Application of microbial preparation in preparation of product for improving poultry production performance

Abstract

The invention relates to application of a microbial preparation in preparation of a product for improving poultry production performance. Strain in the microbial preparation comprises pediococcus acidilactici PA153 and bacillus coagulans BC66. The classification name of the pediococcus acidilactici PA153 is pediococcus acidilactici, the preservation number of the pediococcus acidilactici PA153 is CCTCC (China Center For Type Culture Collection) No: M2023230, and the preservation date of the pediococcus acidilactici PA153 is March 2, 2023. The bacillus coagulans BC66 is classified and named as bacillus coagulans, the preservation number of the bacillus coagulans BC66 is CGMCC (China General Microbiological Culture Collection Center) No.21879, and the preservation date of the bacillus coagulans BC66 is March 8, 2021. The pediococcus acidilactici PA153 and the bacillus coagulans BC66 provided by the invention are combined for use and are mutually matched for synergistic interaction, so that the production performance of the poultry can be improved, and meanwhile, the pediococcus acidilactici PA153 and the bacillus coagulans BC66 can also be used for remarkably regulating the immunocompetence, promoting the development of immune organs, slowing down the atrophy speed of bursa of Fabricius and improving the humoral immune level of the poultry.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms and relates to the application of a microbial preparation in preparing a product for improving the production performance of poultry. Background Art

[0002] Production performance indicators are related to the economic benefits of breeding. How to improve animal production performance is crucial in the process of livestock and poultry breeding. Antibiotics are often used in livestock and poultry feed as a feed additive that resists disease, promotes growth, and improves production indicators. They are used to promote animal growth and reduce the occurrence of diseases. However, problems such as abuse and illegal additions have gradually been exposed, bringing irreversible harms such as bacterial resistance, pathogenic microorganisms causing diseases, and animal body residues. On the other hand, with the improvement of people's living standards and the enhancement of health awareness, the requirements for the quality of livestock products are also constantly increasing. The quality directly affects the price of livestock products.

[0003] In this context, seeking alternative antibiotic products that can improve animal production performance and livestock product quality has become a research hotspot in recent years. Among them, the application of probiotics has brought a good solution. Probiotics can be used not only alone, but also in combination. They can optimize intestinal health, enhance disease resistance and immunity, improve nutrient utilization, improve animal production performance and livestock product quality, and improve economic benefits.

[0004] Pediococcus acidilactici belongs to the genus Pediococcus, which can directly ferment and degrade carbohydrates to produce lactic acid. It has multiple biological activities such as lowering blood sugar and lipids, anti-inflammatory, regulating autoimmunity, and promoting body development. It is a probiotic that is beneficial to the health of animal bodies. Pediococcus acidilactici can colonize in the intestines of animals, have an antagonistic effect on pathogenic microorganisms, competitively inhibit pathogenic microorganisms, enhance the immune function of animal bodies, produce beneficial metabolites, activate the activity of acid proteases, participate in the body's metabolism, and prevent the production of harmful substances.

[0005] Bacillus coagulans is a probiotic of the genus Bacillus, which has many important biological characteristics and functions. In the intestines of animals, Bacillus coagulans can regulate the balance of intestinal flora, inhibit the growth of harmful bacteria, reduce the production of intestinal endotoxins, and enhance the intestinal barrier function and prevent the invasion of pathogenic microorganisms through interaction with the intestinal mucosa. In addition, Bacillus coagulans can also participate in the body's immune regulation process, stimulate the development and function of the immune system, and enhance the immunity and disease resistance of animals.

[0006] Different strains of Pediococcus acidilactici or Bacillus coagulans have been reported to have different probiotic functions, such as regulating intestinal flora, enhancing animal resistance, etc. Pediococcus acidilactici and Bacillus coagulans from different sources have different biological functionalities, so it is of great significance to develop a microbial preparation to improve the production performance of poultry and reduce the incidence and mortality of poultry. Summary of the invention

[0007] In view of the shortcomings of the prior art, the present invention aims to provide an application of a microbial preparation in the preparation of a product for improving the production performance of poultry. The microbial preparation can promote the development of immune organs, significantly improve the survival rate, egg production rate and egg weight of poultry, and improve the production performance of poultry.

