Synbiotic enzyme as well as preparation method and application thereof

Abstract

The invention relates to a synbiotic enzyme as well as a preparation method and application thereof, and belongs to the technical field of biological agriculture. The synbiotic enzyme is prepared from garlic, brown sugar, radix astragali, fructus schisandrae chinensis, fructus chebulae, lactobacillus hirsuti liquid and water. The synbiotic enzyme disclosed by the invention has a broad-spectrum antibacterial effect, and has a relatively strong inhibition effect on bacterial pathogenic bacteria such as vibrio alginolyticus, listonella anguillarum, edwardsiella ictaluri, aeromonas hydrophila, vibrio parahaemolyticus, aeromonas veronii and vibrio cholerae; the composition can be used for preventing and treating hemorrhagic ascites disease, necrotic canker and other aquaculture diseases caused by bacteria and pathogenic bacteria in aquaculture, can improve the feed conversion rate in aquaculture, and improves the growth and reproduction capability, immunity and culture benefits of aquatic animals.

Description

Technical Field

[0001] The present invention relates to the field of bio-agricultural technology, and in particular to a synbiotic enzyme and a preparation method and application thereof. Background Art

[0002] With the increasing degree of aquaculture intensification, the accumulation of large amounts of leftover bait, aquatic animal excrement and debris has led to the deterioration of the aquaculture water environment, which is very likely to induce the occurrence and spread of aquaculture diseases.

[0003] Enzymes are products made from plants, animals, fungi, and other raw materials through microbial fermentation, containing specific bioactive ingredients such as polysaccharides and oligosaccharides, peptides, amino acids, and vitamins. Studies have shown that feeding enzymes to livestock and poultry can improve their digestibility, enhance their immunity, reduce morbidity, and reduce environmental pollution. However, research reports on the use of enzymes in aquaculture are scarce. Summary of the Invention

[0004] The purpose of the present invention is to provide a synbiotic enzyme and its preparation method and application, so as to solve the problems of low aquaculture feed utilization rate and frequent diseases in the middle and late stages of aquaculture in the prior art.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The present invention provides a synbiotic enzyme comprising the following components: garlic, brown sugar, astragalus root, schisandra chinensis, terminalia chebula, Lactobacillus hilgensis liquid, and water;

[0007] The mass ratio of the garlic, brown sugar, astragalus, schisandra chinensis and terminalia chebula is 50-120:20-40:5-20:5-20:5-20;

[0008] The mass volume ratio of the garlic and Lactobacillus hilgridis liquid is 50-120 g:30-60 mL;

[0009] The mass volume ratio of the garlic to water is 50-120 g:700-900 mL.

[0010] Preferably, the Lactobacillus hilgardii bacterial liquid is prepared from Lactobacillus hilgardii RX strain;

[0011] The Lactobacillus hilgardii RX strain is deposited in the General Microbiology Center of the China Culture Collection Administration, located at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The deposit date is April 10, 2020, and the deposit number is CGMCC No. 19559.

[0012] Preferably, the initial live bacteria concentration in the Lactobacillus hilgridis bacterial solution is 0.5 to 2.5×10 9 CFU / mL.

[0013] The present invention also provides a method for preparing the synbiotic enzyme, comprising the following steps:

[0014] (1) Grinding and oxidizing garlic for 20 to 40 minutes to obtain garlic paste;

[0015] (2) Mix garlic paste, brown sugar, astragalus, schisandra chinensis, terminalia chebula, Lactobacillus hilgensis liquid and water, and ferment for 7 to 30 days to obtain synbiotic enzyme.

[0016] Preferably, the grinding speed in step (1) is 2500-3000 rpm, and the grinding time is 2-5 min.

[0017] Preferably, the fermentation temperature in step (2) is 25-35°C.

[0018] The present invention also provides the synbiotic enzyme and the use of the synbiotic enzyme prepared by the preparation method in preparing preparations for preventing and treating bacterial pathogens.

[0019] Preferably, the bacterial pathogens include one or more of Vibrio alginolyticus, Listonella anguillarum, Edwardsiella ictaluri, Aeromonas hydrophila, Vibrio parahaemolyticus, Aeromonas veronii and Vibrio cholerae.

