CN109879561A

CN109879561A - A method for removing heavy metals, antibiotics and resistance genes in livestock and poultry manure

Info

Publication number: CN109879561A
Application number: CN201910208598.6A
Authority: CN
Inventors: 郑莉; 钟裕健; 许燕滨; 谢光炎; 杜青平; 谢志帆
Original assignee: Guangdong University of Technology
Current assignee: Guangdong University of Technology
Priority date: 2019-03-19
Filing date: 2019-03-19
Publication date: 2019-06-14
Anticipated expiration: 2039-03-19
Also published as: CN109879561B

Abstract

The invention discloses the minimizing technologies of heavy metal, antibiotic and resistant gene in a kind of feces of livestock and poultry, comprising the following steps: step 1: preparing aspergillus niger spore suspension；Step 2: the aspergillus niger spore suspension being subjected to fermented and cultured, obtains tunning；Step 3: feces of livestock and poultry being placed in the tunning, leaching obtains the first product.The minimizing technology can effectively remove heavy metal in feces of livestock and poultry, antibiotic and resistant gene, greatly shorten the Bioleaching time.

Description

The minimizing technology of heavy metal, antibiotic and resistant gene in a kind of feces of livestock and poultry

Technical field

The present invention relates to a huge sum of moneys in environmental protection and technical field of resource comprehensive utilization more particularly to a kind of feces of livestock and poultry Belong to, the minimizing technology of antibiotic and resistant gene.

Background technique

In recent years, China's livestock and poultry breeding industry is fast-developing, and large-scale cultivation level significantly improves, and has ensured meat, egg, milk etc. The effective supply of livestock products, but a large amount of feces of livestock and poultry generated do not obtain reasonable disposition in time and utilize, and become China and work as The a great problem of preceding Control of Agricultural Non-point Source Pollution.2015, it was 10.73 hundred million tons that China's feces of livestock and poultry, which generates total amount, however livestock and poultry Manure resource amount is less than 0.1 hundred million tons.Energy and Fertilizer Transformed are the Main ways that livestock and poultry dung recovery utilizes.But just at present I Under the pattern of state's breeding separation, biogas residue and biogas liquid returning to the field difficulty is big.It is efficiently fast in conjunction with current domestic feces of livestock and poultry utilization of resources situation Fast compost-production organic commercial fertilizer is its trend of industry development.

Heavy metal, antibiotic and resistant gene it is exceeded be restrict feces of livestock and poultry cropland application bottleneck, they are with livestock and poultry Excrement enters soil, will seriously threaten ecological environment security and human health.Heavy metal in feces of livestock and poultry, antibiotic is effectively reduced And resistant gene total amount is the key that feces of livestock and poultry is a large amount of agricultural.

The removal of feces of livestock and poultry heavy metal may be implemented in bioleaching technology, and does not lose containing for feces of livestock and poultry organic matter Amount, thus, bioleaching technology is to remove the effective way of sludge heavy-metal, is with a wide range of applications.However, biology drip The leaching period is long, and extraction time is 15 days or so, and soaks low efficiency to the drip of heavy metal, while bioleaching is in feces of livestock and poultry Whether there is or not removal effects to have not been reported for antibiotic and resistant gene.

Summary of the invention

In view of this, it is an object of the present invention to provide heavy metal, antibiotic and resistant genes in a kind of feces of livestock and poultry Minimizing technology, which can effectively remove heavy metal in feces of livestock and poultry, antibiotic and resistant gene, greatly shorten life Object extraction time.

Its specific technical solution is as follows:

The present invention provides the minimizing technology of heavy metal, antibiotic and resistant gene in a kind of feces of livestock and poultry, including it is following Step:

Step 1: preparing aspergillus niger spore suspension；

Step 2: the aspergillus niger spore suspension being subjected to fermented and cultured, obtains tunning；

Step 3: feces of livestock and poultry being placed in the tunning, leaching obtains the first product.

