strain's tolerance to different pH values). Figure 10 ); it can still grow on medium with NaCl content exceeding 4% (the results of strain tolerance to different concentrations of NaCl are shown in Figure 11 ); strain MNH45330 cannot grow in medium with bile salt concentration above 0.1% (the results of strain tolerance to different concentrations of bile salts are shown in Figure 12 ).

[0186] 1.4 Biochemical identification results of strains MNH06277 and MNH45330 1.4.1 Results of biochemical identification of strain MNH06277 by API 50CHL API 50CHL (Mérieux, CN5041010) was used. For specific experimental operations, please refer to the conventional API reagent operation guide. Culture conditions: 37°C, anaerobic. The experimental results are shown in Table 1.

[0187] MNH06277 can utilize ribose RIB, galactose GAL, glucose GLU, fructose FRU, mannose MNE, sorbitol SBE, mannitol MAN, sorbitol SOR, N-acetyl-glucosamine NAG, amygdalin AMY, arbutin ARB, esculin ESC, salicylate SAL, cellobiose CEL, maltose MAL, lactose LAC, sucrose SAC, trehalose TRE, inulin INU, melezitose MLZ, gentiobiose GEN, D-turanose TUR, D-tagatose TAG, L-arabinitol LARL, and gluconate GNT to ferment and produce acid. Therefore, in the process of fermenting or culturing strain MNH06277, ribose RIB, galactose GAL, glucose GLU, fructose FRU, mannose MNE, sorbitol SBE, mannitol MAN, sorbitol SOR, N-acetyl-glucosamine NAG, amygdalin AMY, arbutin ARB, esculin ESC, salicylate SAL, cellobiose CEL, maltose MAL, lactose LAC, sucrose SAC, trehalose TRE, inulin INU, melezitose MLZ, gentiobiose GEN, D-turanose TUR, D-tagatose TAG, L-arabinitol LARL, gluconate GNT, and their derivatives can be used as carbon sources.

[0188]

[0189] 1.4.2 Results of biochemical identification of strain MNH45330 using API 50CHL API 50CHL (Mérieux, CN5041010) was used. For specific experimental operations, please refer to the conventional API reagent operation guide. Culture conditions: 37°C, anaerobic. The experimental results are shown in Table 2.

[0190] MNH45330 can utilize galactose GAL, glucose GLU, fructose FRU, mannose MNE, mannitol MAN, α-methyl-D-glucoside MDG, N-acetyl-glucosamine NAG, amygdalin AMY, arbutin ARB, esculin ESC, salicylate SAL, cellobiose CEL, maltose MAL, lactose LAC, melibiose MEL, sucrose SAC, trehalose TRE, inulin INU, melezitose MLZ, raffinose RAF, starch AMD, gentiobiose GEN, D-turanose TUR, and D-lyxose LYX to ferment and produce acid. Therefore, in the process of fermentation or cultivation of strain MNH45330, galactose GAL, glucose GLU, fructose FRU, mannose MNE, mannitol MAN, α-methyl-D-glucoside MDG, N-acetyl-glucosamine NAG, amygdalin AMY, arbutin ARB, esculin ESC, salicylate SAL, cellobiose CEL, maltose MAL, lactose LAC, melibiose MEL, sucrose SAC, trehalose TRE, inulin INU, melezitose MLZ, raffinose RAF, starch AMD, gentiobiose GEN, D-turanose TUR, D-lyxose LYX, and their derivatives can be used as carbon sources.

[0191]

[0192] 1.5 Antibiotic Minimum Inhibitory Concentration Test for Strains MNH06277 and MNH45330 1.5.1 Antibiotic Minimum Inhibitory Concentration Test for Strain MNH06277 The minimum inhibitory concentration of antibiotics for strain MNH06277 was determined using E-test (purchased from Liofilchem) paper. The test results are shown in Table 3.

[0193]

[0194] The results showed that MNH06277 had an inhibitory effect on imipenem (I / R), ampicillin-sulbactam (SAM), and dopenem (DOR). It is sensitive to amoxicillin (AMC), rifampicin (RD), and ceftriaxone (CRO). It can be seen that MNH06277 is sensitive to most types of antibiotics, and the risk of long-term use of MNH06277 leading to antibiotic resistance in subjects is low.

[0195] 1.5.2 Antibiotic Minimum Inhibitory Concentration Test for Strain MNH45330 The minimum inhibitory concentration of antibiotics for strain MNH45330 was determined using E-test (purchased from Liofilchem) paper. The test results are shown in Table 4.

