CN110540950A - Lactobacillus reuteri 22 and application thereof - Google Patents
Lactobacillus reuteri 22 and application thereof Download PDFInfo
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- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
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- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/173—Reuteri
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- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
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Abstract
Lactobacillus reuteriThe invention discloses a lactobacillus reuteri strain 22 and application thereof. The Lactobacillus reuteri 22 is obtained by separating from healthy chicken flocks, is classified and named as Lactobacillus reuteri (CGMCC No. 17932), and has a preservation number of CGMCC No. 17932. By detecting the probiotic performance, the lactobacillus reuteri 22 is found to have good acid resistance, cholate resistance and adhesion performance and can inhibit the growth of pathogenic bacteria. Through animal experiments, the lactobacillus reuteri 22 does not affect the level of inflammatory factors in a healthy physiological state, but can increase the number of goblet cells and promote the increase of the expression level of related genes such as compact protein, defensin and lysozyme, and can increase the length of intestinal villus and the depth of crypt, and has the potential of maintaining the intestinal mucosal barrier. Therefore, the strain is expected to be developed into probiotics for maintaining intestinal mucosa barriers of chickens and guaranteeing green and healthy breeding of the chickens.
Description
Technical Field
The invention relates to a biological prevention and control technology, in particular to a lactobacillus reuteri strain 22 and application thereof in chicken breeding, wherein oral administration of the lactobacillus reuteri strain 22 can enhance the intestinal immunity function, promote the proliferation of intestinal epithelium and have the potential of maintaining the intestinal barrier steady state.
Background
The genus Lactobacillus (Lactobacillus) is classified as a gram-positive bacterium belonging to the phylum scleromycota (fungi), class Bacili (Bacilli), order Lactobacillales, family Lactobacillaceae (lactobacilleae) in taxonomic order, does not form spores, and can produce lactic acid upon sugar decomposition. The lactobacillus is normal bacteria of human and animal intestinal tracts, and has close relation with the absorption of host nutrient substances and the development of an intestinal tract mucous membrane immune system.
The development of various probiotic properties of lactic acid bacteria is based on the adhesion to intestinal epithelial cells, and if the adhesion capability is poor or the adhesion cannot be realized, only pass-through bacteria can be realized, and the physiological function cannot be fully developed. Host specificity and strain specificity are the characteristics of lactobacillus, that is, lactobacillus from different sources has very different adhesive ability, and strains of different species have different adhesive ability. In addition, the adhesion of the lactic acid bacteria has concentration dependence, and the strength of the adhesion capability is positively correlated with the concentration of the bacteria liquid in a certain range. For example, the adhesion of bifidobacteria has the influence of time and concentration, and the adhesion effect is saturated when the adhesion lasts for 3 hours and the bacterial load is 108 cfu/mL.
Intestinal associated lymphoid tissue (GALT) is the largest lymphoid tissue in the body. Thus, it is an important component of the host's total immune tissue, the gut microflora being an important component of the gut defense barrier, and the increased transport of antigens between the gut mucosae in the absence of the gut microflora. This view is further supported by the intestinal flora inducing specific immune responses both locally and systemically. In addition, the gut flora may promote a constant low level response of the immune system to pathogens. In addition, the intestinal flora promotes the maturation of the intestinal immune system, the content of organism secretory immunoglobulin A is increased in the process of gradual formation of the intestinal flora, and researches show that the translocated flora in the intestinal tract is obviously reduced after the organism immune system is mature.
Under normal physiological conditions, the microbial environment inside the body is in a stable, balanced state. The intestinal homeostasis of the mutualistic symbiosis is closely combined with the health of the organism, and if the intestinal homeostasis is unbalanced, pathogenic bacteria can enter while being deficient, so that the growth of the organism is influenced, and even the organism dies. Research shows that lactobacillus can enhance intestinal mucosa barrier and maintain healthy culture.
Disclosure of Invention
The purpose of the invention is as follows: the first technical problem to be solved by the invention is to provide a lactobacillus reuteri strain 22, the lactobacillus reuteri strain 22 has good tolerance, bacteriostatic effect and excellent adhesion performance, and can enhance the intestinal immunity, promote the intestinal epithelial proliferation and further maintain the intestinal mucosal barrier.
