Compound probiotic preparation for preventing gout in goslings and preparation method thereof

Abstract

The application provides a compound probiotic preparation for preventing gosling gout and a preparation method thereof. Salivaria lactobacillus, plant lactobacillus and xylanolytic bacteroides are scientifically matched, so that the synergistic effect is achieved. The compound probiotic preparation can significantly reduce gosling serum uric acid, reduce the incidence of gosling gout, and improve the growth performance of gosling. The compound probiotic preparation has a good development and application prospect in the prevention and treatment of gosling hyperuricemia and gout drugs.

Description

TECHNICAL FIELD

[0001] The application provides a complex probiotic preparation for preventing gosling gout, and simultaneously discloses a preparation method of the complex probiotic, and belongs to the technical field of livestock disease prevention and treatment preparations. BACKGROUND

[0002] Gout is a metabolic disease shared by human and bird caused by abnormal increase of blood uric acid due to disorder of purine metabolism, formation of hyperuricemia, and deposition of urate in joints, chest and abdominal cavity, and surfaces of various organs. The pathogenic factors of gosling gout are complex. According to the literature, more than 20 factors can cause gosling gout, and new pathogenic factors are continuously discovered. At present, common factors causing gout include infectious factors, nutritional factors, toxic factors and feeding management factors. In general, factors that can cause disorder of purine metabolism and uric acid excretion may cause gout.

[0003] Due to different causes and modes of action of gout, the pathogenesis is different, and there is no product or drug for preventing and treating gosling gout on the market. Therefore, it is urgent to find a new and effective treatment method. Microecological preparation is a kind of live microorganism that can exert beneficial effects on animals by improving the balance of intestinal flora according to the principle of microecology. Microecological preparation can grow and metabolize in the animal body, and the various digestive enzymes produced can improve the digestion and absorption of nutritional components in feed and improve the utilization rate of feed. At present, it has been widely used in feed, agriculture, medicine and health care, food and other fields. In the feed industry, the main applications are lactobacillus, bifidobacterium, bacillus and yeast. However, there is no report on the application of probiotic preparation in preventing gosling hyperuricemia and gout. SUMMARY

[0004] The application aims at the situation that excessive high-protein feed leads to hyperuricemia and gout in goslings, and provides a complex probiotic preparation for preventing hyperuricemia and gout and a preparation method thereof.

[0005] The application provides the following technical scheme: the complex probiotic preparation is prepared by freeze-drying and compounding of Lactobacillus salivarius Lactobacillus salivarius , Lactobacillus plantarum and Bacteroides Bacteroides sp strains, wherein the Lactobacillus salivarius Lactobacillus salivarius and Bacteroides Bacteroides sp strains are different strains isolated from the intestines of healthy and diseased goslings, the compounding ratio (FU / g) of the Lactobacillus salivarius Lactobacillus salivarius , Lactobacillus plantarum and Bacteroides Bacteroides sp strains is 10:1:10, and the compounding Lactobacillus salivarius Lactobacillus salivarius and Bacteroides Bacteroides sp are not less than 1x107 CFU / g, Lactobacillus plantarum is not less than 1x10 8 CFU / g, the balance is a protective agent.

[0006] The compound probiotic preparation for preventing gosling gout provided by the present application is prepared by the following substances in a ratio of 10:1:10 (CFU / g):

[0007] Lactobacillus salivarius (Lactobacillus salivarius) Lactobacillus salivarius ) 10; Lactobacillus plantarum 1; Bacteroides (Bacteroides) Bacteroides sp ) 10;

[0008] The Lactobacillus salivarius is named as: Lactobacillus salivarius Lactobacillus salivarius , preserved in the China General Microbiological Culture Collection Center, located at No. 3, Xibei Road, Beichen, Beijing, China, on September 16, 2022, with a preservation number of CGMCC NO: 25732;

[0009] The Bacteroides is named as: Bacteroides Bacteroides sp , preserved in the China General Microbiological Culture Collection Center on March 28, 2023, located at No. 3, Xibei Road, Beichen, Chaoyang District, Beijing, with a preservation number of CGMCC NO: 26995;

[0010] The Lactobacillus plantarum is purchased from the North Biological Lactobacillus plantarum quantitative strain, with a BNCC number of BNCC364132.

