Mink and fox manure microbial fermentation organic fertilizer and preparation method thereof

Abstract

The invention relates to the technical field of organic fertilizer production, and particularly discloses a mink and fox manure microbial fermentation organic fertilizer and a preparation method thereof. Candida lipolytica, bacillus subtilis, lactobacillus plantarum, azotobacter chroococcum and saccharomyces cerevisiae are compounded according to the volume ratio of 3: 3: 2: 1: 1 to prepare a zymophyte solution, the manure of minks and foxes is mixed with oyster mushroom bran, chestnut shells or corn straw according to a specific proportion, fermentation is started at the temperature of 35 DEG C, and the total fermentation period is 20-30 days. According to the preparation method provided by the invention, the degradation efficiency of grease in feces of minks and foxes is remarkably improved, the nitrogen loss is reduced, the total nitrogen content of the obtained organic fertilizer is greater than or equal to 1.97%, the total phosphorus content is greater than or equal to 0.34%, the coliform group is less than or equal to 98.5 CFU / g, and the standards of NY 884-2012 and GB 8172-1987 are met.

Description

Technical Field

[0001] This invention relates to the field of organic fertilizer production technology, specifically to a method for preparing organic fertilizer made from mink and fox excrement fermented by microorganisms. Background Technology

[0002] Mink and fox manure are characterized by high protein, high fat, and high organic matter content, with crude protein content of 15%–25%, fat content of 8%–12%, and organic matter content ≥60%. Simultaneously, its nitrogen content is 2.5%–4.0%, P2O5 content is 1.5%–2.5%, and K2O content is 1.0%–1.8%, theoretically possessing good potential for organic fertilizer production. However, current mink and fox manure processing faces several technical challenges: the carbon-nitrogen ratio adjustment of commonly used auxiliary materials such as straw and sawdust with mink and fox manure lacks precise design; blindly adding auxiliary materials easily leads to insufficient porosity and poor aeration of the fermentation material, creating an anaerobic environment and requiring a fermentation cycle of over 30 days. Furthermore, traditional composting methods are ineffective in degrading mink and fox manure, resulting in high nitrogen loss rates during fermentation and low nutrient content in the organic fertilizer, failing to meet relevant standards. Therefore, developing a new fermentation scheme for mink and fox manure organic fertilizer is essential. Summary of the Invention

[0003] To develop a novel fermentation scheme for mink and fox manure organic fertilizer, this invention provides a method for preparing mink and fox manure microbial fermented organic fertilizer. The preparation method provided by this invention significantly improves the oil degradation efficiency in mink and fox manure, reduces nitrogen loss, and yields organic fertilizer with total nitrogen ≥1.97%, total phosphorus ≥0.34%, and coliform bacteria ≤98.5 CFU / g, meeting the standards of NY884-2012 and GB 8172-1987.

[0004] This invention provides a method for preparing organic fertilizer from mink and fox feces through microbial fermentation, specifically including the following steps: Animal manure is mixed with agricultural fermentation additives at a mass ratio of 6-10:6-10 to obtain a mixture. Add fermentation liquid to the mixture at a ratio of 25 g to 85 g: 1 mL, add water until the moisture content is 55%, and then carry out fermentation. The first turning of the fermentation pile was carried out when the temperature reached 55°C, and a second turning was carried out on the 7th day. Fermentation should be stopped when the fermentation temperature drops to 25°C and the material turns dark brown, loose and non-sticky. The animal feces were mink feces or fox feces; The agricultural fermentation auxiliary material is oyster mushroom spawn, chestnut husk, or corn stalk. The fermentation broth is prepared by mixing Candida lipolytica broth, Bacillus subtilis broth, Lactobacillus plantarum broth, Azotobacter chrysogenum broth and Saccharomyces cerevisiae broth in a volume ratio of 3:3:2:1:1. The effective viable cell count of the *Candida lipolyticis*, *Bacillus subtilis*, *Lactobacillus plantarum*, *Azotobacter chrysogenum*, and *Saccharomyces cerevisiae* cultures is ≥1×10⁻⁶. 8 CFU / mL.

[0005] The fermentation method provided by this invention significantly improves the degradation efficiency of oils in mink and fox feces, reduces nitrogen loss, and produces organic fertilizer with total nitrogen ≥1.97%, total phosphorus ≥0.34%, and coliform bacteria ≤98.5 CFU / g, which meets the standards of NY 884-2012 and GB 8172-1987.

