阅读说明：本技术 一种大观霉素菌渣抗生素、耐药基因去除方法、大观霉素菌渣堆肥及其制备方法 (Spectinomycin mushroom residue antibiotic, drug-resistant gene removing method, spectinomycin mushroom residue compost and preparation method thereof ) 是由 董坤 王禄山 陈彤 沙国萌 贾传新 李欣 陶晓红 沈建国 于 2021-01-04 设计创作，主要内容包括：本发明涉及抗生素生产副产物处理技术领域,具体涉及一种大观霉素菌渣抗生素、耐药基因去除方法、大观霉素菌渣堆肥及其制备方法,所述去除方法包括将大观霉素菌渣、蘑菇渣和小麦秸秆按照4.5～5.5：1：1的重量比混合均匀得到混合物不翻堆厌氧发酵；厌氧发酵后的堆肥翻堆处理进行好氧发酵；所述大观霉素菌渣堆肥按照厌氧发酵、好氧发酵、陈化缓存的制备方法生产。本发明第一步选择原料并调节配比厌氧发酵富集能够降解抗生素的菌去除抗生素,第二步通过高温杀灭耐药菌并降解耐药基因,实现对抗生素残留及耐药性的有效控制；本发明对于抗生素菌渣的无害化处理及资源化利用进程具有重要意义。(The invention relates to the technical field of antibiotic production byproduct treatment, and particularly relates to a spectinomycin mushroom residue antibiotic, a drug resistance gene removing method, a spectinomycin mushroom residue compost and a preparation method thereof, wherein the removing method comprises the following steps of mixing 4.5-5.5 parts of spectinomycin mushroom residue, mushroom residue and wheat straw: 1: 1, uniformly mixing to obtain a mixture, and performing anaerobic fermentation without turning the mixture; turning the compost after anaerobic fermentation for aerobic fermentation; the spectinomycin bacterial residue compost is produced according to the preparation method of anaerobic fermentation, aerobic fermentation and aging cache. The method comprises the steps of firstly, selecting raw materials, adjusting and proportioning anaerobic fermentation and enriching bacteria capable of degrading antibiotics to remove the antibiotics, and secondly, killing drug-resistant bacteria at high temperature and degrading drug-resistant genes to effectively control antibiotic residues and drug resistance; the invention has important significance for the harmless treatment and resource utilization process of the antibiotic fungi residues.)

1. A method for removing spectinomycin mushroom dreg antibiotics and drug-resistant genes is characterized by comprising the following steps:

(1) anaerobic fermentation: mixing spectinomycin mushroom dregs, mushroom dregs and wheat straws according to the ratio of 4.5-5.5: 1: 1 to obtain a mixture, controlling the water content of the mixture to be 55-65%, performing anaerobic fermentation without turning, naturally raising the fermentation temperature to 45-55 ℃, and maintaining for 5-7 days;

(2) aerobic fermentation: and (2) turning the compost subjected to anaerobic fermentation in the step (1) to perform aerobic fermentation, and naturally raising the fermentation temperature to 65-70 ℃ and maintaining the fermentation temperature for 20-25 days.

2. The removing method of claim 1, wherein the moisture content of the spectinomycin mushroom residue is 70 to 75%, the moisture content of the mushroom residue is 20 to 30%, and the moisture content of the wheat straw is 12 to 18%.

3. The removing method of claim 1, wherein the weight ratio of the spectinomycin mushroom dregs, the mushroom dregs and the wheat straw is 5: 1: 1.

4. the removal method according to claim 1, wherein the pile-turning frequency of the step (2) is one pile-turning every two days.

5. A preparation method of spectinomycin bacterial residue compost is characterized by comprising the following steps:

(1) anaerobic fermentation: mixing spectinomycin mushroom dregs, mushroom dregs and wheat straws according to the ratio of 4.5-5.5: 1: 1 to obtain a mixture, controlling the water content of the mixture to be 55-65%, performing anaerobic fermentation without turning, naturally raising the fermentation temperature to 45-55 ℃, and maintaining for 5-7 days;

(2) aerobic fermentation: turning the compost subjected to anaerobic fermentation in the step (1) to perform aerobic fermentation, naturally raising the fermentation temperature to 65-70 ℃, and maintaining for 20-25 days;

(3) aging and caching: and (3) ageing and caching the compost after aerobic fermentation in the step (2), so that the temperature of the compost is reduced to room temperature, and the ageing and caching time is 15-20 days.

