Lactobacillus amylovora SKLAN202401ZF and its application

Abstract

The present invention relates to the field of microorganisms, and in particular to Lactobacillus amylovora SKLAN202401ZF and applications thereof. Lactobacillus amylovorus ) SKLAN202401ZF, with a deposit number of CGMCC No.30022. This strain can produce the anti-inflammatory metabolite chrysin in the intestines of animals, and has excellent antibacterial and immunomodulatory functions. By adding this strain to piglet feed, the intestinal health of piglets can be significantly improved, the incidence of diarrhea can be reduced, and the feed conversion rate and growth rate can be increased. The application of this strain not only helps to improve the efficiency of pig farming, but also can replace antibiotics, making it suitable for antibiotic-free farming.

Description

Technical Field

[0001] The invention relates to the field of microorganisms, and in particular to Lactobacillus amylovora SKLAN202401ZF and applications thereof. Background Art

[0002] Intestinal inflammation is one of the main problems affecting the health and production performance of piglets. Due to the multiple stress factors faced by weaned piglets during weaning, including dietary changes, environmental changes, weakened immune function and imbalance of microbiota, they are susceptible to bacterial, viral and other infections, leading to local or systemic inflammatory responses. The intestine of piglets is an important part of their digestion and absorption of nutrients. Intestinal inflammation can damage the intestinal mucosa, resulting in impaired digestion and absorption function and slow growth and development of piglets. The high incidence of diarrhea in weaned piglets is closely related to intestinal inflammation, especially when the use of antibiotics in feed is limited, the prevention and control of diarrhea is more challenging. The high incidence of diarrhea in weaned piglets will not only affect their growth and development, but also increase the treatment costs and mortality of farms.

[0003] Lactobacillus amylovora Lactobacillus amylovorus ) is the dominant flora in the intestine of piglets and is also used as a probiotic in fermented foods, but its application research in intestinal health is limited. Chrysin has been shown to have significant anti-inflammatory, antioxidant and immunomodulatory effects. It is naturally present in many plants, honey and propolis. In recent years, studies have shown that some fungi can synthesize chrysin or its analogs through metabolic pathways, thereby exerting anti-inflammatory effects. However, there are no reports on the synthesis of chrysin by bacteria, especially probiotics. Lactobacillus amylovora can not only grow and colonize effectively in the intestine, but also synthesize a variety of biologically active natural compounds through its metabolic activities. However, there are few studies on the relief of intestinal inflammation by Lactobacillus amylovora through the production of chrysin. Therefore, it is of great significance to explore the application of Lactobacillus amylovora that can produce chrysin in the prevention and treatment of intestinal inflammation in piglets, which can not only promote the health of piglets, but also reduce the use of antibiotics and enhance the sustainable development capacity of animal husbandry. Summary of the invention

[0004] The present invention aims to provide a strain of Lactobacillus amylovora (Lactobacillus amylovora) that can produce chrysin. Lactobacillus amylovorus ).

[0005] Another object of the present invention is to provide the above-mentioned Lactobacillus amylovora ( Lactobacillus amylovorus ) application.

[0006] Another object of the present invention is to provide a biological bacterial agent.

[0007] Another object of the present invention is to provide a feed additive.

[0008] The present invention is lactobacillus amylovora Lactobacillus amylovorus ) SKLAN202401ZF, its deposit number is CGMCC No.30022.

[0009] The present invention provides a method for preparing a lactobacillus starch containing Lactobacillus amylovorus ) SKLAN202401ZF. The bacterial agent can be a liquid bacterial agent or a solid bacterial agent, and can be prepared by conventional technical means and by adding auxiliary materials permitted in the field of microbial preparations.

[0010] The present invention provides amylovorax lactobacillus ( Lactobacillus amylovorus ) Application of SKLAN202401ZF for preparing the following preparations:

[0011] Preparations that inhibit pathogenic microorganisms;

[0012] Preparations for promoting animal growth;

[0013] Preparations that enhance intestinal immunity; or

[0014] Preparations that relieve intestinal inflammation.

