Pediococcus acidilactici strain gbw9011, a solid microbial preparation for preventing weaning rabbit diarrhea prepared by the strain and application thereof

Abstract

This invention discloses a strain of *Pediococcus lactis* GBW9011 and its preparation as a solid microbial preparation for preventing diarrhea in weaned rabbits, belonging to the field of microbial preparations. This invention provides a strain of *Pediococcus lactis* GBW9011, with the preservation number CGMCC No. 34454. This invention utilizes *Pediococcus lactis* GBW9011 to ferment and enrich inorganic zinc ions, preparing zinc-rich lactic acid bacteria powder. This powder is then mixed with a compound enzyme preparation, fructooligosaccharides, and compound vitamins to prepare a solid microbial preparation. This solid microbial preparation can prevent diarrhea in weaned rabbits, exerting a synergistic effect through "inhibiting pathogens, strengthening the barrier, aiding digestion, and resisting stress," significantly reducing the incidence of diarrhea in weaned rabbits, increasing weight gain and survival rates, improving digestive enzyme activity, and improving intestinal flora structure. This has significant implications for animal husbandry, including rabbits.

Description

Technical Field

[0001] This invention belongs to the field of microbial preparation technology, specifically relating to a strain of Pediococcus lactis GBW9011 and its preparation of a solid microbial preparation for preventing diarrhea in weaned rabbits and its application. Background Technology

[0002] With increasing consumer demand for healthy and nutritious foods, rabbit meat, as a type of meat that is high in protein, low in fat, and low in cholesterol, has seen a continuous increase in market demand, bringing significant economic benefits to related industries.

[0003] Weaning is a critical stage in the growth of baby rabbits. At this time, their digestive and immune systems are not yet fully developed, making them highly susceptible to intestinal flora imbalance due to factors such as bacterial and viral infections, feed changes, and environmental stress, which can lead to intestinal diseases such as diarrhea, resulting in high mortality rates and severely restricting the economic benefits of rabbit farming.

[0004] Currently, industry solutions for preventing diarrhea in baby rabbits largely focus on traditional management methods and the addition of single-function additives. However, these single-intervention measures have limited targets, their effects are easily affected by interference, and they are difficult to maintain consistently. Furthermore, they cannot comprehensively address the diarrhea problem in weaned baby rabbits caused by multiple factors, including insufficient digestive enzymes, a fragile intestinal barrier, and oxidative stress. Therefore, developing a green, safe, highly effective microbial preparation with synergistic effects on multiple targets for comprehensive prevention of diarrhea in weaned baby rabbits has significant industrial demand and application value. Summary of the Invention

[0005] To overcome the shortcomings of the prior art, the purpose of this invention is to provide a strain of *Pediococcus lactis* GBW9011 and its prepared solid microbial preparation for preventing diarrhea in weaned rabbits and its application. This invention utilizes *Pediococcus lactis* GBW9011 and, based on synergistic bacterial-enzyme design and nutritional fortification, prepares a solid microbial preparation with good stability, which can effectively prevent diarrhea in weaned rabbits, reduce mortality, and has no drug residues or environmental pollution risks.

[0006] To achieve the above-mentioned objectives, the present invention is implemented through the following technical solution:

[0007] This invention provides a strain of Pediococcus acidilactici GBW9011, which is deposited at the China General Microbiological Culture Collection Center (CGMCC) and classified as Pediococcus acidilactici, with accession number CGMCC No. 34454.

[0008] Furthermore, the colonies of the lactic acid cocci GBW9011 are round, milky white, smooth, slightly convex in the middle, and have neat edges. The bacterial cells are spherical, with a diameter of 0.5~1.0 μm, smooth surface, arranged in pairs, and Gram-positive.

[0009] The present invention also provides zinc-rich lactic acid bacteria powder prepared by fermentation of the aforementioned *Pediococcus lactis* GBW9011.

[0010] Furthermore, the number of live bacteria in the zinc-enriched lactic acid bacteria powder is not less than 1.0 × 10⁻⁶. 11 CFU / g, with an organic zinc content of not less than 1 mg / g.