[0011] The preparation method of the compound probiotic preparation for preventing gosling gout provided by the present application comprises the following steps:

[0012] 1) Preparation of Lactobacillus salivarius Lactobacillus salivarius freeze-dried powder:

[0013] (1) Activation: streak Lactobacillus salivarius Lactobacillus salivarius on MRS solid medium, and culture at 37℃ in an anaerobic bag for 48h, then pick single colonies and inoculate in MRS liquid medium, and stand in a 37℃ incubator for 6h;

[0014] (2) Expansion culture: inoculate in MRS liquid medium at a inoculation amount of 1%, and stand in a 37℃ incubator for 6h;

[0015] (3) Centrifugation: after expansion culture, put the bacterial slurry into a centrifuge, and centrifuge at a speed of 5500rpm / min for 8min to separate the bacterial liquid, and then take out the bacterial slurry;

[0016] (4) Freeze-drying: Under aseptic conditions, add the protective agent at a ratio of 2:1 for bacterial solution and protective agent, mix well, and place in the freeze dryer for freeze-drying.

[0017] (5) Grinding: Grind the freeze-dried bacterial mud into powder to obtain Lactobacillus salivarius. Lactobacillus salivarius Freeze-dried bacterial powder.

[0018] The obtained Lactobacillus salivarius Lactobacillus salivarius The live bacteria count in the freeze-dried bacterial powder is not less than 1×10⁻⁶. 7 CFU / g.

[0019] 2) Preparation of freeze-dried Lactobacillus plantarum powder:

[0020] (1) Activation: Lactobacillus plantarum was streaked onto MRS solid medium and cultured in an anaerobic bag at 37°C for 48 hours. Single colonies were picked and inoculated into MRS liquid medium and then placed in a 37°C incubator for 12 hours.

[0021] (2) Expanded culture: Inoculate 1% of the culture into MRS liquid medium and incubate at 37°C for 12 hours.

[0022] (3) Centrifugation: After the expansion culture is completed, put the bacterial sludge into a centrifuge. The centrifuge speed is 5500 rpm / min. Centrifuge for 8 minutes to separate the bacterial liquid. After completion, take out the bacterial sludge.

[0023] (4) Freeze-drying: Under aseptic conditions, add the protective agent at a ratio of 2:1 for bacterial solution and protective agent, mix well, and place in the freeze dryer for freeze-drying.

[0024] (5) Grinding: Grind the freeze-dried bacterial mud into powder to obtain freeze-dried Lactobacillus plantarum powder.

[0025] The viable count of the obtained freeze-dried Lactobacillus plantarum powder is not less than 1×10⁻⁶. 8 CFU / g.

[0026] 3) Bacteroides Bacteroides sp Preparation of freeze-dried powder:

[0027] (1) Activation: Bacteroides Bacteroides sp Streak the bacteria in BHI solid medium and incubate in an anaerobic bag at 37°C for 48 hours. Pick a single colony and inoculate it into BHI liquid medium. Let it stand in a 37°C incubator for 12 hours.

[0028] (2) Expanded culture: Inoculate 1% of the culture into BHI liquid medium and incubate at 37°C for 12 hours.

[0029] (3) centrifugation: after the end of the expansion, the slurry is placed in a centrifuge, the centrifuge speed is 5500 rpm / min, centrifugation is carried out for 8 min to separate the bacterial liquid, and the slurry is taken out after completion;

[0030] (4) freeze-drying: under sterile conditions, the protective agent is added in a ratio of 2:1 of bacterial liquid: protective agent, mixed uniformly, and placed in the plate layer of the freeze dryer for freeze-drying operation;

[0031] (5) grinding: the freeze-dried slurry is ground to powder, that is, the Bacteroides Bacteroides sp freeze-dried powder.

