Probiotic composition for improving chicken production performance and preparation method and application thereof

Abstract

The invention provides a probiotic composition for improving chicken production performance and a preparation method and application thereof, and relates to the technical field of microecological preparations. The probiotic composition comprises lactobacillus plantarum HM-05, lactobacillus casei HM-09 and pediococcus acidilactici PA-19, the preservation number of the lactobacillus plantarum HM-05 is CGMCC (China General Microbiological Culture Collection Center) NO.6739, the preservation number of the lactobacillus casei HM-09 is CGMCC NO.6736, and the preservation number of the pediococcus acidilactici PA-19 is CGMCC NO.19881; the ratio of the viable bacteria of the lactobacillus plantarum HM-05 to the viable bacteria of the lactobacillus casei HM-09 to the viable bacteria of the pediococcus acidilactici PA-19 is (1-5): (0.5-2): (1-10), and the probiotic composition obtained by compounding the viable bacteria of the lactobacillus plantarum HM-05, the viable bacteria of the lactobacillus casei HM-09 and the viable bacteria of the pediococcus acidilactici PA-19 can well inhibit breeding of pathogenic bacteria, improve balance of beneficial microbial communities in intestinal tracts and prevent diseases; the production performance and the health level of the bred chickens are effectively improved.

Description

Technical Field

[0001] This invention relates to the field of microecological preparation technology, specifically to a probiotic composition for improving chicken production performance, its preparation method, and its uses. Background Technology

[0002] Scientific breeding of broiler breeders is a crucial link in improving the quantity and quality of commercial broiler chicks, and is of great significance for ensuring the healthy growth and efficient breeding of broilers in their later stages. Currently, reducing the disease rate and mortality rate of broiler breeders is a common goal pursued by broiler breeder farmers.

[0003] Currently, broiler breeders commonly use antibiotics as additives to prevent disease and promote growth in poultry, achieving significant economic benefits. However, the long-term and excessive use of antibiotics can lead to drug resistance in pathogenic microorganisms, reducing the effectiveness of commonly used antibiotics and exacerbating drug residue problems, indirectly endangering human health.

[0004] Therefore, providing a probiotic composition or microecological preparation that can improve the production performance of farmed chickens to replace the use of antibiotics is a technical problem that urgently needs to be solved. Summary of the Invention

[0005] Specifically addressing the shortcomings of existing technologies, this invention provides a probiotic composition for improving chicken production performance. The probiotic composition comprises *Lactobacillus plantarum* HM-05, *Lactobacillus casei* HM-09, and *Pediococcus lactis* PA-19. The preservation number of *Lactobacillus plantarum* HM-05 is CGMCC NO.6739, that of *Lactobacillus casei* HM-09 is CGMCC No.6736, and that of *Pediococcus lactis* PA-19 is CGMCC No.19881. The live bacteria ratio of *Lactobacillus plantarum* HM-05, *Lactobacillus casei* HM-09, and *Pediococcus lactis* PA-19 is 1–5:0.5–2:1–10. This invention, by combining live bacteria of *Lactobacillus plantarum* HM-05, *Lactobacillus casei* HM-09, and *Pediococcus lactis* PA-19, effectively inhibits the growth of pathogenic bacteria, improves the balance of beneficial intestinal flora, prevents diseases, and thus effectively improves the production performance and health level of farmed chickens.

[0006] The objective of this invention is achieved through the following technical solution:

[0007] In a first aspect, the present invention provides a probiotic composition for improving chicken production performance, characterized in that the probiotic composition comprises Lactobacillus plantarum HM-05, Lactobacillus casei HM-09 and Pediococcus lactis PA-19;

[0008] The preservation number of the Lactobacillus plantarum HM-05 is CGMCC NO.6739;

[0009] The preservation number of the Lactobacillus casei HM-09 is CGMCC No. 6736;

[0010] The preservation number of the lactic acid cocci PA-19 is CGMCC No. 19881;

[0011] The live bacteria ratio of *Lactobacillus plantarum* HM-05, *Lactobacillus casei* HM-09, and *Pediococcus lactis* PA-19 is 1–5:0.5–2:1–10.

[0012] In some specific embodiments of the present invention, the live bacteria ratio of Lactobacillus casei HM-09, Lactobacillus plantarum HM-05, and Pediococcus lactis PA-19 is 2:1:5.

[0013] In some specific embodiments of the present invention, the *Lactobacillus plantarum* HM-05, the *Lactobacillus casei* HM-09, and the *Pediococcus lactis* PA-19 respectively further include a protectant, forming *Lactobacillus plantarum* HM-05 inoculant, *Lactobacillus casei* HM-09 inoculant, and *Pediococcus lactis* PA-19 inoculant;

[0014] The mass ratio of Lactobacillus plantarum HM-05 to the freeze-drying protectant is 1:5-10;

[0015] The mass ratio of Lactobacillus casei HM-09 to the freeze-drying protectant is 1:5-10;

[0016] The mass ratio of the lactic acid cocci PA-19 to the freeze-drying protectant is 1:5 to 10.

[0017] In some specific embodiments of the present invention, the protective agent comprises the following components at mass concentrations: 30-35 g / L skim milk powder, 15-20 g / L demineralized whey powder, 15-20 g / L trehalose, 3-4 g / L vitamin C, and 0.05-0.08 g / L lecithin, with distilled water as the solvent.

[0018] In some specific embodiments of the present invention, the viable count of the *Lactobacillus plantarum* HM-05 is ≥2.0 × 10⁻⁶. 11 CFU / mL, the viable count of the *Lactobacillus casei* HM-09 is ≥2.0 × 10⁻⁶. 11 CFU / mL, the viable count of the lactic acid cocci PA-19 is ≥2.0 × 10⁻⁶. 11 CFU / mL.

[0019] In a second aspect, the present invention provides a microecological preparation comprising the probiotic composition described in the first aspect and a dilution carrier; the mass ratio of the probiotic composition to the dilution carrier is 1-3:1-8; the dilution carrier is one or more of skim milk powder, maltodextrin, corn cob powder, and glucose.

