An ecological technology method for efficiently reducing antibiotic-resistant genes in cow dung

Abstract

The present application relates to the field of biotechnology science, and more particularly to an ecological technology method for efficiently reducing antibiotic-resistant genes in cow dung; by introducing a certain amount of angelica residue and cow dung to improve the self-pollution control ability of earthworms, the antibiotic-resistant gene pollutants in the cow dung are efficiently eliminated, the removal rate of beta-lactam antibiotic-resistant genes can be increased to more than 80%, the removal rate of sulfonamide and tetracycline-resistant genes also reaches 60%-70%, and the reduction of mobile transmission elements is also increased from about 30% of the conventional conversion method to more than 60%, and further, the present application can also make full use of traditional Chinese medicine residue and cow dung waste, so as to achieve the purposes of waste-to-resource, harmless treatment and safe resource utilization.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology science, and in particular to an ecological technology method for efficiently reducing antibiotic resistance genes in bovine manure. Background Technology

[0002] With the rapid development of large-scale and intensive dairy farming, a large number of antibiotics are widely used in dairy farming, resulting in the production and accumulation of a large number of antibiotic-resistant bacteria (ARBs) and antibiotic resistance genes (ARGs) in dairy farm manure. If subsequent treatment or utilization is not handled properly, it can lead to the accumulation and secondary transmission of livestock-derived ARGs and ARBs in the receiving environment, which will further aggravate the pollution of ARGs in the environment and the risks they bring, and may ultimately endanger human health through the food chain.

[0003] The treatment of dairy farm manure is a crucial step in controlling the entry of livestock-derived aeroid glycosides (ARGs) into the environment. Traditional composting, a conventional cattle manure resource utilization technology, can effectively kill pathogens in animal manure and reduce various ARGs to some extent (removal rate ranges from 3% to 56%). Anaerobic fermentation, another important method for utilizing livestock manure as a resource and fertilizer, can also reduce the abundance of some ARGs. Besides studies focusing on ARG changes during aerobic or anaerobic composting, current research also examines the removal effect of combined aerobic and anaerobic composting. Aerobic-anaerobic two-phase composting effectively reduces the abundance of some ARGs and potential pathogens, with relative abundances of ermB and tetK decreasing by 39.68% and 72.19%, respectively. While these traditional manure treatment methods can remove ARGs to some extent, the effects are unstable and limited, still possessing certain limitations and posing potential subsequent environmental pollution problems.

[0004] In response to the above problems, how to achieve the harmless and resource-based treatment of livestock manure has become a key issue that urgently needs to be addressed for the green development of agriculture.

[0005] Cow manure contains antibiotic resistance genes. Vermicomposting can reduce some of these genes, but vermicomposting still leaves some residual genes, posing a risk for future use. To address this issue, and considering that traditional Chinese medicine residue contains abundant bioactive substances, trace elements, and medicinal antibacterial components, this invention proposes a method for the co-conversion of traditional Chinese medicine residue and cow manure by earthworms. Summary of the Invention

[0006] The purpose of this invention is to overcome the shortcomings of the prior art and provide an ecological technology method that uses earthworms to transform specific Chinese medicine residues and cow dung into a compound substrate, which can quickly and efficiently remove ARGs from cow dung, improve the removal rate of ARGs from cow dung, shorten the treatment time of ARGs from cow dung, and make it easy to operate and control.

[0007] The technical problem solved by this invention is achieved through the following technical solution:

[0008] An ecological technology method for efficiently reducing antibiotic resistance genes in bovine manure includes the following steps:

[0009] S1. Enhancement pretreatment of Angelica Root Residue: The Angelica Root Residue was fumigated with 40% sucrose water for 6 hours, turning it over every hour. The fumigated Angelica Root Residue was then air-dried and crushed for later use.

[0010] S2. Pretreatment of cow manure substrate: Collect fresh cow manure, adjust the moisture content, and spray and add Bifidobacterium breve liquid, Bacillus coagulans liquid, and Candida utilis liquid in sequence. Mix evenly and compost naturally for 15 days at an ambient temperature of 23℃±2℃. Crush and set aside for later use.

[0011] S3. Mix the Angelica dregs collected and processed with cow dregs at a dry weight ratio of 3:100 to obtain a mixture of Chinese medicine residue and cow dregs.