[0015] Preferably, the pathogen is a bacterium, more preferably a Gram-negative bacterium.

[0016] As an embodiment of the present invention, the pathogenic microorganism is Escherichia coli or Salmonella.

[0017] According to the animal feed additive of the present invention, it contains the above-mentioned Lactobacillus amylovora ( Lactobacillus amylovorus ) SKLAN202401ZF or Lactobacillus amylovora ( Lactobacillus amylovorus )SKLAN202401ZF is prepared, specifically, containing Lactobacillus amylovora ( Lactobacillus amylovorus )The bacteria obtained by fermentation of SKLAN202401ZF.

[0018] The present invention also provides the above-mentioned Lactobacillus amylovora ( Lactobacillus amylovorus )Application of SKLAN202401ZF in the production of chrysin.

[0019] The beneficial effects of the present invention are:

[0020] The present invention provides a novel amylovorax lactobacillus ( Lactobacillus amylovorus )SKLAN202401ZF, which can produce chrysin in the intestines of animals and has excellent antibacterial and immunomodulatory functions. By adding this strain to piglet feed, the intestinal health of piglets can be significantly improved, the incidence of diarrhea can be reduced, and the feed conversion rate and growth rate can be increased. The application of this strain not only helps to improve the efficiency of pig farming, but also can replace antibiotics and is suitable for antibiotic-free farming.

[0021] Compared with the existing amylovora bacteria, the amylovora bacteria SKLAN202401ZF disclosed in the present application has significant differences in the following aspects:

[0022] 1. Pathogenic bacteria inhibition effect: The strain showed significant inhibitory effect on Salmonella and Escherichia coli;

[0023] 2. It has a strong ability to colonize in animals and has a strong ability to tolerate artificial bile salts, artificial gastric juice and artificial intestinal juice. It is particularly suitable for long-term effects in animals, as well as improving animal intestinal health, alleviating intestinal inflammation after E. coli infection, reducing diarrhea rates and improving animal growth performance;

[0024] 3. The bacteria can synthesize and secrete chrysin, which has significant anti-inflammatory effects, in the animal intestine during its metabolism, which makes the bacteria show unique advantages in relieving intestinal inflammation. Although most other lactobacilli have certain anti-inflammatory effects, they do not have the ability to synthesize or secrete metabolites with strong anti-inflammatory effects such as chrysin;

[0025] 4. Multi-pathway anti-inflammatory effect: The bacteria relieve intestinal inflammation by metabolizing chrysin and other related metabolites. Chrysin inhibits pro-inflammatory cytokines (such as IL-1β and NLRP3) and enhances intestinal barrier function, thereby achieving a more comprehensive anti-inflammatory effect. In contrast, many other amyloliquefaciens may only work by promoting the growth of beneficial bacteria or directly competing to inhibit pathogens, and the diversity and depth of their anti-inflammatory effects are relatively limited;

[0026] 5. Wide applicability: This bacterium relieves intestinal inflammation by self-metabolizing chrysin, can adapt to different intestinal environments (including the intestinal environment of piglets and mice), and shows good therapeutic effects in a variety of intestinal diseases. Although other lactobacilli are used in certain intestinal diseases, their effects are often limited by strain differences, host specificity and environmental factors. The characteristics of this bacterium in metabolizing chrysin enable it to provide more stable and reliable effects in a variety of clinical applications. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 For the Lactobacillus amylovora in Example 2 of the present invention ( Lactobacillus amylovorus ) Growth curve of SKLAN202401ZF;

[0028] Figure 2A and Figure 2B are respectively Lactobacillus amylovora ( Lactobacillus amylovorus ) Colony morphology of SKLAN202401ZF and oil microscopic examination after malachite green staining;

[0029] Figure 3 This is a graph showing the weight changes of mice in the experiment in Example 3 of the present invention;