[0011] The zinc-enriched lactic acid bacteria powder is prepared by inoculating the seed liquid of *Pediococcus lactis* GBW9011 into a culture medium containing zinc sulfate and fermenting it. The zinc ion concentration in the culture medium containing zinc sulfate is 500~750 mg / L.

[0012] Furthermore, the components and contents of the zinc sulfate-containing culture medium include: zinc sulfate 500-750 mg / L, peptone 10-15 g / L, beef extract 8-12 g / L, yeast extract 4-8 g / L, glucose 20-30 g / L, dipotassium hydrogen phosphate 2-5 g / L, triammonium citrate 2-5 g / L, sodium acetate 2-5 g / L, manganese sulfate 50-80 mg / L, and Tween 1-3 g / L, and the pH of the culture medium is 6.0-6.4.

[0013] Furthermore, the fermentation method includes fermentation in a fermenter, where the pressure of the tank is controlled at 0.05 MPa to 0.1 MPa using pure nitrogen gas, and the rotation speed is 80 to 100 r / min.

[0014] The present invention also provides a solid microbial preparation for preventing diarrhea in weaned rabbits, the solid microbial preparation comprising the zinc-enriched lactic acid bacteria powder, the components of the solid microbial preparation comprising, by weight, 40-50 parts of zinc-enriched lactic acid bacteria powder, 30-40 parts of compound enzyme preparation, 10-20 parts of fructooligosaccharides and 5-10 parts of compound vitamins.

[0015] The present invention also provides a method for preparing the aforementioned solid microbial preparation, the method specifically comprising the following steps:

[0016] (1) The seed culture of Pyrococcus lactis GBW9011 was inoculated into a medium containing zinc sulfate, fermented, and then freeze-dried to obtain zinc-enriched lactic acid bacteria powder;

[0017] (2) Mix and stir the compound enzyme to obtain the compound enzyme preparation;

[0018] (3) After sucrose is catalyzed by fructosyltransferase, it is decolorized, desalted, concentrated, and separated by chromatography. After secondary concentration, it is dried to obtain fructooligosaccharides.

[0019] (4) Mix and stir the vitamins to obtain a multivitamin;

[0020] (5) The zinc-rich lactic acid bacteria powder, compound enzyme preparation, fructooligosaccharide and compound vitamin are mixed to prepare a solid microbial preparation that has the effect of preventing diarrhea in weaned rabbits.

[0021] Furthermore, the complex enzyme preparation includes at least one of xylanase, hemicellulase, β-glucanase, protease, amylase, lipase, and glucose oxidase.

[0022] Furthermore, the enzyme activities of the xylanase, hemicellulase, β-glucanase, protease, amylase, lipase, and glucose oxidase are not less than 10,000 U / g, 3,000 U / g, 1,000 U / g, 15,000 U / g, 5,000 U / g, 2,000 U / g, and 2,000 U / g, respectively.

[0023] Furthermore, the mass ratio of xylanase, hemicellulase, β-glucanase, protease, amylase, lipase and glucose oxidase is 8~12:10~15:8~12:15~20:10~15:5~8:5~8.

[0024] Furthermore, the conditions for the fructosyltransferase to catalyze sucrose include: adding the fructosyltransferase enzyme solution to a sterile sucrose solution with a mass concentration of 50%~70%, with the amount of fructosyltransferase added being 5~10 U / mL based on the volume of the sterile sucrose solution; adjusting the pH of the system to 5.5~6.5 with acetic acid / sodium acetate; controlling the temperature of the fermenter to 50~60℃; and reacting for 12~18 h.

[0025] Furthermore, the purity of the oligofructose is not less than 95%.

[0026] Furthermore, the vitamins include at least one of vitamin A, vitamin D3, vitamin E, vitamin C, and vitamin K3.

[0027] Furthermore, the mass ratio of vitamin A, vitamin D3, vitamin E, vitamin C and vitamin K3 is 30~40:15~20:15~20:20~30:10~15.

[0028] Furthermore, the purity of the above vitamins is no less than 98%.

[0029] The present invention also provides the application of the zinc-enriched lactic acid bacteria powder or the solid microbial preparation described herein in the preparation of feed additives for preventing diarrhea in weaned rabbits.

[0030] Furthermore, based on the mass of the feed, the feed contains a solid microbial preparation at a mass content of 1 kg / t to 2 kg / t. The solid microbial preparation is mixed into the feed of weaned rabbits and fed continuously for 30 to 45 days.