The second technical problem to be solved by the present invention is to provide a probiotic, which comprises said lactobacillus reuteri 22.
The third technical problem to be solved by the invention is to provide the application of the lactobacillus reuteri 22 or the probiotics in chicken breeding.
The invention finally aims to solve the technical problem of providing the chicken feed.
The technical scheme is as follows: in order to solve the technical problems, the technical scheme adopted by the invention is as follows: the Lactobacillus reuteri 22 is classified and named as Lactobacillus reuteri, is preserved in China general microbiological culture Collection center on 6-14 th month in 2019 with the preservation number of CGMCC No. 17932; and (4) storage address: xilu No.1 Hospital No. 3, Beijing, Chaoyang, North.
The present disclosure also includes a probiotic comprising said lactobacillus reuteri 22.
The invention also comprises the application of the lactobacillus reuteri 22 or the probiotics in chicken breeding.
Wherein the daily addition amount of Lactobacillus reuteri 22 is 2.5 × 106-2 × 107CFU/g chicken.
The invention also comprises the application of the lactobacillus reuteri 22 or the probiotics in improving one or more of the length of small intestine villi, the depth of small intestine crypts, the expression level of tight junction protein, the expression level of antibacterial peptide or the expression level of lysozyme.
The invention also comprises the application of the lactobacillus reuteri 22 or the probiotics in increasing the number of intestinal stem cells and the number of goblet cells.
The invention also comprises the application of the lactobacillus reuteri 22 or the probiotics in one or more of promoting intestinal IgA secretion, reducing inflammatory factors and improving defensin expression level.
The invention further comprises a chicken feed which comprises the lactobacillus reuteri 22 or the probiotics.
Has the advantages that: compared with the prior art, the invention has the following characteristics and advantages:
1. The differences among lactobacillus species are large, and the phenotypic characters, biochemical reactions and physiological characteristics have obvious differences. The lactobacillus reuteri 22 is separated from the intestinal tract of chicken, is chicken-origin lactobacillus, is proved to have good acid resistance, cholate resistance and adhesion performance by screening, and can inhibit the growth of pathogenic bacteria.
2. There is currently little research on lactobacillus reuteri on poultry animals chickens. The lactobacillus reuteri 22 screened by the invention can stimulate the proliferation of chick intestinal stem cells, the number of chick intestinal crypt proliferation cells, the depth of crypts and the high-level expression of proliferation related genes (PCNA and c-Myc), thereby promoting the healthy development of early intestinal tracts of poultry. The phenomenon and the action mechanism thereof are reported for the first time, and the research result has important developmental research value and practical cultivation significance for nutrition absorption and early pathogenic microorganism infection.
3. The lactobacillus reuteri 22 disclosed by the invention can enhance the intestinal mucosal immune function of the chicks, such as induction of increase of the number of goblet cells, expression level of the claudin and the level of antibacterial peptide (AvBD2, AvBD9 and lysozyme), and is also the action mechanism of the chicken-derived lactobacillus reuteri on the related mucosal immune function on the chicks, which is reported for the first time by the invention.
4. The lactobacillus reuteri 22 is obtained by first separation and identification; the strain is gram-positive and rod-shaped, and has good probiotic performance; oral administration of lactobacillus reuteri 22 can enhance intestinal immunity, increase the length of intestinal villi and the depth of intestinal crypt, maintain intestinal mucosa homeostasis, and the lactobacillus reuteri can be developed into probiotics for preventing enteritis.
Drawings
FIG. 1: gram staining observation picture of lactobacillus optical microscope;
FIG. 2: the lactobacillus has acid resistance and bile salt resistance, and antibacterial and adhesive performance result graphs; A-B: the strain with the number 22 is lactobacillus reuteri 22, and has good tolerance performance under the condition that the pH value is 3 and 0.3% of bile salt; C-D: the lactobacillus reuteri 22 can better adhere to Caco-2 cells; E-F: the strain with the number of 8 is the lactobacillus reuteri 22, and can inhibit the growth of intestinal pathogenic bacteria of Escherichia coli E-coli, salmonella pullorum, and salmonella typhimurium.