[0012] S4. Mix the angelica residue after the enhancement pretreatment and the cow manure after the pretreatment at a dry weight ratio of 3:100 to prepare a pre-compound mixture. Adjust the moisture content of the pre-compound mixture to 65%-70% and place it in the earthworm conversion device.

[0013] S5. Earthworm conversion process: Inoculate the pre-mixed compound with cow manure-domesticated earthworms at a density of 150 earthworms / kg. Maintain the conversion environment temperature at 20-28℃. After inoculation, place the mixture for 15-20 days to carry out earthworm conversion of the manure. During the conversion process, replenish water every 1-2 days and spray with 3% sucrose water every 10 days to maintain the moisture content of the pile at 65%-70%.

[0014] S6. After the earthworm conversion is completed, the earthworm conversion process of Angelica dahurica residue and cow dung is finished. The earthworms and earthworm castings are then removed from the conversion device for further processing and utilization.

[0015] Preferably, in step S1, the pretreatment of Angelica dahurica residue for enhancing efficacy specifically includes the following steps:

[0016] A1. Fumigate the Angelica dregs with 40% sucrose solution for 6 hours, turning them over every hour;

[0017] A2. Allow the fumigated Angelica dregs to air dry naturally or place them in an oven at 20-35℃ for air drying;

[0018] A3. The dried Angelica dahurica residue is then pulverized to a particle size of 1-2 cm.

[0019] Preferably, in step S2, the spraying amount of Bifidobacterium breve liquid, Bacillus coagulans liquid, and Candida utilis liquid is all based on 5 mL of bacterial solution per 1 kg of feces diluted with 100 mL of pure water.

[0020] Preferably, adjusting the moisture content specifically includes:

[0021] B1. Use a moisture meter to monitor the moisture content of the pre-mixed material of Angelica dahurica residue and cow dung in real time.

[0022] B2. If the moisture content of the mixture is between 65% and 70%, no adjustment is needed; simply place it into the earthworm conversion device. If the moisture content is less than 65%, add water to maintain the moisture content of the mixture at 65%-70%.

[0023] Preferably, in step S5, the earthworm transformation process specifically includes the following steps:

[0024] C1. The earthworms used are Eisenia fetidae, which has a high reproduction rate. They are domesticated in a cow dung environment for 30 days. After domestication, adult earthworms with reproductive clitellum and strong health and activity with an average weight of 0.4g are selected.

[0025] C2. Inoculate domesticated earthworms into a mixture of Angelica sinensis residue and cow dung in a conversion device at an inoculation density of 150 earthworms / kg.

[0026] C3. After the earthworm inoculation is completed, place the conversion device for 15-20 days, and control the temperature at 20-28℃ to carry out the earthworm decomposition and conversion of cow dung.

[0027] C4. During the earthworm transformation process, water should be added every 1-2 days to keep the moisture content of the mixture of Angelica dahurica residue and cow dung at 65%-70%.

[0028] C5. During the conversion process, spray a 3% sucrose solution every 10 days to maintain the moisture content of the pile at 65%-70% until the conversion is completed.

[0029] The advantages and positive effects of this invention are:

[0030] This invention introduces a certain amount of Angelica sinensis residue mixed with cow dung to enhance the earthworm's ability to autonomously control pollution, thereby efficiently removing drug-resistant gene pollutants from cow dung. The removal rate of β-lactam antibiotic resistance genes can be increased to over 80%, and the removal rate of sulfonamide and tetracycline resistance genes can reach 60-70%. The reduction of their main transmission vectors (integrins) is also increased from about 30% in conventional conversion methods to over 60%. Furthermore, this invention can make full use of Angelica sinensis residue and cow dung waste to achieve the goals of turning waste into treasure, harmless treatment, and safe resource utilization.

[0031] This invention utilizes earthworms to co-convert medicinal herb residues and cow dung, making full use of both angelica root residues and cow dung to achieve the goals of turning waste into treasure, harmless treatment, and resource utilization. Furthermore, this invention has fewer influencing factors and is easier to control during the treatment process, further realizing the high-value resource utilization of manure. It not only improves the quality of the conversion products but also has a significant treatment effect.