[0006] Furthermore, the number of viable bacteria in the fermentation broth is ≥1×10⁻⁶. 8 CFU / mL.

[0007] Furthermore, the mass ratio of the mixture is mink feces: oyster mushroom spawn = 6:10.

[0008] Furthermore, the ratio of the fermentation liquid in the mixture is 85 g: 1 mL.

[0009] Furthermore, the agricultural fermentation auxiliary material is pulverized and passed through a 2 mm sieve before use.

[0010] Furthermore, the initial temperature of the fermentation is 35°C, and the total fermentation time is 20 to 30 days.

[0011] The present invention also provides a microbial fermented organic fertilizer made from mink and fox feces, wherein the organic fertilizer is prepared by any of the above steps.

[0012] Furthermore, the organic fertilizer is used to improve seed germination rate, root length, or germination index.

[0013] Furthermore, the seeds are uncoated radish seeds.

[0014] Compared with the prior art, the beneficial effects of the present invention are as follows: This invention adds oyster mushroom spawn, chestnut husks or corn stalks as fermentation auxiliary materials. By precisely proportioning the fermentation auxiliary materials and mink or fox feces, the porosity of the fermentation materials is increased. Combined with high-temperature turning and temperature control, the fermentation cycle is shortened to 20 to 30 days.

[0015] This invention employs a compound fermentation agent composed of Candida lipolyticis, Bacillus subtilis, Lactobacillus plantarum, Azotobacter chrysogenum, and Saccharomyces cerevisiae. Candida lipolyticis degrades oils, Bacillus subtilis controls ammonia and retains nitrogen, and Lactobacillus plantarum assists in odor suppression. These three agents exhibit a synergistic effect. When combined with Azotobacter chrysogenum and Saccharomyces cerevisiae in the fermentation liquid, it can minimize nutrient loss, eliminate odors, and improve nutrient availability. This is the most effective strategy for converting high-nitrogen and high-phosphorus waste such as mink and fox into high-quality organic fertilizer. The resulting organic fertilizer has a total nitrogen content ≥1.97%, a total phosphorus content ≥0.34%, and a coliform count ≤98.5 CFU / g, meeting the standards of NY 884-2012 and GB 8172-1987. Detailed Implementation

[0016] The specific embodiments of the present invention are described in detail below, but it should be understood that the scope of protection of the present invention is not limited to the specific embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. Unless otherwise specified, the experimental methods described in the embodiments of the present invention are conventional methods, and the materials and reagents used in the following embodiments are commercially available unless otherwise specified.

[0017] Example 1: A microbial fermentation organic fertilizer made from mink and fox feces and its preparation method *Candida lipolyticis*, *Bacillus subtilis*, *Lactobacillus plantarum*, *Azotobacter chrysogenum*, and *Saccharomyces cerevisiae* were purchased from Gray Algae Biotechnology Co., Ltd. The product codes for *Candida lipolyticis* were HZB227907, *Bacillus subtilis* HZB111503, *Lactobacillus plantarum* HZB115936, *Azotobacter chrysogenum* HZB112096, and *Saccharomyces cerevisiae* HZB854912. After activation according to the manufacturer's instructions, each strain was cultured in the recommended culture medium to obtain a viable count of 1 × 10⁶ cells / year. 8 The bacterial culture was prepared at a concentration of CFU / mL. The fermentation broth was prepared according to a volume ratio of *Candida lipolyticis*: *Bacillus subtilis*: *Lactobacillus plantarum*: *Azotobacter chrysogenum*: *Saccharomyces cerevisiae* = 3:3:2:1:1, ensuring a total viable count of 1 × 10⁻⁶ CFU / mL. 8 CFU / mL.

[0018] The oyster mushroom substrate is waste material from oyster mushroom cultivation. It was taken from the edible fungus cultivation laboratory of Tangshan Normal University and was crushed and passed through a 2 mm sieve before use.

[0019] Following a mink feces:oyster mushroom substrate mixing ratio of 6:10, 637.50 g of mink feces and 1062.5 g of pulverized oyster mushroom substrate were mixed thoroughly, followed by the addition of 20 mL of fermentation liquid and 300 mL of distilled water to bring the moisture content of the fermented material to 55%. Fermentation was then carried out at an initial temperature of 35℃. When the temperature reached 55℃ on the third day of fermentation, the fermentation pile was turned over. The high temperature of 55℃ was maintained until the 12th day of fermentation, at which point the temperature naturally decreased. The pile was turned over again on the 7th day of fermentation. On the 14th day of fermentation, the fermentation temperature dropped back to 25℃. Fermentation ended when the fermented material turned dark brown, loose, and non-sticky, with a total fermentation time of 20 days.