FIG. 3: lactobacillus reuteri 22 protects the intestinal mucosal barrier; con group: negative control, oral PBS; lactobacillus reuteri group 22: inoculating 2 × 108CFU of Lactobacillus reuteri 22; A-C: lactobacillus reuteri 22 slightly reduces the level of inflammatory factors; d: lactobacillus reuteri 22 may protect the intestinal villus structure; E-G: lactobacillus reuteri 22 can slightly increase body weight and improve the expression level of the claudin gene.
FIG. 4: lactobacillus reuteri 22 increases the number of goblet cells; grouping is as in FIG. 3; A-B: PAS staining showed that lactobacillus reuteri 22 was able to increase the number of goblet cells.
FIG. 5: lactobacillus reuteri 22 is capable of promoting cell proliferation at intestinal crypts; grouping is as in FIG. 3; A-B: HE staining showed that lactobacillus reuteri 22 increased the depth of intestinal crypts; C-D: lactobacillus reuteri 22 increases the number of PCNA + cells at the intestinal crypts; E-G: lactobacillus reuteri 22 promotes the increase of the expression level of proliferation-related genes and stem cell marker genes.
FIG. 6: lactobacillus reuteri 22 can improve the intestinal mucosa immunity level; grouping is as in FIG. 3; A-D: lactobacillus reuteri 22 promotes intestinal IgA secretion, and increases defensin and lysozyme expression levels.
Detailed Description
The invention is further illustrated by the following specific examples and figures. The methods used in the following examples are conventional reagents and conventional methods unless otherwise specified.
Example 1 preliminary screening of the origin and strains of the samples, identification of probiotic properties and strains
1. Preliminary screening of the origin of the samples and the strains
The sample is from a healthy chicken, the abdominal cavity is opened after slaughtering, two ends of duodenum are tied by a sterilized cotton rope, the duodenum is immediately placed into precooled physiological saline after being cut off, the duodenum is sent to a laboratory for strain separation within 30min, the bacteria separation operation is carried out in a super clean bench, the intestine section is longitudinally cut off by using sterile scissors, then the content on the surface of the intestine is washed by using sterile PBS buffer solution (pH7.4, 0.01M), then the mucous membrane of the intestinal wall is slightly scraped by a blade to be about 0.5g, the intestine is placed in a triangular cone bottle filled with glass beads for full oscillation, after the intestine is placed at room temperature for 2-3min, 100 mu l of suspension is taken and coated on the surface of MRS agar culture medium (pH5.8), and the intestine is cultured for 24 h. The suspected colonies were picked and gram-stained, and 26 strains were streaked and purified on MRS agar plates as shown in FIG. 1, which is an optical microscopic image of the suspected Lactobacillus.
2. Acid-resistant and bile-resistant salt
Respectively selecting 26 strains of lactobacillus which are preliminarily screened and purified, wherein the serial numbers are respectively 1-26, activating in an MRS liquid culture medium, taking 1mL of bacterial liquid when the lactobacillus is cultured to a logarithmic growth phase, centrifuging at 5000rpm for 5min, respectively adding 1mL of PBS with pH3.0 and 0.01M and 1mL of PBS solution with 0.3 percent of bile salt (pH7.4 and 0.01M), placing in a bacterial incubator for treatment for 2h, centrifuging and resuspending. Culturing in 5mL MRS liquid culture medium at a ratio of 2% (v/v) for 16h, detecting OD600 with a microplate reader, and obtaining the ratio of the OD600 value of the acid/cholate treatment group to the OD600 value of the blank group, namely the relative survival rate, as shown in FIG. 2A-B, the results show that 8 strains of bacteria numbered 1, 3, 6, 9, 10, 14, 15 and 22 have better acid and cholate resistance.
3. Cell adhesion assay
1) Plate paving: caco-2 cells (purchased from Guangzhou Jinie Europe Biotech, China) were digested and then blown into a cell suspension in DMEM without double antibody, and the cell suspension was uniformly spread on a cell culture plate on which cell slide was placed until the cells grew.
