Composite probiotic preparation for preventing gout of goslings and preparation method thereof

Abstract

According to the compound probiotic preparation for preventing gout of the goslings and the preparation method of the compound probiotic preparation, lactobacillus salivarius, lactobacillus plantarum and bacteroides xylans are scientifically matched, so that a synergistic effect is achieved; the serum uric acid of the goslings can be remarkably reduced, the gout incidence rate of the goslings is reduced, and the growth performance of the goslings is improved; the compound probiotic preparation has good development and application prospects in medicines for preventing and treating gosling hyperuricemia and gout.

Description

Technical Field

[0001] The invention provides a composite probiotic preparation for preventing gout in goslings and also discloses a preparation method of the composite probiotic preparation, belonging to the technical field of livestock disease prevention and treatment preparations. Background Art

[0002] Gout is a metabolic disease common to humans and poultry caused by abnormal increase of blood uric acid due to purine metabolism disorder, forming hyperuricemia, and then depositing urate in the joints, chest and abdominal cavity and the surface of various organs. The cause of gout in goslings is relatively complex. According to literature reports, there are more than 20 factors that can cause gout in goslings, and new pathogenic factors are constantly being discovered. At present, common factors that cause gout include infectious diseases, nutritional factors, toxic factors, and feeding and management factors. In general, any factors that can cause purine metabolism disorders and uric acid excretion disorders may lead to the occurrence of gout.

[0003] Due to the different causes and modes of action of gout, the pathogenesis is also different, and there are no products or drugs specifically used to prevent and treat gout in goslings on the market. Therefore, it is urgent to find new and effective treatment methods. Probiotic preparations are living microorganisms that use the principles of microecology to exert a favorable effect on animals by improving the balance of intestinal flora. Probiotic preparations can grow and metabolize in the animal body, and the various digestive enzymes produced can improve the body's digestion and absorption of nutrients in feed and improve feed utilization. At present, it has been widely used in various fields such as feed, agriculture, medical care and food. Lactobacillus, bifidobacterium, bacillus and yeast are mainly used in the feed industry. However, there are no reports on the use of probiotic preparations in preventing hyperuricemia and gout in goslings. Summary of the invention

[0004] The purpose of the present invention is to provide a composite probiotic preparation for preventing hyperuricemia and gout and a preparation method thereof in view of the situation that goslings develop hyperuricemia and gout due to excessive feeding of high-protein feed.

[0005] The present invention provides the following technical solution: the composite probiotic preparation is prepared by freeze-drying Lactobacillus salivarius, Lactobacillus plantarum and Bacteroides xylanolyticus strains, wherein the Lactobacillus salivarius and Bacteroides xylanolyticus strains are differential strains isolated from the intestines of sick and healthy goslings, and the composite ratio (FU / g) of the Lactobacillus salivarius, Lactobacillus plantarum and Bacteroides xylanolyticus strains is 10:1:10, and the composite Lactobacillus salivarius and Bacteroides xylanolyticus strains are not less than 1×10 7 CFU / g, Lactobacillus plantarum not less than 1×10 8 CFU / g, the remainder is protective agent.

[0006] The composite probiotic preparation for preventing gout in goslings of the present invention is prepared from the following substances in a ratio of 10:1:10 (CFU / g): Lactobacillus salivarius 10; Lactobacillus plantarum 1; Bacteroides xylanolyticus 10; The described Lactobacillus salivarius is named as: Lactobacillus salivarius Lactobacillus salivarius ; Deposited in the General Microbiological Management Center of China Committee for Microbiological Culture Collection, the deposit address is No. 3, No. 1 Beichen West Road, Beijing, China, the deposit date is September 16, 2022, and the deposit number is CGMCC NO: 25732;

[0007] The xylanolytic Bacteroides is named as: Bacteroides sp ; On March 28, 2023, the depositor was at the General Microbiology Center of China Microorganism Culture Collection Administration, the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC NO: 26995; Lactobacillus plantarum was purchased from Beina Biotechnology's Lactobacillus plantarum quantitative strain, with the BNCC number being BNCC364132.