[0008] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0009] In the first aspect, the present invention provides an application of a microbial preparation in the preparation of a product for improving the production performance of poultry, wherein the strains in the microbial preparation include Pediococcus acidilactici PA153 and Bacillus coagulans BC66; the classification name of Pediococcus acidilactici PA153 is Pediococcus acidilactici, the preservation number is CCTCC No: M2023230, and the preservation date is March 2, 2023; the classification name of Bacillus coagulans BC66 is Bacillus coagulans, the preservation number is CGMCC No.21879, and the preservation date is March 8, 2021.

[0010] The combination of the lactic acid Pediococcus PA153 and the Bacillus coagulans BC66 provided by the present invention can cooperate with each other to synergistically enhance the synergy, thereby improving the production performance of poultry. At the same time, it can also significantly regulate the immune ability, promote the development of immune organs, slow down the atrophy speed of the bursa of Fabricius, and improve the humoral immunity level of poultry.

[0011] Preferably, the ratio of the bacterial counts of Pediococcus acidilactici PA153 and Bacillus coagulans BC66 is (1-4):1.

[0012] Among them, the specific point values ​​1 to 4 can be selected as 1, 1.5, 2, 2.5, 3, 3.5, 4, etc., and other specific point values ​​within the numerical range can be selected, which will not be described one by one here.

[0013] Based on the potential interaction between the two strains in improving poultry production performance, when the two strains meet the above-mentioned specific ratio relationship, the production performance of poultry can be better improved.

[0014] Preferably, the total bacterial count in the microbial preparation is not less than 1×10 8 CFU / mL or 1×108 CFU / g, of which no less than 1×10 8 CFU / mL can be 1×10 8 CFU / mL, 1×10 9 CFU / mL, 1×10 10 CFU / mL, 1×10 11 CFU / mL, 1×10 12 CFU / mL, 1×10 13 CFU / mL, etc., not less than 1×10 8 CFU / g can be 1×10 8 CFU / g, 1×10 9 CFU / g, 1×10 10 CFU / g, 1×10 11 CFU / g, 1×10 12 CFU / g, 1×10 13 CFU / g, etc. Other specific point values ​​within this numerical range can be selected and will not be described here one by one.

[0015] Preferably, the microbial preparation further comprises a protective agent.

[0016] Preferably, the protective agent comprises any one of lactose, xylo-oligosaccharides, butter powder, Tween-80, mannitol, dextrin, gum arabic, sucrose, sorbitol, polyvinyl pyrrolidone or sodium glutamate, or a combination of at least two thereof.

[0017] Preferably, the microbial preparation is in the form of a solution or a powder.

[0018] The dosage form of the microbial preparation involved in the present invention is not limited, including the most commonly used solutions, powders, or further prepared capsules, tablets or granules.

[0019] Preferably, the poultry comprises chicken, duck or goose.

[0020] In a second aspect, the present invention provides a poultry feed, which includes corn, peanut meal, sesame meal, millet bran, bran and a microbial preparation; the strains in the microbial preparation include Pediococcus acidilactici PA153 and Bacillus coagulans BC66; the classification name of Pediococcus acidilactici PA153 is Pediococcus acidilactici, the preservation number is CCTCC No: M2023230, and the preservation date is March 2, 2023; the classification name of Bacillus coagulans BC66 is Bacillus coagulans, the preservation number is CGMCC No.21879, and the preservation date is March 8, 2021.

[0021] Preferably, the ratio of the bacterial counts of Pediococcus acidilactici PA153 and Bacillus coagulans BC66 is (1-4):1.

[0022] Preferably, the total bacterial count in the microbial preparation is not less than 1×10 8 CFU / mL or 1×10 8 CFU / g, of which no less than 1×10 8 CFU / mL can be 1×10 8 CFU / mL, 1×10 9 CFU / mL, 1×10 10 CFU / mL, 1×10 11 CFU / mL, 1×10 12 CFU / mL, 1×10 13 CFU / mL, etc., not less than 1×10 8 CFU / g can be 1×10 8 CFU / g, 1×10 9 CFU / g, 1×10 10 CFU / g, 1×10 11 CFU / g, 1×10 12 CFU / g, 1×10 13 CFU / g, etc. Other specific point values ​​within this numerical range can be selected and will not be described here one by one.