[0020] The present invention also provides the synbiotic enzyme and the use of the synbiotic enzyme prepared by the preparation method in preparing a preparation for preventing and / or treating hemorrhagic ascites in yellow catfish.

[0021] The present invention also provides the synbiotic enzyme and the use of the synbiotic enzyme prepared by the preparation method in preparing a preparation for preventing and / or treating necrotic ulcer disease in yellow catfish.

[0022] The present invention has the following technical effects and advantages:

[0023] (1) The synbiotic enzyme of the present invention has the characteristics of good fermentation performance and stable function. Existing enzymes are mostly naturally fermented. However, the present invention, by inoculating Lactobacillus hilgardii RX strain, not only greatly shortens the preparation cycle of the synbiotic enzyme, but also facilitates the stable function of the synbiotic enzyme in aquatic animals;

[0024] (2) The synbiotic enzyme of the present invention has the characteristics of comprehensive functions. The synbiotic enzyme of the present invention uses garlic, schisandra chinensis, terminalia chebula and astragalus as fermentation raw materials. Garlic contains allicin and alliin, which have certain antibacterial effects; schisandra chinensis and terminalia chebula have been proven to have strong antibacterial effects; astragalus contains bioactive ingredients such as astragalus polysaccharides, astragaloside, flavonoids, etc., which can improve the immunity of aquatic animals; Lactobacillus hilgardii RX strain also has strong antibacterial effects and probiotic functions against a variety of bacterial pathogens. Therefore, the synbiotic enzyme of the present invention can not only provide a high-quality carbon source for probiotics, promote their growth and reproduction, and thus improve their probiotic function, but also the various bioactive ingredients contained in the synbiotic enzyme can also improve the immunity of aquatic animals, and have an antibacterial effect on various bacterial pathogens such as Vibrio alginolyticus, Listonella anguillarum, Edwardsiella ictaluri, Aeromonas hydrophila, Vibrio parahaemolyticus, Aeromonas vernix and Vibrio cholerae, and can be used to prevent and treat aquaculture diseases such as hemorrhagic ascites and necrotizing ulcer disease caused by bacterial pathogens in aquaculture;

[0025] (3) The synbiotic enzyme of the present invention has an outstanding feeding effect. The synbiotic enzyme of the present invention has a high content of total acid and total phenol, and has a strong feeding effect. It can make aquatic animals feed more actively, reduce the amount of residual bait, and increase the feed conversion rate, thereby promoting the growth and development of aquatic animals, improving aquaculture benefits, and contributing to the healthy and sustainable development of aquaculture. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 It is the antibacterial effect of synbiotic enzymes on various bacterial pathogens;

[0027] Figure 2 These are individuals of yellow catfish suffering from hemorrhagic ascites disease;

[0028] Figure 3 These are individuals suffering from necrotic ulcer disease in yellow catfish.

[0029] Preservation Instructions

[0030] Lactobacillus hilgardii is the Lactobacillus hilgardii RX strain, which is deposited in the General Microbiology Center of the China Culture Collection Administration, located at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The deposit date is April 10, 2020, and the deposit number is CGMCC No. 19559. DETAILED DESCRIPTION

[0031] The present invention provides a synbiotic enzyme comprising the following components: garlic, brown sugar, astragalus root, schisandra chinensis, terminalia chebula, Lactobacillus hilgensis liquid, and water;

[0032] The mass ratio of the garlic, brown sugar, astragalus, schisandra chinensis and terminalia chebula is 50-120:20-40:5-20:5-20:5-20, preferably 100:30:10:10:10;

[0033] The mass volume ratio of the garlic and Lactobacillus hilgeni liquid is 50-120 g:30-60 mL, preferably 100 g:50 mL;

[0034] The mass volume ratio of the garlic to water is 50-120 g:700-900 mL, preferably 100 g:750 mL.

[0035] In the present invention, the Lactobacillus hilgardii bacterial liquid is prepared from the Lactobacillus hilgardii RX strain;

[0036] The Lactobacillus hilgardii RX strain is deposited in the General Microbiology Center of the China Culture Collection Administration, located at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The deposit date is April 10, 2020, and the deposit number is CGMCC No. 19559.