[0008] The preparation method of the composite probiotic preparation for preventing gout in goslings according to the present invention comprises the following steps: 1) Preparation of Lactobacillus salivarius freeze-dried powder: (1) Activation: Streak Lactobacillus salivarius on MRS solid medium, culture in an anaerobic bag at 37°C for 48 h, pick a single colony and inoculate it into MRS liquid medium, and place it in a 37°C incubator for 6 h; (2) Expansion culture: Inoculate 1% of the culture medium into MRS liquid culture medium and place in a 37°C incubator for 6 h. (3) Centrifugation: After the expansion is completed, the bacterial sludge is placed in a centrifuge at a speed of 5500 rpm / min and centrifuged for 8 minutes to separate the bacterial liquid. After completion, the bacterial sludge is removed; (4) Freeze-drying: Add protective agent at a ratio of 2:1 under sterile conditions, mix well and place on the plate layer of the freeze dryer for freeze-drying; (5) Grinding: Grinding the freeze-dried bacterial slurry into powder to obtain freeze-dried Lactobacillus salivarius powder.

[0009] The viable bacterial count of the obtained freeze-dried Lactobacillus salivarius powder is not less than 1×107 CFU / g.

[0010] 2) Preparation of Lactobacillus plantarum freeze-dried powder: (1) Activation: Streak Lactobacillus plantarum onto MRS solid medium, culture in an anaerobic bag at 37°C for 48 h, pick a single colony and inoculate it into MRS liquid medium, and place it in a 37°C incubator for 12 h; (2) Expansion culture: Inoculate 1% of the culture medium into MRS liquid culture medium and place in a 37°C incubator for 12 h. (3) Centrifugation: After the expansion is completed, the bacterial sludge is placed in a centrifuge at a speed of 5500 rpm / min and centrifuged for 8 minutes to separate the bacterial liquid. After completion, the bacterial sludge is removed; (4) Freeze-drying: Add protective agent in a ratio of 2:1 under sterile conditions, mix well and place on the plate layer of the freeze dryer for freeze-drying; (5) crushing: grinding the freeze-dried bacterial mud into powder, thereby obtaining freeze-dried Lactobacillus plantarum powder;

[0011] The viable bacterial count of the obtained freeze-dried Lactobacillus plantarum powder is not less than 1×108 CFU / g;

[0012] 3) Preparation of lyophilized powder of Bacteroides xylanolyticus: (1) Activation: Streak Bacteroides xylanisolans on BHI solid medium, culture in an anaerobic bag at 37°C for 48 h, pick a single colony and inoculate it into BHI liquid medium, and place it in a 37°C incubator for 12 h; (2) Expansion culture: Inoculate 1% of the culture medium into BHI liquid culture medium and place in a 37°C incubator for 12 h. (3) Centrifugation: After the expansion is completed, the bacterial sludge is placed in a centrifuge at a speed of 5500 rpm / min and centrifuged for 8 minutes to separate the bacterial liquid. After completion, the bacterial sludge is removed; (4) Freeze-drying: Add protective agent in a ratio of 2:1 under sterile conditions, mix well and place on the plate layer of the freeze dryer for freeze-drying; (5) pulverizing: grinding the freeze-dried bacterial sludge into powder to obtain freeze-dried bacterial powder of Bacteroides xylanolyticus;

[0013] The viable bacterial count of the obtained Bacteroides xylanolyticus freeze-dried powder is not less than 1×107 CFU / g;

[0014] 4) Evenly mixing 10 Lactobacillus salivarius, 1 Lactobacillus plantarum, and 10 Bacteroides xylanolyticus (CFU / g) prepared in steps 1 to 3 to obtain the composite probiotic preparation of the present invention.