[0032] The obtained Bacteroides Bacteroides sp freeze-dried powder has a viable bacterial count of not less than 1x10 7 CFU / g.

[0033] 4) uniformly mixing the Lactobacillus salivarius Lactobacillus salivarius 10, Lactobacillus plantarum 1 and Bacteroides Bacteroides sp 10 (CFU / g) prepared in steps 1-3, to obtain the complex probiotic preparation of the present application.

[0034] The positive effects of the present application are that the Lactobacillus salivarius Lactobacillus salivarius , Lactobacillus plantarum and Bacteroides Bacteroides sp are scientifically compounded to achieve a synergistic effect; compared with the use of Lactobacillus salivarius Lactobacillus salivarius , Lactobacillus plantarum and Bacteroides Bacteroides sp alone or in combination, the use of the complex bacterial suspension can more efficiently reduce uric acid levels and gout incidence, indicating that the combined use of the three probiotics has a greater effect than the use of single or two-strain combinations.

[0035] The 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 complex probiotic preparation has a good development and application prospect in the prevention and treatment of gosling hyperuricemia and gout drugs. BRIEF DESCRIPTION OF DRAWINGS

[0036] Figure 1 is a comparison chart of serum uric acid values of goslings in normal protein diet group, high protein diet group, high protein diet + Lactobacillus salivarius bacterial suspension, high protein diet + Lactobacillus plantarum bacterial suspension, high protein diet + Bacteroides bacterial suspension, high protein diet + Lactobacillus salivarius and Lactobacillus plantarum bacterial suspension, high protein diet + Lactobacillus salivarius and Bacteroides bacterial suspension, high protein diet + Lactobacillus plantarum and Bacteroides bacterial suspension, and high protein diet + complex bacterial suspension group;

[0037] Figure 2The gout incidence rate comparison chart of the normal protein diet group, the high protein diet group, the high protein diet + saliva lactobacillus bacteria suspension group, the high protein diet + plant lactobacillus bacteria suspension group, the high protein diet + bactobacteria bacteria suspension group, the high protein diet + saliva lactobacillus and plant lactobacillus bacteria suspension group, the high protein diet + saliva lactobacillus and bactobacteria bacteria suspension group, the high protein diet + plant lactobacillus and bactobacteria bacteria suspension group and the high protein diet + compound bacteria suspension group;

[0038] Figure 3 The serum uric acid value and gout incidence rate comparison chart of the normal protein diet group, the high protein diet group and the high protein diet + compound probiotic preparation group of gosling geese;

[0039] Figure 4 The total weight gain and feed-meat ratio comparison chart of the normal protein diet group and the normal protein diet + compound probiotic preparation group of gosling geese. DETAILED DESCRIPTION

[0040] The present application is further exemplarily described by the following examples, which do not limit the present application in any way, and any modification or change made by those skilled in the art to the present application without departing from the technical solutions of the present application will fall within the scope of the claims of the present application.

[0041] Example 1

[0042] Salivary lactobacillus Lactobacillus salivarius Preparation of freeze-dried powder:

[0043] MRS broth medium: 10.0 g / L of proteose peptone, 8.0 g / L of beef powder, 4.0 g / L of yeast powder, 20.0 g / L of glucose, 2.0 g / L of potassium phosphate dibasic, 2.0 g / L of ammonium citrate dibasic, 5.0 g / L of sodium acetate, 0.2 g / L of magnesium sulfate, 0.04 g / L of manganese sulfate, 1.0 g / L of Tween 80, pH 5.7±0.2;

[0044] MRS solid medium: 10.0 g / L of proteose peptone, 8.0 g / L of beef powder, 4.0 g / L of yeast powder, 20.0 g / L of glucose, 2.0 g / L of potassium phosphate dibasic, 2.0 g / L of ammonium citrate dibasic, 5.0 g / L of sodium acetate, 0.2 g / L of magnesium sulfate, 0.04 g / L of manganese sulfate, 14.0 g / L of agar, 1.0 g / L of Tween 80, pH 6.5±0.2;