[0020] Example 2: A microbial fermentation organic fertilizer made from mink and fox feces and its preparation method The fermentation broth was prepared in the same manner as in Example 1; the chestnut husks were crushed and passed through a 2 mm sieve before use.

[0021] Following a mink feces:cheskin mixture ratio of 10:6, 312.50 g of mink feces and 187.50 g of crushed chestnut pods were mixed thoroughly, followed by the addition of 20 mL of fermentation culture liquid and 500 mL of distilled water to bring the moisture content of the fermented material to 55%. Fermentation was then carried out at an initial temperature of 35℃. When the temperature reached 55℃ on the third day of fermentation, the fermentation pile was turned over. The high temperature of 55℃ was maintained until the 12th day of fermentation, at which point the temperature naturally decreased. The pile was turned over again on the 7th day of fermentation. On the 14th day of fermentation, the fermentation temperature dropped back to 25℃. Fermentation ended when the fermented material turned dark brown, loose, and non-sticky, with a total fermentation time of 25 days.

[0022] Example 3: A microbial fermentation organic fertilizer made from mink and fox feces and its preparation method The preparation of the fermentation broth was the same as in Example 1; the treatment of chestnut pods was the same as in Example 2.

[0023] Following a blue fox feces:cheskin mixture ratio of 10:8, 277.00 g of blue fox feces and 223.00 g of crushed chestnut pods were mixed thoroughly, followed by the addition of 20 mL of fermentation culture liquid and 500 mL of distilled water to bring the moisture content of the fermented material to 55%. Fermentation was then carried out at an initial temperature of 35℃. On the third day of fermentation, when the temperature reached 55℃, the fermentation pile was turned over. The high temperature of 55℃ was maintained until the 12th day of fermentation, at which point the temperature naturally decreased. On the 7th day of fermentation, the pile was turned over again. On the 14th day of fermentation, the fermentation temperature dropped back to 25℃. Fermentation ended when the fermented material turned dark brown, loose, and non-sticky, with a total fermentation time of 30 days.

[0024] Example 4: A microbial fermentation organic fertilizer made from mink and fox feces and its preparation method The fermentation liquid was prepared in the same way as in Example 1; the corn stalks were crushed and passed through a 2 mm sieve before use.

[0025] Following a mink feces:corn stalk mixing ratio of 10:8, 277.00 g of mink feces and 223.00 g of pulverized corn stalks were mixed thoroughly, followed by the addition of 20 mL of fermentation liquid and 500 mL of distilled water to bring the moisture content of the fermented material to 55%. Fermentation was then carried out at an initial temperature of 35℃. On the third day of fermentation, when the temperature reached 55℃, the fermentation pile was turned over. The high temperature of 55℃ was maintained until the 12th day of fermentation, at which point the temperature naturally decreased. On the 7th day of fermentation, the pile was turned over again. On the 14th day of fermentation, the fermentation temperature dropped back to 25℃. Fermentation ended when the fermented material turned dark brown, loose, and non-sticky, with a total fermentation time of 25 days.

[0026] Example 5: A microbial fermentation organic fertilizer made from mink and fox feces and its preparation method The preparation of the fermentation liquid was the same as in Example 1; the treatment of the corn stalks was the same as in Example 4.

[0027] Following a mixing ratio of blue fox feces to corn stalks of 10:6, 312.50 g of blue fox feces and 187.50 g of crushed corn stalks were mixed thoroughly, followed by the addition of 20 mL of fermentation liquid and 500 mL of distilled water to bring the moisture content of the fermented material to 55%. Fermentation was then carried out at an initial temperature of 35℃. When the temperature reached 55℃ on the third day of fermentation, the fermentation pile was turned over. The high temperature of 55℃ was maintained until the 12th day of fermentation, at which point the temperature naturally cooled down. The pile was turned over again on the 7th day of fermentation. On the 14th day of fermentation, the temperature dropped back to 25℃. Fermentation ended when the fermented material turned dark brown, loose, and non-sticky, with a total fermentation time of 25 days.