[0015] The positive effect of the present invention is that Lactobacillus salivarius, Lactobacillus plantarum and Bacteroides xylanolyticus are scientifically combined to achieve a synergistic effect; compared with the use of Lactobacillus salivarius, Lactobacillus plantarum and Bacteroides xylanolyticus alone or in combination of two, taking the composite bacterial suspension can more effectively reduce the uric acid level and the incidence of gout, indicating that the combined use of the three probiotics is more effective than the effect of a single strain or a combination of two strains.

[0016] The preparation of the present invention can significantly reduce serum uric acid in goslings, reduce the incidence of gout in goslings, and improve the growth performance of goslings. Therefore, the composite probiotic preparation has a good development and application prospect in the prevention and treatment of hyperuricemia and gout in goslings. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 This is a comparison chart of serum uric acid values ​​of goslings in the normal protein diet group, high protein diet group, high protein diet + Lactobacillus salivarius suspension, high protein diet + Lactobacillus plantarum suspension, high protein diet + Bacteroides xylanolyticus suspension, high protein diet + Lactobacillus salivarius and Lactobacillus plantarum suspension, high protein diet + Lactobacillus salivarius and Bacteroides xylanolyticus suspension, high protein diet + Lactobacillus plantarum and Bacteroides xylanolyticus suspension, and high protein diet + composite bacteria suspension group; Figure 2 It is a comparison chart of gout incidence in goslings in the normal protein diet group, high protein diet group, high protein diet + Lactobacillus salivarius suspension, high protein diet + Lactobacillus plantarum suspension, high protein diet + Bacteroides xylanolyticus suspension, high protein diet + Lactobacillus salivarius and Lactobacillus plantarum suspension, high protein diet + Lactobacillus salivarius and Bacteroides xylanolyticus suspension, high protein diet + Lactobacillus plantarum and Bacteroides xylanolyticus suspension, and high protein diet + compound bacteria suspension group; Figure 3 This is a comparison chart of serum uric acid levels and gout incidence in goslings in the normal protein diet group, high protein diet group, and high protein diet + compound probiotic preparation group; Figure 4 This is a comparison chart of the total weight gain and feed-to-meat ratio of goslings in the normal protein diet group and the normal protein diet + compound probiotic preparation group. DETAILED DESCRIPTION

[0018] The present invention is further described by way of the following examples, which do not limit the present invention in any way. Without departing from the technical solution of the present invention, any modification or change made to the present invention that is easily implemented by a person of ordinary skill in the art will fall within the scope of the claims of the present invention.

[0019] Example 1 Preparation of a composite probiotic preparation for preventing gout in goslings

[0020] Preparation of Lactobacillus salivarius freeze-dried powder: MRS broth medium: peptone 10.0 g / L, beef powder 8.0 g / L, yeast powder 4.0 g / L, glucose 20.0 g / L, dipotassium hydrogen phosphate 2.0 g / L, diammonium hydrogen citrate 2.0 g / L, sodium acetate 5.0 g / L, magnesium sulfate 0.2 g / L, manganese sulfate 0.04 g / L, Tween 80 1.0 g / L, pH 5.7 ± 0.2; MRS solid medium: peptone 10.0g / L, beef powder 8.0g / L, yeast powder 4.0g / L, glucose 20.0g / L, dipotassium hydrogen phosphate 2.0g / L, diammonium hydrogen citrate 2.0g / L, sodium acetate 5.0g / L, magnesium sulfate 0.2g / L, manganese sulfate 0.04g / L, agar 14.0g / L, Tween 80 1.0g / L, pH 6.5±0.2 Activation: Streak Lactobacillus salivarius on MRS solid medium, culture in an anaerobic bag at 37°C for 48 hours, pick a single colony and inoculate it into MRS liquid medium, and place it in a 37°C incubator for 6 hours; Expanded culture: inoculate 1% of the culture medium into MRS liquid culture medium and place in a 37°C incubator for 6 hours; Centrifugation: After the expansion is completed, the bacterial sludge is placed in a centrifuge at a speed of 5500 rpm / min and centrifuged for 8 minutes to separate the bacterial liquid. After completion, the bacterial sludge is taken out; Freeze drying: Add protective agent at a ratio of 2:1 under sterile conditions, mix well and place on the plate layer of the freeze dryer for freeze drying; Crushing: Grind the freeze-dried bacterial mud into powder to obtain freeze-dried Lactobacillus salivarius powder.