[0045] Activation: the salivary lactobacillus Lactobacillus salivarius was streaked on the MRS solid medium and cultured at 37℃ for 48h in an anaerobic bag, and a single colony was inoculated in the MRS liquid medium and placed in a 37℃ incubator for 6h;

[0046] Expansion culture: inoculate into MRS liquid medium at 1% inoculation amount, and stand in a 37°C incubator for 6h;

[0047] Centrifugation: after the expansion culture, put the bacterial slurry into a centrifuge, the centrifuge speed is 5500rpm / min, centrifuge for 8min to separate the bacterial liquid, and then take out the bacterial slurry;

[0048] Freeze-drying: under sterile conditions, add the protective agent at a ratio of 2:1 of bacterial liquid to protective agent, mix well, and then place in the plate layer of the freeze dryer for freeze-drying operation;

[0049] Grinding: grind the freeze-dried bacterial slurry into powder, and then obtain the Lactobacillus salivarius Lactobacillus salivarius Freeze-dried bacterial powder.

[0050] The obtained Lactobacillus salivarius Lactobacillus salivarius The viable count of the freeze-dried bacterial powder is not less than 1×10 7 CFU / g.

[0051] Preparation of Lactobacillus plantarum freeze-dried powder:

[0052] MRS broth medium: 10.0g / L proteose peptone, 8.0g / L beef powder, 4.0g / L yeast powder, 20.0g / L glucose, 2.0g / L potassium phosphate dibasic, 2.0g / L diammonium hydrogen citrate, 5.0g / L sodium acetate, 0.2g / L magnesium sulfate, 0.04g / L manganese sulfate, 1.0g / L Tween 80, pH 5.7±0.2;

[0053] MRS solid medium: 10.0g / L proteose peptone, 8.0g / L beef powder, 4.0g / L yeast powder, 20.0g / L glucose, 2.0g / L potassium phosphate dibasic, 2.0g / L diammonium hydrogen citrate, 5.0g / L sodium acetate, 0.2g / L magnesium sulfate, 0.04g / L manganese sulfate, 14.0g / L agar, 1.0g / L Tween 80, pH 6.5±0.2;

[0054] Activation: streak the Lactobacillus plantarum on the MRS solid medium, and culture in an anaerobic bag at 37°C for 48h, then pick a single colony and inoculate in the MRS liquid medium, and stand in a 37°C incubator for 12h;

[0055] Expansion culture: inoculate into MRS liquid medium at 1% inoculation amount, and stand in a 37°C incubator for 6h;

[0056] Centrifugation: after the expansion culture, put the bacterial slurry into a centrifuge, the centrifuge speed is 5500rpm / min, centrifuge for 8min to separate the bacterial liquid, and then take out the bacterial slurry;

[0057] Freeze-drying: under sterile conditions, the protective agent is added in a ratio of 2:1 of bacteria solution: protective agent, mixed and placed in the plate layer of the freeze dryer for freeze-drying operation;

[0058] Crushing: the freeze-dried bacteria mud is ground into powder, and Lactobacillus plantarum freeze-dried bacteria powder is obtained.

[0059] The obtained Lactobacillus plantarum freeze-dried bacteria powder has a viable bacterial count of not less than 1×10 8 CFU / g.

[0060] Bacteroides Bacteroides sp Preparation of freeze-dried powder:

[0061] BHI broth medium: 10.0 g / L of tryptone, 17.5 g / L of beef heart infusion powder, 5.0 g / L of sodium chloride, 2.0 g / L of glucose, 2.5 g / L of potassium phosphate dibasic, 2.5 g / L of diammonium hydrogen citrate, pH 7.4±0.2;

[0062] BHI solid medium: 10.0 g / L of tryptone, 17.5 g / L of beef heart infusion powder, 5.0 g / L of sodium chloride, 2.0 g / L of glucose, 2.5 g / L of potassium phosphate dibasic, 2.5 g / L of diammonium hydrogen citrate, 13.5 g / L of agar, pH 7.4±0.2.

