Application of selenium-enriched lactobacillus reuteri in prevention and treatment of enteropathogenic bacteria infection and promotion of growth of chicks
The selenium-rich Lactobacillus reuteri obtained through selenium-rich culture is used to prevent and treat intestinal pathogenic bacteria infection in poultry and promote the growth of chicks, solving the problems of disease and bacterial infection in poultry breeding industry, and achieving safe and effective antibacterial and growth-promoting effects.
Patent Information
- Application Number
- CN202411914843.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-24
- Publication Date
- 2025-03-25
AI Technical Summary
In the poultry farming industry, as the scale of breeding expands and becomes intensive, poultry diseases and bacterial infections are serious. Traditional antibiotic treatment leads to bacterial resistance problems, and it is necessary to find safe and effective alternatives.
Lactobacillus reuteri was used to prevent and treat intestinal pathogenic bacteria infection and promote chick growth by placing Lactobacillus reuteri in culture medium with sodium selenite content of 5 to 7 mg/mL.
Lactobacillus reuteri selenium-rich Lactobacillus significantly promotes the growth of chickens, effectively inhibits the growth of pathogens, reduces the risk of bacterial infection in chicken flocks, improves breeding benefits, and has the advantages of being safe and without side effects.
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Figure CN119679832A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of microorganisms, and particularly relates to application of selenium-enriched Lactobacillus reuteri in preventing and treating intestinal pathogenic bacteria infection and promoting the growth of chicks. Background Art
[0002] In recent years, the poultry farming industry has developed rapidly around the world. However, with the expansion and intensification of farming scale, poultry diseases and bacterial infections have become increasingly serious, such as E. coli and Salmonella, which lead to growth stunting and frequent diseases. Although traditional antibiotic treatment can control the disease to a certain extent, long-term use of antibiotics can lead to bacterial resistance, which in turn threatens public health. Therefore, it is crucial to find safe and effective alternatives. Probiotics, as a natural and safe microbial preparation, have been widely used in animal farming.
[0003] Lactobacillus reuteri is an important probiotic that is widely found in the intestines of humans and animals. Studies have shown that Lactobacillus reuteri has good growth-promoting and antibacterial effects. However, there are few studies on the application of Lactobacillus reuteri in chickens. Currently, there are few known antibacterial species of Lactobacillus reuteri, and its antibacterial effect is limited. Summary of the invention
[0004] In view of the above technical problems, the present invention provides the use of selenium-enriched Lactobacillus reuteri in preventing and treating intestinal pathogenic bacteria infection and promoting the growth of chicks.
[0005] The present invention adopts the following technical solution:
[0006] Selenium-enriched Lactobacillus reuteri is used in preventing and treating intestinal pathogenic bacteria infection and promoting the growth of chicks. The selenium-enriched Lactobacillus reuteri is obtained by selenium-enriched culture of Lactobacillus reuteri (Lactobacillus reuteri) with a preservation number of CGMCC No.31975. The selenium-enriched culture is to place the Lactobacillus reuteri in a culture medium with a sodium selenite content of 5-7 mg / mL for culture. The present invention evaluates the antibacterial effect on four intestinal pathogenic bacteria (Vibrio parahaemolyticus, Escherichia coli K99, Vibrio cholerae, Salmonella typhimurium) and four probiotics (Lactococcus lactis, Lactobacillus plantarum, Lactobacillus casei, Lactobacillus delbrueckii), takes double-resistance penicillin-streptomycin as a positive control, and sets MRS culture medium as a control group. The results show that the metabolites of Lactobacillus reuteri all show an inhibitory effect on pathogenic bacteria, have a slight inhibitory effect on Lactococcus lactis, and have no inhibitory effect on the remaining probiotics, indicating that Lactobacillus reuteri can be prepared into a safe antibacterial agent or a preparation for preventing chicken bacterial infection.