[0020] Thirdly, the present invention provides a method for preparing the probiotic composition described in the first aspect, the method comprising: S1 activating the *Lactobacillus plantarum* HM-05, the *Lactobacillus casei* HM-09, and the *Pediococcus lactis* PA-19 respectively, inoculating them into MRS liquid culture medium, and obtaining *Lactobacillus plantarum* HM-05 seed culture, *Lactobacillus casei* HM-09 seed culture, and *Pediococcus lactis* PA-19 seed culture through primary seed culture and secondary seed culture;

[0021] S2. The *Lactobacillus plantarum* HM-05 seed culture, the *Lactobacillus casei* HM-09 seed culture, and the *Pediococcus lactis* PA-19 seed culture are respectively inoculated into a fermenter containing fermentation medium, fermented and cultured under certain conditions, and the pH of the fermentation broth is controlled at 5.6-6.2 to obtain *Lactobacillus plantarum* HM-05 fermentation broth, *Lactobacillus casei* HM-09 fermentation broth, and *Pediococcus lactis* PA-19 fermentation broth;

[0022] S3. After centrifuging the fermentation broth of Lactobacillus plantarum HM-05, the fermentation broth of Lactobacillus casei HM-09, and the fermentation broth of Pediococcus lactis PA-19, Lactobacillus plantarum HM-05 cells, Lactobacillus casei HM-09 cells, and Pediococcus lactis PA-19 cells are obtained. These cells are then mixed with a freeze-drying protectant and dried to obtain Lactobacillus plantarum HM-05 inoculum, Lactobacillus casei HM-09 inoculum, and Pediococcus lactis PA-19 inoculum. These inoculum are then mixed to obtain a probiotic composition.

[0023] In some specific embodiments of the present invention, the viable count of *Lactobacillus plantarum* HM-05 in the fermentation broth is ≥1.0 × 10⁻⁶. 10 CFU / mL; the viable count of Lactobacillus casei HM-09 in the fermentation broth is ≥1.0×10⁻⁶ CFU / mL. 10 CFU / mL; the viable count of *Pediococcus lactis* PA-19 in the fermentation broth is ≥1.0 × 10⁻⁶ CFU / mL. 10 CFU / mL.

[0024] In some specific embodiments of the present invention, the fermentation culture medium comprises the following components at the following mass concentrations: sucrose 50-80 g / L, yeast powder 20-40 g / L, soybean peptone 8-20 g / L, MgSO4·7H2O 1.5-2.0 g / L, MnSO4·5H2O 0.08-0.12 g / L, and Tween-80 0.8-1.0 g / L, with distilled water as the solvent;

[0025] The acidic solution includes one or more of hydrochloric acid, citric acid, and dilute sulfuric acid; the alkaline solution includes one or more of sodium hydroxide, potassium hydroxide, and disodium hydrogen phosphate.

[0026] Fourthly, the present invention provides the use of the probiotic composition of the first aspect and the microecological preparation of the second aspect in the preparation of products that can improve the production performance of farmed chickens, wherein the chickens include broiler chickens or egg-laying chickens.

[0027] The beneficial effects achieved by this invention are as follows:

[0028] 1. The probiotic composition obtained by combining live bacteria of Lactobacillus plantarum HM-05, Lactobacillus casei HM-09 and Pediococcus lactis PA-19 can effectively inhibit the growth of pathogenic bacteria, improve the balance of beneficial intestinal flora, prevent diseases, and thus effectively improve the production performance and health level of farmed chickens.

[0029] 2. The probiotic composition and microecological preparation provided by this invention can significantly improve the average egg weight, the rate of qualified eggs hatched and the egg production rate of broiler breeders, and significantly reduce the mortality rate, deformed egg rate, broken egg rate, double-yolk egg rate and unqualified egg rate of broiler breeders, thereby improving the economic benefits of broiler breeder farming. It does not produce drug resistance, has high safety, no drug residues and low toxicity.

[0030] Information on the preservation of biological materials:

[0031] Lactobacillus plantarum HM-05, taxonomically named Lactobacillus plantarum, was deposited on October 29, 2012, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 6739, located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing.

[0032] Lactobacillus casei HM-09 was deposited on October 29, 2012, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 6736. The deposit address is No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing.

[0033] Lactobacillus casei HM-10 was deposited on October 29, 2012, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 6737. The deposit address is No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing.

[0034] Lactobacillus plantarum HM-20 was deposited on October 29, 2012, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 6744. The deposit address is No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing.

[0035] Lactobacillus plantarum KT-Lp9, taxonomically named Lactobacillus plantarum, was deposited on September 8, 2016, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 12950, ​​located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing.

[0036] Lactobacillus plantarum Ps-8, taxonomically named Lactobacillus plantarum, was deposited on October 28, 2011, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 5359, located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing.

[0037] Lactobacillus plantarum HM-10, taxonomically named Lactobacillus plantarum, was deposited on October 29, 2012, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 6741, located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing.

[0038] Lactobacillus plantarum LP-11, taxonomically named Lactobacillus plantarum, was deposited on May 10, 2019, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 17750, located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing. Attached Figure Description

[0039] To more clearly illustrate the technical solution of this application, the drawings used in the embodiments will be briefly introduced below. Obviously, for those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0040] Figure 1 A statistical chart showing the mortality rate of female chickens in different groups of broiler breeders;

[0041] Figure 2 Statistical chart showing the mortality rate of roosters in different groups of broiler breeders;

[0042] Figure 3 A statistical chart showing the rate of deformed eggs in different groups of broiler breeders;

[0043] Figure 4 Statistical chart showing the egg breakage rate of different groups of broiler breeders;

[0044] Figure 5 Statistical chart showing the rate of double-yolk eggs in different groups of broiler breeders;

[0045] Figure 6 A statistical chart showing the average weight of eggs in different meat varieties;

[0046] Figure 7 Statistical charts showing the egg production rates of different groups of broiler breeders;

[0047] Figure 8 Statistical chart showing the hatching success rate of broiler breeder chickens in different groups;

[0048] Figure 9 Statistical chart showing the rate of substandard eggs hatched from different groups of broiler breeders;

[0049] Figure 10 A statistical chart showing the weekly changes in the mortality rate of female chickens in different groups of broiler breeders;

[0050] Figure 11 A statistical chart showing the weekly changes in the mortality rate of roosters in different groups of broiler breeders;

[0051] Figure 12 A statistical chart showing the weekly changes in the rate of deformed eggs in different groups of broiler breeders;

[0052] Figure 13 A statistical chart showing the weekly changes in egg breakage rate of different groups of broiler breeders;