[0031] Furthermore, the microbial preparation can reduce the diarrhea rate of weaned rabbits and increase the weight gain rate and survival rate.

[0032] Compared with the prior art, the advantages and technical effects of the method of the present invention are:

[0033] 1. This invention provides a zinc-enriched lactic acid bacteria powder, which enriches inorganic zinc ions through fermentation with *Pediococcus lactis* GBW9011, while simultaneously providing active lactic acid bacteria with colonization and antibacterial effects, as well as easily absorbed organic zinc. *Pediococcus lactis* GBW9011 exhibits excellent acid and bile salt resistance, enabling it to colonize the intestines and inhibit pathogens. The enriched zinc exists in an organic form with high bioavailability, directly repairing the intestinal mucosa and enhancing barrier function. The two work synergistically to prevent diarrhea from both the "microbial community regulation" and "physical barrier" perspectives. The solid microbial preparation prepared by this invention can efficiently and synergistically prevent diarrhea in weaned rabbits.

[0034] 2. This invention utilizes a compound enzyme preparation to precisely break down non-starch polysaccharides (xylan, β-glucan, etc.) that are difficult for weaned rabbits to digest, decompose proteins, starches, and fats, supplement insufficient endogenous enzymes, reduce chyme viscosity, and reduce the fermentation substrate of harmful bacteria in the posterior intestinal tract, thus ensuring intestinal health from the perspective of "reducing intestinal burden"; glucose oxidase catalyzes glucose and oxygen to produce gluconic acid and hydrogen peroxide, consuming oxygen while creating an acidic environment, which is conducive to the colonization of zinc-rich lactic acid bacteria and can further inhibit pathogenic bacteria.

[0035] 3. This invention provides fructooligosaccharides, which, as a highly effective prebiotic, selectively promote the proliferation of lactic acid bacteria and beneficial indigenous bacteria in the gut, forming a synbiotic effect of "probiotics + prebiotics" with zinc-rich lactic acid bacteria, resulting in a more lasting effect of stabilizing the gut microbiota.

[0036] 4. This invention provides a complex vitamin that, by providing key antioxidant vitamins (VE, VC) and vitamins related to mucosal health and blood clotting function (VA, VD, VK3), alleviates oxidative damage caused by weaning stress, supports epithelial cell repair, and safeguards intestinal health from the perspective of "nutritional fortification and antioxidation".

[0037] 5. The solid microbial preparation of this invention features a scientifically formulated composition that, through a synergistic effect of "inhibiting pathogens, strengthening the barrier, aiding digestion, and resisting stress," significantly reduces the incidence of diarrhea in weaned rabbits. By improving intestinal health and increasing the digestibility and absorption of feed nutrients, it promotes the growth of young rabbits, reduces mortality, and enhances breeding efficiency. All ingredients are nationally permitted feed additives, free of antibiotics and high-dose inorganic zinc emissions, making it green and safe. Furthermore, its solid dosage form facilitates long-distance transportation and conventional storage, giving it high value for promotion and application. Attached Figure Description

[0038] Figure 1 This image shows colonies and cells of *Pediococcus lactis* GBW9011 on MRS medium.

[0039] Figure 2 Image of zinc-enriched cells of *Pediococcus lactis* GBW9011.

[0040] Figure 3 The appearance of solid microbial preparations.

[0041] Figure 4 This is the test site for Example 5.

[0042] Figure 5 This image shows the diarrhea of ​​weaned rabbits in the control group in Example 5 and an anatomical diagram of the intestines of the diarrheal rabbits.

[0043] Figure 6 The dilution curves represent the levels of amplicon sequence variants in the cecal microbiota of weaned rabbits, with the horizontal axis representing sequencing volume and the vertical axis representing the level of amplicon sequence variants. Detailed Implementation

[0044] The technical solution of the present invention will be further described in detail below with reference to specific embodiments, but the present invention is not limited to the embodiments. Unless otherwise specified, the experimental methods in the following embodiments are conventional methods. Unless otherwise specified, the experimental materials, reagents, instruments, etc. used in the following embodiments are commercially available products.