6. The method according to claim 5, wherein the moisture content of the spectinomycin mushroom residue is 70 to 75%, the moisture content of the mushroom residue is 20 to 30%, and the moisture content of the wheat straw is 12 to 18%;

the weight ratio of the spectinomycin mushroom dregs, the mushroom dregs and the wheat straws is 5: 1: 1;

and (3) turning the pile once every two days at the pile turning frequency of the step (2).

7. The method of claim 5, further comprising the steps of:

(4) crushing and screening: and (4) crushing and screening the compost aged and cached in the step (3), mixing powdery fertilizer with the grain diameter of more than 4mm serving as an auxiliary material with the mixture obtained in the step (1), and fermenting again.

8. The method of claim 5, further comprising the steps of:

(5) granulation and/or packaging: and packaging the powdery fertilizer to obtain a finished product, or granulating the powdery fertilizer and then packaging to obtain the finished product.

9. The method of claim 5, wherein steps (1), (2) and (3) further comprise collecting gas during fermentation and aging.

10. A spectinomycin mushroom residue compost, which is produced by the production method according to any one of claims 5 to 9.

Technical Field

The invention relates to the technical field of antibiotic production byproduct treatment, and particularly relates to a spectinomycin mushroom residue antibiotic, a method for removing drug resistance genes, a spectinomycin mushroom residue compost and a preparation method thereof.

Background

Antibiotics, one of the most important medical findings in the 20 th century, play an irreplaceable role in protecting the health of people. Spectinomycin is a very important aminoglycoside antibiotic, is mainly used for treating uncomplicated gonorrhea and penicillin allergy, and has wide application. In the process of antibiotic production, a large amount of antibiotic dregs, which are a by-product, are generated, and the quantity of the antibiotic dregs reaches millions of tons every year, so that the safe treatment of the dregs becomes a serious problem at present. The antibiotic dregs contain mycelium, residual culture medium, fermentation metabolites and residual antibiotics. At present, most enterprises treat the antibiotic fungi residues by landfill and incineration, so that a large amount of resources are wasted, a large amount of land is occupied in the landfill process, and the environment is polluted. Meanwhile, toxic gases such as nitrogen dioxide and dioxin are generated in the incineration process and a large amount of electric energy is needed, which causes serious pressure on antibiotic production enterprises.

Aerobic composting is a reasonable way of resource utilization of wastes. However, the previous researches mostly focus on materials such as livestock manure, sludge and the like, and the researches on the solid-state fermentation process of the antibiotic fungi residues are less. CN108503399A discloses a treatment method for efficiently removing antibiotics in livestock and poultry manure, which comprises the steps of mixing crop wastes, fresh livestock and poultry manure and an inoculum for anaerobic fermentation and composting, wherein the inoculum refers to anaerobic fermentation residues at the bottom layer of an anaerobic fermentation tank or a methane tank, firstly, the antibiotic content in the livestock and poultry manure is obviously lower than that in antibiotic residues, and the method does not clearly show the change of drug resistance generated by microorganisms due to antibiotic residues, and meanwhile, the prior art does not describe a specific action mechanism for removing antibiotics.

Based on the above, a method for effectively removing antibiotic residues and drug resistance of spectinomycin mushroom dregs is needed, which has profound significance for the development of antibiotic production enterprises.

Disclosure of Invention

Aiming at the technical problems of resource waste, land occupation, environment pollution and high energy cost existing in the conventional treatment mode of antibiotic fungi residues, the invention provides a method for removing spectinomycin fungi residues antibiotics and drug-resistant genes, spectinomycin fungi residues compost and a preparation method thereof, wherein a two-step fermentation process of mixing the spectinomycin fungi residues with wheat straws and mushroom residues is adopted, the bacteria capable of degrading the antibiotics in the fungi residues are enriched in the first step of anaerobic fermentation to remove the antibiotics, and the drug-resistant genes are degraded by exogenous and endogenous DNA enzymes through high-temperature sterilization of the drug-resistant bacteria in the second step; meanwhile, by selecting spectinomycin bacterial residue compost raw materials and adjusting the proportion, the effective control of antibiotic residues and drug resistance is further realized; the invention has important significance for the harmless treatment and resource utilization process of the antibiotic fungi residues.

In a first aspect, the invention provides a method for removing spectinomycin mushroom dreg antibiotics and drug-resistant genes, which comprises the following steps:

(1) anaerobic fermentation: mixing spectinomycin mushroom dregs, mushroom dregs and wheat straws according to the ratio of 4.5-5.5: 1: 1 to obtain a mixture, controlling the water content of the mixture to be 55-65%, performing anaerobic fermentation without turning, naturally raising the fermentation temperature to 45-55 ℃, and maintaining for 5-7 days;

(2) aerobic fermentation: and (2) turning the compost subjected to anaerobic fermentation in the step (1) to perform aerobic fermentation, and naturally raising the fermentation temperature to 65-70 ℃ and maintaining the fermentation temperature for 20-25 days.