[0030] Figure 4 It shows that the starch lactobacillus ( Lactobacillus amylovorus ) Effects of SKLAN202401ZF on intestinal inflammation in mice, A: HE staining of paraffin sections, B: and mouse colon;

[0031] Figure 5 It shows that in Example 3, Lactobacillus amylovora ( Lactobacillus amylovorus ) Effects of SKLAN202401ZF on intestinal inflammation in mice, including: A: colon length, B: colon crypt depth, C: colon paraffin section inflammation score, D: colon paraffin section intestinal epithelial injury score;

[0032] Figure 6 is the amylovorax lactobacillus ( Lactobacillus amylovorus ) Colonization of SKLAN202401ZF in mouse colon;

[0033] Figure 7 This is the VIP analysis of differential metabolites in mouse colon chyme in Example 3 of the present invention, wherein A: L.am + E.coli Group and E.coli Analysis of VIP values ​​of metabolites in different groups, B: L.am + E.coli Analysis of VIP values ​​of differential metabolites between the group and CON group;

[0034] Figure 8 This is an analysis of the weight changes and inflammation conditions of the mice in Example 4 of the present invention, wherein A: mouse weight, B: relative expression of IL-1β in mouse colon, and C: relative expression of NLRP3 in mouse colon inflammatory body.

[0035] Figure 9 This is an analysis of the weight change and diarrhea of ​​piglets in Example 5 of the present invention, wherein A: piglet weight, B: piglet diarrhea rate.

[0036] Lactobacillus amylovora Lactobacillus amylovorus ) SKLAN202401ZF, deposited on March 13, 2024 in the General Microbiology Center of China Microorganism Culture Collection Administration (referred to as CGMCC, address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, Postal Code 100101), and classified as Lactobacillus amylovora Lactobacillus amylovorus , the deposit number is CGMCC No.30022. DETAILED DESCRIPTION

[0037] The technical scheme of the present invention is described clearly and completely below in conjunction with specific embodiments. Lactobacillus amylovorus ) SKLAN202401ZF, all reagents and raw materials used were commercially available.

[0038] Example 1 Isolation and identification of Lactobacillus amylovorus SKLAN202401ZF strain

[0039] Isolated from the colonic digesta of healthy 38-day-old Du×Chang×Da piglets fed with the traditional Chinese medicine baicalin.

[0040] (1) Colonic chyme collection: Ten healthy and strong 38-day-old piglets of the Du×Chang×Da type that had been fed with a diet containing 100 mg / kg baicalin for 2 weeks were selected and slaughtered to separate the colons of the piglets. The intestinal segments were opened and 1 g of colonic chyme was collected from each piglet. The chyme was added to 20 mL of sterile saline containing 20% ​​glycerol and vortexed to mix. The chyme was then dispensed into sterile cryovials, 1.5 mL per tube, and quickly frozen in liquid nitrogen and stored in a -80°C refrigerator.

[0041] (2) Strain isolation: Take 1 mL of the piglet colon chyme preserved in step (1), dilute it 10-fold with sterile saline, and evenly spread 0.1 mL of the dilution on MRS agar medium. Incubate at 37°C for 48 h. Inoculate the grown colonies onto new MRS agar medium by streaking and purify them for 48 h.

[0042] (3) Strain enrichment: The purified strain was inoculated into sterile MRS broth medium using a sterile inoculation loop and placed in a shaker. After culturing at 37°C for 48 h, 16S rDNA identification was performed and the remaining portion was diluted with physiological saline containing 20% ​​glycerol and stored in a -80°C refrigerator.

[0043] (4) 16S rDNA identification: The enriched bacterial solution was subjected to colony PCR amplification using bacterial universal primers, and then 16S rDNA sequencing was performed for identification. The 16S rDNA sequence of each strain was compared with the 16S rDNA sequences of all bacteria that had been determined in the database on NCBI. Based on the sequencing results, microbiological characteristics, and physiological and biochemical properties, 4 strains were identified as Lactobacillus amylovora. The strain selected in the present invention had a similarity of 99.73% with Lactobacillus amylovora and was identified as Lactobacillus amylovora.