[0021] The viable count of the obtained freeze-dried Lactobacillus salivarius powder is not less than 1×10 7 CFU / g.

[0022] Preparation of Lactobacillus plantarum freeze-dried powder: MRS broth medium: peptone 10.0 g / L, beef powder 8.0 g / L, yeast powder 4.0 g / L, glucose 20.0 g / L, dipotassium hydrogen phosphate 2.0 g / L, diammonium hydrogen citrate 2.0 g / L, sodium acetate 5.0 g / L, magnesium sulfate 0.2 g / L, manganese sulfate 0.04 g / L, Tween 80 1.0 g / L, pH 5.7 ± 0.2; MRS solid medium: peptone 10.0g / L, beef powder 8.0g / L, yeast powder 4.0g / L, glucose 20.0g / L, dipotassium hydrogen phosphate 2.0g / L, diammonium hydrogen citrate 2.0g / L, sodium acetate 5.0g / L, magnesium sulfate 0.2g / L, manganese sulfate 0.04g / L, agar 14.0g / L, Tween 80 1.0g / L, pH 6.5±0.2 Activation: Streak Lactobacillus plantarum on MRS solid medium, culture in an anaerobic bag at 37°C for 48 hours, pick a single colony and inoculate it into MRS liquid medium, and place it in a 37°C incubator for 12 hours; Expanded culture: inoculate 1% of the culture medium into MRS liquid culture medium and place in a 37°C incubator for 12 hours; Centrifugation: After the expansion is completed, the bacterial sludge is placed in a centrifuge at a speed of 5500 rpm / min and centrifuged for 8 minutes to separate the bacterial liquid. After completion, the bacterial sludge is taken out; Freeze drying: Add protective agent in a ratio of 2:1 under sterile conditions, mix well and place on the plate layer of the freeze dryer for freeze drying; Crushing: Grind the freeze-dried bacterial mud into powder to obtain freeze-dried Lactobacillus plantarum powder.

[0023] The viable count of the obtained freeze-dried Lactobacillus plantarum powder is not less than 1×10 8 CFU / g.

[0024] Preparation of Bacteroides xylanolyticus lyophilized powder: BHI broth medium: tryptone 10.0 g / L, beef heart extract powder 17.5 g / L, sodium chloride 5.0 g / L, glucose 2.0 g / L, dipotassium hydrogen phosphate 2.5 g / L, diammonium hydrogen citrate 2.5 g / L, pH 7.4 ± 0.2; BHI solid medium: tryptone 10.0 g / L, beef heart extract powder 17.5 g / L, sodium chloride 5.0 g / L, glucose 2.0 g / L, dipotassium hydrogen phosphate 2.5 g / L, diammonium hydrogen citrate 2.5 g / L, agar 13.5 g / L, pH 7.4±0.2.

[0025] Activation: Streak the xylanolytic Bacteroides on BHI solid medium, culture in an anaerobic bag at 37°C for 48 hours, pick a single colony and inoculate it into BHI liquid medium, and place it in a 37°C incubator for 12 hours; Expanded culture: inoculate 1% of the culture medium into BHI liquid culture medium and place in a 37°C incubator for 12 hours; Centrifugation: After the expansion is completed, the bacterial sludge is placed in a centrifuge at a speed of 5500 rpm / min and centrifuged for 8 minutes to separate the bacterial liquid. After completion, the bacterial sludge is taken out; Freeze drying: Add protective agent in a ratio of 2:1 under sterile conditions, mix well and place on the plate layer of the freeze dryer for freeze drying; Crushing: Grind the freeze-dried bacterial mud into powder to obtain the freeze-dried bacterial powder of Bacteroides xylanolyticus.