[0023] Preferably, the mass percentage of the microbial preparation in the poultry feed is 0.1-2%, for example, it can be 0.1%, 0.5%, 1%, 1.5%, 2%, etc. Other specific point values ​​within this numerical range can be selected and will not be described here one by one.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] The present invention uses Pediococcus acidilactici PA153 and Bacillus coagulans BC66 in combination, and the two cooperate with each other to synergistically enhance the efficiency, thereby improving the production performance of poultry, specifically in terms of increasing egg weight, egg production rate, and qualified egg rate, and reducing the mortality rate; meanwhile, the present invention can also significantly regulate the immune ability, promote the development of immune organs, slow down the atrophy speed of the bursa of Fabricius, and improve the humoral immunity level of poultry.

[0026] The PA153 strain involved in the present invention is classified and named Pediococcus acidilactici, the preservation number is CCTCC No: M2023230, the preservation date is March 2, 2023, and the preservation address is Wuhan University, Wuhan City, Hubei Province.

[0027] The BC66 strain involved in the present invention is classified and named Bacillus coagulans, the preservation number is CGMCC No.21879, the preservation date is March 8, 2021, and the preservation address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. DETAILED DESCRIPTION

[0028] The technical solution of the present invention is further described below by specific implementation methods. It should be understood by those skilled in the art that the embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0029] The culture medium formula involved in the following content is as follows:

[0030] Glucose yeast medium: 10 g glucose, 5 g peptone, 2 g yeast extract, 0.5 g potassium monohydrogen phosphate, 5 g sodium chloride, 2.5 g potassium chloride, and 1 L water.

[0031] MRS liquid culture medium: tryptone 10 g, yeast extract 5 g, glucose 20 g, dipotassium hydrogen phosphate 2 g, ammonium citrate 2 g, sodium acetate 5 g, magnesium sulfate 0.2 g, manganese sulfate 0.05 g, distilled water 1000 mL, Tween-80 1 mL.

[0032] RPMI1640 medium: 2.0 g sodium bicarbonate, 6.8 g sodium chloride, 0.4 g potassium chloride, 0.2 g potassium dihydrogen phosphate, 1.0 g glucose, 0.11 g calcium chloride dihydrate, 0.49 g magnesium sulfate heptahydrate, 0.14 g disodium hydrogen phosphate heptahydrate, 2.0 g L-glutamine, 0.015 g phenol red, 3.0 g pancreatic casein digest, 0.5 g yeast extract, 0.2 g vitamin B, and about 950 mL of distilled water.

[0033] The PA153 strain involved in the following content is classified and named Pediococcus acidilactici, the preservation number is CCTCC No: M2023230, and the preservation date is March 2, 2023.

[0034] The BC66 mentioned in the following content is classified as Bacillus coagulans, with a deposit number of CGMCC No.21879 and a deposit date of March 8, 2021.

[0035] Example 1

[0036] Functional testing of Pediococcus acidilactici PA153 and Bacillus coagulans BC66

[0037] (1) Evaluation of strain growth rate and acid production rate:

[0038] The activated seed liquid of lactic acid Pediococcus PA153 and Bacillus coagulans BC66 were transferred to glucose yeast medium and MRS liquid medium respectively, and cultured at 37℃ for 48h. Samples were taken every 4 hours to determine the OD600 and pH value of the culture liquid. Each sample was repeated in three groups. The growth curve of the strain was drawn with the culture time as the horizontal axis and OD600 as the vertical axis. The results showed that lactic acid Pediococcus PA153 entered the logarithmic growth phase at 8h and the stable phase at 24h. The fermentation cycle was short and the growth rate was fast. The pH curve dropped rapidly from 0 to 10h, and the pH dropped slowly after 10 to 24h. After 24h, the pH was maintained at about 4.0. Bacillus coagulans BC66 entered the logarithmic growth phase at 7h and the stable phase after 23h. The fermentation cycle was relatively short and the growth rate was fast. The pH curve dropped rapidly from 0 to 8h, and the pH dropped slowly after 8 to 24h. After 24h, the pH was maintained at about 4.5.