[0053] Figure 14 A statistical chart showing the weekly changes in the rate of double-yolk eggs in different groups of broiler breeders;

[0054] Figure 15 A statistical chart showing the weekly changes in the average weight of eggs from different meat varieties;

[0055] Figure 16 A statistical chart showing the weekly changes in egg production rate of different groups of broiler breeders;

[0056] Figure 17 A statistical chart showing the weekly changes in the hatching rate of qualified eggs from different groups of broiler breeders;

[0057] Figure 18 A statistical chart showing the weekly changes in the rate of substandard eggs hatched from different groups of broiler breeders. Detailed Implementation

[0058] The present invention will be further described in detail below with reference to specific embodiments. The following embodiments are not intended to limit the present invention, but only to illustrate the present invention. Unless otherwise specified, the experimental methods used in the following embodiments are generally performed under conventional conditions. Unless otherwise specified, the materials and reagents used in the following embodiments are commercially available.

[0059] The strains used in this application are described below:

[0060] Lactobacillus plantarum HM-05 was derived from a naturally fermented feed sample from Inner Mongolia. The sequence obtained was compared with the nucleic acid sequence in GeneBank, and the results showed that the strain was Lactobacillus plantarum.

[0061] Lactobacillus casei HM-09 is described in the text of patent application publication number CN103911328A;

[0062] Pediococcus lactis PA-19 is described in the text of patent application publication number CN116769625A;

[0063] Escherichia coli, purchased from Hunan Keai Medical Equipment Co., Ltd., product number ST1030;

[0064] Salmonella, purchased from Shanghai Center for Biotechnology Preservation, catalog number CMCC 50023;

[0065] Clostridium perfringens was purchased from Qiyi Biotechnology (Shanghai) Co., Ltd., product number ATCC13124.

[0066] The reagents used in this application are described below:

[0067] MRS solid culture medium was purchased from Guangdong Huankai Microbial Technology Co., Ltd.

[0068] Bacterial genomic DNA extraction kit, purchased from Tiangen Biotech (Beijing) Co., Ltd.;

[0069] LB medium was purchased from Shanghai Shenqi Biotechnology Co., Ltd.

[0070] Salmonella culture medium was purchased from Qingdao Haibo Company.

[0071] BCP medium was purchased from Qingdao Haibo Biotechnology Co., Ltd.

[0072] The MRS liquid culture medium is prepared as follows: 10g peptone, 10g beef extract, 5g yeast extract, 20g glucose, 2g triammonium citrate, 5g sodium acetate, 0.1g magnesium sulfate, 0.05g manganese sulfate, 2g dipotassium hydrogen phosphate, 1mL Tween 80, add 1000mL distilled water, adjust the pH to 6.2-6.4, and sterilize at 121℃ for 15min.

[0073] Example 1: Obtaining Lactobacillus plantarum HM-05

[0074] (1) Screening of Lactobacillus plantarum HM-05

[0075] Using naturally fermented feed from Inner Mongolia as a sample, 1.0 g of the naturally fermented feed sample was added to 9 mL of PBS buffer (concentration 0.15 mol / L, pH 7.4), and vortexed to obtain a well-mixed sample. 0.5 mL of the well-mixed sample was added to 4.5 mL of PBS buffer to obtain a 10⁻¹ dilution. Then, 0.5 mL of the 10⁻¹ dilution was added to... -1 Dilute the buffer in 4.5 mL of PBS buffer to obtain 10 -2 Diluent, follow these steps to obtain 10 -3 10 -4 10 -5 10 -6 10 -7 Diluent; Pipette 100 μL of serially diluted solution onto MRS solid medium, 10 -4 10 -5 10 -6 10 -7 Two plates were used for each gradient, and the culture was anaerobic at 37°C for 72 hours to obtain colonies. Colonies with typical characteristics of Lactobacillus casei were selected from MRS solid medium based on their shape, size, edge, and transparency. Colonies were picked up with an inoculation loop and streaked onto MRS solid medium, and anaerobically cultured at 37°C for 72 hours to obtain purified single colonies. The purified single colonies were picked and inoculated into 4 mL of MRS liquid medium, and anaerobically cultured at 37°C for 24 hours to obtain bacterial suspensions. The bacterial strains corresponding to each bacterial suspension were numbered and subjected to strain identification and analysis to obtain strain HM-05.

[0076] (2) Identification of Lactobacillus plantarum HM-05

[0077] 10 μL of bacterial culture of strain HM-05 was taken, and DNA was extracted using a bacterial genomic DNA extraction kit to obtain a lysis buffer. Using the universal bacterial primers 27F and 1492R (27F: 5′-AGAGTTTGATCCTGGCTCAG-3′, 1492R: 5′-TACGACTTAACCCCAATCGC-3′) as shown in SEQ ID NO.5 and SEQ ID NO.6, respectively, the lysis buffer was used as a template for amplification to obtain the 16S rRNA of strain HM-05. The 16S rDNA of strain HM-05 was sequenced using the NCBI Blasten program, and the results showed that this strain is *Lactobacillus plantarum*, and it was named *Lactobacillus plantarum* HM-05. The bacterial culture of Lactobacillus plantarum HM-05 was placed in a 20% (v / v) glycerol aqueous solution (the volume ratio of glycerol aqueous solution to bacterial culture was 1:1) and deposited at the China General Microbiological Culture Collection Center, with the accession number CGMCC No. 6739.

[0078] Example 2: Characteristics of Lactobacillus plantarum HM-05

[0079] (1) Gastric juice resistance properties

[0080] After adjusting the pH of sterile PBS buffer (concentration 0.15mol / L, pH 7.4) to 2.5 with 1mol / L hydrochloric acid solution, 3.5g / L pepsin was added to the PBS buffer, and then the mixture was filtered through a 0.22μm microporous membrane to sterilize it and prepare simulated gastric juice. A bacterial suspension containing 1×108CFU / mL of live Lactobacillus plantarum HM-05 was mixed with the simulated gastric juice at a volume ratio of 1:1 to obtain a mixed solution.