[0032] Angelica sinensis is a commonly used blood-tonifying herb. Its residue is still rich in nutrients, which can enhance earthworms' immunity and disease resistance, promote their metabolism, regulate intestinal flora, and improve their transformation ability. Furthermore, adding Angelica sinensis residue enriches the cellulose content of the material, improves its palatability, changes the number and type of bacteria in the material, and ultimately promotes earthworm growth, thus having the unique advantage of enhancing earthworms' ability to control pollution.

[0033] Using 40% sucrose solution to fumigate the Angelica dahurica residue can stimulate the release of polysaccharide components and active factors in the residue. On the other hand, it can also increase the energy intake of earthworms by introducing sucrose disaccharide factors, thereby improving earthworm growth, increasing body weight, and ultimately enhancing earthworm conversion efficiency and pollution control capabilities.

[0034] The purpose of spraying with Bifidobacterium breve solution, Bacillus coagulans solution, and Candida utilis is to accelerate the composting process and improve the earthworms' conversion and pollution control capabilities. Bifidobacterium breve solution can alleviate allergic discomfort, reduce damage to the earthworm's intestinal mucosa from bacteria or viruses, accelerate metabolism, and enhance immunity. Bacillus coagulans contains probiotic components that promote intestinal peristalsis. Additionally, Candida utilis solution is rich in vitamin B and protein, providing earthworms with some of the necessary nutrients, offering nutritional support, improving intestinal microbial balance, and promoting digestion and absorption. It also accelerates the composting and maturation of cow manure. Detailed Implementation

[0035] The present invention will be further described in detail below through specific embodiments. The following embodiments are merely descriptive and not limiting, and should not be used to limit the scope of protection of the present invention.

[0036] An ecological technology method for efficiently reducing antibiotic resistance genes in bovine manure includes the following steps:

[0037] S1. Enhancement pretreatment of Angelica Root Residue: The Angelica Root Residue was fumigated with 40% sucrose water for 6 hours, turning it over every hour. The fumigated Angelica Root Residue was then air-dried and crushed for later use.

[0038] S2. Pretreatment of cow manure substrate: Collect fresh cow manure, adjust the moisture content, and spray and add Bifidobacterium breve liquid, Bacillus coagulans liquid, and Candida utilis liquid in sequence. Mix evenly and compost naturally for 15 days at an ambient temperature of 23℃±2℃. Crush and set aside for later use.

[0039] S3. Mix the Angelica dregs collected and processed with cow dregs at a dry weight ratio of 3:100 to obtain a mixture of Chinese medicine residue and cow dregs.

[0040] S4. Mix the angelica residue after the enhancement pretreatment and the cow manure after the pretreatment at a dry weight ratio of 3:100 to prepare a pre-compound mixture. Adjust the moisture content of the pre-compound mixture to 65%-70% and place it in the earthworm conversion device.

[0041] S5. Earthworm conversion process: Inoculate the pre-mixed compound with cow manure-domesticated earthworms at a density of 150 earthworms / kg. Maintain the conversion environment temperature at 20-28℃. After inoculation, place the mixture for 15-20 days to carry out earthworm conversion of the manure. During the conversion process, replenish water every 1-2 days and spray with 3% sucrose water every 10 days to maintain the moisture content of the pile at 65%-70%.

[0042] S6. After the earthworm conversion is completed, the earthworm conversion process of Angelica dahurica residue and cow dung is finished. The earthworms and earthworm castings are then removed from the conversion device for further processing and utilization.

[0043] Unless otherwise specified, all reagents and consumables used in the following examples are commercially available.

[0044] Example 1

[0045] An ecological technology method for efficiently reducing antibiotic resistance genes in bovine manure includes the following steps:

[0046] S1. Enhancement pretreatment of Angelica dahurica residue: Fumigate the Angelica dahurica residue with 40% sucrose water for 6 hours, turning it over once every hour. After fumigation, air dry the Angelica dahurica residue naturally or put it in an oven (20-35℃) to air dry, and crush it to a particle size of 1-2cm for later use.

[0047] S2. Pretreatment of cow dung substrate: Collect fresh cow dung, adjust the moisture content, and spray with a solution containing Bifidobacterium shortieum (containing no less than 10 bacteria per ml). 9Bacillus coagulans solution (containing no less than 10 bacteria per ml) 8 ), Candida utilis broth (containing no less than 10 bacteria per ml), 8 For each manure sample, the dosage of bacteria is 5 ml of bacterial solution diluted with 100 ml of pure water. Mix well and allow to compost naturally for 15 days at an ambient temperature of 23℃±2℃. Then crush the mixture for later use.