[0039] (2) Evaluation of strain tolerance to artificial gastrointestinal fluid:

[0040] (2.1) Preparation of artificial gastric juice: Prepare 0.85% physiological saline, adjust the pH value to 3.0 with dilute hydrochloric acid, add 0.3% pepsin, and filter with a microporous filter membrane to sterilize after fully dissolving.

[0041] (2.2) Preparation of artificial intestinal fluid: prepare 0.85% physiological saline, adjust the pH value to 8.0 with sodium hydroxide, then add 0.1% trypsin, fully dissolve it and filter it with a microporous filter membrane for sterilization.

[0042] (2.3) Prepare bile salt culture medium: add 0.3% porcine bile salt and 0.2% sodium thioglycolate to MRS culture medium, adjust the pH to 6.0, sterilize at 118°C for 15 min and set aside.

[0043] The lactic acid Pediococcus PA153 and Bacillus coagulans BC66 strains were activated to the stable period, and 0.1 mL of the sample was added to 9.9 mL of artificial gastric juice, artificial intestinal juice and bile salt culture medium, respectively, and incubated at 37 ° C for 2.5 h. The in vitro simulation passed through the gastrointestinal environment, and the number of viable bacteria was collected and measured after the end to evaluate the survival of the strain. Survival rate % = Nt / N0×100%, where N0 represents the initial viable bacteria count of the strain (CFU / mL), and Nt represents the viable bacteria count (CFU / mL) after gastrointestinal simulation. The results are shown in Table 1.

[0044] Table 1

[0045]

[0046]

[0047] (3) Evaluation of strain stability:

[0048] Take 10 respectively 11 The PA153 and Bacillus coagulans BC66 bacterial liquids with a moisture content of ≤5% CFU / g were placed in a constant temperature incubator at 37°C for 3 weeks, and samples were taken every week to detect the content of live bacteria. The results are shown in Table 2, which show that after 3 weeks, the survival rate of Pediococcus acidilactici PA153 was above 82%, and the survival rate of Bacillus coagulans BC66 was above 98%.

[0049] Table 2

[0050]

[0051] (4) Evaluation of the strain’s ability to produce extracellular polysaccharides

[0052] Extracellular polysaccharides can improve the intestinal mucosal barrier function, which is beneficial to the absorption of nutrients, promote the growth and egg production of hens, improve the production performance of hens, and enhance the body's resistance and reduce the mortality rate. The ability of strains to produce extracellular polysaccharides directly affects the production performance of poultry. The evaluation method is as follows:

[0053] (4.1) Drawing the standard curve of extracellular polysaccharide

[0054] Take 0.1g of glucose, dissolve it in distilled water and make up to 100mL, accurately transfer 10mL, and make up to 100mL again. At this time, the final concentration of glucose is 0.10mg / mL, which is the glucose standard solution. Accurately pipette 0.1, 0.2, 0.4, 0.6, 0.8, 1.0, 1.5, 2.0mL of glucose standard solution into a 10mL stoppered test tube, add distilled water to 2.00mL, and take another empty test tube and add 2.00mL of distilled water as a control. Accurately add 1mL of 60g / L phenol solution (freshly prepared) and 5mL of concentrated sulfuric acid to each test tube, mix well, place for 5min, heat in a 60℃ water bath for 30min to develop color, cool in a cold water bath for 15min to room temperature, and measure OD490. Draw a standard curve with glucose content as the horizontal axis and OD490 as the vertical axis, and the formula for the standard curve is y=0.0053x-0.0142.

[0055] (4.2) Determination of extracellular polysaccharide content

[0056] The activated seed solutions of Pediococcus acidilactici PA153 and Bacillus coagulans BC66 were inoculated into MRS medium respectively and cultured at 37℃ and 200r / min for 24h. The extracellular polysaccharide production capacity of the bacteria was compared and determined according to the phenol-sulfuric acid method combined with the standard curve.