[0063] Activation: the Bacteroides Bacteroides sp Streaked on BHI solid medium, cultured at 37℃ for 48h in an anaerobic bag, and single colonies were picked and inoculated in BHI liquid medium and placed in a 37℃ incubator for 12h;

[0064] Expansion culture: inoculated in BHI liquid medium at a 1% inoculation amount and placed in a 37℃ incubator for 12h;

[0065] Centrifugation: after the expansion culture, the bacteria mud was placed in a centrifuge, the centrifuge speed was 5500rpm / min, and the bacteria solution was separated by centrifugation for 8min, and then the bacteria mud was taken out;

[0066] Freeze-drying: under sterile conditions, the protective agent is added in a ratio of 2:1 of bacteria solution: protective agent, mixed and placed in the plate layer of the freeze dryer for freeze-drying operation;

[0067] Crushing: the freeze-dried bacteria mud is ground into powder, and Bacteroides Bacteroides sp freeze-dried bacteria powder is obtained.

[0068] The obtained Bacteroides Bacteroides sp freeze-dried bacteria powder has a viable bacterial count of not less than 1×10 7 CFU / g.

[0069] The positive effects of the present application are proved by the following experimental examples

[0070] Experimental Example 1: Effects of bacterial suspension on serum uric acid and gout incidence in goslings

[0071] 1. Preparation of bacterial suspension

[0072] Lactobacillus salivarius Lactobacillus salivarius Lactobacillus plantarum and Bacteroides Bacteroides sp Centrifuge, discard the supernatant, add sterile physiological saline, and prepare a bacterial suspension (1×10⁻⁶). 9 CFU / mL); according to viable count (1×10⁻⁶) 9 The bacterial suspensions were prepared in a 1:1 ratio (CFU / mL) as follows: *Lactobacillus salivarius* + *Lactobacillus plantarum* suspension, *Lactobacillus salivarius* + *Bacteroides salivarius* suspension, and *Lactobacillus plantarum* + *Bacteroides salivarius* suspension. The compound bacterial suspensions were prepared according to a viable count (1×10⁻⁶ 9 A compound bacterial suspension of Lactobacillus salivarius, Lactobacillus plantarum, and Bacteroides was prepared in a 1:1:1 ratio (CFU / mL).

[0073] 2. Laboratory animals

[0074] One-day-old goslings were raised on wire mesh beds with heating lamps. The temperature was maintained at 32±1℃ for the first 7 days, decreasing by 2.5±0.5℃ each week, with a final temperature of 26℃. The animal room was kept clean and well-ventilated at all times.

[0075] 3. Experimental Grouping

[0076] Ninety healthy 1-day-old goslings were randomly divided into 9 groups of 10 each. 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 suspension intervention group, high protein diet + Lactobacillus salivarius and Lactobacillus plantarum suspension intervention group, high protein diet + Lactobacillus salivarius and Bacteroides suspension intervention group, high protein diet + Lactobacillus plantarum and Bacteroides suspension intervention group, and high protein diet + compound bacterial suspension intervention group.

[0077] 4. Experiment Content

[0078] (1) Normal protein diet group: fed a normal protein diet;

[0079] (2) High-protein diet group: fed a high-protein diet;

[0080] (3) High-protein diet + Lactobacillus salivarius suspension intervention group: fed a high-protein diet, and Lactobacillus salivarius suspension (1×10) 9 Administer CFU / mL to goslings via gavage;

[0081] (4) High-protein diet + Lactobacillus plantarum suspension intervention group: fed with a high-protein diet, and Lactobacillus plantarum suspension (1×10 9CFU / mL) were given by gavage to goslings;

[0082] (5) High protein diet + Bacteroides suspension intervention group: high protein diet was fed, and Bacteroides suspension (1 x 10 9 CFU / mL) were given by gavage to goslings;