[0007] Selenium-enriched probiotics are a special type of probiotics, which not only have the functions of general probiotics, but are also rich in selenium. Selenium is an essential trace element for animal growth and immune function, and has multiple biological functions such as anti-oxidation, immunity enhancement and anti-tumor. The selenium-enriched Lactobacillus reuteri obtained by culturing the Lactobacillus reuteri with selenium in the present invention has a stronger effect of promoting chicken growth and preventing bacterial infection.
[0008] Furthermore, the prevention is reflected in alleviating the slow growth of chicks caused by the intestinal pathogenic bacteria infection.
[0009] Furthermore, the prevention is manifested in enhancing the cellular immune response of the chicks.
[0010] Furthermore, the selenium-enriched culture is based on MRS liquid culture medium, with the addition of 15-25 mg / mL CaCO3, 5-15 mg / mL glutathione and 1-2 mg / mL ammonium citrate, and 5-7 mg sodium selenite is added to each 1 mL of the basic culture medium.
[0011] Furthermore, 1-10 g of the Lactobacillus reuteri is inoculated per 100 ml of the basic culture medium.
[0012] Furthermore, the sodium selenite is added 4 to 8 hours after inoculating the Lactobacillus reuteri.
[0013] Furthermore, the temperature of the selenium-enriched culture is 36-37° C., and the time is 22-24 hours.
[0014] The present invention has the following beneficial effects:
[0015] The Lactobacillus reuteri in the present invention has excellent selenium resistance. The selenium-enriched Lactobacillus reuteri obtained by selenium-enriched culture can not only significantly promote the growth of chickens, but also effectively inhibit the growth of pathogens, reduce the risk of bacterial infection in chickens, and improve breeding efficiency. At the same time, since Lactobacillus reuteri YG 01 is enriched with selenium in a natural way, the use of selenium supplements is avoided, and it has the advantages of safety and no side effects, providing a new, safe and effective solution for the poultry breeding industry.
[0016] Storage Instructions:
[0017] Bacteria species name: Lactobacillus reuteri;
[0018] Latin name: Lactobacillus reuteri;
[0019] Strain number: YG 01;
[0020] Depository: China General Microbiology Center of Microbiological Culture Collection Administration (CGMCC for short);
[0021] Address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing;
[0022] Deposit date: September 14, 2024;
[0023] Deposit number: CGMCC No.31975. BRIEF DESCRIPTION OF THE DRAWINGS
[0024] Figure 1 This is the Gram staining result of Lactobacillus reuteri YG 01, 1000×.
[0025] Figure 2 The status of individual chickens (A) and chicken groups (B) in experimental group I.
[0026] Figure 3 The status of individual chickens (A) and chicken groups (B) in Experiment II group.
[0027] Figure 4 The status of individual chickens (A) and chicken groups (B) in Experiment III group.
[0028] Figure 5 The status of individual chickens (A) and chicken groups (B) in Experimental Group IV.
[0029] Figure 6 The status of individual chickens (A) and chicken groups (B) in experimental group V.
[0030] Figure 7 The status of individual chickens (A) and chicken groups (B) in experimental group VI.
[0031] Figure 8 The status of a single chicken (A) and a flock of chickens (B) in the blank control group.
[0032] Fig. 9 For weight gain.
[0033] Fig.10 The figure shows the changes in body weight after infection.
[0034] Fig.11 Clinical scoring.
[0035] Fig.12 is the serum SOD level of each group.
[0036] Fig.13 is the serum CAT level of chickens in each group.
[0037] Fig.14 is the serum IL-10 level of chickens in each group.
[0038] Fig.15 Serum IL-4 levels of chickens in each group.
[0039] Fig.16 is the serum IL-6 level of chickens in each group.
[0040] Fig.17 is the serum TNF-α level of chickens in each group.
[0041] In the above figure: K99 represents experimental group III, K99+LR represents experimental group I, K99+Se represents experimental group II, S represents experimental group VI, S+LR represents experimental group IV, S+Se represents experimental group V, and Control represents the blank control group. DETAILED DESCRIPTION
[0042] The present invention is described in detail below in conjunction with the accompanying drawings and specific examples, but should not be construed as limiting the present invention. Unless otherwise specified, the technical means used in the following examples are conventional means well known to those skilled in the art, and the materials, reagents, etc. used in the following examples, unless otherwise specified, can be obtained from commercial sources.