[0028] Comparative Example 1: A microbial fermentation organic fertilizer made from mink and fox feces and its preparation method The fermentation broth was prepared according to a volume ratio of Bacillus subtilis: Lactobacillus plantarum: Azotobacter chrysogenum: Saccharomyces cerevisiae = 3:2:1:1, ensuring that the total effective viable count in the fermentation broth was 1×10⁻⁶. 8 CFU / mL.

[0029] The organic fertilizer fermentation steps are the same as in Example 1.

[0030] Comparative Example 2: A microbial fermentation organic fertilizer made from mink and fox feces and its preparation method Fermentation broth was prepared according to the volume ratio of *Candida lipolyticis*: *Bacillus subtilis*: *Azotobacter chrysogenum*: *Saccharomyces cerevisiae* = 3:3:1:1, ensuring that the total effective viable count in the fermentation broth was 1 × 10⁻⁶. 8 CFU / mL.

[0031] The organic fertilizer fermentation steps are the same as in Example 1.

[0032] Comparative Example 3: A microbial fermentation organic fertilizer made from mink and fox feces and its preparation method Fermentation broth was prepared according to a volume ratio of *Candida lipolyticis*: *Lactobacillus plantarum*: *Azotobacter chrysococcus*: *Saccharomyces cerevisiae* = 3:2:1:1, ensuring a total viable count of 1 × 10⁻⁶ cells in the broth. 8 CFU / mL.

[0033] The organic fertilizer fermentation steps are the same as in Example 1.

[0034] Comparative Example 4: A microbial fermentation organic fertilizer made from mink and fox feces and its preparation method No fermentation liquid was added; the remaining organic fertilizer fermentation steps were the same as in Example 1.

[0035] To verify the preparation effect of the organic fertilizer of the present invention, the contents of relevant substances in the organic fertilizers prepared in Examples 1 to 5 and Comparative Examples 1 to 4 of the present invention were determined, and the organic fertilizer prepared in Example 1 was used to conduct a seed germination test.

[0036] I. Test Reagents 40% NaOH solution: Weigh 40 g of NaOH and dissolve it in 100 mL of water.

[0037] 2% boric acid solution: Weigh 20 g of boric acid, dissolve it in water, and dilute to 1000 mL.

[0038] Nitrogen determination mixed indicator: Weigh 0.5 g of bromocresol green powder and 0.1 g of methyl red powder, and dissolve them in 100 mL of 95% ethanol.

[0039] Boric acid-indicator mixture: Slowly add 20 mL of 2% boric acid solution to an Erlenmeyer flask, add 2 drops of nitrogen-distillation mixed indicator, and observe the liquid turn blue-purple.

[0040] 0.05 mol / L hydrochloric acid standard solution: Measure 4.5 mL of concentrated hydrochloric acid solution and transfer it to a 1000 mL volumetric flask. Slowly add water to dilute to 1000 mL, mix thoroughly, and then standardize with borax.

[0041] 0.02 mol / L hydrochloric acid standard solution: Measure 1.7 mL of concentrated hydrochloric acid solution and transfer it to a 1000 mL volumetric flask. Slowly add water to dilute to 1000 mL, mix thoroughly, and then standardize with sodium carbonate solution.

[0042] Ammonium vanadate colorimetric reagent: Weigh 1.25 g of ammonium metavanadate and slowly add it to 250 mL of concentrated nitric acid solution. Separately, take 25 g of ammonium molybdate, add it to 400 mL of water and heat to dissolve it. After cooling to room temperature, pour this solution into the above solution, and then add water to make up to 1000 mL. Store in the dark.

[0043] Phosphorus standard solution: Take an appropriate amount of potassium dihydrogen phosphate, dry it in an oven at 105℃ for 1 h, and after cooling, accurately weigh 0.2195 g of potassium dihydrogen phosphate, dissolve it in an appropriate amount of water, transfer it to a 1000 mL volumetric flask, add 3 mL of concentrated nitric acid solution, dilute with water to 1000 mL, and shake well to obtain a 50 μg / mL phosphorus standard solution.

[0044] Bromothymol blue indicator: Weigh 0.1 g of bromothymol blue and dissolve it in 50 mL of water. Add 20 mL of ethanol, mix well, and dilute with water to 100 mL.

[0045] Ammonia nitrogen mixed indicator: Weigh 200 mg of methyl red powder and dissolve it in 100 mL of ethanol. Separately, take 100 mg of methylene blue powder and dissolve it in 100 mL of ethanol. Mix the above methyl red solution and methylene blue solution at a volume ratio of 2:1 and set aside.