[0026] The number of viable bacteria of the obtained freeze-dried Bacteroides xylanolyticus powder is not less than 1×10 7 CFU / g.

[0027] The positive effects of the present invention are demonstrated by the following experimental examples:

[0028] Experimental Example 1 1. Preparation of bacterial suspension Lactobacillus salivarius, Lactobacillus plantarum and Bacteroides xylanolyticus were centrifuged separately, the supernatant was discarded, and sterile saline was added to prepare a bacterial suspension (1×10 9 CFU / mL); according to the number of viable bacteria (1×10 9 CFU / mL) was prepared into a suspension of Lactobacillus salivarius + Lactobacillus plantarum, a suspension of Lactobacillus salivarius + Bacteroides xylanolyticus, and a suspension of Lactobacillus plantarum + Bacteroides xylanolyticus in a ratio of 1:1; the composite bacterial suspension was prepared according to the number of viable cells (1×10 9 CFU / mL) was prepared in a 1:1:1 ratio of Lactobacillus salivarius + Lactobacillus plantarum + Bacteroides xylanolyticus composite bacterial suspension.

[0029] 2. Experimental Animals One-day-old goslings were kept on a mesh bed with a warm lamp. The temperature was 32±1°C for the first 7 days, and then decreased by 2.5±0.5°C every week to a final temperature of 26°C. The animal room was kept clean and ventilated.

[0030] 3. Experimental Grouping Ninety one-day-old healthy goslings were randomly divided into 9 groups, with 10 goslings in each group. The 9 groups were: normal protein diet group, high protein diet group, high protein diet + Lactobacillus salivarius suspension intervention group, high protein diet + Lactobacillus plantarum suspension intervention group, high protein diet + Bacteroides xylanolyticus suspension intervention group, high protein diet + Lactobacillus salivarius and Lactobacillus plantarum suspension intervention group, high protein diet + Lactobacillus salivarius and Bacteroides xylanolyticus suspension intervention group, high protein diet + Lactobacillus plantarum and Bacteroides xylanolyticus suspension intervention group, high protein diet + composite bacteria suspension intervention group.

[0031] 4. Experimental content (1) Normal protein diet group: fed with normal protein diet; (2) High-protein diet group: fed with a high-protein diet; (3) High-protein diet + Lactobacillus salivarius suspension intervention group: The rats were fed a high-protein diet and the Lactobacillus salivarius suspension (1×10 9 CFU / mL) was administered intragastrically to goslings; (4) High-protein diet + Lactobacillus plantarum suspension intervention group: The rats were fed a high-protein diet and the Lactobacillus plantarum suspension (1×10 9 CFU / mL) was administered intragastrically to goslings; (5) High-protein diet + Bacteroides xylanolyticus suspension intervention group: The rats were fed a high-protein diet and the Bacteroides xylanolyticus suspension (1×10 9 CFU / mL) was administered intragastrically to goslings; (6) High-protein diet + Lactobacillus salivarius and Lactobacillus plantarum suspension intervention group: Goslings were fed a high-protein diet and Lactobacillus salivarius and Lactobacillus plantarum suspension (1:1) was gavaged; (7) High-protein diet + Lactobacillus salivarius and Bacteroides xylanisolvens suspension intervention group: Goslings were fed a high-protein diet and Lactobacillus salivarius and Bacteroides xylanisolvens suspension (1:1) was gavaged; (8) High-protein diet + Lactobacillus plantarum and Bacteroides xylanisolvens suspension intervention group: Goslings were fed a high-protein diet and Lactobacillus plantarum and Bacteroides xylanisolvens suspension (1:1) was gavaged; (9) High-protein diet + compound bacterial suspension intervention group: Goslings were fed a high-protein diet and the compound bacterial suspension (1:1:1) was gavaged into the stomach; (10) The experimental period was 28 days, during which the incidence of gout in goslings was observed and the data of gout goslings were recorded; (11) On day 29, the goslings were killed, blood was collected by cardiac bleeding, and serum uric acid (Serum UA) was measured.