[0081] The mixture was incubated at 37℃, and samples were taken at 0 and 3 hours of incubation. The samples taken at different time points were serially diluted 10-fold with sterile physiological saline to obtain diluted solutions. The diluted solutions were spread on MRS solid medium and incubated at 37℃ for 48 hours. The number of colonies on the MRS solid medium was counted to calculate the survival rate of Lactobacillus plantarum HM-05. The calculation results are shown in Table 1. Wherein, survival rate = [N1 / N0] × 100% (N0 is the number of viable bacteria at 0h, and N1 is the number of viable bacteria after treatment with simulated intestinal and gastric fluids for different times).

[0082] (2) Resistance to intestinal fluid

[0083] Adjust the pH of sterile PBS buffer (0.15 mol / L, pH 7.4) to 8.0 using 0.1 mol / L sodium hydroxide solution. Then, add 0.1% (w / v, g / 100 mL) trypsin and 1.8% (w / v, g / 100 mL) ox bile salts to the PBS buffer, and filter through a 0.22 μm microporous membrane to sterilize, preparing simulated intestinal fluid. Collect *Lactobacillus plantarum* HM-05 treated in simulated gastric fluid for 3 h in Experiment 1, wash twice with PBS buffer by centrifugation, and mix with an equal volume of simulated intestinal fluid to obtain a mixture. Incubate the mixture at 37 °C. Samples were taken at 4h and 8h of culture. Samples from different time points were serially diluted 10-fold with sterile physiological saline to obtain diluents. These diluents were spread onto MRS solid medium and incubated at 37℃ for 48h. The number of colonies on the MRS solid medium was then counted to calculate the survival rate of *Lactobacillus plantarum* HM-05. The results are shown in Table 1. Survival rate = [N1 / N0] × 100% (N0 is the number of viable bacteria at 0h, and N1 is the number of viable bacteria after treatment with simulated intestinal and gastric fluids for different times). The survival rates of *Lactobacillus plantarum* HM-05 after treatment with simulated gastric and intestinal fluids for different times are shown in Table 1.

[0084] Table 1. Survival rate of Lactobacillus plantarum HM-05 after treatment with simulated gastric and intestinal fluids for different time periods.

[0085]

[0086] (3) Antibacterial properties

[0087] Antibacterial experiments were conducted using the Oxford cup method. 20 mL of sterilized MRS solid medium, cooled to 50°C, was mixed with 200 μL of bacterial suspensions of *Escherichia coli*, *Salmonella*, and *Clostridium perfringens* (each with a viable bacterial concentration of 1 × 10⁶ CFU / mL). The mixture was then poured into plates. After the MRS solid medium containing the enteric pathogens cooled and solidified, 8 mm diameter holes were punched in the plates. 100 μL of bacterial suspension containing 1 × 10⁸ CFU / mL *Lactobacillus plantarum* HM-05 was added to each hole. The plates were then incubated at 4°C for 12 h for diffusion, followed by incubation at 37°C for 48 h. After incubation, the size of the inhibition zone was observed. The diameter of the inhibition zone was measured using calipers (two significant figures). The results are shown in Table 2.

[0088] Table 2. Antibacterial properties of Lactobacillus plantarum HM-05 against different enteropathogenic bacteria.

[0089] strain Escherichia coli O517:H7 (mm) Salmonella (mm) Clostridium perfringens (mm) HM-05 26.55±1.08 20.72±1.62 25.38±1.15

[0090] As shown in Tables 1 and 2, *Lactobacillus plantarum* HM-05 exhibits good acid and bile salt resistance, and also demonstrates broad-spectrum inhibitory properties against three pathogenic bacteria: *Escherichia coli*, *Salmonella*, and *Clostridium perfringens*. This indicates that *Lactobacillus plantarum* HM-05 possesses excellent gastrointestinal mucosal colonization ability, significantly inhibiting *Escherichia coli*, *Salmonella*, and *Clostridium perfringens* in the intestines, improving the host's intestinal flora, and enhancing immunity. Adding it to the daily drinking water system of broiler breeders can effectively inhibit the growth of pathogenic bacteria, improve the balance of beneficial intestinal flora, prevent diseases, and thus effectively improve the production performance and health level of broiler breeders.

[0091] Example 3: A probiotic composition

[0092] This embodiment provides a probiotic composition comprising live bacteria of Lactobacillus plantarum HM-05 obtained in Experimental Example 1, live bacteria of Lactobacillus casei HM-09 with accession number CGMCC NO.6736, and live bacteria of Pediococcus lactis PA-19 with accession number CGMCC No.19881.

[0093] The preparation method of the probiotic composition is as follows:

[0094] (1) Live bacteria culture: The preserved bacterial solutions of Lactobacillus plantarum HM-05, Lactobacillus casei HM-09 and Pediococcus lactis PA-19 were thawed in a water bath at 37°C for 30 min and then vortexed to mix, so as to obtain the bacterial solutions of Lactobacillus plantarum to be activated, Lactobacillus casei to be activated and Pediococcus lactis to be activated.

[0095] Take 1 mL of the unactivated bacterial suspensions of *Lactobacillus plantarum*, *Lactobacillus casei*, and *Pediococcus lactis* and inoculate them into MRS liquid medium, respectively. Activate and culture at 33℃ and 80 rpm for 24 h to obtain activated bacterial suspensions of *Lactobacillus plantarum*, *Lactobacillus casei*, and *Pediococcus lactis*. Use an inoculation loop to streak the activated bacterial suspensions of *Lactobacillus plantarum*, *Lactobacillus casei*, and *Pediococcus lactis* onto solid slant agar plates, and then culture on slant plates at 33℃ for 24 h to obtain activated slant cells of *Lactobacillus plantarum*, *Lactobacillus casei*, and *Pediococcus lactis*.

[0096] One loopful of activated slant culture of Lactobacillus plantarum, Lactobacillus casei, and Pediococcus lactis was inoculated into MRS liquid medium and seed cultured at 33℃ and 80 rpm for 24 h to obtain primary seed culture of Lactobacillus plantarum, Lactobacillus casei, and Pediococcus lactis.

[0097] Primary seed cultures of *Lactobacillus plantarum*, *Lactobacillus casei*, and *Pediococcus lactis* were inoculated into MRS liquid medium at an inoculation rate of 3% of the total volume of the MRS liquid medium. The cultures were then incubated at 33°C and 80 rpm for 24 hours to obtain secondary seed cultures of *Lactobacillus plantarum*, *Lactobacillus casei*, and *Pediococcus lactis*.