[0044] Example 2 Detection of characteristics of Lactobacillus amylovorus SKLAN202401ZF

[0045] 1. Growth curve determination

[0046] Lactobacillus amylovora Lactobacillus amylovorus ) SKLAN202401ZF was cultured until the concentration reached 8×10 6 CFU / mL aspirate the bacterial solution and inoculate it into MRS broth at a volume of 1% of the culture medium, and place it in a shaker at 37°C. Use an enzyme-labeled instrument to measure the OD value of the culture medium every 2 hours. 600 The result shows ( Figure 1 )Lactobacillus amylovora Lactobacillus amylovorus ) SKLAN202401ZF strain reached the logarithmic growth phase after 4 hours and entered the plateau phase after 6 hours, with the maximum number of viable bacteria reaching 8×10 6 CFU / mL.

[0047] 2. Morphology and Colony Observation

[0048] Smear the bacteria on a glass slide and dry it, then stain it with 1% malachite green for 3 minutes. After staining, slowly rinse it with deionized water to remove the excess staining solution, dry the slide, and observe and take pictures under an oil lens of a microscope. Lactobacillus amylovorus ) SKLAN202401ZF shape Figure 2B The bacterial solution was spread on MRS agar medium and cultured at 37℃ for 48h before observing the colony morphology. Figure 2A .

[0049] Lactobacillus amylovora Lactobacillus amylovorus ) The microbiological characteristics of SKLAN202401ZF are as follows:

[0050] Colony morphology: Single colony is round, milky white, uniform in color, with neat edges and smooth surface. Lactobacillus amylovorus ) SKLAN202401ZF bacteria are rod-shaped, non-motile, and have no flagella.

[0051] Growth characteristics: In MRS broth medium, placed in a 37°C shaker culture, it enters the logarithmic growth phase after 4 hours and enters the plateau phase after 6 hours. The maximum number of viable bacteria is 8×10 6 CFU / mL.

[0052] 3. Antibacterial ability test

[0053] Two pathogenic bacteria were selected as pathogen indicator bacteria: Escherichia coli ( Escherichia coli ) BW25113 and Salmonella typhimurium ( Salmonella typhimurium )ATCC14028.

[0054] The antibacterial activity of the strains was determined by the Oxford cup method. Escherichia coli) BW25113 and Salmonella typhimurium ( Salmonella typhimurium )ATCC14028 were inoculated into LB broth medium and cultured in a 37°C incubator for 12-18 hours. Escherichia coli ) BW25113 and Salmonella typhimurium ( Salmonella typhimurium )ATCC14028 strain concentration was adjusted to 1×10 8 CFU / mL, take 200μl of each, evenly spread on LB agar medium, put the sterilized Oxford cup evenly on the LB medium, add 100μl of activated bacterial solution with a concentration of 8×10 6 CFU / mL of bacterial solution with MRS broth. Place in a 37°C incubator for 12 hours. Use a vernier caliper to measure the size of the inhibition zone and evaluate the antibacterial effect. The results show (Table 1) that compared with the other three lactobacilli, Lactobacillus amylovora Lactobacillus amylovorus ) SKLAN202401ZF can better inhibit the growth of Salmonella and Escherichia coli.

[0055] Table 1 Inhibitory ability of Lactobacillus amylovora SKLAN202401ZF against pathogens

[0056] Strain number Salmonella Escherichia coli SKLAN202419ZF 0.17 0.32 SKLAN202401ZF 0.22 0.38 SKLAN202406ZF 0.20 0.30 SKLAN202404ZF 0.15 0.27

[0057] Note: The above values ​​are the diameters of the inhibition zones, in cm.