[0032] 5. Experimental results like Figure 1 The following table shows the serum uric acid values ​​of goslings in the normal protein diet group, high protein diet group, high protein diet + Lactobacillus salivarius, high protein diet + Lactobacillus planturum, high protein diet + Bacteroides xylanolyticus, high protein diet + Lactobacillus salivarius and Lactobacillus planturum, high protein diet + Lactobacillus salivarius and Bacteroides xylanolyticus, high protein diet + Lactobacillus planturum and Bacteroides xylanolyticus, high protein diet + Lactobacillus plantarum and Bacteroides xylanolyticus, and high protein diet + Bacteria mixture groups. UA) comparison chart), the serum uric acid level of goslings in the high-protein diet group was significantly higher than that of the normal diet group; the serum uric acid levels of the groups using Lactobacillus salivarius, Lactobacillus plantarum and Bacteroides xylanolyticus alone or in combination were different from those of the modeling group, but the difference in serum uric acid level in the group taking the composite bacterial suspension was more significant, indicating that taking the composite bacterial suspension can more effectively reduce the uric acid level of goslings. Figure 2As shown in the results, the gout incidence rate in the modeling group was about 40%, the gout incidence rate in the group using Lactobacillus salivarius, Lactobacillus plantarum and Bacteroides xylanolyticus alone or in combination was between 24% and 35%, and the gout incidence rate in the group taking the composite bacterial suspension was about 20%, indicating that taking the composite bacterial suspension can more effectively reduce the occurrence of gout in goslings. In summary, the combined use of the three probiotics is more effective than the effect of a single or two-combination strain.

[0033] Experimental Example 2 Effects of compound probiotic preparation on serum uric acid and gout incidence in goslings

[0034] 1. Preparation of compound probiotic preparations The freeze-dried powder of Lactobacillus salivarius, the freeze-dried powder of Lactobacillus plantarum and the freeze-dried powder of Bacteroides xylanolyticus were mixed in a ratio of 10:1:10 to complete the preparation of the composite probiotic preparation.

[0035] 2. Experimental animals are the same as those in Experimental Example 1.

[0036] 3. Experimental Grouping Thirty one-day-old healthy goslings were randomly divided into three groups, with 10 goslings in each group. The three groups were: normal protein diet group, high protein diet group and high protein diet + compound probiotic preparation intervention group.

[0037] 4. Experimental content (1) Normal protein diet group: fed with normal protein diet; (2) High-protein diet group: fed with a high-protein diet; (3) High-protein diet + compound probiotic preparation intervention group: The rats were fed a high-protein diet and a compound probiotic preparation (1×10 8 CFU / g); (4) The experimental period was 28 days, during which the incidence of gout in goslings was observed and the data of gout goslings were recorded; (5) On day 29, the goslings were killed, blood was collected by cardiac bleeding, and serum uric acid (Serum UA) was measured.

[0038] 5. Experimental results like Figure 3 As shown in the results, the serum uric acid level of goslings in the high-protein diet group was significantly higher than that in the normal diet group; compared with the high-protein diet group, the serum uric acid level of the high-protein diet + compound probiotic preparation intervention group was significantly reduced, and the incidence of gout in goslings was reduced. This shows that the compound probiotic preparation can reduce the serum uric acid level of goslings and reduce the incidence of gout in goslings.

[0039] Experimental Example 3 Effect of compound probiotic preparation on growth performance of goslings

[0040] 1. The preparation of the composite probiotic preparation is the same as that of Experimental Example 2.