2) And (3) incubation: gently wash with sterile PBS along the walls of the cell wells, leaving only adherent cells. Respectively taking 1mL of 108cfu/mL of the acid-resistant and bile-salt-resistant screened 8 strains of lactic acid bacteria, centrifuging at 5000rpm for 5min, then resuspending the bacteria by using an equal volume of DMEM, adding the bacteria into cell pores, and co-culturing for 2h in a cell culture box.
3) Fixing: washing with sterile PBS buffer (pH7.4, 0.01M) retained only adherent bacteria. Methanol fixation for 30min, gram staining, microscopic examination. Dripping cedar oil, randomly selecting visual fields under an oil lens, uniformly distributing the visual fields, calculating the adhesion quantity of lactic acid bacteria on the surfaces of all Caco-2 cells in each visual field, calculating an average value to represent the adhesion capacity of bacteria, and taking a picture by using imaging software matched with a computer, wherein the result shows that 8 strains tested have obvious difference on the adhesion performance of Caco-2, the highest adhesion capacity is No. 22 bacteria, and the lowest adhesion capacity is No.1 bacteria, as shown in figures 2C-D.
4. Plate bacteriostasis test
The experiment was carried out by Oxford cup punch plate diffusion method, in LB solid medium agar was not solidified, to which different concentrations of E.coli (106cfu/mL, 105cfu/mL, 104cfu/mL) (from BNCC125987, North Nano biology Inc.), Salmonella Pullorum ATCC 9120(106cfu/mL, 105cfu/mL, 104cfu/mL) (presented by Zhang Wei of Nanjing university of agriculture), Salmonella typhimurium SL1344(106cfu/mL, 105cfu/mL, 104cfu/mL) (presented by professor Liu forest, university of Beijing university of microbiology) were added, the bacterial liquid and the medium were mixed and added to a sterile plate 15cm in diameter, and when the medium was not completely solidified, Oxford cup was placed in a place set in advance and not shaken. And (3) after the culture medium is completely solidified, adding 150 mu l of 8 strains of lactobacillus bacteria liquid screened at 108cfu/mL into holes generated in the Oxford cup, adding 8 strains of lactobacillus bacteria with corresponding numbers of 1, 3, 6, 9, 10, 14, 15 and 22 into 1-8 holes on the flat plate, placing the flat plate in a incubator for culture, and calculating the actual area of the inhibition zone by using a digital display electronic vernier caliper, wherein the results show that the inhibition effect of the 8 strains on escherichia coli is not biologically different, but the definition of the inhibition zone of the total 4 strains of 10, 14, 15 and 22 is obviously higher than that of other strains, particularly the inhibition effect of the No. 22 strain and the No. 15 strain on Salmonella typhimurium is obviously different from that of other groups. However, the bacterial strains No. 15 and No. 22 have very outstanding bacteriostatic effects on the effect of inhibiting the Salmonella Pullorum (Salmonella Pullorum ATCC 9120), and particularly, the bacterial strain No. 22 has a large and obvious bacteriostatic zone and a very good bacteriostatic effect.
5. Lactobacillus specific PCR identification
Selecting a single colony of the No. 22 strain with excellent probiotic performance in the test to an MRS culture medium (a lactobacillus selective culture medium purchased from Qingdao Haibo organism, China), extracting bacterial genomes by using a bacterial genome DNA extraction kit (Tiangen, China) after amplification culture, and performing PCR amplification on the extracted bacterial genomes by using a pair of 16S universal primers 27F and 1492R.
27F：AGA GTT TGA TCC TGG CTC AG
1492R：TAC GGY TAC CTT GTT ACG ACT T
The above primers were synthesized by Shanghai Biotech, and the reaction was carried out using a 25. mu.l system (12.5. mu.l BU-Taq 2 × Master PCR mix (Takara, China), 2. mu.l template DNA (bacterial genome), 1. mu.l each of upstream and downstream primers, 9.5. mu.l ddH2O) in a Biometra PCR apparatus (Bio-Rad, USA) according to the following reaction procedure: pre-denaturation at 95 ℃ for 3min, 30s at 94 ℃ and 1min at 72 ℃ for 30 cycles in total, and final extension at 72 ℃ for 15 min. Sequencing the reaction product to obtain SEQ ID NO: 1 Sequence was found to be identical to Lactobacillus reuteri strain DFM 316S ribosomal RNA gene, partial Sequence, Sequence ID: the MN128548.1 similarity reaches 100 percent, thereby determining and naming the strain as the Lactobacillus reuteri 22, the strain is preserved in the China general microbiological culture Collection center with the preservation number of CGMCC No. 17932.
