Animal waste viscous degradation strain and application thereof
By using Bacillus stercoris A-14 strain to degrade the stickiness of animal feces, the problems of high difficulty in fecal treatment and high equipment investment have been solved, achieving efficient reduction of stickiness and volume and improvement of porosity in fecal bedding, thus promoting resource utilization.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-01-07
- Publication Date
- 2026-04-17
AI Technical Summary
Existing technologies for treating animal feces are difficult due to their high viscosity, and the water washing and separation process is expensive, time-consuming, and labor-intensive, which affects the processing efficiency.
Bacillus stercoris A-14 strain was used as a biodegrading agent for animal feces. Through aerobic fermentation, the sticky substances in the feces were degraded to prepare recycled animal feces bedding, which reduced viscosity and bulk density and increased porosity.
It significantly reduces the viscosity of cow manure bedding by 22.54%, decreases the bulk density by 28.15%, and increases the porosity by 22.18%, improving the physical properties of manure, providing a more comfortable lying environment, and promoting resource utilization.
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Abstract
Description
Technical Field
[0001] This invention relates to a strain of animal feces that degrades stickiness and its applications, belonging to the field of microbial technology. Background Technology
[0002] With the continuous improvement of the quality of life of Chinese residents, the demand for dairy and meat products is constantly increasing, driving the livestock industry towards large-scale development. The livestock industry generates a large amount of manure, and animal manure, due to its high water content, fine texture, and strong viscosity, is significantly more difficult to treat, becoming a bottleneck restricting the green development of the livestock industry. Currently, the technology for preparing recycled animal manure bedding mainly relies on pretreatment processes such as washing and solid-liquid separation followed by aerobic fermentation. Although this method can reduce viscosity through physical means, the multi-stage washing and separation process requires huge investment in equipment and is time-consuming and labor-intensive, affecting processing efficiency. Therefore, developing bacterial strains that can effectively degrade the viscosity of animal manure, and using bacterial agents to effectively reduce its viscosity and bulk density, thereby improving the fluffiness of recycled bedding, has become an urgent task to overcome technical bottlenecks and promote the resource utilization of animal manure. Summary of the Invention
[0003] The technical problem to be solved by the present invention is to overcome the defects of the prior art and provide a strain of animal feces adhesive degradation strain that promotes the preparation of animal feces recycled bedding.
[0004] The technical solution provided by this invention is as follows:
[0005] A bacterial strain capable of degrading sticky substances from animal feces has been deposited on July 7, 2025, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO.35116 and classified as *Bacillus stercoris*, located in Beijing. This strain is able to grow and degrade sticky substances from animal feces using them as the sole carbon source.
[0006] The present invention also provides an animal fecal adhesive degradation agent, wherein the agent contains the above-mentioned animal fecal adhesive degradation strain.
[0007] The present invention also provides a method for preparing the above-mentioned animal feces adhesive degrading microbial agent, the preparation method comprising the following steps:
[0008] The bacterial culture of the above-mentioned strains was inoculated into the fermentation medium at an inoculum rate of 2-4 vol%, and cultured in a constant temperature shaker at 35-38℃ and a rotation speed of 160-200 r / min for 20-28 h. The bacterial count was then adjusted to 1-5 × 10⁻⁶. 9 CFU / mL bacterial agent.
[0009] Furthermore, the culture medium is prepared as follows: 8-12 mL / L of mixed salt ion solution, 6-10 g / L of animal fecal viscous substance, 1.5-2.5 g / L of (NH4)2SO4, pH 7.1-7.3, sterilized at 115-120 ℃ for 20-30 min.
[0010] Furthermore, the mixed salt ion solution contains MgSO4·7H2O 40~60 g / L, FeSO4·7H2O 0.8~1.2 g / L, K2HPO4 60~80 g / L, and KH2PO4 25~35 g / L.
[0011] Furthermore, the method for extracting viscous substances from animal feces includes: soaking air-dried, pulverized, and sieved animal feces powder with anhydrous ethanol at a ratio of 1g:4-6mL, allowing it to stand to remove lipids, then filtering, repeatedly filtering the filter residue, and drying the filter residue at room temperature. The animal feces powder is then mixed with ultrapure water at a ratio of 1g:25-35mL, ultrasonically treated, and thoroughly shaken at 45-55℃ and 160-200 r / min for 4-6 hours, followed by filtration and concentration. Anhydrous ethanol is added, and the mixture is allowed to stand overnight at 4℃, centrifuged again, and the precipitate is freeze-dried to obtain the animal feces extract.
[0012] The present invention also provides the application of the above-mentioned microbial agent with the ability to degrade the viscosity of animal feces in the preparation process of animal feces recycled bedding, which can significantly reduce the viscosity and bulk density of animal feces and increase its porosity, making the fermented animal feces soft and fluffy.