[0037] In the present invention, the initial live bacteria concentration in the Lactobacillus hilgeni bacterial solution is 0.5 to 2.5×10 9 CFU / mL, preferably 2×10 9 CFU / mL.

[0038] In the present invention, the Lactobacillus hilgardii is the Lactobacillus hilgardii RX strain, which is deposited in the General Microbiology Center of the China Culture Collection Administration, with the address at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The deposit date is April 10, 2020, and the deposit number is CGMCC No. 19559.

[0039] In the present invention, the method for preparing the Lactobacillus hilgeni bacterial liquid comprises the following steps: inoculating Lactobacillus hilgeni into MRS culture medium for culturing to obtain the Lactobacillus hilgeni bacterial liquid.

[0040] In the present invention, the MRS culture medium uses water as a solvent and includes the following components in the following concentrations: 10-20 g / L of brown sugar, preferably 15 g / L; 5-15 g / L of yeast extract, preferably 8 g / L; 2-10 g / L of sodium acetate, preferably 5 g / L; 1-5 g / L of diammonium hydrogen citrate, preferably 2 g / L; 0.01-0.05 g / L of manganese sulfate, preferably 0.02 g / L; and 1-4 g / L of potassium dihydrogen phosphate, preferably 2 g / L.

[0041] In the present invention, the culture temperature is 25-40° C., preferably 35° C.; the culture time is 18-36 h, preferably 24 h.

[0042] The present invention also provides a method for preparing the synbiotic enzyme, comprising the following steps:

[0043] (1) grinding and oxidizing garlic for 20 to 40 minutes, preferably 30 minutes, to obtain garlic paste;

[0044] (2) Mix garlic paste, brown sugar, astragalus root, schisandra chinensis, terminalia chebula, Lactobacillus hilgensis liquid and water, and ferment for 7 to 30 days, preferably 14 days, to obtain synbiotic enzyme.

[0045] In the present invention, step (1) further comprises peeling the garlic and then soaking the garlic, wherein the soaking time is 2 to 4 hours, preferably 3 hours.

[0046] In the present invention, the grinding speed in step (1) is 2500-3000 rpm, preferably 2900 rpm; the grinding time is 2-5 min, preferably 3 min.

[0047] In the present invention, the fermentation temperature in step (2) is 25-35°C, preferably 30°C.

[0048] The present invention also provides the synbiotic enzyme and the use of the synbiotic enzyme prepared by the preparation method in preparing preparations for preventing and treating bacterial pathogens.

[0049] In the present invention, the bacterial pathogens include one or more of Vibrio alginolyticus, Listonella anguillarum, Edwardsiella ictaluri, Aeromonas hydrophila, Vibrio parahaemolyticus, Aeromonas veronii and Vibrio cholerae.

[0050] The present invention also provides the synbiotic enzyme and the use of the synbiotic enzyme prepared by the preparation method in preparing a preparation for preventing and / or treating hemorrhagic ascites in yellow catfish.

[0051] The present invention also provides the synbiotic enzyme and the use of the synbiotic enzyme prepared by the preparation method in preparing a preparation for preventing and / or treating necrotic ulcer disease in yellow catfish.

[0052] The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0053] Among the strains of the present invention, Lactobacillus hilgardii RX strain is deposited in the General Microbiology Center of China National Committee for Culture Collection of Microorganisms; Vibrio alginolyticus, Listonella anguillarum, Edwardsiella ictaluri, Aeromonas hydrophila, and Vibrio parahaemolyticus are from Tianjin Fisheries Research Institute; Aeromonas veronii and Vibrio cholerae are from Tianjin Academy of Agricultural Sciences;

[0054] Among the reagents of the present invention, Folin phenol reagent was purchased from Beijing Solaibao Technology Co., Ltd., and gallic acid was purchased from Tianjin Guangfu Fine Chemical Research Institute;

[0055] In the instruments of the present invention, the JM-F50 colloid mill was purchased from Wenzhou Qiangzhong Machinery Technology Co., Ltd., the pH meter was purchased from Mettler-Toledo of Switzerland, and the Oxford cup (Φ8nm×6nm×10nm) was purchased from Shanghai Sanshe Industrial Co., Ltd.