[0043] Example 1: Isolation and identification of Lactobacillus reuteri
[0044] A strain of Lactobacillus reuteri was isolated from pig feces. The Lactobacillus reuteri was streaked onto an MRS agar plate and cultured at 37°C for 24 hours. Milky white colonies with protrusions or flattened in the center were observed to grow, with regular edges, smooth, shiny, opaque, and soft texture. Gram staining results showed that it was Gram-positive and the bacteria were rod-shaped, see Figure 1 .
[0045] After sequencing and homology comparison, the homology between the strain and the 16S ribosomalRNA gene of Lactobacillus reuteri strain 455 was as high as 100%, indicating that the strain was Lactobacillus reuteri. The strain was subjected to selenium resistance acclimation treatment to obtain Lactobacillus reuteri with strong tolerance to sodium selenite, named YG 01. YG 01 can still grow normally at a sodium selenite concentration of 8 mg / ml.
[0046] Example 2: Selenium-enriched culture transformation of Lactobacillus reuteri YG 01
[0047] 6 mg / mL sodium selenite was added to MRS liquid culture medium as a basic selenium-enriched culture medium, the inoculation amount of Lactobacillus reuteri YG 01 was 5 g / 100 ml, sodium selenite was added at the 8th hour, 20 mg / mL CaCO3, 15 mg / mL glutathione and 2 mg / mL ammonium citrate were added to the culture medium, and the culture was cultured at 37°C for 24 hours to obtain selenium-enriched Lactobacillus reuteri.
[0048] Example 3: Antibacterial test of Lactobacillus reuteri YG 01
[0049] The antibacterial effect of the filtered sterilized supernatant (fine-tuned / adjusted to pH 7.0) after culturing Lactobacillus reuteri for 24 hours was evaluated on four pathogenic bacteria and four probiotics. The double-resistant penicillin-streptomycin was used as the positive control, and MRS culture medium was set as the control group. The results are shown in Table 1. The metabolites of Lactobacillus reuteri showed an inhibitory effect on pathogenic bacteria, a slight inhibitory effect on Lactococcus lactis, and no inhibitory effect on the remaining probiotics.
[0050] Table 1 Antibacterial test results (diameter of antibacterial ring / mm)
[0051]
[0052]
[0053] Example 4: Effects of Selenium-Enriched Lactobacillus reuteri on Chicken Growth and Resistance to Bacterial Infection
[0054] One-day-old chicks were divided into seven groups (10 in each group). The amount of oral bacteria was calculated in CFU / g (chicken weight). The specific grouping and operation were as follows:
[0055] In the experimental group I, the oral dose was 1×10 9 CFU / g of Lactobacillus reuteri, oral infection with 1×10 8 CFU / g of E. coli K99, the status of the infected chickens Figure 2 shown.
[0056] Experimental group II, oral dose of 1×10 9 CFU / g of selenium-enriched Lactobacillus reuteri, and oral infection with 1×10 8 CFU / g of E. coli K99, the status of the infected chickens Figure 3 shown.
[0057] In the experimental group III, 1 mL of sterile PBS was orally administered for 42 consecutive days and then 1×10 8 CFU / g of E. coli K99, the status of the infected chickens Figure 4 shown.
[0058] Experimental group IV, oral dose of 1×10 9 CFU / g Lactobacillus reuteri, oral infection 1×10 8 CFU / g of Salmonella typhimurium, the status of the infected chickens Figure 5 shown.
[0059] Experimental group V, oral dose of 1×10 9 CFU / g of selenium-enriched Lactobacillus reuteri, oral infection 1×10 8 CFU / g of Salmonella typhimurium, the status of the infected chickens Figure 6 shown.