[0058] 4. Bile salt tolerance test

[0059] Take 100uL of enriched cultured Lactobacillus amylovora ( Lactobacillus amylovorus ) SKLAN202401ZF bacterial solution (concentration of 8×10 6 CFU / mL) was added to 900uL of 0.3% bile salt solution, incubated in a 37°C incubator for 1 hour, and 100uL of bacterial solution was spread on a plate and counted. The results are shown in Table 2. Compared with other lactobacilli, Lactobacillus amylovora Lactobacillus amylovorus ) SKLAN202401ZF has stronger bile salt tolerance, with a survival rate of 11.1% in 0.3% bile salt.

[0060] Table 2 Bile salt tolerance of Lactobacillus amylovora SKLAN202401ZF

[0061] Strain number Survival rate in 0.3% bile salts, % SKLAN202419ZF 10.2 SKLAN202401ZF 11.1 SKLAN202406ZF 7.0 SKLAN202404ZF 10.1 .

[0062] 5. Artificial gastric fluid resistance test

[0063] Take 10uL of enriched cultured Lactobacillus amylovora ( Lactobacillus amylovorus) SKLAN202401ZF bacterial solution (concentration of 8×10 6 CFU / mL) was added to 990uL of artificial gastric juice with a pH value of 1.2, and cultured in a 37°C incubator for 2.5 hours. 100uL of the bacterial solution was spread on a plate and counted. The results are shown in Table 3. In artificial gastric juice with a pH value of 1.2, Lactobacillus amylovora ( Lactobacillus amylovorus ) The survival rates of SKLAN202401ZF and 202419 were higher than those of the other two lactobacilli strains.

[0064] Table 3 Resistance of Lactobacillus amylovora to artificial gastric juice

[0065] Strain number Survival rate in artificial gastric juice, % SKLAN202419ZF 0.42 SKLAN202401ZF 0.41 SKLAN202406ZF 0.01 SKLAN202404ZF 0.18 .

[0066] 6. Artificial intestinal fluid resistance test

[0067] Take 10uL of enriched cultured Lactobacillus amylovora ( Lactobacillus amylovorus ) SKLAN202401ZF bacterial solution (concentration of 8×10 6 CFU / mL) were added to 990uL of artificial intestinal fluid with a pH value of 4, and cultured in a 37°C incubator for 4 hours. 100uL of the bacterial solution was spread on a plate and counted. The results are shown in Table 4. After 4 hours in artificial intestinal fluid, Lactobacillus amylovora ( Lactobacillus amylovorus ) The survival rate of SKLAN202401ZF was higher than that of the other three strains, reaching 79.4%.

[0068] Table 4 Resistance of Lactobacillus amylovora to artificial intestinal fluid

[0069] Strain number Survival rate in artificial intestinal fluid, % 202419 25.0 202401 79.4 202406 12.6 202404 19.1 .

[0070] 7. Metabolite Detection

[0071] Lactobacillus amylovora Lactobacillus amylovorus ) SKLAN202401ZF was cultured until the concentration reached 8×10 6 CFU / mL was obtained by centrifugation, and clean cells were obtained after washing 3 times with normal saline. The cells were used for non-targeted metabolomics to detect their metabolite components. The results are shown in Table 5. Lactobacillus amylovora SKLAN202401ZF can produce metabolites such as lactic acid, isocitrate, methylmalonic acid, indoleacrylic acid, and taurocholic acid, which have been reported to have anti-inflammatory and antibacterial effects. In addition, Lactobacillus amylovora SKLAN202401ZF also produces chrysin, which has not been reported in previous literature.

[0072] Table 5 Metabolites of Lactobacillus amylovora SKLAN202401ZF

[0073] Metabolites Relative abundance Chrysin 119.89 Isocitrate 369230 lactic acid 172165 Methylmalonic acid 87198 Indole acrylic acid 88366 Taurocholic acid 36409 .

[0074] Example 3 Effects of Lactobacillus amylovorus SKLAN202401ZF on intestinal inflammation in mice

[0075] In the following experiments, Lactobacillus amylovora Lactobacillus amylovorus ) The concentration of SKLAN202401ZF bacterial solution is 8×10 6 CFU / mL.