[0041] 2. Experimental animals are the same as in Example 1.

[0042] 3. Experimental groups: Twenty one-day-old healthy goslings were randomly divided into two groups, with 10 in each group. The two groups were normal protein diet group and normal protein diet + compound probiotic preparation intervention group. 4. Experimental content (1) Normal protein diet group: fed with normal protein diet; (2) Normal diet + compound probiotic preparation intervention group: fed with normal protein diet and compound probiotic preparation (1×10 8 CFU / g); (3) Weigh the body weights of the two groups of goslings every two days in the morning and record their food consumption; (4) The experimental period is 28 days.

[0043] 5. Experimental results like Figure 4 As shown in the figure, compared with the normal diet group, the total weight gain (Total weight gain) of the normal protein diet + compound probiotic preparation intervention group increased, and the feed conversion ration decreased, indicating that the compound probiotic preparation can improve the feed conversion rate and enhance the growth performance of goslings.

[0044] in conclusion:

[0045] Serum UA of nine groups of goslings is compared in Figure 1 The comparison of gout incidence in nine groups of goslings is shown in Figure 2 Serum UA and gout incidence of the three groups of goslings are compared in Figure 3 ; The total weight gain and feed conversion ratio of goslings in the normal protein diet group and the normal protein diet + compound probiotic preparation group are compared in Figure 4. As can be seen from the figure, although the serum uric acid of the group using Lactobacillus salivarius, Lactobacillus plantarum and Bacteroides xylanolyticus alone or in combination is different from that of the modeling group, the difference in serum uric acid in the group taking the composite bacterial suspension is more significant, indicating that taking the composite bacterial suspension can more effectively reduce the uric acid level of goslings. The gout incidence rate of gout in the modeling group was about 40%, the gout incidence rate of Lactobacillus salivarius, Lactobacillus plantarum and Bacteroides xylanolyticus alone or in combination was between 24% and 35%, and the gout incidence rate of the group taking the composite bacterial suspension was about 20%, indicating that taking the composite bacterial suspension can more effectively reduce the occurrence of gout in goslings. And taking a composite probiotic preparation can significantly reduce the serum uric acid of goslings, reduce the incidence of gout in goslings, and improve the growth performance of goslings. Therefore, the composite probiotic preparation has a good development and application prospect in the prevention and treatment of hyperuricemia and gout drugs in goslings.

Claims

1. A composite probiotic preparation for preventing gout in goslings, characterized in that Prepared from the following substances in mass ratio: 10 parts of Lactobacillus salivarius; 1 part of Lactobacillus plantarum; 10 parts of Bacteroides xylanolyticus; The described Lactobacillus salivarius is named as: Lactobacillus salivarius Lactobacillus salivarius ; Deposited in the General Microbiological Management Center of China Committee for Microbiological Culture Collection, the deposit address is No. 3, No. 1 Beichen West Road, Beijing, China, the deposit date is September 16, 2022, and the deposit number is CGMCC NO: 25732; The xylanolytic Bacteroides is named as: Bacteroides sp ; On March 28, 2023, the depositor was at the General Microbiology Center of China Microorganism Culture Collection Administration, the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC NO: 26995; Lactobacillus plantarum was purchased from Beina Biotechnology's Lactobacillus plantarum quantitative strain, with the BNCC number being BNCC364132.