Example 2 testing of the Effect of oral administration of Lactobacillus reuteri 22 on the intestinal tract of chicks
Selecting newborn healthy white feather broilers (the body weight is about 44g) of 3 days old, and 20 white feather broilers per group, dividing the newborn healthy white feather broilers into a blank group and a control group, and orally feeding the blank group of chicks with stomach-filling sterile 200 mu l PBS (pH7.4, 0.01M) for 7 days; in the group of Lactobacillus reuteri 22, each chick was inoculated with 200. mu.l of 109CFU/ml of Lactobacillus reuteri 22 in succession and sampled on day 8. The content of Lipopolysaccharide (LPS) in serum is detected by an ELISA method, meanwhile, inflammatory factors TNF-alpha and IL-1 beta of jejunal tissues are detected, the results of figure 3 show that the level of inflammatory factors is slightly reduced after the treatment of the lactobacillus reuteri 22 (figures 3A-C), the weight is increased (figure 3E), the expression level of genes related to the tight junction protein is increased (figures 3F-G) and the HE staining result shows that the intestinal villus is intact after the treatment of the lactobacillus reuteri 22 (figure 3D) compared with a negative control Con group.
Periodic acid Schiff staining (PAS) was used to detect the number of goblet cells in the intestine. Cells that appeared red under ordinary light microscopy (i.e., goblet cells) were counted, and the results in fig. 4 show a significant increase in the number of goblet cells after treatment with lactobacillus reuteri 22 compared to the negative control Con group (fig. 4A-B). Changes in intestinal crypt depth were shown by HE staining, increasing crypt depth in the lactobacillus reuteri 22 treated group (fig. 5A-B), while lactobacillus reuteri 22 was able to significantly increase the number of proliferating cells at crypts by counting the number of intestinal PCNA + cells (fig. 5C-D). The genes c-Myc, PCNA, Lgr5 and Bmi1 were detected by fluorescent quantitative PCR, and the expression levels of the genes were consistently increased after treatment with Lactobacillus reuteri 22 (FIGS. 5E-G). The results show that the lactobacillus reuteri 22 can regulate and control the development of intestinal stem cells of the chicks and has the capacity of obviously promoting the proliferation of intestinal epithelium.
IgA in jejunal tissue was detected by ELISA, and the results showed that there was a tendency for increasing the secretory immunoglobulin IgA of jejunum in the Lactobacillus reuteri 22-treated group (FIG. 6A). The expression levels of the jejunal defensins AvBD2, AvBD9, Lysozyme gene were detected by fluorescent quantitative PCR, and the expression level of the defensin in the Lactobacillus reuteri 22-treated group was slightly increased as compared to the negative control Con group (FIGS. 6B-D).