[0098] Secondary seed cultures of *Lactobacillus plantarum*, *Lactobacillus casei*, and *Pediococcus lactis* were inoculated into fermenters containing fermentation medium at an inoculation rate of 3% of the total fermentation medium volume. Fermentation was carried out at 33℃, 80 rpm, and 0.8 L / min for 12 h to obtain *Lactobacillus plantarum*, *Lactobacillus casei*, and *Pediococcus lactis* fermentation broths. (In the *Lactobacillus plantarum* fermentation broth, the viable count of *Lactobacillus plantarum* HM-05 was 5.2 × 10⁻⁶.) 10 CFU / mL; The viable count of Lactobacillus casei HM-09 in the Lactobacillus casei fermentation broth was 6.7 × 10⁻⁶. 10 CFU / mL; The viable count of Pediococcus lactis PA-19 in the fermentation broth was 8.4 × 10⁻⁶ CFU / mL. 10 CFU / mL); During fermentation, the pH of the fermentation broth was controlled at 6.0 using a 0.2% (w / v, g / 100mL) citric acid solution;

[0099] (2) Post-processing steps: The fermentation broths of Lactobacillus plantarum, Lactobacillus casei, and Pediococcus lactis were centrifuged at 8000 rpm for 5 min, the supernatant was discarded, and the precipitate was collected to obtain Lactobacillus plantarum cells, Lactobacillus casei cells, and Pediococcus lactis cells.

[0100] (3) Preparation of microbial agents: *Lactobacillus plantarum* cells, *Lactobacillus casei* cells, and *Pediococcus lactis* cells were mixed with a preservative at a mass ratio of 1:5, and then freeze-dried at -50℃ for 30 min to obtain *Lactobacillus plantarum* freeze-dried powder, *Lactobacillus casei* freeze-dried powder, and *Pediococcus lactis* freeze-dried powder (the viable count of *Lactobacillus plantarum* HM-05 in the *Lactobacillus plantarum* freeze-dried powder was 4.9 × 10⁻⁶). 11 CFU / mL; The viable count of Lactobacillus casei HM-09 in the lyophilized Lactobacillus casei powder was 5.1 × 10⁻⁶ CFU / mL. 11 CFU / mL; The viable count of Pediococcus lactis PA-19 in the lyophilized powder was 9.3 × 10⁻⁶ CFU / mL. 11 CFU / mL);

[0101] (4) Compound preparation of microbial agents: The freeze-dried powders of Lactobacillus casei, Lactobacillus plantarum, and Pediococcus lactis were mixed in a mass ratio of 2:1:5 to obtain a probiotic composition.

[0102] The fermentation medium consists of 80 g / L sucrose, 20 g / L yeast extract, 8 g / L soybean peptone, 1.5 g / L MgSO4·7H2O, 0.08 g / L MnSO4·5H2O, and 1.0 g / L Tween-80, with water as the solvent.

[0103] The protective agent contains 35g / L skim milk powder, 15g / L demineralized whey powder, 15g / L trehalose, 3g / L vitamin C, and 0.05g / L lecithin, with distilled water as the solvent.

[0104] Example 4: A microecological preparation

[0105] The microecological preparation comprises the probiotic composition of Example 3 and a dilution carrier; the dilution carrier is maltodextrin. In the compound lactic acid bacteria microecological preparation, the total number of viable bacteria of *Lactobacillus plantarum* HM-05, *Lactobacillus casei* HM-09, and *Pediococcus lactis* PA-19 is 1.5 × 10⁻⁶. 9 CFU / g.

[0106] Example 5: Effects of probiotic preparations on broiler chicken production performance

[0107] Arbor Acrylonitrile AA+ parent broiler breeders, aged 191 days, were divided into two groups: an experimental group of 23,347 hens and 2,446 roosters, and a control group of 23,429 hens and 2,458 roosters. The experimental group received the microecological preparation described in Example 4, while the control group did not. Both groups were raised and managed in the same manner. The experiment lasted 28 days, corresponding to ages from 191 to 218 days.

[0108] The method of adding the probiotic preparation is as follows: add it through the drinking water dosing system, stir thoroughly to dissolve, and the dosage is 1.0 kg / 10,000 birds / day. During the experiment, the number of dead hens and roosters, the number of eggs laid (deformed eggs, broken eggs, double-yolk eggs, eggs that hatch successfully, and eggs that fail to hatch), the number of eggs laid, and the egg weight were recorded daily. At the end of the experiment, the mortality rate of hens and roosters, the rate of deformed eggs, the rate of broken eggs, the rate of double-yolk eggs, the rate of eggs that hatch successfully, the rate of eggs that fail to hatch, the laying rate, and the average egg weight were calculated for statistical analysis of production performance indicators. The statistical results are shown in […]. Figures 1-18 See Tables 3 and 4.

[0109] Depend on Figure 1 The statistical results of the culling rate of hens showed that during the 4-week experimental period, the culling rates of hens in the experimental group and the control group were 0.03±0.01% and 0.04±0.02%, respectively, with a significant difference between the groups (P<0.05). This indicates that adding a compound lactic acid bacteria microecological preparation to the daily drinking water system of broiler breeders can significantly reduce the culling rate of hens.

[0110] Depend on Figure 2The statistical results of the rooster mortality rate showed that during the 4-week experimental period, the mortality rates of roosters in the experimental group and the control group were 0.04±0.01% and 0.08±0.01%, respectively, with a highly significant difference between the groups (P<0.01). This indicates that adding a compound lactic acid bacteria microecological preparation to the daily drinking water system of broiler breeders can significantly reduce the rooster mortality rate.

[0111] Depend on Figure 3 The statistical results of the abnormal egg rate showed that during the 4-week experimental period, the abnormal egg rates in the experimental group and the control group were 0.49±0.23% and 0.97±0.38%, respectively, with a highly significant difference between the groups (P<0.01). This demonstrates that adding a compound lactic acid bacteria microecological preparation to the daily drinking water system of broiler breeders can significantly reduce the abnormal egg rate.

[0112] Depend on Figure 4 The statistical results of egg breakage rate showed that during the 4-week trial period, the egg breakage rates of the experimental group and the control group were 0.24±0.11% and 0.62±0.46%, respectively, with a highly significant difference between the groups (P<0.01). This indicates that adding a compound lactic acid bacteria microecological preparation to the daily drinking water system of broiler breeders can significantly reduce the egg breakage rate.