[0048] S3. Mix the Angelica dregs collected and processed with cow dregs at a dry weight ratio of 3:100 to obtain a mixture of Chinese medicine residue and cow dregs.

[0049] S4. Mix the angelica residue after the enhancement pretreatment and the cow manure after the pretreatment at a dry weight ratio of 3:100 to prepare a pre-compound mixture. Adjust the moisture content of the pre-compound mixture to 65%-70% and place it in the earthworm conversion device.

[0050] S5. Earthworm Conversion Process: Inoculate Eisenia fetidae, which has been domesticated with cow manure for 30 days, into the pre-mixed compound at a density of 150 earthworms / kg. Maintain the conversion environment temperature at 20-28℃. After inoculation, place the mixture for 15-20 days to carry out earthworm conversion of the manure. During the conversion process, replenish water every 1-2 days and spray with 3% sucrose water every 10 days to maintain the moisture content of the pile at 65%-70%. The selected Eisenia fetidae are healthy, active adult earthworms with an average weight of 0.4g and possessing a reproductive clitellum.

[0051] S6. After the earthworm conversion is completed, the earthworm conversion process of Angelica dahurica residue and cow dung is finished. The earthworms and earthworm castings are then removed from the conversion device for further processing and utilization.

[0052] This invention introduces a certain amount of Angelica sinensis residue mixed with cow dung to enhance the earthworm's ability to autonomously control pollution, thereby efficiently removing drug-resistant gene pollutants from cow dung. The removal rate of β-lactam antibiotic resistance genes can be increased to over 80%, and the removal rate of sulfonamide and tetracycline resistance genes can reach 60-70%. The reduction of mobile transmission elements is also increased from about 30% in conventional conversion methods to over 60%. Furthermore, this invention can make full use of Chinese medicinal residue and cow dung waste to achieve the goals of turning waste into treasure, harmless treatment, and safe resource utilization.

[0053] Example 2

[0054] Based on Example 1, but with the difference of designing a control experiment to verify the effect of the combination of Angelica sinensis residue and cow dung on earthworm growth and activity, specifically including the following steps:

[0055] S1. Enhancement pretreatment of Angelica dahurica residue: Fumigate the Angelica dahurica residue with 40% sucrose water for 6 hours, turning it over once every hour. After fumigation, air dry the Angelica dahurica residue naturally or put it in an oven (20-35℃) to air dry, and crush it to a particle size of 1-2cm for later use.

[0056] S2. Pretreatment of cow dung substrate: Collect fresh cow dung, adjust the moisture content, and spray with a solution containing Bifidobacterium shortieum (containing no less than 10 bacteria per ml). 9 Bacillus coagulans solution (containing no less than 10 bacteria per ml) 8 ), Candida utilis broth (containing no less than 10 bacteria per ml), 8 For each manure sample, the dosage of bacteria is 5 ml of bacterial solution diluted with 100 ml of pure water. Mix well and allow to compost naturally for 15 days at an ambient temperature of 23℃±2℃. Then crush the mixture for later use.

[0057] S3. Weigh out 2 portions of pretreated pure cow dung equal to the amount of the mixture. Collect the pretreated Angelica dung residue and mix it with cow dung at a dry weight ratio of 3:100 to obtain 1 portion of the mixture of Chinese medicine residue and cow dung. Adjust the moisture content to 65%-70% and put them into three identical earthworm conversion devices, numbered a, b, and c respectively.

[0058] S4. Transfer the a, b, and c conversion devices together to a temperature-controlled greenhouse at 20-28℃;

[0059] S5. a) Earthworms are not inoculated in the conversion device, and natural degradation is carried out; b) Earthworms are inoculated in the conversion device, and conventional conversion is carried out; c) The material of Angelica dahurica residue and cow dung in the conversion device is converted and digested according to the operation steps described in Example 1.

[0060] S6. Using a and b as control groups and c as experimental group, the mortality rate and weight of earthworms in the three conversion devices a, b, and c were measured respectively. The obtained data were sorted, analyzed, and the earthworm activity index in each group was calculated to determine the effect of different treatments on earthworm activity, as shown in Table 1.