[0046] II. Test Methods 1. Determination of total nitrogen content using the Kjeldahl method 1.1 Sample digestion Accurately weigh 2.0 g of fermented organic fertilizer sample and transfer it to a dry 100 mL digestion tube. Slowly add 10 mL of concentrated sulfuric acid solution, shake well, and secure the digestion tube with clips on a digestion rack. Place the rack in a digestion furnace and digest at 220 V for about 10 minutes. When the white smoke in the digestion tube has mostly dissipated and the digestion liquid in the tube turns dark brown, remove the tube and rack together and place it on a cooling rack. After slightly cooling, slowly add 5 mL of 30% hydrogen peroxide solution to the digestion tube and return it to the digestion furnace. Adjust the voltage to 165 V and continue heating for 3 minutes. Observe the color of the liquid in the digestion tube. If it is not transparent, carefully add a few more drops of 30% hydrogen peroxide solution. Once the liquid in the digestion tube becomes transparent, remove it and cool it to room temperature. Add distilled water to bring the volume to 100 mL, and transfer 10 mL to a new digestion tube, labeling it for later use.

[0047] 1.2 Distillation Place the digestion tube containing 10 mL of pre-digested liquid into the left side of the nitrogen, phosphorus and calcium analyzer. Press the alkali addition button to add 40% NaOH solution to about 40 mL. Prepare a 250 mL conical flask and add 20 mL of boric acid-indicator mixture to it. Then attach it to the lower end of the condenser tube and press the steam button to start heating and distillation. Stop distillation when the volume of distilled liquid is about 150 mL.

[0048] 1.3 Titration Titrate the distillate with a 0.05 mol / L standard hydrochloric acid solution. Stop titrating when the liquid color just changes from blue to transparent, and record the volume of standard hydrochloric acid solution consumed.

[0049] 1.4 Blank Test Replace the sample digest with 10 mL of water, and perform distillation and titration according to the above steps, recording the volume of hydrochloric acid standard solution consumed.

[0050] 1.5 Calculate the total nitrogen content in the sample. The total nitrogen content in the sample is expressed as a mass fraction and is calculated using the following formula: N(%)=c(V-V0)×0.014×D×100 / m; In the formula: c: Molar concentration of the standard solution, in mol / L; V: The volume of hydrochloric acid standard solution consumed during sample titration, in mL; V0: The volume of hydrochloric acid standard solution consumed in the blank test, in mL; 0.014: Molar mass of nitrogen, in g / mol; m: Mass of the sample, in grams; D: Dividing factor, fixed volume / divided volume, which is 100 / 10 in this experiment.

[0051] 2. Determination of total phosphorus content by molybdenum yellow colorimetric method 2.1 Sample digestion The digestion steps are the same as in 1.1.

[0052] 2.2. Plotting the phosphorus standard curve Accurately transfer 1.0 mL, 2.0 mL, 3.0 mL, 4.0 mL, 6.0 mL, 8.0 mL, 10.0 mL, and 12.0 mL of phosphorus standard solution into 50 mL volumetric flasks. Add 10 mL of ammonium vanadomolybdate colorimetric reagent to each flask, dilute with water to 50 mL, shake thoroughly, and let stand for 10 min. Using the 0 mL solution as a reference, measure the absorbance of each solution at 400 nm using a spectrophotometer with a 10 mm cuvette, and plot the phosphorus standard solution curve.

[0053] 2.3 Determination of phosphorus content in samples Take 10 mL of the sample digest into a 50 mL volumetric flask, add 10 mL of ammonium vanadate colorimetric reagent, dilute with water to 50 mL, shake thoroughly, and let stand for 10 min. Using 0 mL of the solution as a reference, measure the absorbance of the sample solution at 400 nm wavelength using a spectrophotometer with a 10 mm cuvette. Determine the phosphorus content in the sample using a standard curve.

[0054] 2.4 Calculate the total phosphorus content in the sample. The total phosphorus content in the sample is expressed as a mass fraction and is calculated using the following formula: P(%) = a × 10 -6 ×V / mV1; In the formula: m: Mass of the sample, in grams; V: The final volume of the sample after digestion, in mL; V1: The volume of sample digestion liquid transferred during colorimetric determination, in mL; a: The phosphorus content of the colorimetric sample solution is obtained from the standard curve, in μg / 50mL; 10 -6 : The coefficient for converting μg to g.