[0196]

[0197] The results showed that MNH45330 was sensitive to imipenem, ampicillin-sulbactam, dopenem, ertapenem, meropenem, piperacillin-tazobactam, and ceftriaxone. It can be seen that MNH45330 is sensitive to most types of antibiotics, and the risk of long-term use of MNH45330 leading to antibiotic resistance in subjects is low.

[0198] 1.6 Identification of strains MNH06277 and MNH45330 1.6.1 Amplification of 16S rRNA genes of strains MNH06277 and MNH45330 Fresh cultures of strains MNH06277 and MNH45330 were taken to extract genomic DNA of the strains, and the extracted genomic DNA of the strains was used as a template for 16S rRNA gene amplification.

[0199] The primer pairs used for PCR of 16S rRNA gene are: 27F: 5'-AGAGTTTGATCMTGGCTCAG-3' (SEQ ID NO: 1); 1492R: 5'-TACGGYTACCTTGTTACGACTT-3' (SEQ ID NO: 2).

[0200] The PCR reaction procedure is as follows: Pre-denaturation: 94℃, 4 min; (denaturation: 94℃, 50 sec, annealing: 52℃, 40 sec; extension: 72℃, 70sec), 36 cycles; final extension: 72℃, 10 min.

[0201] 1.6.2 16S rRNA gene sequencing of strains MNH06277 and MNH45330 The PCR product was purified and sequenced by Bioengineering to obtain the 16S rRNA gene.

[0202]

[0203] 1.6.3 Identification results of strains MNH06277 and MNH45330 The 16S rRNA gene sequences measured above, such as those shown in SEQ ID NO: 3 and SEQ ID NO: 4, were used to perform strain 16S rRNA gene analysis using the NCBI Basic Local Alignment Search Tool to confirm the strain classification information.

[0204] The measured sequences were compared with the data in GenBank by BLAST analysis. The results showed that the strain with the highest similarity to MNH06277 was Lacticaseibacillus paracasei , the similarity is 100%, so strain MNH06277 is Lacticaseibacillus paracasei strains planted; the strain with the highest similarity to MNH45330 is Lactobacillus crispatus The similarity is 100%, so strain MNH45330 is Lactobacillus crispatus Planted strains.

[0205] MNH06277 and MNH45330 were compared with the Lacticaseibacillus paracasei and Lactobacillus crispatus The 16S rRNA gene sequences of related strains of the genus were compared, and phylogenetic trees were constructed respectively.

[0206] The sequences of MNH06277 and MNH45330 were aligned with those of the model strains with high 16S rRNA gene sequence similarity in the NCBI database, and then a phylogenetic tree was constructed using the software MEGA 5 (the phylogenetic tree was constructed using the maximum likelihood method). Figure 13 , 14 Only the Bootstrap values ​​greater than 50% are displayed in the developmental tree nodes.

[0207] From the phylogenetic tree ( Figure 13 , 14 ) It can be seen that strain MNH06277 and Lacticaseibacillus Fungal Lacticaseibacillus paracasei Get together for Lacticaseibacillus paracasei The strain MNH45330 and Lactobacillus Fungal Lactobacillus crispatus Get together for Lactobacillus crispatus New plants planted.

[0208] The genomes of MNH06277 and MNH45330 were extracted and sequenced, and then tested by the gtdb method. The strain with the highest similarity to the MNH06277 genome was: Lacticaseibacillus paracasei(GCF_000829035.1), with an average nucleotide similarity (ANI) of 98.2% and an alignment score (AF) of 88%. Therefore, MNH06277 can be identified as Lacticaseibacillus paracasei The strain with the highest genome similarity to MNH45330 is: Lactobacillus crispatus (GCF_002088015.1), with an average nucleotide similarity (ANI) of 97.4% and an alignment score (AF) of 86%. Therefore, MNH45330 can be identified as Lactobacillus crispatus .

[0209] 1.7 Genome analysis of strains MNH06277 and MNH45330 1.7.1 Genome analysis of strain MNH06277 The genome of strain MNH06277 was fragmented by ultrasound, with a fragment length range of ~350bp, and then an Illumina sequencing library was constructed using a standard DNA library construction kit (NEB UltraTM). The constructed sequencing library was sequenced with NovaSeq (Illumina) for double-end 150bp. The sequencing obtained 0.89Gbp of data, of which Q20 accounted for 94.82%.