2. The method for preparing a composite probiotic preparation for preventing gout in goslings according to claim 1, comprising the following steps: 1) Preparation of Lactobacillus salivarius freeze-dried powder: (1) Activation: Streak Lactobacillus salivarius on MRS solid medium, culture in an anaerobic bag at 37°C for 48 h, pick a single colony and inoculate it into MRS liquid medium, and place it in a 37°C incubator for 6 h; (2) Expansion culture: Inoculate 1% of the culture medium into MRS liquid culture medium and place in a 37°C incubator for 6 h. (3) Centrifugation: After the expansion is completed, the bacterial sludge is placed in a centrifuge at a speed of 5500 rpm / min and centrifuged for 8 minutes to separate the bacterial liquid. After completion, the bacterial sludge is removed; (4) Freeze-drying: Add protective agent at a ratio of 2:1 under sterile conditions, mix well and place on the plate layer of the freeze dryer for freeze-drying; (5) crushing: grinding the freeze-dried bacterial mud into powder to obtain freeze-dried Lactobacillus salivarius powder; The viable bacterial count of the obtained freeze-dried Lactobacillus salivarius powder is not less than 1×107 CFU / g; 2) Preparation of Lactobacillus plantarum freeze-dried powder: (1) Activation: Streak Lactobacillus plantarum onto MRS solid medium, culture in an anaerobic bag at 37°C for 48 h, pick a single colony and inoculate it into MRS liquid medium, and place it in a 37°C incubator for 12 h; (2) Expansion culture: Inoculate 1% of the culture medium into MRS liquid culture medium and place in a 37°C incubator for 12 h. (3) Centrifugation: After the expansion is completed, the bacterial sludge is placed in a centrifuge at a speed of 5500 rpm / min and centrifuged for 8 minutes to separate the bacterial liquid. After completion, the bacterial sludge is removed; (4) Freeze-drying: Add protective agent in a ratio of 2:1 under sterile conditions, mix well and place on the plate layer of the freeze dryer for freeze-drying; (5) crushing: grinding the freeze-dried bacterial mud into powder, thereby obtaining freeze-dried Lactobacillus plantarum powder; The viable bacterial count of the obtained freeze-dried Lactobacillus plantarum powder is not less than 1×108 CFU / g; 3) Preparation of lyophilized powder of Bacteroides xylanolyticus: (1) Activation: Streak Bacteroides xylanisolans on BHI solid medium, culture in an anaerobic bag at 37°C for 48 h, pick a single colony and inoculate it into BHI liquid medium, and place it in a 37°C incubator for 12 h; (2) Expansion culture: Inoculate 1% of the culture medium into BHI liquid culture medium and place in a 37°C incubator for 12 h. (3) Centrifugation: After the expansion is completed, the bacterial sludge is placed in a centrifuge at a speed of 5500 rpm / min and centrifuged for 8 minutes to separate the bacterial liquid. After completion, the bacterial sludge is removed; (4) Freeze-drying: Add protective agent in a ratio of 2:1 under sterile conditions, mix well and place on the plate layer of the freeze dryer for freeze-drying; (5) pulverizing: grinding the freeze-dried bacterial sludge into powder to obtain freeze-dried bacterial powder of Bacteroides xylanolyticus; 4) 10 parts of Lactobacillus salivarius, 1 part of Lactobacillus plantarum, and 10 parts of Bacteroides xylanolyticus prepared in steps 1 to 3 were mixed evenly and freeze-dried in vacuum.

3. The method according to claim 2, characterized in that The culture conditions of the Lactobacillus salivarius are: temperature 37° C., anaerobic culture, inoculation volume 1%, culture time 6 hours, and viable bacteria count greater than 109 CFU / mL.

4. The method according to claim 2, characterized in that: The culture conditions of the plant lactobacillus are: temperature 37° C., anaerobic culture, inoculation amount 1%, culture time 12 h, and viable bacteria count greater than 1010 CFU / mL.

5. The method according to claim 2, characterized in that: The culture conditions of the xylanolytic Bacteroides are: temperature 37° C., strict anaerobic culture, inoculation amount 1%, culture time 12 h, and viable bacterial count greater than 109 CFU / mL.

6. The method according to claim 2, characterized in that: The content of Lactobacillus plantarum in the composite probiotic preparation after compounding is not less than 1×108 CFU / g, and the content of Lactobacillus salivarius and Bacteroides xylanolyticus after compounding is not less than 1×107 CFU / g.

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