[0055] 3. Determination of ammonia nitrogen content by distillation-neutralization titration method 3.1 Pretreatment of the distillation apparatus Add 250 mL of water to a Kjeldahl flask, then add 0.25 g of light magnesium oxide and a few glass beads. Heat and distill until the distillate is free of ammonia. Stop distillation and discard the residue in the flask.

[0056] 3.2 Sample distillation Weigh 2.0 g of fermented organic fertilizer sample and place it in a volumetric flask. Add 250 mL of water and mix thoroughly to prepare a water sample. Adjust the pH of the water sample to 7.0. Transfer 50 mL of 2% boric acid solution into a receiving flask, ensuring that the outlet of the condenser is submerged below the surface of the boric acid solution. Measure 250 mL of the water sample and place it in a new flask. Add 2 drops of bromothymol blue indicator, 0.25 g of magnesium oxide, and several glass beads. Immediately connect the nitrogen bulb and the condenser to begin heating and distillation. Adjust the distillation rate to approximately 10 mL / min. Stop distillation when the volume of the distillate reaches 200 mL.

[0057] 3.3 Titration Prepare an Erlenmeyer flask, transfer all the distillate into it, add 2 drops of mixed indicator, and titrate with 0.02 mol / L hydrochloric acid standard solution. Observe that the distillate color changes from green to light purple, which is the titration endpoint. Record the volume of hydrochloric acid standard solution consumed.

[0058] 3.4 Blank Test Replace the water sample with 250 mL of water and perform distillation and titration as described above, recording the volume of hydrochloric acid standard solution consumed. This data serves as a blank control.

[0059] 3.5 Calculate the ammonia nitrogen content in the sample. The concentration of ammonia nitrogen in the water sample is calculated using the following formula: ρN=(V1-V0)×C×14.01×1000 / V; In the formula: ρN: The concentration of ammonia nitrogen in the water sample, expressed as N, in mg / L; V: Sample volume, in mL; V1: The volume of standard hydrochloric acid solution consumed in the measurement of the sample, in mL; V0: The volume of standard hydrochloric acid solution consumed in the blank test, in mL; C: Concentration of the hydrochloric acid solution used in the titration, in mol / L; 14.01: Atomic weight of nitrogen, in g / mol.

[0060] The ammonia nitrogen concentration calculated using the above formula can be used to calculate the mass fraction of ammonia nitrogen in the sample based on the volume of the water sample.

[0061] 4. Determination of Escherichia coli and coliform colony count using the BE203 Escherichia coli count test strip method. 4.1 Sample Processing Weigh 25 g of fermented organic fertilizer sample and place it in a sampling container or homogenizing cup containing 225 mL of sterile phosphate buffer diluent to make a 1:10 sample homogenate. Use a 1 mL sterile pipette to draw 1 mL of the 1:10 sample homogenate and inject it into a test tube containing 9 mL of diluent. Shake well to make a 1:100 sample homogenate. Dilute the homogenate in this way.

[0062] 4.2 Sample inoculation and culture Each sample was inoculated using three different inoculation amounts, with five paper discs inoculated for each amount, for a total of 15 paper discs. Immediately after inoculation, the paper discs were placed in a constant temperature incubator and incubated at 45°C for 24 hours.

[0063] 4.3 Result Interpretation If red spots or halos appear on the paper and the surrounding area turns yellow, it is a positive result; If the entire piece of paper turns yellow without any red spots or red tinges, it is a positive result; If the paper turns yellow without any red spots or redness, it is considered negative. If red spots or halos appear on the purple background of the paper, but the surrounding area does not turn yellow, it is considered negative.

[0064] If the paper shows no change, it is negative.

[0065] 4.4 Result Calculation For counting, select test strips with colony counts greater than 15 and less than 150. If the colony counts of both dilutions are greater than 15 and less than 150, multiply the average colony count by the dilution factor to obtain the number of coliform colony-forming units per milliliter or gram of sample. If the colony counts on all test strips at all dilutions are less than 15, multiply the average colony count on the test strip at the lowest dilution by the dilution factor to obtain the number of coliform colony-forming units per milliliter or gram of sample. If no colonies grow on the test strips at all dilutions, multiply the value less than 1 by the lowest dilution factor to obtain the number of coliform colony-forming units per milliliter or gram of sample. If the colony counts of all dilutions are greater than 150, multiply the average colony count on the test strip at the highest dilution by the dilution factor to obtain the number of coliform colony-forming units per milliliter or gram of sample. When counting colonies on a test strip greater than 150, you can also count the colonies in one or two representative squares. After converting the colony count to the number in a single square, multiply by 20 to get the estimated colony count on the test strip. 20 is the area of ​​the filter paper area. The results are expressed in CFU / g.