[0083] (6) High protein diet + Lactobacillus salivarius and Lactobacillus plantarum suspension intervention group: high protein diet was fed, and Lactobacillus salivarius and Lactobacillus plantarum suspension (1:1) were given by gavage to goslings;

[0084] (7) High protein diet + Lactobacillus salivarius and Bacteroides suspension intervention group: high protein diet was fed, and Lactobacillus salivarius and Bacteroides suspension (1:1) were given by gavage to goslings;

[0085] (8) High protein diet + Lactobacillus plantarum and Bacteroides suspension intervention group: high protein diet was fed, and Lactobacillus plantarum and Bacteroides suspension (1:1) were given by gavage to goslings;

[0086] (9) High protein diet + complex bacteria suspension intervention group: high protein diet was fed, and complex bacteria suspension (1:1:1) were given by gavage to goslings;

[0087] (10) The experimental period was 28 days, during which gosling gout incidence was observed, and gout gosling data was recorded;

[0088] (11) On the 29th day, goslings were sacrificed, blood was collected by heart puncture, and serum uric acid value (Serum UA) was detected.

[0089] 5、Experimental results

[0090] As Figure 1The serum UA of the high protein diet group was significantly higher than that of the normal diet group; the serum UA of the Lactobacillus salivarius, Lactobacillus planturum and Bacteroides xylanolyticus groups used alone or in combination was different from that of the model group, but the serum UA of the group taking the compound bacteria suspension was more significantly different, indicating that taking the compound bacteria suspension can more efficiently reduce the uric acid level of goslings. Figure 2 As shown, the gout incidence of the model group goslings was about 40%, the gout incidence of the Lactobacillus salivarius, Lactobacillus planturum and Bacteroides xylanolyticus groups used alone or in combination was between 24% and 35%, and the gout incidence of the group taking the compound bacteria suspension was about 20%, indicating that taking the compound bacteria suspension can more effectively reduce the gout incidence of goslings. In summary, the effect of the combined use of the three probiotics is greater than that of single or two-strain combination.

[0091] Example 2 Effect of Compound Probiotic Preparation on Serum Uric Acid and Gout Incidence of Goslings

[0092] 1. Preparation of Compound Probiotic Preparation

[0093] Lactobacillus salivarius Lactobacillus salivarius Freeze-dried bacteria powder, Lactobacillus planturum freeze-dried bacteria powder and Bacteroides xylanolyticus Bacteroides sp Freeze-dried bacteria powder were mixed in a ratio of 10:1:10, i.e. the preparation of compound probiotic preparation was completed.

[0094] 2. Experimental animals are the same as in Example 1.

[0095] 3. Experimental grouping

[0096] Thirty 1-day-old healthy goslings were randomly divided into three groups, 10 in each group, and the three groups were: normal protein diet group, high protein diet group and high protein diet + compound probiotic preparation intervention group.

[0097] 4. Experimental content

[0098] (1) Normal protein diet group: fed with normal protein diet;

[0099] (2) High protein diet group: fed with high protein diet;

[0100] (3) High protein diet + complex probiotic preparation intervention group: fed with high protein diet and complex probiotic preparation (1 x 10 8 CFU / g);

[0101] (4) The experimental period was 28 days, during which the gosling gout incidence was observed, and the gout gosling data was recorded;

[0102] (5) On the 29th day, the goslings were sacrificed, blood was collected by heart puncture, and serum uric acid (Serum UA) was detected.

[0103] 5、Experimental results

[0104] As Figure 3 shown, compared with the normal diet group, the serum uric acid of the high protein diet group was significantly increased; compared with the high protein diet group, the serum uric acid of the high protein diet + complex probiotic preparation intervention group was significantly decreased, and the incidence of gout in goslings was reduced. It shows that the complex probiotic preparation can reduce the serum uric acid level of goslings and reduce the occurrence of gosling gout.

[0105] Example 3 Effect of complex probiotic preparation on growth performance of goslings

[0106] 1、The preparation of complex probiotic preparation is the same as in experimental example 2.

[0107] 2、The experimental animals are the same as in example 1.