[0210] The raw sequencing data of the genome were filtered using fastp (version: 0.20.0) with filtering parameters: "--poly_g_min_len 10 --poly_x_min_len 10 -q 15 -u 40 -n 5 -l 50". The filtered raw data were assembled using SPAdes (version: v3.14.0) with assembly parameters "--isolate --cov-cutoff 10". The total gene length of the genome assembly was 2.90 Mbp, the N50 length was 196.8 kbp, and the GC content was 46.43%.

[0211] The genomic genes were predicted and analyzed using the prokaryotic analysis software genome annotation pipeline prokka (version: 1.14.5) with the parameters "--gcode 11 --evalue 1e-09". A total of 2728 CDS sequences were predicted, with an average CDS sequence length of 909 bp.

[0212] Potential antibiotic resistance genes in the genome were analyzed using the RGI pipeline (version: 4.2.2), with the antibiotic resistance gene database being CARD (version: 3.0.0, https: / / card.mcmaster.ca / analyze / rgi). No resistance genes were found.

[0213] The analysis of potential virulence factors and related genes in the genome was performed using NCBI blastp (version: 2.7.1+) to compare the virulence factor database VFDB (virulence factor database, http: / / www.mgc.ac.cn / cgi-bin / VFs / v5 / main.cgi, updated on September 19, 2019). The detailed comparison results are shown in Table 5.

[0214]

[0215] The analysis of potential primary metabolic gene clusters in the genome was performed using gutSMASH5 (version: 1.0.0). See 6 for detailed alignment results.

[0216]

[0217] 1.7.2 Genome analysis of strain MNH 45330 The genome of strain MNH 45330 was fragmented by ultrasound, with a fragment length range of ~350 bp, and then an Illumina sequencing library was constructed using a standard DNA library construction kit (NEB UltraTM). The constructed sequencing library was sequenced with NovaSeq (Illumina) for double-end 150 bp. 1.43 Gbp of data was obtained by sequencing, of which Q20 accounted for 95.33%.

[0218] The raw sequencing data of the genome were filtered using fastp (version: 0.20.0) with filtering parameters: "--poly_g_min_len 10 --poly_x_min_len 10 -q 15 -u 40 -n 5 -l 50". The filtered raw data were assembled using SPAdes (version: v3.14.0) with assembly parameters "--isolate --cov-cutoff 10". The total gene length of the genome assembly was 2.45 Mbp, the N50 length was 43.3 kbp, and the GC content was 36.51%.

[0219] The genomic genes were predicted and analyzed using the prokaryotic analysis software genome annotation pipeline prokka (version: 1.14.5) with the parameters "--gcode 11 --evalue 1e-09". A total of 2468 CDS sequences were predicted, with an average CDS sequence length of 870 bp.

[0220] Potential antibiotic resistance genes in the genome were analyzed using the RGI pipeline (version: 4.2.2), with the antibiotic resistance gene database being CARD (version: 3.0.0, https: / / card.mcmaster.ca / analyze / rgi). No resistance genes were found.

[0221] The analysis of potential virulence factors and related genes in the genome was performed using NCBI blastp (version: 2.7.1+) to compare the virulence factor database VFDB (virulence factor database, http: / / www.mgc.ac.cn / cgi-bin / VFs / v5 / main.cgi, updated on September 19, 2019). The detailed comparison results are shown in Table 7.

[0222]

[0223] The analysis of potential primary metabolic gene clusters in the genome was performed using gutSMASH5 (version: 1.0.0). The detailed alignment results are shown in Table 8.

[0224]

[0225] Example 2: Strains MNH06277 and MNH45330 promote growth and development in a low-fat, low-protein diet-induced growth retardation mouse model The growth retardation mouse model induced by low-fat and low-protein diet was used to conduct experiments on the improvement of growth and development indicators by MNH06277 and MNH45330. This invention has passed the ethical review of the Animal Ethics Committee of Muen Biotechnology.

[0226] 2.1 Experimental methods 1) Experimental animals: The experimental mice were C57BL / 6J mice, 3 weeks old, purchased from Guangdong Weitonglihua Experimental Animal Technology Co., Ltd.

[0227] 2) Preparation of test materials of strains MNH06277 and MNH45330: After thawing the glycerol cryovials of MNH06277 and MNH45330 strains at 37°C, they were inoculated into MRS plate medium in an anaerobic workstation for activation. The activated strains were inoculated into MRS liquid medium and cultured anaerobically to obtain a sufficient amount of culture. The cultured bacterial solution was concentrated by centrifugation and resuspended in PBS containing 25% glycerol and 0.05% L-Cys HCl to obtain the purity and viable count (2×10 9 CFU / mL) of the test substances that meet the requirements of animal experiments.