[0076] 1. Test methods

[0077] Thirty-six 21-day-old female ICR mice were randomly divided into three groups: control group (CON) and intestinal inflammation group ( E. coli )、Lactobacillus amylovora Lactobacillus amylovorus ) SKLAN202401ZF group ( L.am + E. coli ). L.am + E. coli Mice in this group were gavaged with 0.2 mL of Lactobacillus amylovorax daily. Lactobacillus amylovorus ) SKLAN202401ZF, administered intragastrically for 3 weeks. E. coli Group and L.am + E. coli On the 22nd, 23rd, and 24th days of the experiment, mice in the group were injected intraperitoneally with a concentration of 2×10 9 CFU / mL of Escherichia coli ( Escherichia coli ) 0.1mL, 0.2mL again after 24 hours, and 0.3mL injected after 48 hours to construct an intestinal inflammation model. The control group was injected with an equal amount of normal saline. The mice ate and drank water normally during the experiment. The weight of the mice was recorded at the beginning and end of the experiment.

[0078] 2. Test results

[0079] (1) Changes in mouse weight

[0080] The weight changes of mice during the whole experimental period were as follows Figure 3 As shown in the figure, there was no significant difference in the body weight of mice in each group within 3 weeks before the experiment. L.am + E. coli The body weight of mice in the CON group was significantly higher than E. coli Group( p <0.05).

[0081] (2) Colon length and crypt depth

[0082] like Figure 4Figure B and Figure 5 As shown in Figure A, L.am + E. coli The colon length of mice in the CON group was significantly higher than that in the E. coli Group. H&E staining was used to measure the depth of mouse colon crypts. The results showed L.am + E. coli The colon crypt depth of mice in the CON group was significantly higher than that in the E. coli Group( p <0.05, Figure 5 Figure B in the figure).

[0083] (3) Colon morphology and pathological scoring

[0084] like Figure 4 As shown in Figure A, E. coli The colon of mice in the control group showed severe inflammatory cell infiltration and epithelial damage. Pathological scores showed that L.am + E. coli The colonic inflammation scores and epithelial injury scores of mice in the CON group were significantly lower than those in the E. coli Group( p <0.05, Figure 5 Figure C and Figure 5 Figure D in the figure).

[0085] (4) Colonization effect of Lactobacillus amylovora SKLAN202401ZF in mouse colon

[0086] Using Lactobacillus amylovora Lactobacillus amylovorus ) SKLAN202401ZF sequence was used to design a probe and perform fluorescence in situ hybridization to detect the colonization of Lactobacillus amylovora SKLAN202401ZF in mouse colon. Figure 6 As shown, after gavage with Lactobacillus amylovora SKLAN202401ZF L.am + E. coli The colonization of Lactobacillus amylovora in the mice group was significantly higher than that in the control group and E. coli.

[0087] (5) Analysis of differential metabolites in mouse colon

[0088] like Figure 7 As shown in Figure A, L.am + E. coli and E. coli Chrysin had the highest VIP value among the differential metabolites in the colonic chyme of mice in the L.am + E. coli group was significantly upregulated ( p <0.05). Figure 7 As shown in Figure B, L.am + E. coliCompared with the CON group, the abundance of chrysin was also significantly upregulated ( p <0.05).

[0089] Example 4 Effects of Lactobacillus amylovorus SKLAN202401ZF on Growth Performance and Diarrhea Rate in Piglets

[0090] 1. Test methods

[0091] Thirty-six 21-day-old female ICR mice were randomly divided into three groups: control group (CON) and intestinal inflammation group ( E. coli ), chrysin group + E. coli ). + E. coli The mice in group A were gavaged with 10 mg / kg body weight of chrysin every day for 3 consecutive weeks. E. coli Group and chrysin + E. coli On the 22nd, 23rd, and 24th days of the experiment, mice in the group were injected intraperitoneally with 2×10 9 CFU / mL of Escherichia coli ( Escherichia coli ) 0.1mL, 0.2mL again after 24 hours, and 0.3mL injected after 48 hours to construct an intestinal inflammation model. The control group was injected with an equal amount of normal saline. The mice ate and drank water normally during the experiment. The weight of the mice was recorded at the beginning and end of the experiment.