[0057] 4 mL of PA153 and BC66 bacterial liquid were taken respectively, 2 mL of trichloroacetic acid (TCA) with a mass fraction of 30% was added, and the mixture was centrifuged at 4000 r / min for 15 min to remove protein. The supernatant was collected, and 3 volumes of anhydrous cold ethanol were added to precipitate overnight. The precipitate was collected by centrifugation at 4000 r / min for 20 min, and dissolved in distilled water to obtain an exopolysaccharide aqueous solution. The exopolysaccharide aqueous solution obtained was treated according to the phenol-sulfuric acid method, and the absorbance value was determined by OD490. According to the standard curve, the exopolysaccharide content of lactic acid Pediococcus PA153 was 2232 mg / L, and the exopolysaccharide content of Bacillus coagulans BC66 was 1982 mg / L, indicating that lactic acid Pediococcus PA153 and Bacillus coagulans BC66 both had excellent exopolysaccharide production capabilities, were beneficial to the absorption of nutrients in chickens, and promoted the production and egg-laying ability of hens.

[0058] Example 2

[0059] Immunomodulatory activity test of Pediococcus acidilactici PA153 and Bacillus coagulans BC66

[0060] (1) Test method:

[0061] (1.1) Bacterial culture:

[0062] Pediococcus acidilactici PA153 and Bacillus coagulans BC66 were inoculated into MRS liquid medium, cultured at 30°C for 18 h, centrifuged at 3000 r / min and 4°C for 10 min, and then the cells were collected and washed twice with an equal volume of sterile saline. The cells were centrifuged under the same conditions and suspended in an equal volume of sterile saline. The concentration of the bacterial solution was adjusted to 1×10 by hemocytometer counting. 8 CFU / mL was used to prepare the test bacterial solution.

[0063] (1.2) Mouse gavage test:

[0064] In this experiment, 40 SPF-grade BALB / C female mice with an average body weight of (20±2) g (from Hubei Provincial Center for Disease Control and Prevention) were selected and divided into 4 groups (PA153 strain treatment group, BC66 strain treatment group, PA153+BC66 strain treatment group and control group), with 10 mice in each group.

[0065] The mice in the PA153 strain treatment group were gavage-administered with 1 mL of the above-mentioned concentration of 1×10 8 CFU / mL of PA153 test bacteria solution; mice in the BC66 strain treatment group were gavaged with 1 mL of the above concentration of 1×10 8 CFU / mL of BC66 test bacteria solution; mice in the PA153+BC66 strain treatment group were gavaged with 0.5 mL of the above concentration of 1×10 8CFU / mL of PA153 test bacteria solution and 0.5mL of the above concentration of 1×10 8 CFU / mL of BC66 test bacteria solution; the mice in the control group were gavaged with 1 mL of normal saline once a day for 21 consecutive days.

[0066] (2) Evaluation of immunomodulatory activity

[0067] (2.1) Assessment of macrophage phagocytic function

[0068] 21 days after oral administration of bacterial solution, the carbon clearance test was used to detect the phagocytic function of macrophages. The mice were placed in a fixator and injected with 20% India ink 10mL / kg through the tail. 20μL of blood was collected from the orbital vein 1min and 10min after injection, and added to 2mL 0.1% Na2CO3 solution and shaken well. The optical density values ​​OD1 and OD2 of t1 and t2 were measured at 680nm by spectrophotometry, and the clearance index K was calculated. K=(lg OD1 -lg OD2 ) / (t2-t1), where t1 and t2 are the time between the two injections of ink, and OD1 and OD2 are the absorbances of blood samples measured at different time points.

[0069] 21 days after oral administration of bacterial solution, the mice were weighed and then killed by cervical dislocation. The liver and spleen were removed and rinsed with saline. The surface moisture was absorbed by filter paper. The liver and spleen weights of mice in each test group and control group were weighed, and the phagocytic index α was calculated. α=(M 体质量 ×K 1 / 3 ) / (M 肝质量 +M 脾质量 ×K 1 / 3 ), and the results are shown in Table 3.