[0108] 3、Experimental grouping:

[0109] Twenty 1-day-old healthy Sanhua goslings were randomly divided into 2 groups, 10 in each group, and the 2 groups were: normal protein diet group, normal protein diet + complex probiotic preparation intervention group

[0110] 4、Experimental content

[0111] (1) Normal protein diet group: fed with normal protein diet;

[0112] (2) Normal white diet + complex probiotic preparation intervention group: fed with normal protein diet and complex probiotic preparation (1 x 10 8 CFU / g);

[0113] (3) The body weight of goslings in the two groups was weighed every two days in the morning, and the feed consumption was recorded;

[0114] (4) The experimental period was 28 days.

[0115] 5、Experimental results

[0116] AsFigure 4 As shown, compared with the normal diet group, the total weight gain of the normal protein diet + compound probiotic preparation intervention group increased and the feed conversion ratio decreased, indicating that the compound probiotic preparation can improve feed conversion rate and enhance the growth performance of goslings.

[0117] in conclusion

[0118] A comparison of serum uric acid (UA) levels in nine groups of goslings is shown in the figure. Figure 1 The incidence of gout was compared among the nine groups of goslings. Figure 2 The comparison of serum uric acid (UA) and incidence of gout among the three groups of goslings is shown in the figure below. Figure 3 The comparison of total weight gain and feed conversion ratio of goslings in the normal protein diet group and the normal protein diet + compound probiotic preparation group is shown in the figure. Figure 4 As shown in the figure, Lactobacillus salivarius can be used alone or in combination. Lactobacillus salivarius Although there were differences in serum uric acid levels between the *Lactobacillus plantarum* and *Bacteroides* groups and the model group, the difference was more significant in the group receiving the compound bacterial suspension, indicating that the compound bacterial suspension was more effective in reducing uric acid levels in goslings. The incidence of gout in the model group was approximately 40%. *Lactobacillus salivarius* was used alone or in combination. Lactobacillus salivarius Lactobacillus plantarum and Bacteroides Bacteroides sp The incidence of gout in the control group was between 24% and 35%, while the incidence in the group receiving the compound probiotic suspension was around 20%, indicating that the compound probiotic suspension was more effective in reducing the incidence of gout in goslings. Furthermore, the compound probiotic preparation significantly reduced serum uric acid levels in goslings, decreased the incidence of gout, and improved their growth performance. Therefore, the compound probiotic preparation shows promising potential for development and application in the prevention and treatment of hyperuricemia and gout in goslings.

Claims

1. A compound probiotic preparation for preventing gout in goslings, characterized in that: Prepared from the following substances in a mass ratio: Lactobacillus salivarius (Lactobacillus salivarius) Lactobacillus salivarius ) 10 parts; Lactobacillus plantarum (Lactobacillus plantarum) Lactobacillus plantarum ) 1 part; Bacteroides (Bacteroides) Bacteroides sp ) 10 parts; The Lactobacillus salivarius is named as: Lactobacillus salivarius Lactobacillus salivarius and was preserved in the China General Microbiological Culture Collection Center, located at No. 1, Xibei Road, Beichen, Beijing, China, on September 16, 2022, with a preservation number of CGMCC NO: 25732. The Bacteroides is named as: Bacteroides sp. Bacteroides sp The Bacteroides was preserved in China General Microbiological Culture Collection Center, located at No. 1, Xibaheyi Road, Chao Yang District, Beijing, on March 28, 2023, and the preservation number is CGMCC NO: 26995. The Lactobacillus plantarum is purchased from Beina Biological Lactobacillus plantarum quantitative strain, and the BNCC number is BNCC364132.