[0228] 3) Negative control: Use PBS containing 25% glycerol and 0.05% L-Cys HCl as the negative control.

[0229] 4) Positive control: Bifidobacterium lactis CGMCC No.20847. After thawing the glycerol cryopreserved tube of Bifidobacterium lactis CGMCC No.20847 (disclosed in patent CN112980725B) at 37°C, inoculate it into PYG medium in an anaerobic workstation for activation. Inoculate the activated strain into PYG liquid medium and culture it anaerobically to obtain a sufficient amount of culture. After centrifuging and concentrating the cultured bacterial solution, resuspend the bacterial strain in PBS containing 25% glycerol and 0.05% L-Cys HCl to obtain the purity and viable count (2×10 9 CFU / mL) is a positive control substance that meets the requirements of animal experiments.

[0230] 5) Experimental process: After the quarantine period, 40 C57BL / 6J male mice weighing 13g-16g were selected for 3-week-old mice. They were randomly divided into 5 groups according to their weight, 8 mice / group, namely NCD-Control group, LFD-Control group, MNH06277 group, MNH45330 group, and positive control group. The NCD-Control group was fed with maintenance feed (Cat. No.: PD24043001, Changzhou Shuyi Shuer Biotechnology Co., Ltd.), and the other four groups were fed with low-fat and low-protein feed (Cat. No.: PD24043003, Changzhou Shuyi Shuer Biotechnology Co., Ltd.). Drug administration began after grouping (D1). The NCD-Control group and the LFD-Control group were given negative control products, the positive control group was gavaged with positive control products, the MNH06277 group was gavaged with MNH06277, and the MNH45330 group was gavaged with MNH45330. The drugs were administered orally once a day, 200 μL each time, for a total of 35 days. During the experiment, mice were free to drink water and eat, and a 12h / 12h day and night cycle was adopted. During the experiment, a general clinical observation was performed once after each administration. The endpoint of this experiment was the day after the end of administration (D36). The end point of the experiment was anatomical sampling according to the protocol, and the results of each anatomical data and serum test data were summarized and analyzed. All data were expressed in the form of Mean±SD, and GraphPad Prism software was used for drawing and statistical analysis. For pairwise comparisons, the t test (Student's t test) analysis method was used. No significance is indicated; significant differences are indicated by *, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

[0231] Food intake was determined by weighing the remaining amount of feed.

[0232] The weight of internal organs such as the liver and spleen, the length of the large intestine, and the length and weight of various bones and muscles are measured through dissection.

[0233] Method for detecting serum insulin-like growth factor-1: Mouse serum was collected and detected using Mouse IGF-1 ELISA Development Kit (PeproTech).

[0234] The immune cell detection method is to take 0.1g of mouse spleen tissue, wash the surface with PBS and then crush the tissue, suspend the crushed spleen tissue in PBS, filter through a 70μm filter membrane, and prepare a mouse spleen single cell suspension. Take a small amount of the mouse spleen single cell suspension prepared above, stain it, and complete the machine detection in the flow cytometer to accurately count the number of mouse spleen immune cells.

[0235] The method for determining the content of short-chain fatty acids in cecal contents is as follows: weigh about 100 mg of cecal contents and place it in a 2 mL EP tube, add 1 ml of ether, vortex mix immediately, extract short-chain fatty acids by ultrasonication for 10 min, centrifuge for 10 min to remove precipitation, pass the supernatant through a 0.22 μm organic filter membrane, transfer the supernatant to a gas phase vial, wrap it with a sealing film and store it in a -20 ° C refrigerator, waiting for determination. Use a DB-FFAP column and a mass spectrometer to separate and detect each short-chain fatty acid.

[0236] 2.2 Experimental Results See also Figures 15 - 17 The results showed that MNH06277 and MNH45330 could significantly increase the length of the large intestine, spleen and kidney weight of developmentally retarded mice, indicating that MNH06277 and MNH45330 can promote the development of the spleen, kidney, and / or intestine, and have the use of promoting organ development.

[0237] See also Figures 18 - 23 The results showed that MNH06277 and MNH45330 could significantly promote the body weight, body length, tibia length, femur length, soleus muscle and / or gastrocnemius muscle weight of the low-fat, low-protein diet-induced growth retardation mouse model, indicating that MNH06277 and MNH45330 are useful for promoting height (body length), weight, bone, and / or muscle development.