[0061] Table 1. Survival rate and weight data of earthworms

[0062]

[0063] As can be seen from Table 1, compared with the conventional cow dung-earthworm conversion, the Angelica sinensis-cow dung-earthworm conversion is more conducive to the growth and survival of earthworms, and plays a certain role in promoting the growth and survival of earthworms.

[0064] Example 3

[0065] Based on Example 1, but with the difference, a control experiment was designed to verify the effect and influence of combining Angelica sinensis residue with cow manure on earthworm transformation to reduce drug-resistant genes in cow manure. The specific steps included:

[0066] S1. Enhancement pretreatment of Angelica dahurica residue: Fumigate the Angelica dahurica residue with 40% sucrose water for 6 hours, turning it over once every hour. After fumigation, air dry the Angelica dahurica residue naturally or put it in an oven (20-35℃) to air dry, and crush it to a particle size of 1-2cm for later use.

[0067] S2. Pretreatment of cow dung substrate: Collect fresh cow dung, adjust the moisture content, and spray with a solution containing Bifidobacterium shortieum (containing no less than 10 bacteria per ml). 9 Bacillus coagulans solution (containing no less than 10 bacteria per ml) 8 ), Candida utilis broth (containing no less than 10 bacteria per ml), 8 For each manure sample, the dosage of bacteria is 5 ml of bacterial solution diluted with 100 ml of pure water. Mix well and allow to compost naturally for 15 days at an ambient temperature of 23℃±2℃. Then crush the mixture for later use.

[0068] S3. Weigh out 2 portions of pretreated pure cow dung equal to the amount of the mixture. Collect the pretreated Angelica dung residue and mix it with cow dung at a dry weight ratio of 3:100 to obtain 1 portion of the mixture of Chinese medicine residue and cow dung. Adjust the moisture content to 65%-70% and put them into three identical earthworm conversion devices, numbered d, e, and f respectively.

[0069] S4. Transfer the d, e, and f conversion devices together to a temperature-controlled greenhouse at 20-28℃;

[0070] S5 and d are converted without earthworms and undergo natural degradation; e is converted with earthworms and undergoes conventional conversion; f is converted and digested in the material of angelica residue and cow dung according to the operation steps described in Example 1.

[0071] S6. Using d and e as control groups and f as the experimental group, the content of antibiotic resistance genes in the vermicompost of the three transformation devices (d, e, and f) was measured. The obtained data were collected, analyzed, and the removal rate of mobile genetic elements in the vermicompost of each group was calculated to determine the removal effect of different treatments on the vectors of antibiotic resistance gene transmission in vermicompost. The results are shown in Table 2.

[0072] Table 2. Removal rate data of drug resistance genes and transmission elements.

[0073]

[0074] As can be seen from the table, the earthworm transformation method using Angelica dahurica residue-cow dung is superior to conventional earthworm transformation and natural degradation treatment of cow dung in removing β-lactam, tetracycline, and sulfonamide resistance genes and mobile transmission element integrase genes. The introduction of a certain amount of Angelica dahurica residue can not only fatten earthworms and improve transformation efficiency, but also make up for the poor transformation effect of cow dung alone.

[0075] Example 4

[0076] Based on Example 1, but with the difference that,

[0077] A controlled experiment was designed to verify the effect and influence of combining Angelica sinensis residue with cow manure on earthworm transformation to reduce drug-resistant genes in cow manure. The specific steps included:

[0078] S1. Enhancement pretreatment of Angelica dahurica residue: Fumigate the Angelica dahurica residue with 40% sucrose water for 6 hours, turning it over once every hour. After fumigation, air dry the Angelica dahurica residue naturally or put it in an oven (20-35℃) to air dry, and crush it to a particle size of 1-2cm for later use.

[0079] S2. Pretreatment of cow manure substrate: Collect fresh cow manure, adjust the moisture content, mix it evenly, and compost it naturally for 15 days at an ambient temperature of 23℃±2℃. Then crush it for later use.

[0080] S3. Weigh out 2 portions of pretreated pure cow dung equal to the amount of the mixture. Collect the pretreated Angelica dung residue and mix it with cow dung at a dry weight ratio of 3:100 to obtain 1 portion of the mixture of Chinese medicine residue and cow dung. Adjust the moisture content to 65%-70% and put them into two identical earthworm conversion devices, labeled g and h respectively.