[0066] 5. Odor Evaluation The odor of the prepared organic fertilizer was evaluated, and the deodorization effect of the fermentation liquid during the organic fertilizer fermentation process was assessed by the type of odor and whether there was an irritating odor.

[0067] 6. Seed germination test 10.00 g of the organic fertilizer prepared in Example 1 was weighed and placed in a 250 mL Erlenmeyer flask. 100 mL of distilled water was added at a solid-liquid ratio of 1:10. The flask was placed in a reciprocating horizontal shaker at a frequency of 100 times / min and an amplitude of 40 mm, and extracted at 25°C for 1 h. After extraction, the mixture was allowed to stand for 0.5 h. The supernatant was then filtered through qualitative filter paper, and the filtrate was collected to obtain the organic fertilizer extract, which served as the treatment group. Distilled water was used as the control group. Uncoated radish seeds were selected as the test seeds. Radish L., germination rate ≥90%, purchased from Tangshan Lotus Seed Industry Co., Ltd.

[0068] Two layers of qualitative filter paper were placed in a petri dish, and 10 seeds were evenly placed on each dish. The treatment group received 10 mL of organic fertilizer extract, while the control group received 10 mL of distilled water. Each treatment was replicated in triplicate. After treatment, the petri dishes were placed in an incubator at 25℃ and incubated in the dark for 48 hours. Water was added daily to keep the filter paper moist due to evaporation. The following parameters were measured after the incubation period:

[0069] Germination rate: final number of germinated seeds / total number of seeds tested × 100%, where the germination standard is radicle breaking through seed coat ≥ 2 mm; Root length: Measure the length of the taproot of each seedling using vernier calipers; Germination index: GI = Σ(Gt / Dt); In the formula, Σ is the summation symbol, which is to add up the Gt / Dt results for each day during the experiment; Gt is the number of newly germinated seeds on day t, which only counts the number of seeds that meet the germination standard on that day, rather than the cumulative number; Dt is the corresponding germination day, such as day 1, day 2, ... day t.

[0070] III. Test Results The results of the determination of the relevant substance content of the organic fertilizers prepared in Examples 1 to 5 of this invention are shown in Table 1.

[0071] To further determine whether the organic fertilizers prepared in each embodiment meet the standards for bio-organic fertilizers, the content determination results of relevant substances in the organic fertilizers prepared in Examples 1 to 5 of this invention were compared and analyzed with the national standards NY 884-2012 "Bio-organic Fertilizers" and GB 8172-1987 "Standards for Agricultural Use of Urban Waste". The results are shown in Table 2.

[0072] Table 1. Determination of content of organic fertilizer-related substances The analysis results show that the organic fertilizers prepared in Examples 1 to 5 of this invention have total nitrogen and total phosphorus contents that are higher than the standard limits for total nitrogen and total phosphorus in GB 8172-1987 "Control Standard for Agricultural Use of Urban Waste", meaning that the total nitrogen and total phosphorus contents are qualified; the coliform counts are lower than the standard limits for coliform counts in NY 884-2012 "Bio-organic Fertilizer", meaning that the coliform counts are qualified. Considering the total nitrogen, total phosphorus, and coliform parameters, the organic fertilizer prepared in Example 1 of this invention has balanced properties and can be considered the optimal product.

[0073] Table 2 Comparative Analysis of Organic Fertilizer Related Substance Content with National Standards Note: "—" indicates that this column of literature does not contain the standard limit value of the index content in this row.

[0074] The results of the determination of relevant substance content and odor evaluation of the organic fertilizers prepared in Example 1 and Comparative Examples 1 to 4 of this invention are shown in Table 3.