[0056] Example 1: Preparation of Synbiotic Ferment

[0057] (1) Purple garlic was soaked in water for 3 h and then peeled. 100 g of the peeled garlic was ground using a JM-F50 colloid mill at 2900 rpm for 3 min and then oxidized in air for 30 min to obtain garlic paste.

[0058] (2) 15 g of brown sugar, 8 g of yeast extract, 5 g of sodium acetate, 2 g of diammonium citrate, 0.02 g of manganese sulfate, and 2 g of potassium dihydrogen phosphate were added to 1 L of water and sterilized at 121°C for 20 min to obtain MRS medium. Lactobacillus shilgardii RX strain was inoculated into the MRS medium at a 5% inoculum volume and cultured at 35°C for 24 h to obtain a Lactobacillus shilgardii bacterial solution with an initial viable bacterial concentration of 2 × 10 9 CFU / mL;

[0059] (3) Take 100 g of garlic paste, 30 g of brown sugar, 10 g of astragalus, 10 g of schisandra chinensis, 10 g of terminalia chebula and 750 mL of water and mix them evenly, inoculate 50 mL of Lactobacillus hilgridis culture liquid, and ferment at 30°C for 14 days to obtain synbiotic enzyme.

[0060] Comparative Example 1: Preparation of Synthetic Ferment

[0061] The synbiotic enzyme was prepared according to the method described in Example 1, except that, in this comparative example, the fermentation time after inoculation with the Lactobacillus hilgris liquid was 7 days.

[0062] Comparative Example 2: Preparation of Synthetic Ferment

[0063] The synbiotic enzyme was prepared according to the method described in Example 1, except that, in this comparative example, the fermentation time after inoculation with the Lactobacillus hilgris liquid was 30 days.

[0064] Experimental Example 1: Determination of fermentation index of synbiotic enzyme

[0065] (1) Determination of effective viable bacterial count: 1 mL of the synbiotic enzyme prepared in Example 1 and Comparative Examples 1-2 was taken and thoroughly mixed with 9 mL of sterile water, and then graded diluted to obtain concentrations of 10 -5 , 10 -6 , 10 -7 1 mL of each dilution was inoculated into MRS medium, and the effective viable bacterial count of each synbiotic enzyme was determined after culturing at 30°C for 2 days. The results are shown in Table 1.

[0066] (2) Drawing of standard curve: 0.1 g of gallic acid was diluted with 100 mL of distilled water to obtain a gallic acid solution, which was then diluted with distilled water to obtain standard solutions with gallic acid contents of 6.25 mg / mL, 12.5 mg / mL, 25 mg / mL, 50 mg / mL, 100 mg / mL, and 200 mg / mL, respectively. Distilled water was used as a blank solution. 1.0 mL of each standard solution was mixed with 1.0 mL of 0.25 mol / L Folin phenol reagent and allowed to stand for 3 min. 2.0 mL of 20% Na2CO3 solution was then added and mixed, and allowed to stand for 30 min. The mixture was centrifuged at 3500 rpm for 3 min, and the supernatant was collected. The absorbance of each supernatant and blank solution was measured at 760 nm, and a standard curve was drawn.

[0067] (3) Determination of total phenol content: 8 mL of each of the synbiotic enzymes prepared in Example 1 and Comparative Examples 1-2 were taken and centrifuged at 8000 rpm for 10 min to obtain a supernatant. The absorbance of each supernatant at 760 nm was determined using the method described in step (2), and the total phenol content of each synbiotic enzyme was calculated based on the standard curve. The results are shown in Table 1.

[0068] (4) pH determination: The pH of each synbiotic enzyme was measured using a Mettler pH meter;

[0069] (5) Determination of total acid content: Add 3 drops of phenolphthalein to each supernatant remaining after determining the total phenol content, and determine the total acid content of each synergistic enzyme according to the method disclosed in "GB12456-2021 National Food Safety Standard Determination of Total Acid in Food". The results are shown in Table 1.