[0045] Example 1: Screening, isolation and identification of Pediococcus lactis GBW9011

[0046] 1. Isolation, screening and purification of *Pediococcus lactis* GBW9011

[0047] The bacterial suspension from the collected samples was serially diluted and spread onto MRS agar medium. After multiple separations and purifications, a single colony was obtained, named GBW9011, and stored.

[0048] The colonies of strain GBW9011 cultured on MRS solid medium for 40 h were... Figure 1 As shown, its colonies Figure 1As shown in the left-middle image, the cells are round, milky white, smooth, slightly convex in the middle, and have neat edges; these are bacterial cells of strain GBW9011 cultured in MRS liquid medium for 18 h. Figure 1 As shown in the middle right figure, the bacteria are spherical, with a diameter of 0.5-1.0 μm, smooth surface, arranged in pairs, and Gram-positive.

[0049] 2. Molecular identification of Pietrococcus lactis GBW9011

[0050] Using the DNA of strain GBW9011 as a template, 16S rRNA universal primers were used for amplification. The amplified fragments were sequenced, and the 16S rDNA sequencing results of strain GBW9011 were compared with the sequences in NCBI. The results showed that strain GBW9011 had the highest homology with Pediococcus acidilactici. Therefore, strain GBW9011 was identified as Pediococcus acidilactici.

[0051] 3. Preservation of *Pediococcus lactis* GBW9011 strain

[0052] The selected strain, *Pediococcus acidilactici* GBW9011, was deposited at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, on May 8, 2025. The accession number for *Pediococcus acidilactici* is CGMCC No. 34454.

[0053] Example 2: Investigating the zinc ion enrichment method of Pyrococcus lactis GBW9011

[0054] This embodiment explores the zinc ion enrichment method of *Pediococcus lactis* GBW9011, specifically including the following:

[0055] 1. Sample preparation

[0056] (1) Preparation of seed culture of Pediococcus lactis GBW9011

[0057] Single colonies of *Pediococcus lactis* GBW9011 cultured on MRS solid medium were picked, inoculated into MRS liquid medium, and anaerobically cultured at 37°C for 18 h to serve as seed culture of *Pediococcus lactis* GBW9011.

[0058] (2) Sample preparation

[0059] A 2% seed culture of *Pediococcus lactis* GBW9011 was inoculated into 5 L of MRS medium with a zinc ion concentration of 500 mg / L and cultured anaerobicly at 37 °C for 18 h. Wet cells were then obtained by centrifugation at 8000 × g for 20 min at 4 °C. The wet cells were washed twice with sterile ultrapure water, centrifuged again under the same conditions, and the precipitate was collected as sample 1.

[0060] A 2% seed culture of *Pediococcus lactis* GBW9011 was inoculated into 5 L of MRS medium and anaerobically cultured at 37 °C for 18 h. Subsequently, wet cells were obtained by centrifugation at 8000 × g for 20 min at 4 °C. The wet cells were then resuspended in sterile physiological saline with a zinc ion concentration of 500 mg / L, stirred at 200 r / min for 30 min, washed twice with sterile ultrapure water, and centrifuged under the same conditions. The precipitate was collected as sample 2.

[0061] 2. Sample testing

[0062] Accurately weigh 2 g of Sample 1 and Sample 2 into beakers, add 45 ml of distilled water, adjust the pH to 6.5 with hydrochloric acid or sodium hydroxide, bring the volume to 50 mL, stir at 500 r / min for 10 min, centrifuge at 8000 r / min at room temperature for 15 min, and use the supernatant to detect the content of water-soluble zinc (i.e., inorganic zinc) on the cell surface. Add 45 ml of 10 mmol / L EDTA solution to the precipitate, adjust the pH to 6.5 with hydrochloric acid or sodium hydroxide, bring the volume to 50 mL, stir at 500 r / min for 10 min, centrifuge at 8000 r / min at room temperature for 15 min, and use the supernatant to detect the content of zinc bound to cell wall polysaccharides and proteins (i.e., cell wall bound zinc), and use the precipitate to determine the content of zinc bound to organic macromolecules or small molecules in cells (i.e., intracellular bound zinc).