The present invention has no special limitation to the source of inoculation bacterium, can make by oneself, or general commercially available.The present invention is excellent Black-koji mould is isolated in choosing from the feces of livestock and poultry of domestication, and through morphology and ITS sequencing identification, obtains black-koji mould.It is black Indigenous bacterium of the aspergillus in feces of livestock and poultry, it is adaptable in feces of livestock and poultry, there is certain resistance.

In the present invention, resistant gene is antibiotics resistance gene (antibioticresistancegenes, ARGs).

In the present invention, the tunning be organic acid (gluconic acid, oxalic acid and citric acid), Aspergillus Niger acid amount compared with It is excellent；First product is to eliminate heavy metal, antibiotic and resistant gene feces of livestock and poultry.

In the present invention, after fermented and cultured described in step 2, before obtaining tunning, further includes: filtering.

It is described to leach to leach indirectly in the present invention, i.e., it is leached using the organic acid of Aspergillus Niger；The indirect leaching Out specifically: the tunning containing feces of livestock and poultry is placed in constant temperature oscillator and is vibrated.It is described to be placed in feces of livestock and poultry The concentration of the dissolved oxygen of the mixture formed into the tunning is 3~4mg/L, more preferably 3mg/L or 4mg/L.

Preferably, the temperature of the leaching is 28~30 DEG C, more preferably 30 DEG C, time of the leaching be 20h~ 22h, more preferably 20h, the shaking speed of the leaching are 160~200r/min, more preferably 180r/min or 200r/min.

Preferably, the concentration of aspergillus niger spore is 10 in the aspergillus niger spore suspension⁷~10⁸A/mL, more preferably 10⁷A/mL or 10⁸A/mL.

Preferably, the culture medium of the fermented and cultured is sterile sucrose culture medium.

Preferably, in the sterile sucrose culture medium sucrose concentration be 100~120g/L, more preferably 100g/L, 110g/L or 120g/L.

Preferably, in the fermented and cultured, the inoculum density of the aspergillus niger spore suspension is 4 × 10⁵~5 × 10⁵ A/mL, more preferably 4 × 10⁵A/mL or 5 × 10⁵A/mL.

Preferably, the time of the fermented and cultured is 12~15 days, more preferably 12 days, 13 days or 15 days, the fermentation The temperature of culture is 28~30 DEG C, more preferably 30 DEG C.

Preferably, concentration of the feces of livestock and poultry described in step 3 in the tunning is 40~50g/L, wherein fermentation Product is the solvent of feces of livestock and poultry, more preferably 40g/L or 50g/L.

Preferably, the preparation aspergillus niger spore suspension specifically:

Aspergillus niger is seeded in after being cultivated in sucrose culture medium and is dissolved in water, obtains the aspergillus niger spore suspension；

The time of the culture is 6~7 days, more preferably 6 days or 7 days.

As can be seen from the above technical solutions, the invention has the following advantages that

The present invention provides the minimizing technology of heavy metal, antibiotic and resistant gene in a kind of feces of livestock and poultry, including it is following Step: step 1: preparing aspergillus niger spore suspension；Step 2: the aspergillus niger spore suspension being subjected to fermented and cultured, is obtained Tunning；Step 3: feces of livestock and poultry being placed in the tunning, leaching obtains the first product.The minimizing technology can be with Heavy metal in feces of livestock and poultry, antibiotic and resistant gene are effectively removed, the Bioleaching time is greatly shortened, foreshortens to 20h.It should In minimizing technology, aspergillus niger is using organic matter as carbon source, and by the metabolism secretion organic acid of itself, one side organic acid can To provide H⁺, so that heavy metal is more with the presence of free state, another aspect acid ion can with complex, Promote dissolution.Thus, there is apparent facilitation for heavy metal, antibiotic and the ARGs in removal feces of livestock and poultry.Experiment knot Fruit shows that minimizing technology provided by the invention is not less than 64% to the removal rate of heavy metal Cu in feces of livestock and poultry, to heavy metal Zn Removal rate be not less than 80%, 43% is not less than to the removal rate of heavy metal As.Not to Enrofloxacin removal rate in feces of livestock and poultry Lower than 68%, 40% or less the total abundance of 69%, ARGs is reduced in feces of livestock and poultry is not less than to Ciprofloxacin removal rate.