Further, the moisture content of the spectinomycin mushroom dregs is 70% -75%, the moisture content of the mushroom dregs is 20% -30%, and the moisture content of the wheat straws is 12% -18%.

Further, the weight ratio of the spectinomycin mushroom dregs, the mushroom dregs and the wheat straw is 5: 1: 1.

further, the pile turning frequency of the step (2) is that pile turning is carried out once every two days.

In a second aspect, the invention also provides a preparation method of the spectinomycin bacterial residue compost, which comprises the following steps:

(1) anaerobic fermentation: mixing spectinomycin mushroom dregs, mushroom dregs and wheat straws according to the ratio of 4.5-5.5: 1: 1 to obtain a mixture, controlling the water content of the mixture to be 55-65%, performing anaerobic fermentation without turning, naturally raising the fermentation temperature to 45-55 ℃, and maintaining for 5-7 days;

(2) aerobic fermentation: turning the compost subjected to anaerobic fermentation in the step (1) to perform aerobic fermentation, naturally raising the fermentation temperature to 65-70 ℃, and maintaining for 20-25 days;

(3) aging and caching: and (3) ageing and caching the compost after aerobic fermentation in the step (2), so that the temperature of the compost is reduced to room temperature, and the ageing and caching time is 15-20 days.

Further, the moisture content of the spectinomycin mushroom dregs is 70% -75%, the moisture content of the mushroom dregs is 20% -30%, and the moisture content of the wheat straws is 12% -18%.

Further, the weight ratio of the spectinomycin mushroom dregs, the mushroom dregs and the wheat straw is 5: 1: 1.

further, the pile turning frequency of the step (2) is that pile turning is carried out once every two days.

Further, the preparation method also comprises the following steps:

(4) crushing and screening: and (4) crushing and screening the compost aged and cached in the step (3), mixing powdery fertilizer with the grain diameter of more than 4mm serving as an auxiliary material with the mixture obtained in the step (1), and fermenting again.

Further, the preparation method also comprises the following steps:

(5) granulation and/or packaging: and packaging the powdery fertilizer to obtain a finished product, or granulating the powdery fertilizer and then packaging to obtain the finished product.

Further, the steps (1), (2) and (3) of the preparation method also comprise collecting gas in the fermentation and aging processes.

In a third aspect, the invention provides a spectinomycin bacterial residue compost produced by the preparation method.

The beneficial effect of the invention is that,

firstly, adopting a non-turning anaerobic fermentation mode in the earlier stage of composting, modifying amino or hydroxyl of spectinomycin by using modification enzymes, such as acetyl transferase, phosphotransferase, nucleotidyl transferase and the like, contained in spectinomycin residues and having the function of degrading the aminoglycoside antibiotics secreted by microorganisms with spectinomycin, wherein the modification enzymes exist under the microaerobic condition, the amino or hydroxyl of the spectinomycin is modified, the spectinomycin is passivated, and the spectinomycin is inactivated, so that the content of the antibiotics is reduced, and meanwhile, because the spectinomycin producing bacteria are mesophilic bacteria, the temperature in the anaerobic fermentation process can be naturally raised to 45-55 ℃, and the spectinomycin producing bacteria can be inactivated at the temperature;

because the microorganism with antibiotic degradation function is also antibiotic drug-resistant bacteria, drug resistance can be caused to diffuse among microorganisms of the same species and different species by virtue of the transfer original, so that the control on the drug-resistant genes is particularly important, the oxygen in the compost is increased by turning the compost in the second step of the invention, the composting temperature is increased, the antibiotic drug-resistant bacteria are killed, and the structure of the drug-resistant genes is destroyed, according to the detection on the drug-resistant genes and the related transfer original, the abundance of the drug-resistant genes is gradually reduced along with the progress of the composting process, and the dual control on the antibiotic residue and the drug-resistant genes in the bacterial residues is realized;

the spectinomycin mushroom residues, the mushroom residues and the wheat straws are matched for use, the ventilation and warming effects of the mixture are improved by using the characteristic that the mushroom residues and the wheat straws belong to loose materials, the carbon-nitrogen ratio of the mixture is adjusted by using the characteristic that the wheat straws are rich in lignocellulose and can be used as a carbon source, and the water content of the mixture is adjusted by using the characteristic that the mushroom residues and the wheat straws are low in water content until the water content requirement of the compost at the initial stage is met;

the raw materials used by the invention are spectinomycin bacterial residues in weight ratio: mushroom dregs: 4.5-5.5 parts of wheat straw: 1: 1, the consumption of spectinomycin mushroom dregs is the most, the consumption of auxiliary materials (mushroom dregs and wheat straws) is less, the composting cost is reduced, and the mushroom dregs and the wheat straws are cheap and easily available raw materials, so that great convenience is brought to the production of fertilizers; the composting characteristics are combined under the proportion, so that the anaerobic fermentation and the aerobic fermentation can be carried out, and the temperature is raised to a proper temperature.