[0070] Table 1 Fermentation index determination results of synbiotic enzymes

[0071] Synbiotic enzymes Cultivation time (d) <![CDATA[Number of viable bacteria (10 9 CFU / mL)]]> Total phenol content (g / L) pH Total acid content (g / L) Example 1 7 2.15 0.37 4.43 13.46 Comparative Example 1 14 10.23 3.54 3.92 16.45 Comparative Example 2 30 1.36 4.48 3.82 23.56

[0072] The results showed that the standard curve regression equation of gallic acid was y = 31.377x-3.4936, R 2 =0.998;

[0073] With the extension of culture time, the effective viable bacterial count in the synbiotic enzyme showed a trend of first increasing and then decreasing, with the highest effective viable bacterial count on the 14th day of culture. The total phenol content and total acid content showed a continuous upward trend, while the pH showed a continuous downward trend. This indicates that the optimal culture period for preparing the synbiotic enzyme is 7 to 14 days.

[0074] Experimental Example 2: Antibacterial Effect of Synbiotic Enzyme

[0075] Vibrio alginolyticus, Listonella anguillarum, Edwardsiella ictaluri, Aeromonas hydrophila, Vibrio parahaemolyticus, Aeromonas veronii, and Vibrio cholerae were inoculated into LB culture medium, respectively, and cultured at 30°C and 150 rpm for 18 h to obtain bacterial pathogen solutions; 0.1 mL of each bacterial pathogen solution was applied to LB culture medium, an Oxford cup was placed on each LB culture medium, 0.2 mL of the synbiotic enzyme prepared in Example 1 was injected into each Oxford cup, and the culture was carried out at 30°C for 48 h. The diameter of the inhibition zone of each LB culture medium was measured. The results are as follows: Figure 1 and as shown in Table 2.

[0076] Table 2 Antibacterial results of synbiotic enzymes on bacterial pathogens

[0077] bacterial pathogens Original Source Diameter of inhibition zone (mm) Vibrio alginolyticus Semi-laevis tongue sole 16.3 Listonella anguillarum Largemouth Bass 13.8 Edwardsiella ictaluri Snakehead 17.5 Aeromonas hydrophila Koi 16.1 Vibrio parahaemolyticus Whiteleg shrimp 15.9 Aeromonas vermiformis Intestinal tract of yellow catfish with hemorrhagic ascites 14.1 Vibrio cholerae Necrotic ulcer disease on the skin of yellow catfish 15.3

[0078] The results show that the synbiotic enzyme of the present invention has a broad-spectrum antibacterial effect, and has a strong inhibitory effect on Vibrio alginolyticus, Listonella anguillarum, Edwardsiella ictaluri, Aeromonas hydrophila, Vibrio parahaemolyticus, Aeromonas welchii and Vibrio cholerae. Among them, the diameters of the inhibition zones of three Vibrio species, namely Vibrio alginolyticus, Vibrio parahaemolyticus and Vibrio cholerae, are all above 15 mm. It also has an outstanding antibacterial effect on bacterial pathogens isolated from the intestines of yellow catfish with hemorrhagic ascites and the epidermis of yellow catfish with necrotizing ulcer disease.

[0079] Experimental Example 3: Application of Synbiotic Enzyme in Treating Hemorrhagic Ascites in Pelteobagrus fulvidraco

[0080] In August 2024, fish in ponds at a yellow catfish breeding base in Ninghe District, Tianjin, were found to have decreased food intake, a distended abdomen, a red and swollen anus, and increased mucus. Autopsies revealed large amounts of blood and fluid accumulation in the abdominal cavity, and a transparent, thinned intestinal wall, typical symptoms of hemorrhagic ascites. The primary bacterial pathogens isolated and identified from the fish were Aeromonas hydrophila and Edwardsiella ictaluri.

[0081] 1000 mL of the synbiotic enzyme prepared in Example 1 was mixed evenly with 20 kg of yellow catfish feed. After continuous feeding for 5 days in the yellow catfish breeding pond, the yellow catfish's feeding condition gradually returned to normal and the abdominal distension gradually disappeared.

[0082] Experimental Example 4: Application of Synbiotic Enzyme in Treating Necrotic Ulcer Disease in Pelteobagrus fulvidraco

[0083] In August 2024, severe ulcers were discovered in yellow catfish (Petrio fulvidraco) fish ponds at a breeding base in Ninghe District, Tianjin. Autopsies revealed pale, pale livers and enlarged kidneys with hemorrhagic spots, typical manifestations of necrotizing ulcer disease. Vibrio cholerae was identified as the primary bacterial pathogen isolated from the lesions.