Detailed description of the invention

In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below There is attached drawing needed in technical description to be briefly described, it should be apparent that, the accompanying drawings in the following description is only this Some embodiments of invention without any creative labor, may be used also for those of ordinary skill in the art To obtain other attached drawings according to these attached drawings.

Fig. 1 is the heavy metal removing rate schematic diagram that the embodiment of the present invention 1 to embodiment 3 provides；

Fig. 2 is the antibiotic removal rate schematic diagram that the embodiment of the present invention 1 to embodiment 3 provides；

Fig. 3 is the total abundance schematic diagram of resistant gene that the embodiment of the present invention 1 is provided to embodiment 3.

Specific embodiment

It in order to make the invention's purpose, features and advantages of the invention more obvious and easy to understand, below will be to of the invention real The technical solution applied in example is clearly and completely described, it is clear that the embodiments described below are only present invention a part Embodiment, and not all embodiment.Based on the embodiments of the present invention, those of ordinary skill in the art are not making creation Property labour under the premise of all other embodiment obtained, shall fall within the protection scope of the present invention.

Embodiment 1

The black-koji mould of preservation is inoculated into sucrose culture medium and cultivates 6d, with spore under sterile washing in sterile conical flask Concussion shakes up, and is configured to aspergillus niger spore suspension, and spore concentration is about 1.0 × 10⁷A/mL.Above-mentioned spore suspension is taken to set Fermentation and acid is carried out in fermentation medium, the sucrose concentration of fermentation medium is 100g/L, and the bacteria concentration that connects of fermentation and acid is 5 ×10⁵A/mL, the temperature of fermentation and acid are 30 DEG C.After 12d, the organic acid for taking fermentation to generate is filtered, and feces of livestock and poultry is pressed 50g/L concentration is added in organic acid, is placed in 30 DEG C, in the constant temperature oscillator of 180rpm, is leached 20h, is kept in reaction vessel Dissolved oxygen reach 4mg/L, leaching obtain the first product feces of livestock and poultry.

Embodiment 2

The black-koji mould of preservation is inoculated into sucrose culture medium and cultivates 7d, with spore under sterile washing in sterile conical flask Concussion shakes up, and is configured to aspergillus niger spore suspension, and spore concentration is about 1.0 × 10⁸A/mL.Above-mentioned spore suspension is taken to set Fermentation and acid is carried out in fermentation medium, the sucrose concentration of fermentation medium is 120g/L, and the bacteria concentration that connects of fermentation and acid is 4 ×10⁵A/mL, the temperature of fermentation and acid are 28 DEG C.After 13d, the organic acid for taking fermentation to generate is filtered, and feces of livestock and poultry is pressed 40g/L concentration is added in organic acid, is placed in 30 DEG C, in the constant temperature oscillator of 180rpm, is leached 22h, is kept in reaction vessel Dissolved oxygen reach 3mg/L, leaching obtain the first product feces of livestock and poultry.

Embodiment 3

The black-koji mould of preservation is inoculated into sucrose culture medium and cultivates 7d, with spore under sterile washing in sterile conical flask Concussion shakes up, and is configured to aspergillus niger spore suspension, and spore concentration is about 1.0 × 10⁸A/mL.Above-mentioned spore suspension is taken to set Fermentation and acid is carried out in fermentation medium, the sucrose concentration of fermentation medium is 110g/L, and the bacteria concentration that connects of fermentation and acid is 3.5×10⁵A/mL, the temperature of fermentation and acid are 28 DEG C.After 15d, the organic acid for taking fermentation to generate is filtered, feces of livestock and poultry It is added in organic acid by 40g/L concentration, is placed in 30 DEG C, in the constant temperature oscillator of 200rpm, leached 20h, keep reaction vessel Interior dissolved oxygen reaches 4mg/L, and leaching obtains the first product feces of livestock and poultry.