[0238] See also Figure 24 The results showed that MNH06277 could significantly increase the serum insulin-like growth factor 1 (IGF-1) content in the growth retardation mouse model induced by a low-fat and low-protein diet, indicating that MNH06277 has the use of promoting growth and development.

[0239] See also Figures 25 - 26The results showed that MNH06277 and MNH45330 did not increase food intake, indicating that MNH06277 and MNH45330 have the function of promoting digestion and absorption when promoting growth and development.

[0240] See also Figures 27 - 29 The results showed that MNH06277 and MNH45330 can significantly promote the development of major immune organs such as the thymus and spleen in the low-fat and low-protein diet-induced growth retardation mouse model; MNH45330 can increase the proportion of T cells, indicating that MNH06277 and MNH45330 have the use of promoting the development of the immune system and enhancing immunity.

[0241] See also Figures 30 - 34 The results showed that MNH45330 can promote the production of short-chain fatty acids in the cecum of a low-fat, low-protein diet-induced growth retardation mouse model, and significantly promote the production of propionic acid, isobutyric acid, 2-methylbutyric acid, and isovaleric acid; MNH06277 can increase the proportion of butyric acid in total short-chain fatty acids. This shows that MNH06277 and / or MNH45330 can promote the production of short-chain fatty acids and have the effect of promoting intestinal health.

[0242] Although the present invention has been disclosed with reference to certain embodiments, it is apparent that modifications and variations may be made without departing from the spirit and scope of the present invention as disclosed herein and as provided in the appended claims. In addition, it should be understood that although all examples disclosed illustrate embodiments of the present invention, they are provided only as non-limiting examples and therefore should not be considered as limiting the various aspects of the present invention thus described. The present invention is intended to have the full scope defined by the language of the present disclosure, the following claims, and any equivalents thereof. Therefore, the drawings and detailed description should be considered illustrative rather than restrictive.

Claims

1. A composition for promoting growth and development, characterized in that: The composition comprises Lactobacillus paracasei ( Lacticaseibacillus paracasei ) MNH06277 and / or Lactobacillus crispatus ( Lactobacillus crispatus ) culture, live bacteria, freeze-dried bacteria or inactivated bacteria of MNH45330, wherein the Lactobacillus paracasei ( Lacticaseibacillus paracasei )The deposit number of MNH06277 is GDMCC No: 65552; The Lactobacillus crispatus ( Lactobacillus crispatus ) The deposit number of MNH45330 is GDMCC No: 65556.

2. The composition according to claim 1, characterized in that The composition further comprises one or more pharmaceutically and / or food acceptable carriers, excipients and / or adjuvants; and / or The composition may further comprise one or more other active agents for promoting growth and development, enhancing immunity, or promoting intestinal health.

3. The composition according to claim 2, characterized in that The other active agents include one or more of probiotics and prebiotics.

4. The composition according to any one of claims 1 to 3, characterized in that The composition is any one of medicine, health care product or food.

5. The composition according to claim 4, characterized in that The health care product or food is any one of a nutritional composition, a food, a candy, a food bar, a food additive, a drink additive, and a dietary supplement.

6. Use of the composition according to any one of claims 1 to 5 in promoting growth and development, enhancing immunity, or promoting intestinal health.

7. Use of the composition according to any one of claims 1 to 5 in the preparation of medicines, health products or foods for promoting growth and development, enhancing immunity, or promoting intestinal health.

8. The use according to claim 6 or 7, characterized in that: The growth promotion includes: promoting at least one of the development of organs, height, weight, bones and muscles.

9. The use according to claim 8, characterized in that: The organ includes at least one of the thymus, spleen, kidney, and intestine; or The bone comprises at least one of a femur and a tibia; or The muscle includes at least one of the soleus muscle and the gastrocnemius muscle.

10. The use according to claim 6 or 7, characterized in that: The medicine, health product or food promotes growth and development by increasing hormone content and / or promoting digestion and absorption; or The medicine, health product or food enhances immunity by promoting the development of the immune system; or The medicine, health product or food promotes intestinal health by increasing short-chain fatty acids and / or improving intestinal flora.

11. The use according to claim 10, characterized in that: The hormones include insulin-like growth factor 1 and / or growth hormone; or The promoting the development of the immune system comprises at least one of promoting the development of immune organs and promoting the generation of immune cells.

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