[0013] Furthermore, the aforementioned animal feces sticky degrading strains or animal feces sticky degrading agents are added to animal feces for aerobic fermentation.
[0014] Furthermore, the inoculum is administered at an inoculation rate of 5-10 vol / wt% of animal feces.
[0015] Furthermore, the initial moisture content of the animal manure during fermentation is 60-70%, and the fermentation time is 8-12 days.
[0016] Beneficial effects
[0017] The *Bacillus stercoris* A-14 strain screened in this invention can thrive in animal feces and exhibits a high capacity for degrading the stickiness of animal feces. When used as a fermentation agent for cow manure, the agent reduces the viscosity of cow manure bedding by 22.54%, decreases bulk density by 28.15%, and increases porosity by 22.18%. The fermentation agent prepared in this invention significantly reduces the viscosity and bulk density of cow manure and promotes porosity, effectively improving the physical properties of cow manure and laying the foundation for its subsequent resource utilization. Attached Figure Description
[0018] Figure 1 Preliminary screening results of the effects of different strains on the degradation of sticky animal feces;
[0019] Figure 2 Results of secondary screening of temperature suitability for different strains;
[0020] Figure 3 Results of pH suitability rescreening for different strains;
[0021] Figure 4 Results of secondary screening of different strains for oligotrophic suitability;
[0022] Figure 5 The identification results of Bacillus stercoris strain A-14;
[0023] Figure 6 The curve showing the effect of animal manure viscosity-degrading bacteria on the viscosity of cow manure during fermentation.
[0024] Figure 7 The curve showing the effect of animal manure viscous degrading bacteria on the bulk density of cow manure during fermentation.
[0025] Figure 8 The curves show the effect of animal manure viscous degrading bacteria on the porosity during cow manure fermentation. Detailed Implementation
[0026] Example 1
[0027] Isolation of animal fecal sticky degrading strain A-14
[0028] 1. Extraction of sticky substances from cow dung: Air-dried, pulverized cow dung powder (passed through a 0.149 mm sieve) was soaked in anhydrous ethanol at a ratio of 1:5 (g / mL). After standing for 12 h to remove lipids, the mixture was filtered. The filter residue was repeatedly filtered and then dried at room temperature. 2 g of cow dung powder was mixed with ultrapure water at a ratio of 1:30 (w / v), sonicated for 5 min, and then thoroughly shaken at 50℃ and 180 r / min for 5 h. The mixture was then filtered through four layers of sterile gauze to remove particulate matter. The filtrate was concentrated to 1 / 4 of its original volume using a rotary evaporator at 50℃. Four times the volume of anhydrous ethanol was added, and the mixture was allowed to stand overnight at 4℃. The precipitate, the cow dung extract, was obtained by centrifugation at 4000 r / min for 20 min and then freeze-dried for storage at -80℃.
[0029] 2. Culture medium: (1) Mixed salt ion solution: MgSO4·7H2O 50.0g / L, FeSO4·7H2O 1.0g / L, K2HPO4 70.0g / L, KH2PO4 30.0g / L.
[0030] (2) Liquid viscous degrading strain culture medium: 10.0 mL / L mixed salt ion solution, 8.0 g / L cow dung viscous substance, 2.0 g / L (NH4)2SO4, pH 7.2, sterilized at 115℃ for 20 min.
[0031] (3) Solid culture medium: Add 18.0 g / L of agar powder to the liquid culture medium.
[0032] 3. Preparation of bacterial suspension for samples: Collect samples of beef cattle manure and surrounding soil, sludge, paddy soil and rhizosphere soil. Weigh 5 g of the sample into an Erlenmeyer flask containing 50 mL of sterile water. Shake thoroughly for 1 h at 37 ℃ and 180 r / min to fully disperse the sample, and let it stand for 30 min.
[0033] 4. Enrichment and Isolation of Bacterial Strains: 1 mL of the sample bacterial suspension was placed in 50 mL of liquid viscous degrading bacterial culture medium and incubated at 37℃ and 180 r / min for 72 h until the viscosity of the medium decreased. This enrichment process was repeated three times. The enriched bacterial suspension was then diluted to 10⁻⁶ with sterile water. -9 Take 10 respectively -7 10 -8 10 -9 100 μL of each dilution was spread onto solid, viscous, degradative culture medium and incubated at 37 °C for 24 h. Single colonies from the plates were picked and streaked onto fresh solid culture plates using the triple-line method, repeating this process until only one strain was visible on the plate. The purified strain was then inoculated into 25% glycerol and stored at -80 °C.