[0113] Depend on Figure 5 The statistical results of the double-yolk egg rate showed that during the 4-week experimental period, the double-yolk egg rates in the experimental group and the control group were 3.09±0.80% and 3.96±0.63%, respectively, with a significant difference between the groups (P<0.05). This indicates that adding a compound lactic acid bacteria microecological preparation to the daily drinking water system of broiler breeders can significantly reduce the double-yolk egg rate.

[0114] Depend on Figure 6 The statistical results of the average egg weight showed that, during the 4-week experimental period, the average egg weight of the experimental group and the control group were 54.62±1.56 g / egg and 53.97±1.25 g / egg, respectively, with a significant difference between the groups (P<0.05). This indicates that adding a compound lactic acid bacteria microecological preparation to the daily drinking water system of broiler breeders can significantly increase the average egg weight.

[0115] Depend on Figure 7 The statistical results of egg production showed that during the 4-week trial period, the egg production rates of the experimental group and the control group were 85.56±5.00% and 85.05±4.15%, respectively, with a significant difference between the groups (P<0.05). This demonstrates that adding a compound lactic acid bacteria microecological preparation to the daily drinking water system of broiler breeders can significantly increase egg production.

[0116] Depend on Figure 8The statistical results of the hatching qualification rate showed that, during the 4-week trial period, the hatching qualification rates of the experimental group and the control group were 91.90±4.29% and 88.67±4.13%, respectively, with a highly significant difference between the groups (P<0.01). This demonstrates that adding a compound lactic acid bacteria microecological preparation to the daily drinking water system of broiler breeders can significantly increase the hatching qualification rate.

[0117] Depend on Figure 9 The statistical results of the hatching failure rate showed that, during the 4-week experimental period, the hatching failure rates in the experimental group and the control group were 4.29±3.84% and 5.79±4.50%, respectively, with a highly significant difference between the groups (P<0.01). This demonstrates that adding a compound lactic acid bacteria microecological preparation to the daily drinking water system of broiler breeders can significantly reduce the hatching failure rate.

[0118] Depend on Figure 10 The weekly mortality rate of hens showed a decreasing trend in the experimental group and an increasing trend in the control group. Furthermore, from week 2 to week 4 of the experiment, the mortality rate of hens in the experimental group was consistently lower than that in the control group.

[0119] Depend on Figure 11 The weekly mortality rate of roosters showed a downward trend in the experimental group and an upward trend in the control group. Furthermore, throughout the four-week trial period, the weekly mortality rate of roosters in the experimental group was consistently lower than that in the control group.

[0120] Depend on Figure 12 The weekly variation of the deformed egg rate showed that the experimental group exhibited an overall downward trend, while the control group showed fluctuations. Furthermore, throughout the 4-week trial period, the weekly deformed egg rate in the experimental group was consistently lower than that in the control group.

[0121] Depend on Figure 13 The weekly changes in egg breakage rate showed that the experimental group exhibited an overall decreasing trend, while the control group showed an increasing trend. Furthermore, throughout the 4-week trial period, the weekly egg breakage rate in the experimental group was consistently lower than that in the control group.

[0122] Depend on Figure 14 The weekly statistics of the double-yolk egg rate showed that the experimental group generally decreased, while the control group showed an increasing trend. Furthermore, throughout the 4-week trial period, the weekly double-yolk egg rate in the experimental group was consistently lower than that in the control group.

[0123] Depend on Figure 15 The statistical results of the weekly changes in average egg weight showed that both groups showed an upward trend. However, during the 4-week experimental period, the average egg weight of the experimental group was higher than that of the control group every week.

[0124] Depend on Figure 16The statistical results of the weekly changes in egg production rate show that both groups showed an upward trend. However, the experimental group's egg production rate was higher than that of the control group from the second to the fourth week of the experiment.

[0125] Depend on Figure 17 The weekly changes in the rate of qualified eggs hatched showed that both groups showed an upward trend. However, during the 4-week trial period, the rate of qualified eggs hatched in the experimental group was higher than that in the control group every week.

[0126] Depend on Figure 18 The weekly changes in the rate of unqualified eggs during incubation showed that both groups showed a downward trend. However, during the 4-week experimental period, the rate of unqualified eggs during incubation in the experimental group was lower than that in the control group.

[0127] The statistical results of broiler breeder production performance at each stage of the experiment are shown in Tables 3 and 4. From the start of the experiment (week 1) to the end of the experiment (week 4), the mortality rate of female hens in the experimental group was significantly lower than that in the control group by 0.01% (P < 0.05), the mortality rate of male hens in the experimental group was significantly lower than that in the control group by 0.04% (P < 0.01), the rate of deformed eggs in the experimental group was significantly lower than that in the control group by 0.48% (P < 0.01), and the rate of broken eggs in the experimental group was significantly lower than that in the control group by 0.38%. The percentage of double-yolk eggs in the experimental group was significantly lower than that in the control group by 0.87% (P<0.05), the average egg weight in the experimental group was significantly higher than that in the control group by 1.20% (P<0.05), the egg production rate in the experimental group was significantly higher than that in the control group by 0.51% (P<0.05), the percentage of qualified eggs hatched in the experimental group was significantly higher than that in the control group by 3.23% (P<0.01), and the percentage of unqualified eggs hatched in the experimental group was significantly lower than that in the control group by 1.50% (P<0.01).

[0128] In summary, adding compound lactic acid bacteria microecological preparations to the daily drinking water system of broiler breeders can significantly improve the average egg weight, the rate of qualified eggs that hatch, and the egg production rate, and significantly reduce the mortality rate of hens, the mortality rate of roosters, the rate of deformed eggs, the rate of broken eggs, the rate of double-yolk eggs, and the rate of unqualified eggs that hatch. Therefore, it can improve the economic benefits of broiler breeder farming to a certain extent.

[0129] Table 3. Statistical results of broiler breeder production performance at each stage during the experimental period.

[0130]

[0131] Table 4. Statistical results of broiler breeder production performance at each stage during the experimental period.

[0132]

[0133]

[0134] Example 6: A probiotic composition

[0135] The probiotic composition was obtained by referring to the probiotic composition preparation method of Example 3. The difference between the probiotic composition and the one in Example 3 is that the mass ratio of Lactobacillus plantarum HM-05, Lactobacillus casei HM-09, and Pediococcus lactis PA-19 is 5:0.5:10.