[0063] 3. Test Results

[0064] The cell image of *Pediococcus lactis* GBW9011 enriched with zinc ions is shown below. Figure 2 As shown in the figure, the results of the test data (Table 1) also indicate that when Pyocortisone lactis GBW9011 is directly inoculated into a culture medium containing zinc ions, the conversion rate of organic zinc is relatively high.

[0065] Table 1. Percentage of zinc content at different bonding sites in the samples

[0066]

[0067] Example 3: Investigating the culture conditions of zinc-enriched lactic acid bacteria

[0068] Experiment 1: 10 L of MRS medium was prepared and divided into 5 groups, with 2 replicates per group. Based on the volume of MRS medium, 0 mg / L, 250 mg / L, 500 mg / L, 750 mg / L, and 1000 mg / L zinc sulfate were added to the medium, respectively. After sterilization and cooling to room temperature, 2% Pediococcus lactis seed culture was inoculated into each group. The cultures were anaerobic at 37°C for 18 h, and then centrifuged at 8000×g for 20 min at 4°C to obtain wet cells. The wet cells were washed twice with sterile ultrapure water and centrifuged under the same conditions. The precipitate was collected, and the zinc content in different groups was detected according to the detection methods for inorganic zinc, cell wall complexed zinc, and intracellular bound zinc described in Example 2. The precipitate was then diluted to an appropriate gradient to detect the viable cell count in each group.

[0069] The results showed that the conversion rate of organic zinc was better when the zinc ion concentration was between 500 and 750 mg / L (Table 2), while the expansion effect was affected when the zinc ion concentration was greater than 750 mg / L (Table 3).

[0070] Table 2. Effects of zinc ion concentration in culture medium on organic zinc conversion rate and viable bacterial count.

[0071]

[0072] Table 3. Effect of zinc ion concentration in culture medium on propagation effect

[0073]

[0074] Example 4: Preparation of a solid microbial preparation for preventing diarrhea in weaned rabbits

[0075] This embodiment provides a method for preparing a solid microbial preparation for preventing diarrhea in weaned rabbits, specifically including the following steps:

[0076] 1. Preparation of zinc-enriched lactic acid bacteria powder: The *Pediococcus lactis* GBW9011 seed culture prepared in Example 2 was inoculated into a fermenter at an inoculation rate of 2% (v / v) and cultured at 37°C for 12-18 hours. Subsequently, the wet cells were obtained by centrifugation at 8000×g for 20 minutes at 4°C. The wet cells were washed twice with sterile ultrapure water, centrifuged under the same conditions, resuspended in 13% skim milk as a freeze-drying protectant, freeze-dried, and then pulverized through an 80-mesh sieve to obtain active zinc-enriched lactic acid bacteria powder. The viable count of the zinc-enriched lactic acid bacteria powder is ≥1.0×10⁻⁶. 11 CFU / g, organic zinc content ≥1 mg / g.

[0077] 2. Preparation of compound enzyme preparation: Xylanase (10000 U / g), hemicellulase (3000 U / g), β-glucanase (1000 U / g), protease (15000 U / g), amylase (5000 U / g), lipase (2000 U / g), and glucose oxidase (2000 U / g) are mixed in a mass ratio of 8:10:8:15:10:5:5. Under low temperature (≤25℃) and low humidity (relative humidity ≤50%) conditions, the mixture is stirred in a mixer at a speed of 300 r / min for 30 minutes. The coefficient of variation of the mixing uniformity is ≤5%.

[0078] 3. Preparation of Fructose-Oligosaccharides: Fructosyltransferase solution is added to a 50%-70% (w / v) sterile sucrose solution. The amount of fructosyltransferase added is 5-10 U / mL, based on the volume of the sterile sucrose solution. The pH of the system is adjusted to 5.5-6.5 using acetic acid / sodium acetate. The fermentation tank temperature is controlled at 50-60℃, and the reaction is carried out for 12-18 h. After cooling to room temperature, the raw material solution is first decolorized by passing it through a column filled with activated carbon, then desalted by passing it through cation exchange and anion exchange columns respectively. It is then transferred to an evaporator and concentrated to obtain a concentrate with a dry matter content of 30%-50%. The concentrate is continuously pumped into a simulated moving bed (SMB) system to obtain high-purity frucose-oligosaccharide syrup. This syrup is then concentrated again in an evaporator to a dry matter content of 60%-70%, and spray-dried to obtain frucose-oligosaccharides with a purity ≥95%.