[0034] Example 2
[0035] Initial screening of the degradation effect of the strain:
[0036] (1) Preparation of bacterial suspension: The 17 strains of animal fecal viscous degrading strains were inoculated into 50 mL of liquid viscous degrading strain culture medium and placed in a constant temperature shaker at 37 ℃ and 180 r / min for 24 h.
[0037] (2) Viscosity degradation effect: 2 vol% bacterial suspension was inoculated into liquid viscous degradation strain culture medium and incubated in a constant temperature shaker at 37℃ and 180 r / min for 72 h. The viscosity change was measured using a rotational viscometer. The experimental results are as follows: Figure 1As shown, strains A-14, A-31, Z-26, Z-7, A-1, Z-15, A-18, Z-11, and Z-22, with a fecal viscosity degradation rate greater than 20%, were selected as highly efficient strains for fecal viscosity degradation. Further tests were conducted on the adaptability of the strains to the growth environment. Among them, the fecal viscosity degradation rates of A-14, A-31, and Z-26 were 35.65%, 33.66%, and 32.69%, respectively.
[0038] Example 3
[0039] Environmental adaptability rescreening of strains:
[0040] (1) Adaptability to different temperatures: 2 vol% bacterial suspension was inoculated into liquid culture medium and placed at 10, 20, 30, 40 and 50 ℃ respectively. The suspension was then shaken in a constant temperature shaker at 180 r / min for 24 h. The OD of the bacterial suspension was measured using an enzyme-linked immunosorbent assay (ELISA) reader. 600 Values. All other conditions were exactly the same, with three parallels set for each temperature gradient. Results are as follows: Figure 2 As shown, the animal fecal adhesive degrading strains with strong temperature adaptability were screened out as A-14, A-31, Z-22, Z-7, and Z-26.
[0041] (2) Adaptability to different pH values: The pH values of the liquid culture medium were adjusted to 4, 5, 6, 7, 8, and 9 using HCl and NaOH solutions, respectively. 2 vol% bacterial suspension was inoculated into the liquid culture medium and cultured at 37℃ and 180 r / min for 24 h using a constant temperature shaker. The OD values of the bacterial suspension were measured using an enzyme-linked immunosorbent assay (ELISA) reader. 600 Values. All other conditions were exactly the same, with three replicates set for each pH. Results are as follows: Figure 3 As shown, the strains that exhibit strong pH adaptability and are capable of degrading the stickiness of animal feces were selected as A-31, Z-26, A-14, Z-7, and Z-22.
[0042] (3) Adaptability to different oligotrophic conditions: Liquid culture medium was prepared and diluted with sterile water to 100, 500, 1000, 2000 and 5000 times respectively. 2 vol% bacterial suspension was then inoculated into the liquid culture medium and cultured in a constant temperature shaker at 37℃ and 180 r / min for 24 h. The OD of the bacterial suspension was measured using an enzyme-linked immunosorbent assay (ELISA) reader. 600 Values. All other conditions were exactly the same, with three replicates for each oligotrophic nutrient. Results are as follows: Figure 4 As shown, the strains that exhibit strong adaptability to oligotrophic conditions and are capable of degrading the stickiness of animal feces were screened out as A-14, A-31, Z-26, Z-7, and A-18.
[0043] Based on the initial screening results of the strain's degradation effect and the secondary screening results of its environmental suitability, strain A-14, which is highly efficient at degrading the stickiness of animal feces, was selected.
[0044] Example 4
[0045] Strain identification: Strawberry A-14 was inoculated onto solid culture medium using the streak plating method and cultured at 37 °C for 48 h. Colony morphology was observed after Gram staining and observation of morphological structure using an optical microscope (10 × 100). The 16S rDNA sequence of the strain was also detected and compared with the NCBI database. A phylogenetic tree was constructed using sequences with high homology. The cell morphology photographs and phylogenetic tree are shown below. Figure 6 As shown. The colony characteristics of strain A-14 are: large, round, smooth-edged, dull white, Gram-positive, as... Figure 5 As shown.
[0046] Preservation of biological material samples:
[0047] A-14 has been deposited at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, with accession number CGMCC NO.35116 and deposit date of July 7, 2025.
[0048] The 16S rRNA sequence of Bacillus stercoris strain A-14:
[0049]
[0050] Example 5
[0051] Verification experiment on the application effect of animal fecal adhesive degradation bacteria: The A-14 strain was prepared with a bacterial count of 1×10⁻⁶. 9 A microbial agent of CFU / mL was used as experimental group S1, added to cow manure with a moisture content of 65% at an inoculum rate of 7.5 vol / wt%. The mixture was placed in a plastic container and fermented at 37 ℃ for 10 days. A control group (CK) without added microbial agent was used. All other conditions were identical. Each combination was replicated in triplicate. Samples were taken on days 1, 3, 5, 7, and 10 to measure bulk density and porosity, and the manure was turned and aerated. Changes in cow manure viscosity were measured using a rotational viscometer.