[0070] Table 3

[0071] Strain / Indicator Clearance Index K Phagocytic index α PA153 strain treatment group 0.085 3.98 BC66 strain treatment group 0.074 3.45 PA153+BC66 strain treatment group 0.115 5.08 Blank group 0.045 2.24

[0072] (2.2) Detection of mouse serum IgG and related cytokine levels

[0073] After 21 days of gavage treatment, the eyeballs were removed and blood was collected in EP tubes. When serum was precipitated, the supernatant was collected by centrifugation (3000r / min, 15min), and stored in a -80°C refrigerator. The levels of related cytokines and IgA and IgG in mouse serum were detected using ELISA detection kits. The results are shown in Table 4.

[0074] Table 4

[0075]

[0076] (2.3) Lymphocyte transformation test

[0077] After 21 days of oral administration, the eyeballs of the experimental mice were removed and the spleen was washed with Hank's solution and crushed into cytoplasmic bodies on a 200-mesh cell sieve. The cell concentration was adjusted to 3×10 6 / mL, take a 96-well culture plate, divide the well plate into two areas, namely the well area with ConA (concanavalin) added and the well area without ConA added, take the spleen cytoplasmic fluid of each group of mice and inject 1mL into each well, each test group has 24 wells with and without ConA added, the concentration of ConA added is 7.5μg / mL, and 75μL is added to each well. The well plate is placed in a 37℃, 5% CO2 incubator for 72h. 4h before the end, 0.7mL of supernatant is aspirated from each well, 0.7mL of RPMI1640 culture medium without calf serum is added, and 50μL / well of thiazolyl blue (5mg / mL) is added at the same time, and culture is continued for 4h. Afterwards, 1mL of triple solution (prepared by SDS, isobutanol, and hydrochloric acid) is added to each well to completely dissolve the purple crystals. The OD value is measured with an enzyme-linked immunosorbent assay at a wavelength of 595nm. Analyze and compare the differences between each test group and the control group. The stimulation index SI calculation formula is as follows: Stimulation index SI = OD value (ODconA + / ODconA - ), and the results are shown in Table 5.

[0078] Table 5

[0079]

[0080] (2.4) Measurement of immune organ index

[0081] The mice were weighed and then killed by cervical dislocation. The thymus and spleen were removed and weighed after fat removal. The thymus index (the ratio of thymus mass to body mass) and spleen index (the ratio of spleen mass to body mass) of each experimental group and the control group were calculated. The results are shown in Table 6.

[0082] Table 6

[0083]

[0084] It can be seen from the test data in Tables 3 to 6 that the immune organ index, lymphocyte transformation ability, macrophage phagocytosis ability and related cytokine levels of the PA153+BC66 strain treatment group were higher than those of the control group and the single bacteria treatment group, indicating that the combination of Pediococcus acidilactici PA153 and Bacillus coagulans BC66 can significantly enhance the immune system function of chickens, thereby reducing the negative impact of the disease on production performance and improving production quality.

[0085] Example 3

[0086] Effects of testing composite bacteria (Pediococcus acidilactici PA153 and Bacillus coagulans BC66) on immunomodulatory ability and growth performance

[0087] (1) Experimental groups:

[0088] Two hundred newborn Roman pink laying hens were evenly divided into 8 groups, with 25 chickens in each group. The 8 groups were PA153+BC66 high-dose group 1 (the ratio of the bacterial counts of PA153 and BC66 was 1:1), PA153+BC66 high-dose group 2 (the ratio of the bacterial counts of PA153 and BC66 was 4:1), PA153+BC66 low-dose group (the ratio of the bacterial counts of PA153 and BC66 was 1:1), PA153 group, BC66 group, commercially available Pediococcus acidilactici + commercially available Bacillus coagulans group (the ratio of the bacterial counts of commercially available Pediococcus acidilactici and commercially available Bacillus coagulans was 1:1, the commercially available Pediococcus acidilactici was CGMCC NO.21345, and the commercially available Bacillus coagulans was CCTCC NO.M 2015273), immunized control group and blank control group.