Embodiment 4

Performance detection is carried out to the first product that embodiment 1 to 3 provides.

It please refers to Fig.1 to Fig.3, the experimental results showed that, the minimizing technology that embodiment 1 provides is to heavy metal Cu in feces of livestock and poultry Removal rate 70%, to heavy metal Zn removal rate be 80%, to Enrofloxacin removal rate in feces of livestock and poultry be 68%, to cyclopropyl sand Star removal rate is 38% that the total abundance of 69%, ARGs is reduced in raw material.

The minimizing technology that embodiment 2 provides is 70% to the removal rate of heavy metal Cu in feces of livestock and poultry, is gone to heavy metal Zn It is 70% to Enrofloxacin removal rate in feces of livestock and poultry except rate is 82%, is the total abundance of 71%, ARGs to Ciprofloxacin removal rate 35% be reduced in feces of livestock and poultry.

The minimizing technology that embodiment 3 provides is 67% to the removal rate of heavy metal Cu in feces of livestock and poultry, is gone to heavy metal Zn It is 70% to Enrofloxacin removal rate in feces of livestock and poultry except rate is 80%, is the total abundance of 69%, ARGs to Ciprofloxacin removal rate 36% be reduced in feces of livestock and poultry.

The above, the above embodiments are merely illustrative of the technical solutions of the present invention, rather than its limitations；Although referring to before Stating embodiment, invention is explained in detail, those skilled in the art should understand that: it still can be to preceding Technical solution documented by each embodiment is stated to modify or equivalent replacement of some of the technical features；And these It modifies or replaces, the spirit and scope for technical solution of various embodiments of the present invention that it does not separate the essence of the corresponding technical solution.

Claims

1. a method for removing heavy metals, antibiotics and resistance genes in livestock manure, is characterized in that, comprises the following steps:

Step 1: prepare Aspergillus niger spore suspension;

Step 2: fermenting and culturing the Aspergillus niger spore suspension to obtain a fermentation product;

Step 3: placing livestock and poultry manure in the fermentation product, and leaching to obtain the first product.

2 . The removal method according to claim 1 , wherein the concentration of Aspergillus niger spores in the Aspergillus niger spore suspension is 10 ⁷ to 10 ⁸ /mL. 3 .

3. The removal method according to claim 1, wherein the medium of the fermentation culture is a sterile sucrose medium.

4 . The removal method according to claim 3 , wherein the concentration of sucrose in the sterile sucrose medium is 100-120 g/L. 5 .

5 . The removing method according to claim 1 , wherein in the fermentation culture, the inoculation concentration of the Aspergillus niger spore suspension is 4-5×10 ⁵ cells/mL. 6 .

6 . The removal method according to claim 1 , characterized in that, the time of the fermentation and cultivation is 12-15 days, and the temperature of the fermentation and cultivation is 28-30° C. 7 .

7 . The removal method according to claim 1 , wherein the concentration of the livestock and poultry manure in the fermentation product in step 3 is 40-50 g/L. 8 .

8 . The removal method according to claim 1 , wherein the temperature of the leaching is 28-30° C., the time of the leaching is 20h-22h, and the rotating speed of the shaking table for the leaching is 160-180r/min. 9 . .

9 . The removal method according to claim 1 , wherein the concentration of dissolved oxygen in the mixture formed by placing livestock and poultry manure in the fermentation product is 3-4 mg/L. 10 .

10. removal method according to claim 1, is characterized in that, described preparation Aspergillus niger spore suspension is specially:

Inoculate the Aspergillus niger on the sucrose medium for cultivation and dissolve in water to obtain the Aspergillus niger spore suspension;

The culturing time is 6-7 days.