Sequence listing
<110> Nanjing university of agriculture
<120> lactobacillus reuteri 22 and application thereof
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1403
<212> DNA
<213> Lactobacillus reuteri (Lactobacillus reuteri)
<400> 1
taggccaccg actttgggcg ttacaaactc ccatggtgtg acgggcggtg tgtacaaggc 60
ccgggaacgt attcaccgcg gcatgctgat ccgcgattac tagcgattcc gacttcgtgt 120
aggcgagttg cagcctacag tccgaactga gaacggcttt aagagattag cttactctcg 180
cgagcttgcg actcgttgta ccgtccattg tagcacgtgt gtagcccagg tcataagggg 240
catgatgatc tgacgtcgtc cccaccttcc tccggtttgt caccggcagt ctcactagag 300
tgcccaactt aatgctggca actagtaaca agggttgcgc tcgttgcggg acttaaccca 360
acatctcacg acacgagctg acgacgacca tgcaccacct gtcattgcgt ccccgaaggg 420
aacgccttat ctctaaggtt agcgcaagat gtcaagacct ggtaaggttc ttcgcgtagc 480
ttcgaattaa accacatgct ccaccgcttg tgcgggcccc cgtcaattcc tttgagtttc 540
aaccttgcgg tcgtactccc caggcggagt gcttaatgcg ttagctccgg cactgaaggg 600
cggaaaccct ccaacaccta gcactcatcg tttacggcat ggactaccag ggtatctaat 660
cctgttcgct acccatgctt tcgagcctca gcgtcagttg cagaccagac agccgccttc 720
gccactggtg ttcttccata tatctacgca ttccaccgct acacatggag ttccactgtc 780
ctcttctgca ctcaagtcgc ccggtttccg atgcacttct tcggttaagc cgaaggcttt 840
cacatcagac ctaagcaacc gcctgcgctc gctttacgcc caataaatcc ggataacgct 900
tgccacctac gtattaccgc ggctgctggc acgtagttag ccgtgacttt ctggttggat 960
accgtcactg cgtgaacagt tactctcacg cacgttcttc tccaacaaca gagctttacg 1020
agccgaaacc cttcttcact cacgcggtgt tgctccatca ggcttgcgcc cattgtggaa 1080
gattccctac tgctgcctcc cgtaggagta tggaccgtgt ctcagttcca ttgtggccga 1140
tcagtctctc aactcggcta tgcatcatcg ccttggtaag ccgttacctt accaactagc 1200
taatgcaccg caggtccatc ccagagtgat agccaaagcc atctttcaaa caaaagccat 1260
gtggcttttg ttgttatgcg gtattagcat ctgtttccaa atgttatccc ccgctccggg 1320
gcaggttacc tacgtgttac tcacccgtcc gccactcact ggtgatccat cgtcaatcag 1380
gtgcaagcac catcaatcag ttg 1403
<210> 2
<211> 20
<212> DNA
<213> upstream primer 27F (artificial sequence)
<400> 2
agagtttgat cctggctcag 20
<210> 3
<211> 22
<212> DNA
<213> downstream primer 1492R (Artificial sequence)
<400> 3
tacggytacc ttgttacgac tt 22
Claims (8)
1. The lactobacillus reuteri 22 is characterized in that the preservation number of the lactobacillus reuteri 22 is CGMCC number 17932, and the lactobacillus reuteri is preserved in China general microbiological culture Collection center (CGMCC) in 2019, 6 and 14.
2. A probiotic, characterized in that it comprises lactobacillus reuteri 22 according to claim 1.
3. Use of the lactobacillus reuteri 22 of claim 1 or the probiotic of claim 2 in chicken breeding.
4. The use according to claim 3, wherein the daily dosage of Lactobacillus reuteri 22 is 2.5 x 106-2 x 107CFU/g chicken.
5. Use of lactobacillus reuteri 22 according to claim 1 or a probiotic according to claim 2 for increasing one or more of the length of intestinal villi, the depth of intestinal crypts, the expression level of claudin, the expression level of antimicrobial peptides or the expression level of lysozyme.
6. Use of lactobacillus reuteri 22 according to claim 1 or a probiotic according to claim 2 for increasing the number of intestinal stem cells, the number of goblet cells.
7. Use of lactobacillus reuteri 22 according to claim 1 or a probiotic according to claim 2 for one or more of promoting intestinal IgA secretion, reducing inflammatory factors, increasing defensin expression levels.
8. A chicken feed comprising lactobacillus reuteri 22 of claim 1 or a probiotic of claim 2.
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| CN112358999B (en) * | 2020-11-26 | 2022-04-08 | 中国农业大学 | Lactobacillus reuteri and application thereof |
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| CN114107088A (en) * | 2021-10-20 | 2022-03-01 | 山西大学 | Lactobacillus reuteri LRSY523 and application thereof |
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| CN114836358B (en) * | 2022-06-28 | 2023-02-24 | 中国农业科学院北京畜牧兽医研究所 | Lactobacillus reuteri SXDT-32 and application thereof |
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