[0084] 1000 mL of the synbiotic enzyme prepared in Example 1 was mixed evenly with 20 kg of yellow catfish feed and continuously fed to the yellow catfish culture pond once daily. Simultaneously, the culture pond was sprayed with 2000 mL per meter of water depth per acre, every other day. After 3 days, the necrotizing ulcer disease in the yellow catfish was under control, with no new diseased fish appearing. After 7 days, the yellow catfish's feeding and activity returned to normal.

[0085] As can be seen from the above examples, the present invention provides a synbiotic enzyme and its preparation method and application. The synbiotic enzyme of the present invention has a broad-spectrum antibacterial effect and has a strong inhibitory effect on bacterial pathogens such as Vibrio alginolyticus, Listonella anguillarum, Edwardsiella ictaluri, Aeromonas hydrophila, Vibrio parahaemolyticus, Aeromonas vernix and Vibrio cholerae. It can be used to prevent and treat aquaculture diseases such as hemorrhagic ascites and necrotizing ulcer disease caused by bacterial pathogens in aquaculture. At the same time, it can improve the feed conversion rate in aquaculture, and improve the growth and reproduction capacity, immunity and breeding efficiency of aquatic animals.

[0086] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A synbiotic enzyme, characterized in that The invention comprises the following components: garlic, brown sugar, astragalus root, schisandra chinensis, terminalia chebula, Lactobacillus hilarii liquid and water; The mass ratio of the garlic, brown sugar, astragalus, schisandra chinensis and terminalia chebula is 50-120:20-40:5-20:5-20:5-20; The mass volume ratio of the garlic and Lactobacillus hilgridis liquid is 50-120 g:30-60 mL; The mass volume ratio of the garlic to water is 50-120 g:700-900 mL.

2. The synbiotic enzyme according to claim 1, characterized in that The Lactobacillus hilgardii bacterial liquid is prepared from the Lactobacillus hilgardii RX strain; The Lactobacillus hilgardii RX strain is deposited in the General Microbiology Center of the China Culture Collection Administration, located at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The deposit date is April 10, 2020, and the deposit number is CGMCC No. 19559.

3. The synbiotic enzyme according to claim 2, characterized in that The initial live bacteria concentration in the Lactobacillus hilarii bacterial solution is 0.5 to 2.5×10 9 CFU / mL.

4. The method for preparing the synbiotic enzyme according to any one of claims 1 to 3, wherein: The steps include: (1) Grinding and oxidizing garlic for 20 to 40 minutes to obtain garlic paste; (2) Mix garlic paste, brown sugar, astragalus, schisandra chinensis, terminalia chebula, Lactobacillus hilgensis liquid and water, and ferment for 7 to 30 days to obtain synbiotic enzyme.

5. The preparation method according to claim 4, characterized in that The grinding speed in step (1) is 2500-3000 rpm, and the grinding time is 2-5 min.

6. The preparation method according to claim 5, characterized in that The fermentation temperature in step (2) is 25-35°C.

7. Use of the synbiotic enzyme according to any one of claims 1 to 3 or the synbiotic enzyme prepared by the preparation method according to any one of claims 3 to 6 in the preparation of preparations for preventing and treating bacterial pathogens.

8. The use according to claim 7, characterized in that The bacterial pathogens include one or more of Vibrio alginolyticus, Listonella anguillarum, Edwardsiella ictaluri, Aeromonas hydrophila, Vibrio parahaemolyticus, Aeromonas veronii and Vibrio cholerae.

9. Use of the synbiotic enzyme described in any one of claims 1 to 3 or the synbiotic enzyme prepared by the preparation method described in any one of claims 3 to 6 in the preparation of a preparation for preventing and / or treating hemorrhagic ascites in yellow catfish.

10. Use of the synbiotic enzyme according to any one of claims 1 to 3 or the synbiotic enzyme prepared by the preparation method according to any one of claims 3 to 6 in the preparation of a preparation for preventing and / or treating necrotic ulcer disease in yellow catfish.

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