[0081] S4. Transfer the g and h conversion devices together to a temperature-controlled greenhouse at 20-28℃;

[0082] In step S2 of the S5 and g conversion devices, do not spray other bacterial solutions; otherwise, follow the same operating steps as described in Example 1. The material of Angelica dahurica residue and cow dung compounded in the h conversion device is converted and digested according to the operating steps described in Example 1.

[0083] S6. Using g as the control group and h as the experimental group, the content of antibiotic resistance genes in the vermicompost of both g and h transformation devices was measured. The obtained data were collected, analyzed, and the removal rate of mobile genetic elements in the vermicompost of each group was calculated to determine the removal effect of different treatments on the vectors of antibiotic resistance gene transmission in vermicompost. The results are shown in Table 3.

[0084] Table 3. Data on earthworm activity and removal rate of drug resistance genes / transmission elements.

[0085]

[0086] As can be seen from the table, the earthworm transformation method using Angelica dahurica residue and cow dung with the addition of exogenous bacteria not only benefits the survival and growth of earthworms, but also has a better effect on removing resistance genes of β-lactams, tetracyclines, and sulfonamides, as well as integrase genes of mobile transmission elements than the control group without the addition of exogenous bacteria. It is evident that spraying a certain amount of Bifidobacterium breve liquid, Bacillus coagulans liquid, and Candida utilis can help improve the transformation and pollution control capabilities of earthworms.

[0087] Example 5

[0088] Based on Example 1, but with the difference that,

[0089] A controlled experiment was designed to verify the effect and influence of fumigation with Angelica sinensis residue on earthworm transformation and reduction of drug-resistant genes in cow manure. The specific steps included:

[0090] S1. Pretreatment of Angelica Root Residue: Crush the air-dried Angelica Root Residue into particles with a diameter of 1-2 cm and set aside.

[0091] S2. Pretreatment of cow manure substrate: Collect fresh cow manure, adjust the moisture content, and spray in sequence with Bifidobacterium breve solution (containing no less than 109 bacteria per ml), Bacillus coagulans solution (containing no less than 108 bacteria per ml), and Candida utilis solution (containing no less than 108 bacteria per ml). The spraying amount is 5 ml of bacterial solution per 1 kg of manure diluted with 100 ml of pure water. Mix well and allow to compost naturally for 15 days at an ambient temperature of 23℃±2℃. Crush and set aside for later use.

[0092] S3. Weigh out 2 portions of pretreated pure cow dung equal to the amount of the mixture. Collect the pretreated Angelica dung residue and mix it with cow dung at a dry weight ratio of 3:100 to obtain 1 portion of the mixture of Chinese medicine residue and cow dung. Adjust the moisture content to 65%-70% and put them into two identical earthworm conversion devices, numbered i and j respectively.

[0093] S4. Transfer the i and j conversion devices together to a temperature-controlled greenhouse at 20-28℃;

[0094] In step S1 of the S5 and i conversion devices, the Angelica dahurica residue is not fumigated, and the other steps are the same as those described in Example 1; the Angelica dahurica residue and cow dung compound material in the j conversion device are converted and digested according to the steps described in Example 1.

[0095] S6. Using i as the control group and j as the experimental group, the content of antibiotic resistance genes in the vermicompost of both transformation devices i and j was measured. The obtained data were collected, analyzed, and the removal rate of mobile genetic elements in the vermicompost of each group was calculated to determine the removal effect of different treatments on the vectors of antibiotic resistance gene transmission in vermicompost. The results are shown in Table 4.

[0096] Table 4. Data on earthworm activity and removal rate of drug resistance genes / transmission elements.

[0097]

[0098] As can be seen from the table, the earthworm transformation method using Angelica dregs-cow dung compounded with fumigated Angelica dregs reduced earthworm mortality and increased earthworm weight to some extent. In addition, its removal effect on β-lactam, tetracycline, and sulfonamide resistance genes and mobile transmission element integrase genes was better than that of the control group without fumigation, indicating that the fumigation of Angelica dregs also plays an important role in enhancing and reducing earthworm transformation of cow dung ARGs.

[0099] Although embodiments of the invention have been disclosed for illustrative purposes, those skilled in the art will understand that various substitutions, variations, and modifications are possible without departing from the spirit and scope of the invention and the appended claims. Therefore, the scope of the invention is not limited to the contents disclosed in the embodiments.