[0052] like Figure 6-8 As shown, during the experiment, the viscosity of the cow dung bedding in the S1 group was significantly lower than that in the CK group (P < 0.001), showing a rapid decreasing trend. The viscosity degradation rate reached 22.54% on day 10 of fermentation, while the control group (CK) only had a degradation rate of 9.91%, representing a 12.63% increase compared to the CK group. Regarding bulk density, the S1 group was significantly lower than the CK group during the experiment (P < 0.01), showing a rapid decreasing trend. The bulk density reduction rate reached 28.15% on day 10, an increase of 12.06% compared to the CK group. During the experiment, the porosity of the cow dung bedding in the S1 group was significantly higher than that in the CK group (P < 0.05), showing a rapid increasing trend. The porosity increase rate reached 22.18% on day 10, an increase of 5.75% compared to the CK group. Experimental results show that adding a cow manure viscosity-degrading microbial agent during cow manure fermentation can significantly reduce the viscosity and bulk density of fermented cow manure and increase its porosity, thereby making the fermented cow manure bedding softer and fluffier, providing better comfort for cattle, helping to improve their rest quality, and thus improving production performance. The animal manure viscosity-degrading microbial agent described in this invention can grow using the sticky substances of animal manure as the sole carbon source. The prepared recycled bedding contains only two components: cow manure and the microbial agent, and can be directly fermented without the addition of auxiliary materials, demonstrating good practicality and economic value.
[0053] The microbial agent prepared by this invention has a significant ability to reduce viscosity, bulk density and increase porosity of animal feces, effectively improving the physical properties of feces and making the treated fecal bedding soft and fluffy, providing a more comfortable lying environment for animals, and laying the foundation for subsequent resource utilization.
Claims
1. A strain of bacteria that degrades the stickiness of animal feces, characterized in that, The strain was deposited on July 7, 2025, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO.35116, classified as Bacillus stercoris, and located in Beijing.
2. A biodegrading agent for animal feces, characterized in that, The microbial agent contains the animal feces-degrading bacterial strain as described in claim 1.
3. The method for preparing the animal feces adhesive degrading bacterial agent according to claim 2, characterized in that, The preparation method includes the following steps: The cultured bacterial suspension according to claim 1 is inoculated into the fermentation medium at an inoculum rate of 2-4 vol%, and cultured in a constant temperature shaker at a temperature of 35-38 ℃ and a rotation speed of 160-200 r / min for 20-28 h, adjusting the bacterial count to 1-5 × 10⁻⁶. 9 CFU / mL bacterial agent.
4. The method for preparing the animal feces adhesive degrading bacterial agent according to claim 3, characterized in that, The fermentation medium is prepared as follows: 8-12 mL / L of mixed salt ion solution, 6-10 g / L of animal fecal viscous substance, 1.5-2.5 g / L of (NH4)2SO4, pH 7.1-7.3, sterilized at 115-120 ℃ for 20-30 min.
5. The method for preparing the animal feces adhesive degrading bacterial agent according to claim 3, characterized in that, The mixed salt ion solution comprises: MgSO4·7H2O 40~60 g / L, FeSO4·7H2O 0.8~1.2 g / L, K2HPO4 60~80 g / L, and KH2PO4 25~35 g / L.
6. The culture medium according to claim 3, characterized in that, The method for extracting viscous substances from animal feces includes: soaking air-dried, pulverized, and sieved animal feces powder with anhydrous ethanol at a ratio of 1g:4-6mL, allowing it to stand to remove lipids, then filtering, repeatedly filtering the filtered residue, and drying the residue at room temperature; mixing animal feces powder with ultrapure water at a ratio of 1g:25-35mL, ultrasonicating, and then shaking thoroughly at 45-55℃ and 160-200 r / min for 4-6 hours, followed by filtration and concentration; adding anhydrous ethanol, allowing it to stand overnight at 4℃, centrifuging again, and freeze-drying the precipitate to obtain the animal feces extract.
7. The application of the animal feces viscosity-degrading strain of claim 1 or the microbial agent of claim 2 in the preparation of animal feces recycled bedding, wherein the strain or microbial agent is used to reduce the viscosity and bulk density of the animal feces recycled bedding and to increase the porosity of the animal feces recycled bedding.
8. The application according to claim 7, characterized in that, The animal feces adhesive degrading strain of claim 1 or the animal feces adhesive degrading agent of claim 2 is added to animal feces for aerobic fermentation.
9. The application according to claim 7, characterized in that, The inoculum is administered at an inoculation rate of 5-10 vol / wt% of animal feces.
10. The application according to claim 7, characterized in that, The initial moisture content of the animal manure during fermentation is 60-70%, and the fermentation time is 8-12 days.