[0092] 2. Test results

[0093] (1) Changes in mouse weight

[0094] Mouse weight Figure 8 As shown in Figure A, after the experiment, chrysin + E. coli The body weight of mice in the CON group was significantly higher than E. coli Group( p <0.05). This indicates that chrysin can increase the body weight of mice under inflammatory conditions.

[0095] (2) Analysis of colonic inflammatory factor expression

[0096] like Figure 8 As shown in Figures B and C, E. coli The relative expression levels of inflammatory factors IL-1β and inflammasome NLRP3 in the colon of mice in the control group were significantly higher than those in the control group, while chrysin + E. coli The relative expression levels of IL-1β and NLRP3 in the E. coli Group( p<0.05). This indicates that chrysin can significantly alleviate intestinal inflammation in mice.

[0097] Example 5 Lactobacillus amylovora Lactobacillus amylovorus ) Effects of SKLAN202401ZF on growth performance and diarrhea rate of piglets

[0098] In the following experiments, Lactobacillus amylovora Lactobacillus amylovorus ) The concentration of SKLAN202401ZF bacterial solution is 8×10 6 CFU / mL.

[0099] 1. Test methods

[0100] Sixteen 21-day-old weaned piglets were randomly divided into two groups: control group (CON) and Lactobacillus amylovora ( Lactobacillus amylovorus ) SKLAN202401ZF group ( L.am ). L.am Piglets in group A were fed with 8×10 6 CFU / kg of feed was fed for 17 consecutive days, and the control group was fed with basic feed. During the experimental period, the piglets were fed according to the routine feeding and management procedures of the pig farm, and they were free to eat and drink water, and were immunized according to the immunization procedures of the pig farm. The weight of the piglets was recorded at the beginning and end of the experiment.

[0101] 2. Test results

[0102] (1) Piglet weight changes

[0103] like Fig. 9 As shown in Figure A, after the test L.am The weight of piglets in group A was significantly higher than that in group B, indicating that Lactobacillus amylovora Lactobacillus amylovorus ) SKLAN202401ZF can promote the growth of piglets ( p <0.05).

[0104] (2) Diarrhea rate

[0105] like Fig. 9 As shown in Figure B, L.am The diarrhea rate of piglets in group A was significantly lower than that in control group ( p <0.05). Description Lactobacillus amylovora Lactobacillus amylovorus ) SKLAN202401ZF can prevent diarrhea in piglets caused by weaning stress ( p <0.05).

[0106] The above descriptions are only some specific implementation methods of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be easily thought of by any person familiar with the art within the technical scope disclosed in the present invention should be covered within the protection scope of the present invention.

Claims

1. Lactobacillus amylovora Lactobacillus amylovorus ) SKLAN202401ZF, characterized in that: The Lactobacillus amylovora Lactobacillus amylovorus ) The deposit number of SKLAN202401ZF is CGMCC No.30022.

2. A method comprising the lactobacillus starch according to claim 1 ( Lactobacillus amylovorus ) Microbial agent of SKLAN202401ZF.

3. The microbial agent according to claim 2, characterized in that: The microbial agent is a liquid microbial agent or a solid microbial agent.

4. The amylovorax lactobacillus of claim 1 ( Lactobacillus amylovorus ) Application of SKLAN202401ZF for preparing the following preparations: Preparations for inhibiting pathogenic microorganisms, wherein the pathogenic microorganisms are Escherichia coli or Salmonella; Preparations for promoting animal growth; A preparation for treating inflammation of the colon in animals; or Preparations for reducing the rate of diarrhea in animals.

5. An animal feed additive, characterized in that: Containing the amylovora bacteria according to claim 1 ( Lactobacillus amylovorus ) SKLAN202401ZF, or the amylovora bacillus according to claim 1 ( Lactobacillus amylovorus ) SKLAN202401ZF is prepared by fermentation.

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