[0089] (2) Treatment methods for each experimental group:

[0090] Blank control group: fed with conventional broiler feed;

[0091] The immune control group was fed with conventional broiler feed and immunized with one dose of Newcastle disease LaSota attenuated virus vaccine at 7 days of age;

[0092] PA153+BC66 low-dose group: fed with conventional broiler feed, and given 1×10 8 CFU / mL bacterial solution, the oral dose is 1mL / day, and one dose of Newcastle disease LaSota attenuated virus vaccine is administered to the eyes at 7 days of age;

[0093] PA153+BC66 high-dose group 1, PA153+BC66 high-dose group 2, PA153 group, BC66 group, commercially available lactic acid Pediococcus + commercially available Bacillus coagulans group: broiler chickens were fed conventional feed and given 1×10 8 CFU / mL bacterial solution, the oral dose is 2mL / day, and one dose of Newcastle disease LaSota attenuated virus vaccine is immunized in the eyes at 7 days of age.

[0094] (3) Measurement indicators

[0095] (3.1) Determination of immune organ index

[0096] On the 14th day, 5 chickens were randomly selected from each group and weighed. 2 mL of blood was collected from the heart. After slaughter, the spleen, thymus and bursa of Fabricius were taken to determine the immune organ index. The results are shown in Table 7.

[0097] Table 7

[0098]

[0099]

[0100] (3.2) Determination of cytokine and serum antibody levels

[0101] 2 mL of blood was collected from the hearts of 5 chickens randomly in each group, and the contents of cytokine gamma interferon (IFN-γ), interleukin 6 (IL-6), immunoglobulin IgA, and immunoglobulin IgG in the serum on the 14th and 28th days were determined using ELISA kits, and the Newcastle disease (ND) antibody titer was measured using a hemagglutination inhibition test. The results are shown in Table 8.

[0102] Table 8

[0103]

[0104]

[0105] (3.3) Growth performance measurement

[0106] Before the start of the experiment, the initial body weight of 15 Roman pink laying hens in each group without blood sampling was weighed, and the final body weight (final weight) of 15 Roman pink laying hens in each group was weighed on the 49th day. The average daily weight gain, daily feed intake, survival rate and feed-to-weight ratio were calculated. The results are shown in Table 9.

[0107] Table 9

[0108]

[0109] It can be seen from the test data in Tables 7 to 9 that the combination of lactic acid Pediococcus PA153 and Bacillus coagulans BC66 involved in the present invention can improve the organ indexes of spleen, bursa of Fabricius and thymus, indicating that the combination of the two promotes the immune development of these organs and slows down the atrophy rate of bursa of Fabricius. At the same time, the combination of lactic acid Pediococcus PA153 and Bacillus coagulans BC66 can increase the content of IgA, IgG, IFN-γ and IL-6 in serum, indicating that the combination of the two improves the humoral immunity level of chickens and can promote T lymphocyte differentiation. In addition, the combination of lactic acid Pediococcus PA153 and Bacillus coagulans BC66 can also increase the daily weight gain of Roman pink laying hens, reduce the feed-to-meat ratio, and improve the breeding survival rate of Roman pink laying hens. The above test results show that the combination of lactic acid Pediococcus PA153 and Bacillus coagulans BC66 involved in the present invention has excellent immune system regulation ability, reduces the negative impact of diseases on production performance, and thus improves production quality.

[0110] Example 4

[0111] Test the effect of composite bacteria (Pediococcus acidilactici PA153 and Bacillus coagulans BC66) on production performance

[0112] (1) Experimental groups:

[0113] A total of 210 48-week-old Roman pink laying hens were evenly divided into 7 groups, with 30 chickens in each group. The 7 groups were PA153+BC66 high-dose group 1 (the ratio of the bacterial counts of PA153 and BC66 was 1:1), PA153+BC66 high-dose group 2 (the ratio of the bacterial counts of PA153 and BC66 was 4:1), PA153+BC66 low-dose group (the ratio of the bacterial counts of PA153 and BC66 was 1:1), PA153 group, BC66 group, commercially available Pediococcus acidilactici + commercially available Bacillus coagulans group (the ratio of the bacterial counts of commercially available Pediococcus acidilactici and commercially available Bacillus coagulans was 1:1, the commercially available Pediococcus acidilactici was CGMCC NO.21345, and the commercially available Bacillus coagulans was CCTCCNO.M 2015273) and a blank control group.