[0079] 4. Preparation of compound vitamins: Vitamin A, vitamin D3, vitamin E, vitamin C and vitamin K3 are mixed to obtain compound vitamins. The purity of the above vitamins is ≥98%, and their mass ratio is 30:15:15:20:10. The compound vitamins are stirred in a mixer at 300 r / min for 30 minutes under the conditions of avoiding light, low temperature (≤25℃) and low humidity (relative humidity ≤50%). The coefficient of variation of the mixing uniformity is ≤5%.

[0080] 5. Mixing: Mix the prepared zinc-enriched lactic acid bacteria powder, compound enzyme preparation, fructooligosaccharides, and compound vitamins in a mass ratio of 4:3:1:0.5 to obtain a solid microbial preparation.

[0081] The appearance of the prepared solid microbial preparation is as follows: Figure 3 As shown.

[0082] Example 5: Application of solid microbial preparations in rabbit farming

[0083] This embodiment provides the application of the solid microbial preparation prepared in Example 4 in domestic rabbit breeding to explore its effect in preventing diarrhea in weaned rabbits.

[0084] Two hundred and fifty 35-day-old weaned New Zealand White rabbits were randomly divided into five groups (control group, 500 g / t, 1 kg / t, 2 kg / t, and 3 kg / t) according to the principles of equal numbers of males and females, origin, and similar weight. Each group had five replicates, with ten rabbits per replicate. The control group was fed a basal diet, while the experimental groups were fed a basal diet supplemented with 500 g / t, 1 kg / t, 2 kg / t, and 3 kg / t of the solid microbial preparation prepared in Example 4, respectively. Diarrhea and mortality were observed daily, and feed intake was recorded. The experiment lasted for 30 days. After the experiment, the body weight of each group was measured, and the average daily weight gain, feed conversion ratio, diarrhea rate, and mortality rate were calculated. One rabbit was randomly selected from each replicate and slaughtered. The duodenum and cecum were collected and frozen in sterilized cryovials for later testing. The experimental site was as follows: Figure 4 As shown, Figure 5 The image shows the diarrhea status of weaned rabbits in the control group and an anatomical diagram of the intestines of rabbits with diarrhea.

[0085] Table 4 shows that during the experimental period, the average daily weight gain of the experimental group increased by 2.02g, 3.94g, 4.06g, and 3.00g compared to the control group, respectively; the feed conversion ratio of the experimental group decreased by 0.09, 0.18, 0.12, and 0.03 compared to the control group, respectively; the diarrhea rate of the baby rabbits in the experimental group decreased by 10.80%, 16.00%, 15.20%, and 12.40% compared to the control group, respectively; and the mortality rate of the baby rabbits in the experimental group decreased by 9.20%, 13.20%, 14.00%, and 8.00% compared to the control group, respectively.

[0086] Table 5 shows that during the experimental period, the duodenal α-amylase activity in the experimental group was 51.9 U / mgprot, 73.2 U / mgprot, 86.1 U / mgprot, and 86.8 U / mgprot higher than that in the control group, respectively; the duodenal trypsin activity in the experimental group was 152.4 U / mgprot, 188.6 U / mgprot, 219.2 U / mgprot, and 256.7 U / mgprot higher than that in the control group, respectively; and the cecal cellulase activity in the experimental group was 23.8 U / g, 57.4 U / g, 45.1 U / g, and 38.5 U / g higher than that in the control group, respectively.

[0087] Table 6 shows that during the experimental period, the abundance of Firmicutes in the cecum of the experimental group increased by 1.42%, 2.96%, 2.21%, and -1.18% compared to the control group; the abundance of Microbes in the cecum of the experimental group increased by 0.57%, 1.71%, 1.35%, and -1.95% compared to the control group; and the abundance of Proteobacteria in the cecum of the experimental group decreased by 0.24%, 2.29%, 2.03%, and 3.58% compared to the control group. Additionally, combined with... Figure 6The results showed that the dilution curve of amplicon sequence variants in the cecal microbiota of weaned rabbits indicated that the sequencing volume had reached a critical value, and further increasing the sequencing volume would not result in the emergence of new species. Compared with the control group, the addition of solid microbial preparations increased the abundance of cecal microbiota in rabbits, further verifying that solid microbial preparations can improve the intestinal environment of weaned rabbits.