[0060] In the experimental group VI, 1×10 8 CFU / g of Salmonella typhimurium, the status of the infected chickens Figure 7 shown.
[0061] The control group was fed orally with 1 mL of sterile PBS for 42 consecutive days as a blank control group. Figure 8 shown.
[0062] Weight results are as follows Fig. 9 As shown, the weight gain rate of the groups fed with Lactobacillus reuteri and selenium-enriched Lactobacillus reuteri was significantly higher than that of the control group.
[0063] Fig.10 Figure 2 shows the weight changes of each group after bacterial infection. The weight of the groups infected with Escherichia coli K99 and Salmonella decreased, while the weight of the groups fed with Lactobacillus reuteri and selenium-enriched Lactobacillus reuteri showed a normal growth trend.
[0064] An animal clinical scoring table was developed as shown in Table 2, and the infected animals in each group were scored according to the scoring table. The results are as follows: Fig.11 shown.
[0065] Table 2 Animal clinical scores
[0066]
[0067] Chicken serum from each group was collected and the levels of IL-10, IL-4, IL-6, TNF-α, CAT and SOD in the serum were determined by ELISA.
[0068] like Figure 12-Figure 17 The results showed that oral administration of selenium-enriched Lactobacillus reuteri could enhance the body's cellular immune response, and the regulation level was higher than that of the group fed only with Lactobacillus reuteri.
[0069] Although the preferred embodiments of the present invention have been described, those skilled in the art may make additional changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.
[0070] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.
Claims
1. Application of selenium-enriched Lactobacillus reuteri in preventing and treating intestinal pathogenic bacteria infection and promoting growth of chicks, characterized in that: The selenium-enriched Lactobacillus reuteri is obtained by culturing Lactobacillus reuteri with a preservation number of CGMCC No. 31975 through selenium enrichment. The selenium-enriched culture is to culture the Lactobacillus reuteri in a culture medium with a sodium selenite content of 5-7 mg / mL.
2. The use of selenium-enriched Lactobacillus reuteri in preventing and treating intestinal pathogenic bacteria infection and promoting the growth of chicks according to claim 1, characterized in that: The intestinal pathogenic bacteria include Vibrio cholerae, Escherichia coli, Vibrio cholerae and Salmonella typhimurium.
3. The selenium-enriched Lactobacillus reuteri according to claim 2 is used in preventing and treating intestinal pathogenic bacteria infection and promoting the growth of chicks, characterized in that: The prevention is manifested in alleviating the slow growth of chicks caused by the infection of the intestinal pathogenic bacteria.
4. The use of selenium-enriched Lactobacillus reuteri in preventing and treating intestinal pathogenic bacteria infection and promoting the growth of chicks according to claim 1, characterized in that: The prevention is manifested in enhancing the cellular immune response of the chicks.
5. The use of selenium-enriched Lactobacillus reuteri in preventing and treating intestinal pathogenic bacteria infection and promoting the growth of chicks according to claim 1, characterized in that: The selenium-enriched culture is based on MRS liquid culture medium, with 15-25 mg / mL of CaCO3, 5-15 mg / mL of glutathione and 1-2 mg / mL of ammonium citrate added, and 5-7 mg of sodium selenite is added to each 1 mL of the basic culture medium.
6. The use of selenium-enriched Lactobacillus reuteri in preventing and treating intestinal pathogenic bacteria infection and promoting the growth of chicks according to claim 5, characterized in that: 1-10 g of the Lactobacillus reuteri is inoculated per 100 ml of the basic culture medium.
7. The use of selenium-enriched Lactobacillus reuteri in preventing and treating intestinal pathogenic bacteria infection and promoting the growth of chicks according to claim 6, characterized in that: The sodium selenite is added 4 to 8 hours after inoculating the Lactobacillus reuteri.
8. The use of the selenium-enriched Lactobacillus reuteri in preventing and treating intestinal pathogenic bacteria infection and promoting the growth of chicks according to claim 7, characterized in that: The temperature of the selenium-enriched culture is 36-37° C., and the time is 22-24 hours.