[0114] (2) Treatment methods for each experimental group:

[0115] Blank control group: fed with regular egg-laying feed every day for 90 consecutive days;

[0116] PA153+BC66 low-dose group: fed with conventional egg-laying feed, and 1g1×10 8 CFU / g bacterial powder, and continued to eat for 90 days. PA153+BC66 high-dose group 1, PA153+BC66 high-dose group 2, PA153 group, BC66 group, commercially available lactic acid Pediococcus + commercially available Bacillus coagulans group: eat regular egg-laying feed, add 2g 1×10 8 CFU / g bacterial powder, continuous feeding for 90 days.

[0117] After 90 days of feeding, the average egg weight, egg production rate, qualified egg rate and mortality rate of each test group were compared. The results are shown in Table 10.

[0118] Table 10

[0119]

[0120] It can be seen from the test data in Table 10 that the combination of Pediococcus acidilactici PA153 and Bacillus coagulans BC66 involved in the present invention can significantly increase egg weight, egg production rate, qualified egg rate, and reduce mortality rate, and has a certain synergistic effect in improving poultry production performance, which can improve poultry production performance and improve the economic benefits of poultry farming.

[0121] The applicant declares that the present invention illustrates the technical solution of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0122] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0123] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. Use of a microbial preparation in the preparation of a product for improving poultry production performance, characterized in that: The strains in the microbial preparation include Pediococcus acidilactici PA153 and Bacillus coagulans BC66; The classification name of the lactic acid Pediococcus PA153 is Pediococcus acidilactici, the preservation number is CCTCC No: M2023230, and the preservation date is March 2, 2023; The classification name of the Bacillus coagulans BC66 is Bacillus coagulans, the preservation number is CGMCC No.21879, and the preservation date is March 8, 2021.

2. The use according to claim 1, characterized in that: The bacterial count ratio of the lactic acid bacteria PA153 and the Bacillus coagulans BC66 is (1-4):

1.

3. The use according to claim 1 or 2, characterized in that: The total bacterial count in the microbial preparation is not less than 1×10 8 CFU / mL or 1×10 8 CFU / g.

4. The use according to any one of claims 1 to 3, characterized in that: The microbial preparation also includes a protective agent.

5. The use according to claim 4, characterized in that: The protective agent includes any one of lactose, oligoxylose, butter powder, Tween-80, mannitol, dextrin, gum arabic, sucrose, sorbitol, polyvinyl pyrrolidone or sodium glutamate, or a combination of at least two thereof.

6. The use according to any one of claims 1 to 5, characterized in that: The dosage form of the microbial preparation includes solution or powder.

7. The use according to any one of claims 1 to 6, characterized in that: The poultry includes chicken, duck or goose.

8. A poultry feed, characterized in that: The poultry feed includes corn, peanut meal, sesame meal, millet bran, bran and microbial preparation; The strains in the microbial preparation include Pediococcus acidilactici PA153 and Bacillus coagulans BC66; The classification name of the lactic acid Pediococcus PA153 is Pediococcus acidilactici, the preservation number is CCTCC No: M2023230, and the preservation date is March 2, 2023; The classification name of the Bacillus coagulans BC66 is Bacillus coagulans, the preservation number is CGMCC No.21879, and the preservation date is March 8, 2021.

9. The poultry feed according to claim 8, characterized in that: The bacterial count ratio of the lactic acid bacteria PA153 and the Bacillus coagulans BC66 is (1-4):1; Preferably, the total bacterial count in the microbial preparation is not less than 1×10 8 CFU / mL or 1×10 8 CFU / g.

10. The poultry feed according to claim 8 or 9, characterized in that: The mass percentage of the microbial preparation in the poultry feed is 0.1-2%.

Images