[0136] Example 7: A microecological preparation

[0137] The microecological preparation includes the probiotic composition of Example 3 and a dilution carrier, wherein the dilution carrier is maltodextrin. In the microecological preparation, the total number of viable bacteria of *Lactobacillus plantarum* HM-05, *Lactobacillus casei* HM-09, and *Pediococcus lactis* PA-19 is 6.0 × 10⁻⁶. 10 The compound lactic acid bacteria microecological preparation was added to the daily drinking water system of broiler breeders, and the statistical results of the production performance of broiler breeders at each stage are shown in Table 5.

[0138] Table 5. Statistical results of broiler breeder production performance at each stage during the experimental period.

[0139]

[0140] Example 8: A probiotic composition

[0141] The probiotic composition was obtained by referring to the probiotic composition preparation method of Example 3. The difference between the probiotic composition and the one in Example 3 is that the mass ratio of Lactobacillus plantarum HM-05, Lactobacillus casei HM-09, and Pediococcus lactis PA-19 is 1:2:1.

[0142] Example 9: A microecological preparation

[0143] The microecological preparation includes the probiotic composition of Example 3 and a dilution carrier, wherein the dilution carrier is corn cob powder. In the microecological preparation, the total number of viable bacteria of *Lactobacillus plantarum* HM-05, *Lactobacillus casei* HM-09, and *Pediococcus lactis* PA-19 is 7.0 × 10⁻⁶. 10 The compound lactic acid bacteria microecological preparation was added to the daily drinking water system of broiler breeders, and the statistical results of the production performance of broiler breeders at each stage are shown in Table 6.

[0144] Table 6. Statistical results of broiler breeder production performance at each stage during the experimental period.

[0145]

[0146] Comparative Example 1: A probiotic composition

[0147] The probiotic composition was obtained by referring to the probiotic composition preparation method of Example 3. The difference between the probiotic composition and the one in Example 3 is that Lactobacillus plantarum HM-05 was replaced with Lactobacillus plantarum HM-20 (CGMCC No. 6744), and Lactobacillus casei HM-09 was replaced with Lactobacillus casei HM-10 (CGMCC No. 6737).

[0148] Comparative Example 2: A probiotic preparation

[0149] The microecological preparation includes the probiotic composition of Comparative Example 1 and a dilution carrier, wherein the dilution carrier is maltodextrin. Adding the compound lactic acid bacteria microecological preparation to the daily drinking water system of broiler breeders resulted in a 0.25% decrease in egg production, a 0.34% increase in the rate of weak chicks, and a 0.05% increase in the rate of empty shells.

[0150] Comparative Example 3: A probiotic composition

[0151] The probiotic composition was obtained by referring to the probiotic composition preparation method of Example 3. The difference between the probiotic composition and the one in Example 3 is that Lactobacillus plantarum HM-05 was replaced with Lactobacillus plantarum KT-Lp9 (CGMCC No. 12950), and Lactobacillus casei HM-09 was replaced with Lactobacillus plantarum Ps-8 (CGMCC No. 5359).

[0152] Comparative Example 4: A probiotic preparation

[0153] The microecological preparation includes the probiotic composition of Comparative Example 3 and a dilution carrier, wherein the dilution carrier is maltodextrin. Adding the compound lactic acid bacteria microecological preparation to the daily drinking water system of broiler breeders resulted in a 0.32% decrease in egg production, a 0.33% increase in the rate of weak chicks, and a 0.09% increase in the rate of empty shells.

[0154] Comparative Example 5: A probiotic composition

[0155] The probiotic composition was obtained by referring to the probiotic composition preparation method of Example 3. The difference between the probiotic composition and the one in Example 3 is that Lactobacillus plantarum HM-05 was replaced with Lactobacillus plantarum HM-10 (CGMCC No. 6741).

[0156] Comparative Example 6: A probiotic preparation

[0157] The microecological preparation includes the probiotic composition of Comparative Example 5 and a dilution carrier, wherein the dilution carrier is maltodextrin. Adding the compound lactic acid bacteria microecological preparation to the daily drinking water system of broiler breeders resulted in a 0.31% decrease in egg production, a 0.22% increase in the rate of weak chicks, and a 0.03% increase in the rate of empty shells.

[0158] Comparative Example 7: A probiotic composition

[0159] The probiotic composition was obtained by referring to the probiotic composition preparation method of Example 3. The difference between the probiotic composition and the one in Example 3 is that Lactobacillus plantarum HM-05 was replaced with Lactobacillus plantarum LP-11 (CGMCC No. 17750).

[0160] Comparative Example 8: A probiotic preparation

[0161] The microecological preparation includes the probiotic composition of Comparative Example 7 and a dilution carrier, wherein the dilution carrier is maltodextrin. Adding the compound lactic acid bacteria microecological preparation to the daily drinking water system of broiler breeders resulted in a 0.13% decrease in egg production, a 0.15% increase in the rate of weak chicks, and a 0.06% increase in the rate of empty shells.

[0162] Comparative Example 9: A probiotic composition

[0163] The probiotic composition was obtained by referring to the probiotic composition preparation method of Example 3. The difference between the probiotic composition and the one in Example 3 is that the mass ratio of Lactobacillus plantarum HM-05, Lactobacillus casei HM-09 and Pediococcus lactis PA-19 was changed from 2:1:5 to 0:1:5.

[0164] Comparative Example 10: A probiotic preparation

[0165] The microecological preparation includes the probiotic composition of Comparative Example 9 and a dilution carrier, wherein the dilution carrier is maltodextrin. Adding the compound lactic acid bacteria microecological preparation to the daily drinking water system of broiler breeders resulted in a 1.3% decrease in broiler growth rate, a 0.2 increase in feed conversion ratio, and a 0.05% increase in mortality compared to Example 5.

[0166] Comparative Example 11: A probiotic composition

[0167] The probiotic composition was obtained by referring to the probiotic composition preparation method of Example 3. The difference between the probiotic composition and the one in Example 3 is that the mass ratio of Lactobacillus plantarum HM-05, Lactobacillus casei HM-09, and Pediococcus lactis PA-19 was changed from 2:1:5 to 2:0:5.