2. The preparation method of the compound probiotic preparation for preventing gosling gout 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, cultivate at 37℃ in an anaerobic bag for 48h, pick single colonies and inoculate in MRS liquid medium, and stand in a 37℃ incubator for 6h; (2) Expansion culture: inoculate in MRS liquid medium at an inoculation amount of 1%, and stand in a 37℃ incubator for 6h; (3) Centrifugation: after the expansion culture is completed, put the bacterial slurry into a centrifuge, the centrifuge speed is 5500rpm / min, centrifuge for 8min to separate the bacterial liquid, and then take out the bacterial slurry; (4) Freeze-drying: under sterile conditions, add a protective agent in a ratio of 2:1 of bacterial liquid: protective agent, mix well, and then place in the plate layer of a freeze dryer to perform freeze-drying operation; (5) Grinding: grind the freeze-dried bacterial slurry into powder, and Lactobacillus salivarius freeze-dried bacterial powder is obtained; The obtained freeze-dried Lactobacillus salivarius powder has a viable cell count of not less than 1 x 10 7 CFU / g; 2) Preparation of Lactobacillus plantarum freeze-dried powder: (1) Activation: streak Lactobacillus plantarum on MRS solid medium, cultivate at 37℃ in an anaerobic bag for 48h, pick single colonies and inoculate in MRS liquid medium, and stand in a 37℃ incubator for 12h; (2) Expansion culture: inoculate in MRS liquid medium at an inoculation amount of 1%, and stand in a 37℃ incubator for 12h; (3) Centrifugation: after the expansion culture is completed, put the bacterial slurry into a centrifuge, the centrifuge speed is 5500rpm / min, centrifuge for 8min to separate the bacterial liquid, and then take out the bacterial slurry; (4) Freeze-drying: under sterile conditions, add a protective agent in a ratio of 2:1 of bacterial liquid: protective agent, mix well, and then place in the plate layer of a freeze dryer to perform freeze-drying operation; (5) Grinding: grind the freeze-dried bacterial slurry into powder, and Lactobacillus plantarum freeze-dried bacterial powder is obtained; The obtained Lactobacillus plantarum freeze-dried bacterial powder has a viable bacterial count not less than 1 × 10 8 CFU / g; 3) Preparation of Bacteroides sp freeze-dried powder: (1) Activation: streak Bacteroides sp on BHI solid medium, cultivate at 37℃ in an anaerobic bag for 48h, pick single colonies and inoculate in BHI liquid medium, and stand in a 37℃ incubator for 12h; (2) Expansion culture: inoculate in BHI liquid medium at an inoculation amount of 1%, and stand in a 37℃ incubator for 12h; (3) Centrifugation: after the expansion culture is completed, put the bacterial slurry into a centrifuge, the centrifuge speed is 5500rpm / min, centrifuge for 8min to separate the bacterial liquid, and then take out the bacterial slurry; (4) Freeze-drying: under sterile conditions, add a protective agent in a ratio of 2:1 of bacterial liquid: protective agent, mix well, and then place in the plate layer of a freeze dryer to perform freeze-drying operation; (5) Grinding: grind the freeze-dried bacterial slurry into powder, and Bacteroides sp freeze-dried bacterial powder is obtained; 4) Mix 10 parts of Lactobacillus salivarius, 1 part of Lactobacillus plantarum and 10 parts of Bacteroides prepared in steps 1)~3) uniformly, to obtain the composite probiotic preparation.

3. The method of claim 2, wherein, The culture condition of the Lactobacillus salivarius is: temperature 37℃, anaerobic culture, inoculation amount 1%, culture time 6h, and viable bacterial count greater than 10 9 CFU / mL.

4. The method of claim 2, wherein: The culture condition of the Lactobacillus plantarum is: temperature 37℃, anaerobic culture, inoculation amount 1%, culture time 12h, and viable bacterial count greater than 10 10 CFU / mL.

5. The method of claim 2, wherein: The culture condition of the Bacteroides is: temperature 37℃, strict anaerobic culture, inoculation amount 1%, culture time 12h, and viable bacterial count greater than 10 9 CFU / mL.

6. The method of claim 2, wherein: The compounded Lactobacillus plantarum in the complex probiotic preparation is not less than 1 x 10 8 CFU / g, the compounded Lactobacillus salivarius and Bacteroides is not less than 1 x 10 7 CFU / g.

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