[0088] The above results indicate that the solid microbial preparation provided by this invention can effectively prevent diarrhea in weaned rabbits, increase the activity of intestinal digestive enzymes, improve the composition of intestinal microbiota, enhance production performance, and reduce mortality. The recommended addition amount is 1 kg / t to 2 kg / t.

[0089] Table 4. Effects of adding solid microbial preparations on daily weight gain, feed conversion ratio, diarrhea rate, and mortality in New Zealand white rabbits.

[0090]

[0091] Table 5. Effects of adding solid microbial preparations on the activity of intestinal digestive enzymes in New Zealand white rabbits.

[0092]

[0093] Table 6. Effects of adding solid microbial preparations on the cecal microbiota structure of New Zealand white rabbits.

[0094]

[0095] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit them. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions claimed by the present invention.

Claims

1. A strain of *Pediococcus lactis* GBW9011, characterized in that, The *Pediococcus lactis* GBW9011 is deposited at the China General Microbiological Culture Collection Center, and its classification name is *Pediococcus lactis*. Pediococcus acidilactici Its accession number is CGMCC No. 34454.

2. The zinc-rich lactic acid bacteria powder prepared by fermentation of *Pediococcus lactis* GBW9011 as described in claim 1.

3. The zinc-enriched lactic acid bacteria powder according to claim 2, characterized in that, The zinc-enriched lactic acid bacteria powder has a viable count of not less than 1.0 × 10⁻⁶. 11 CFU / g, with an organic zinc content of not less than 1 mg / g.

4. The zinc-enriched lactic acid bacteria powder according to claim 2, characterized in that, The zinc-enriched lactic acid bacteria powder is prepared by inoculating the seed liquid of *Pediococcus lactis* GBW9011 into a culture medium containing zinc sulfate and fermenting it. The zinc ion concentration in the culture medium containing zinc sulfate is 500~750 mg / L.

5. A solid microbial preparation for preventing diarrhea in weaned rabbits, characterized in that, The solid microbial preparation comprises the zinc-enriched lactic acid bacteria powder of claim 2, and the components of the solid microbial preparation include: 40-50 parts by weight of zinc-enriched lactic acid bacteria powder, 30-40 parts by weight of compound enzyme preparation, 10-20 parts by weight of fructooligosaccharide and 5-10 parts by weight of compound vitamins.

6. The method for preparing the solid microbial preparation according to claim 5, characterized in that, The preparation method of the solid microbial preparation specifically includes the following steps: (1) The seed culture of Pyrococcus lactis GBW9011 was inoculated into a medium containing zinc sulfate for fermentation, and then freeze-dried to obtain zinc-rich lactic acid bacteria powder. (2) Mix and stir the compound enzyme to obtain the compound enzyme preparation; (3) After sucrose is catalyzed by fructosyltransferase, it is decolorized, desalted, concentrated, and separated by chromatography. After secondary concentration, it is dried to obtain fructooligosaccharides. (4) Mix and stir the vitamins to obtain a compound vitamin; (5) The zinc-rich lactic acid bacteria powder, compound enzyme preparation, fructooligosaccharide and compound vitamin are mixed to prepare a solid microbial preparation that can prevent diarrhea in weaned rabbits.

7. The preparation method according to claim 6, characterized in that, The compound enzyme preparation includes at least one of xylanase, hemicellulase, β-glucanase, protease, amylase, lipase and glucose oxidase.

8. The preparation method according to claim 6, characterized in that, The vitamins mentioned include at least one of vitamin A, vitamin D3, vitamin E, vitamin C, and vitamin K3.

9. The use of the zinc-enriched lactic acid bacteria powder of claim 2 or the solid microbial preparation of claim 5 in the preparation of feed additives for preventing diarrhea in weaned rabbits.

10. The application according to claim 9, characterized in that, The feed contains 1 kg / t to 2 kg / t of solid microbial preparations by weight of feed. The solid microbial preparations are mixed into the feed of weaned rabbits and fed continuously for 30 to 45 days.

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