[0168] Comparative Example 12: A probiotic preparation

[0169] The microecological preparation includes the probiotic composition of Comparative Example 11 and a dilution carrier, wherein the dilution carrier is maltodextrin. Adding the compound lactic acid bacteria microecological preparation to the daily drinking water system of broiler breeders resulted in a 0.4% decrease in egg production, a 0.2% increase in the rate of weak chicks, and a 0.06% increase in the rate of empty shells.

[0170] The above specific embodiments further illustrate the purpose, technical solution and beneficial effects of this application. It should be understood that the above are only specific embodiments of this application and are not intended to limit the scope of protection of this application. Any modifications, equivalent substitutions, improvements, etc., made on the basis of the technical solution of this application should be included within the scope of protection of this application.

Claims

1. A probiotic composition for improving chicken production performance, characterized in that, The probiotic composition includes Lactobacillus plantarum HM-05, Lactobacillus casei HM-09, and Pediococcus lactis PA-19; The preservation number of the Lactobacillus plantarum HM-05 is CGMCC NO.6739; The preservation number of the Lactobacillus casei HM-09 is CGMCC No. 6736; The preservation number of the lactic acid cocci PA-19 is CGMCC No. 19881; The live bacteria ratio of *Lactobacillus plantarum* HM-05, *Lactobacillus casei* HM-09, and *Pediococcus lactis* PA-19 is 1–5:0.5–2:1–10.

2. The probiotic composition according to claim 1, characterized in that, The live bacteria ratio of Lactobacillus casei HM-09, Lactobacillus plantarum HM-05, and Pediococcus lactis PA-19 is 2:1:

5.

3. The probiotic composition according to claim 1 or 2, characterized in that, The Lactobacillus plantarum HM-05, Lactobacillus casei HM-09 and Pediococcus lactis PA-19 each also include a protectant, forming Lactobacillus plantarum HM-05 inoculum, Lactobacillus casei HM-09 inoculum and Pediococcus lactis PA-19 inoculum; The mass ratio of Lactobacillus plantarum HM-05 to the freeze-drying protectant is 1:5-10; The mass ratio of Lactobacillus casei HM-09 to the freeze-drying protectant is 1:5-10; The mass ratio of the lactic acid cocci PA-19 to the freeze-drying protectant is 1:5 to 10.

4. The probiotic composition according to claim 3, characterized in that, The protective agent comprises the following components at mass concentrations: 30-35 g / L skim milk powder, 15-20 g / L demineralized whey powder, 15-20 g / L trehalose, 3-4 g / L vitamin C, and 0.05-0.08 g / L lecithin, with distilled water as the solvent.

5. The probiotic composition according to claim 3, characterized in that, The viable count of *Lactobacillus plantarum* HM-05 is ≥2.0 × 10⁻⁶. 11 CFU / mL, the viable count of the *Lactobacillus casei* HM-09 is ≥2.0 × 10⁻⁶. 11 CFU / mL, the viable count of the lactic acid cocci PA-19 is ≥2.0 × 10⁻⁶. 11 CFU / mL.

6. A microecological preparation, characterized in that, The microecological preparation comprises the probiotic composition and dilution carrier as described in any one of claims 1 to 5; the mass ratio of the probiotic composition to the dilution carrier is 1 to 3: 1 to 8; the dilution carrier is one or more of skim milk powder, maltodextrin, corn cob powder, and glucose.

7. A method for preparing the probiotic composition according to any one of claims 1 to 5, characterized in that, The method for preparing the probiotic composition includes, in step S1, activating the *Lactobacillus plantarum* HM-05, *Lactobacillus casei* HM-09, and *Pediococcus lactis* PA-19 respectively, inoculating them into MRS liquid culture medium, and obtaining *Lactobacillus plantarum* HM-05 seed culture, *Lactobacillus casei* HM-09 seed culture, and *Pediococcus lactis* PA-19 seed culture through primary seed culture and secondary seed culture; S2. The *Lactobacillus plantarum* HM-05 seed culture, the *Lactobacillus casei* HM-09 seed culture, and the *Pediococcus lactis* PA-19 seed culture are respectively inoculated into a fermenter containing fermentation medium, fermented and cultured under certain conditions, and the pH of the fermentation broth is controlled at 5.6-6.2 to obtain *Lactobacillus plantarum* HM-05 fermentation broth, *Lactobacillus casei* HM-09 fermentation broth, and *Pediococcus lactis* PA-19 fermentation broth; S3. After centrifuging the fermentation broth of Lactobacillus plantarum HM-05, the fermentation broth of Lactobacillus casei HM-09, and the fermentation broth of Pediococcus lactis PA-19, Lactobacillus plantarum HM-05 cells, Lactobacillus casei HM-09 cells, and Pediococcus lactis PA-19 cells are obtained. These cells are then mixed with a freeze-drying protectant and dried to obtain Lactobacillus plantarum HM-05 inoculum, Lactobacillus casei HM-09 inoculum, and Pediococcus lactis PA-19 inoculum. These inoculum are then mixed to obtain a probiotic composition.

8. The method for preparing the probiotic composition according to claim 7, characterized in that, The viable count of *Lactobacillus plantarum* HM-05 in the fermentation broth is ≥1.0 × 10⁻⁶. 10 CFU / mL; the viable count of Lactobacillus casei HM-09 in the fermentation broth is ≥1.0×10⁻⁶ CFU / mL. 10 CFU / mL; the viable count of *Pediococcus lactis* PA-19 in the fermentation broth is ≥1.0 × 10⁻⁶ CFU / mL. 10 CFU / mL.

9. The method for preparing the probiotic composition according to claim 7, characterized in that, The fermentation medium comprises the following components at the following mass concentrations: sucrose 50–80 g / L, yeast extract 20–40 g / L, soybean peptone 8–20 g / L, MgSO4·7H2O 1.5–2.0 g / L, MnSO4·5H2O 0.08–0.12 g / L, and Tween-80 0.8–1.0 g / L, with distilled water as the solvent; The acidic solution includes one or more of hydrochloric acid, citric acid, and dilute sulfuric acid; the alkaline solution includes one or more of sodium hydroxide, potassium hydroxide, and disodium hydrogen phosphate.

10. The use of the probiotic composition according to any one of claims 1 to 5 and the microecological preparation according to claim 6 in the preparation of products capable of improving the production performance of farmed chickens, characterized in that, The chickens mentioned include broiler chickens or egg-laying chickens.

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