Claims

1. An ecological technology method for efficiently reducing antibiotic resistance genes in bovine manure, characterized in that: Includes the following steps: S1. Enhancement pretreatment of Angelica Root Residue: The Angelica Root Residue was fumigated with 40% sucrose water for 6 hours, turning it over every hour. The fumigated Angelica Root Residue was then air-dried and crushed for later use. S2. Pretreatment of cow manure substrate: Collect fresh cow manure, adjust the moisture content, and spray and add Bifidobacterium breve liquid, Bacillus coagulans liquid, and Candida utilis liquid in sequence. Mix evenly and compost naturally for 15 days at an ambient temperature of 23℃±2℃. Crush and set aside for later use. S3. Mix the Angelica dregs collected and processed with cow dregs at a dry weight ratio of 3:100 to obtain a mixture of Chinese medicine residue and cow dregs. S4. Mix the angelica residue after the enhancement pretreatment and the cow manure after the pretreatment at a dry weight ratio of 3:100 to prepare a pre-compound mixture. Adjust the moisture content of the pre-compound mixture to 65%-70% and place it in the earthworm conversion device. S5. Earthworm Conversion Process: Inoculate the pre-mixed compound with cow manure-acclimated earthworms at a density of 150 earthworms / kg. Maintain the conversion environment temperature at 20-28℃. After inoculation, place the mixture for 15-20 days to carry out earthworm conversion of the manure. During the conversion process, replenish water every 1-2 days and spray with 3wt% sucrose solution every 10 days to maintain the moisture content of the pile at 65%-70%. S6. After the earthworm conversion is completed, the earthworm conversion process of Angelica dahurica residue and cow dung is finished. The earthworms and earthworm castings are then removed from the conversion device for further processing and utilization.

2. The ecological technology method for efficiently reducing antibiotic resistance genes in bovine manure according to claim 1, characterized in that: In step S1, the pretreatment of Angelica sinensis residue for enhancing efficacy specifically includes the following steps: A1. Fumigate the Angelica dregs with 40% sucrose solution for 6 hours, turning them over every hour; A2. Allow the fumigated Angelica dregs to air dry naturally or place them in an oven at 20-35℃ for air drying; A3. The dried Angelica dahurica residue is then pulverized to a particle size of 1-2 cm.

3. The ecological technology method for efficiently reducing antibiotic resistance genes in bovine manure according to claim 1, characterized in that: In step S2, the spraying dosage of Bifidobacterium breve solution, Bacillus coagulans solution, and Candida utilis solution is all based on 5 mL of bacterial solution diluted with 100 mL of pure water per kg of feces, ensuring that the bacterial count per kilogram of material is not less than 10. 9 indivual.

4. The ecological technology method for efficiently reducing antibiotic resistance genes in bovine manure according to claim 1, characterized in that: Adjusting the moisture content specifically includes: B1. Use a moisture meter to monitor the moisture content of the pre-mixed material of Angelica dahurica residue and cow dung in real time. B2. If the moisture content of the mixture is between 65% and 70%, no adjustment is needed; simply place it into the earthworm conversion device. If the moisture content is less than 65%, add water to maintain the moisture content of the mixture at 65%-70%.

5. The ecological technology method for efficiently reducing antibiotic resistance genes in bovine manure according to claim 1, characterized in that: In step S5, the earthworm transformation process specifically includes the following steps: C1. The earthworms used are Eisenia fetidae, which has a high reproduction rate. They are domesticated in a cow dung environment for 30 days. After domestication, adult earthworms with reproductive clitellum and strong health and activity with an average weight of 0.4g are selected. C2. Inoculate domesticated earthworms into a mixture of Angelica sinensis residue and cow dung in a conversion device at an inoculation density of 150 earthworms / kg. C3. After the earthworm inoculation is completed, place the conversion device for 15-20 days, and control the temperature at 20-28℃ to carry out the earthworm decomposition and conversion of cow dung; C4. During the earthworm transformation process, water should be added every 1-2 days to keep the moisture content of the mixture of Angelica dregs and cow dung at 65%-70%. C5. During the conversion process, spray a 3% sucrose solution every 10 days to maintain the moisture content of the pile at 65%-70% until the conversion is completed.

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