The content of nitrogen, phosphorus and potassium in the spectinomycin bacterial residue compost provided by the invention is more than 5, and antibiotics and drug resistance genes are effectively reduced.

Drawings

In order to more clearly illustrate the embodiments or technical solutions in the prior art of the present invention, the drawings used in the description of the embodiments or prior art will be briefly described below, and it is obvious for those skilled in the art that other drawings can be obtained based on these drawings without creative efforts.

FIG. 1 is a graph showing the variation of antibiotic content in the composting process of example 3 and comparative examples 1 to 2;

FIG. 2 is a graph showing the changes in absolute abundance of drug-resistant genes and transfer elements in the composting processes of example 3 and comparative examples 1-2;

FIG. 3 is a flow chart of a method for preparing spectinomycin mushroom residue compost in example 4.

Detailed Description

In order to make those skilled in the art better understand the technical solution of the present invention, the technical solution in the embodiment of the present invention will be clearly and completely described below with reference to the drawings in the embodiment of the present invention, and it is obvious that the described embodiment is only a part of the embodiment of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1

A method for removing spectinomycin mushroom dreg antibiotics and drug-resistant genes comprises the following steps:

(1) anaerobic fermentation: mixing spectinomycin mushroom dregs, mushroom dregs and wheat straws according to the proportion of 4.5: 1: 1 to obtain a mixture, wherein the water content of the spectinomycin mushroom residues is 70-75 percent, the water content of the mushroom residues is 20-30 percent, and the water content of the wheat straws is 12-18 percent, after the materials are mixed according to the proportion, the water content of the mixture is kept within the range of 55-65 percent, the mixture is subjected to anaerobic fermentation without turning, and the fermentation temperature is naturally raised to 45-55 ℃ and maintained for 5 days;

(2) aerobic fermentation: and (2) turning the compost after the anaerobic fermentation in the step (1) to perform aerobic fermentation, naturally raising the fermentation temperature to 65-70 ℃ and maintaining for 20 days, wherein the turning frequency is once every two days.

Example 2

A method for removing spectinomycin mushroom dreg antibiotics and drug-resistant genes comprises the following steps:

(1) anaerobic fermentation: mixing spectinomycin mushroom dregs, mushroom dregs and wheat straws according to the proportion of 5.5: 1: 1 to obtain a mixture, wherein the water content of the spectinomycin mushroom residues is 70-75 percent, the water content of the mushroom residues is 20-30 percent, and the water content of the wheat straws is 12-18 percent, after the materials are mixed according to the proportion, the water content of the mixture is kept within the range of 55-65 percent, the mixture is subjected to anaerobic fermentation without turning, and the fermentation temperature is naturally raised to 45-55 ℃ and maintained for 7 days;

(2) aerobic fermentation: and (2) turning the compost after the anaerobic fermentation in the step (1) to perform aerobic fermentation, naturally raising the fermentation temperature to 65-70 ℃ and maintaining for 25 days, wherein the turning frequency is once every two days.

Example 3

A method for removing spectinomycin mushroom dreg antibiotics and drug-resistant genes comprises the following steps:

(1) anaerobic fermentation: mixing spectinomycin mushroom dregs, mushroom dregs and wheat straws according to the ratio of 5: 1: 1 to obtain a mixture, wherein the water content of the spectinomycin mushroom residues is 70-75 percent, the water content of the mushroom residues is 20-30 percent, and the water content of the wheat straws is 12-18 percent, after the materials are mixed according to the proportion, the water content of the mixture is kept within the range of 55-65 percent, the mixture is subjected to anaerobic fermentation without turning, and the fermentation temperature is naturally raised to 45-55 ℃ and maintained for 6 days;

(2) aerobic fermentation: and (2) turning the compost after the anaerobic fermentation in the step (1) to perform aerobic fermentation, naturally raising the fermentation temperature to 65-70 ℃ and maintaining for 22 days, wherein the turning frequency is once every two days.