[0075] Table 3. Results of determination of relevant substance content and odor evaluation of organic fertilizer The results showed that the organic fertilizer prepared in Example 1 was significantly superior to all comparative examples in terms of total nitrogen content, total phosphorus content, and deodorization effect. Furthermore, a comparison between Example 1 and Comparative Examples 1-3 revealed that *Candida lipolyticis*, *Lactobacillus plantarum*, and *Bacillus subtilis* played crucial roles in the fermentation of mink and fox feces to prepare organic fertilizer. *Candida lipolyticis*'s core function was to degrade grease, *Bacillus subtilis*'s core function was to control ammonia and retain nitrogen, and *Lactobacillus plantarum*'s core function was to assist in odor suppression. These three components exhibited a synergistic effect. When combined with *Azotobacter chrysogenum* and *Saccharomyces cerevisiae* in the fermentation broth, it could minimize nutrient loss, eliminate odor, and improve nutrient availability, making it the most effective strategy for converting high-nitrogen and high-phosphorus waste such as mink and fox into high-quality organic fertilizer. The fermentation broth provided by this invention has a reasonable formulation, with significant synergistic effects among the various microbial species. The absence of any core component would lead to a significant decrease in fermentation effect, thus verifying the uniqueness and inventiveness of the fermentation broth formulation of this invention.

[0076] Seed germination test was conducted using the organic fertilizer prepared in Example 1. The results are shown in Table 4. After treatment with the organic fertilizer prepared in this invention, the germination rate, root length and germination index of radish seeds were significantly improved compared with the control group.

[0077] Table 4 Seed germination test results The above results indicate that the organic fertilizer prepared by the microbial fermentation method of mink and fox feces provided by this invention meets the relevant standards for organic fertilizer, can promote the germination and growth of crop seeds, and can be put into production.

[0078] Although preferred embodiments of the invention have been described, those skilled in the art, once they have learned the basic inventive concept, can make other changes and modifications to these embodiments.

[0079] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. Therefore, if these modifications and variations fall within the scope of the claims of this invention and their equivalents, this invention also intends to include these modifications and variations.

Claims

1. A method for preparing a mink, fox feces microbial fermented organic fertilizer, characterized by, Specifically comprising the following steps: mixing animal feces and agricultural fermentation accessories in a mass ratio of 6-10:6-10 to obtain a mixture; adding fermentation bacteria liquid in a ratio of 25 g-85 g:1 mL to the mixture, and supplementing water to a moisture content of 55% before fermentation; first turning pile when the fermentation pile temperature rises to 55℃, and second turning pile on the 7th day; ending fermentation when the fermentation temperature falls to 25℃, and the material is dark brown, loose and non-sticky; the animal feces are mink feces or fox feces; the agricultural fermentation accessories are shiitake mushroom dregs, chestnut pia or corn straw; the fermentation bacteria liquid is obtained by compounding Candida lipolytica bacteria liquid, Bacillus subtilis bacteria liquid, Lactobacillus plantarum bacteria liquid, Azotobacter chroococcum bacteria liquid and Saccharomyces cerevisiae bacteria liquid in a volume ratio of 3:3:2:1:1; The effective viable cell number of the Candida lipolytica bacterial solution, Bacillus subtilis bacterial solution, Lactobacillus plantarum bacterial solution, Azotobacter paspali bacterial solution and Saccharomyces cerevisiae bacterial solution is ≥1×10 8 CFU / mL.

2. The method of claim 1, wherein the mink, fox feces microbial fermented organic fertilizer is characterized by, The effective viable cell number in the fermentation bacterial solution is ≥1×10 8 CFU / mL.

3. The method according to claim 1, wherein the mink and fox feces microbial fermented organic fertilizer is characterized by, the mass ratio of the mixture is mink feces:shiitake mushroom dregs=6:

10.

4. The method of claim 1, wherein the mink and fox feces microbial fermented organic fertilizer is characterized by, the ratio of the fermentation bacteria liquid of the mixture is 85 g:1 mL.

5. The method of claim 1, wherein the mink and fox feces microbial fermented organic fertilizer is characterized by, the agricultural fermentation accessories are crushed before use, and pass through a 2 mm sieve.

6. The method of claim 1, wherein the mink and fox feces microbial fermented organic fertilizer is characterized by, the initial temperature of the fermentation is 35℃, and the total duration of the fermentation is 20-30 days.

7. A mink, fox excrement microbial fermented organic fertilizer, characterized in that, the organic fertilizer is prepared by any one of the steps of claims 1-6.

8. The mink, fox feces microbial fermented organic fertilizer according to claim 7, characterized in that, the organic fertilizer is used to improve seed germination rate, root length or germination index.

9. The mink, fox feces microbial fermented organic fertilizer according to claim 8, characterized in that, the seed is uncoated radish seed.

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