Comparative example 1

A method for removing spectinomycin mushroom dreg antibiotics and drug-resistant genes comprises the following steps:

(1) anaerobic fermentation: mixing spectinomycin mushroom dregs, mushroom dregs and wheat straws according to the proportion of 1: 1: 1 to obtain a mixture, wherein the water content of the spectinomycin mushroom residues is 70-75 percent, the water content of the mushroom residues is 20-30 percent, and the water content of the wheat straws is 12-18 percent, after the materials are mixed according to the proportion, the water content of the mixture is kept within the range of 55-65 percent, the mixture is subjected to anaerobic fermentation without turning, and the fermentation temperature is naturally raised to 45-55 ℃ and maintained for 6 days;

(2) aerobic fermentation: and (2) turning the compost after the anaerobic fermentation in the step (1) to perform aerobic fermentation, naturally raising the fermentation temperature to 65-70 ℃ and maintaining for 22 days, wherein the turning frequency is once every two days.

Comparative example 2

A method for removing spectinomycin mushroom dreg antibiotics and drug-resistant genes comprises the following steps:

(1) anaerobic fermentation: mixing spectinomycin mushroom dregs, mushroom dregs and wheat straws according to the proportion of 3: 1: 1 to obtain a mixture, wherein the water content of the spectinomycin mushroom residues is 70-75 percent, the water content of the mushroom residues is 20-30 percent, and the water content of the wheat straws is 12-18 percent, after the materials are mixed according to the proportion, the water content of the mixture is kept within the range of 55-65 percent, the mixture is subjected to anaerobic fermentation without turning, and the fermentation temperature is naturally raised to 45-55 ℃ and maintained for 6 days;

(2) aerobic fermentation: and (2) turning the compost after the anaerobic fermentation in the step (1) to perform aerobic fermentation, naturally raising the fermentation temperature to 65-70 ℃ and maintaining for 22 days, wherein the turning frequency is once every two days.

The change of antibiotic residues in the composting process of example 3 and comparative examples 1-2 was examined, and the results are shown in fig. 1. It can be seen that the amount of spectinomycin remaining decreases with composting time, with the compost antibiotic of example 3 degrading more rapidly, reducing the antibiotic by more than 90% at the end of the compost.

The drug resistance genes and the abundance of related transfer elements in the composting process of example 3 and comparative examples 1-2 were examined, and the results are shown in fig. 2. As can be seen, the abundance of the drug resistance gene is gradually reduced along with the composting time, which indicates that the two-step fermentation process, particularly the high temperature during aerobic fermentation, effectively controls the drug resistance of the compost.

Example 4

As shown in fig. 3, the preparation method of the spectinomycin bacterial residue compost comprises the following steps:

(1) anaerobic fermentation: mixing spectinomycin mushroom dregs, mushroom dregs and wheat straws according to the proportion of 4.5: 1: 1 to obtain a mixture, wherein the water content of the spectinomycin mushroom residues is 70-75%, the water content of the mushroom residues is 20-30%, and the water content of the wheat straws is 12-18%, the water content of the mixture is kept within the range of 55-65% after the materials are mixed according to the proportion, the mixture is subjected to anaerobic fermentation without turning, the fermentation temperature is naturally raised to 45-55 ℃ and maintained for 6 days, and compost release gas is collected during the period for waste gas treatment;

(2) aerobic fermentation: turning the compost subjected to anaerobic fermentation in the step (1) for aerobic fermentation, naturally raising the fermentation temperature to 65-70 ℃ and maintaining for 22 days, wherein the turning frequency is once every two days, and during the turning, compost release gas is collected for waste gas treatment;

(3) aging and caching: aging and caching the compost after aerobic fermentation in the step (2) to reduce the temperature of the compost to room temperature, wherein the aging and caching time is 18 days, and during the aging and caching time, gas released by the compost is collected to perform waste gas treatment;

(4) crushing and screening: crushing and screening the compost aged and cached in the step (3), mixing powdery fertilizer with the grain diameter of more than 4mm serving as an auxiliary material with the mixture obtained in the step (1), and fermenting again;

(5) granulation and/or packaging: and packaging the powdery fertilizer to obtain a finished product of the spectinomycin bacterial residue compost, or granulating the powdery fertilizer and then packaging to obtain the finished product of the spectinomycin bacterial residue compost.

Although the present invention has been described in detail by referring to the drawings in connection with the preferred embodiments, the present invention is not limited thereto. Various equivalent modifications or substitutions can be made on the embodiments of the present invention by those skilled in the art without departing from the spirit and scope of the present invention, and these modifications or substitutions are within the scope of the present invention/any person skilled in the art can easily conceive of the changes or substitutions within the technical scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.