CN105254346B - The minimizing technology of fluoroquinolone antibiotics in a kind of birds and beasts excrement - Google Patents

Abstract

The invention discloses a method for removing fluoroquinolone antibiotics in poultry feces. The steps include: (1) configuring composting raw materials: making composting raw materials from chicken manure, sawdust, sterile distilled water and high-temperature-resistant bacterial agents, wherein chicken manure is mixed with The mass ratio of sawdust is 5-10:1, the moisture content of compost raw materials is between 55% and 60%, and the high-temperature-resistant bacterial agent is made of Cellulomonas flavinogenes; (2) High-temperature composting: control the frequency of composting so that The temperature is between 45-65°C to achieve high-temperature fermentation; (3) Medium-temperature composting: After high-temperature fermentation, the frequency of manual turning is controlled so that the temperature of the compost product is between 30-45°C to achieve medium-temperature fermentation. The method of the invention fully utilizes the degradation effect of the high-temperature-resistant strains and related microorganisms to remove the fluoroquinolone antibiotics in poultry manure, and reduces the fluoroquinolone antibiotics in the organic fertilizer to a safe amount.

Description

A method for removing fluoroquinolone antibiotics in poultry feces

technical field

The invention relates to a method for removing antibiotics in poultry manure organic fertilizer, in particular to a method for removing six kinds of fluoroquinolone (FQs) antibiotics.

Background technique

Antibiotics are widely used in human medicine, livestock and poultry farming, aquaculture, etc. As animal growth promoters and therapeutic drugs, animal husbandry has become the industry that uses the largest amount of antibiotics. According to statistics, about 50% to 70% of antibiotics consumed each year are used in animal husbandry. After antibiotics enter the animal body, 40% to 90% enter the environment in the form of the mother or metabolites along with the manure of livestock and poultry. As early as 2007, some scholars investigated 143 livestock farms in 8 provinces of China, and found that the survey samples contained high levels of fluoroquinolone antibiotics, among which the concentration of enrofloxacin in pig manure and cow manure was 33.26 mg·kg ^-1 , The concentration of ciprofloxacin was 33.98 mg·kg ^-1 ; the maximum concentration of norfloxacin in chicken manure was 225.45 mg·kg ^-1 , the maximum concentration of ciprofloxacin was 45.59 mg·kg ^-1 , the maximum concentration of enrofloxacin It is 1420.76 mg·kg ^-1 .

Antibiotic residues in the environment threaten the health of animals, plants and humans. Studies have found that quinolone antibiotics have teratogenic effects on the fetus, and are also very harmful to the human nervous system, digestive and absorption system, heart and vascular system. The results of the study show that the heavy use of antibiotics will lead to the development of resistance of pathogenic microorganisms, so that the effective dose of antibiotics to kill bacteria is continuously increasing. For example, after applying cow manure to the garden, the soil resistance to specific antibiotics increases by 70%. In addition, the use of antibiotics will affect the properties of microorganisms in the soil. For example, Ma Yi et al. have shown that enrofloxacin with a content higher than 0.1 μg/g will significantly reduce the richness and diversity of soil microorganisms, thereby reducing their degradation of other organic pollutants. and heavy metals.

Fluoroquinolone antibiotics widely exist in the soil environment, especially in poultry manure, and pose a serious threat to the soil environment, animals, plants, and human health. Therefore, it is urgent to develop related technologies to reduce or remove the concentration of fluoroquinolone antibiotics in the environment. At present, there is a small amount of research on the removal of antibiotics in water, such as ultrasonic degradation, artificial wetland, etc. Ultrasonic degradation is fast and has a wide range of applications, but its energy consumption is high, the cost is high, and the reaction conditions are difficult to control; the artificial wetland is low in cost. It is easy to maintain but it occupies a large area and emits a foul smell. The research on fluoroquinolone antibiotics in poultry feces mainly stays in its detection and analysis methods, and there are few studies on their removal methods.

Contents of the invention

The purpose of the present invention is to break through the technical bottleneck of removing antibiotic pollutants in poultry manure, and propose a method for removing fluoroquinolone antibiotics in poultry manure to reduce its concentration and realize the harmlessness of poultry manure as organic fertilizer.

The technical scheme provided by the invention is: a method for removing fluoroquinolone antibiotics in poultry manure, comprising the steps of:

(1) Composting raw materials: Chicken manure, sawdust, sterile distilled water and high-temperature-resistant bacterial agents are used to make compost raw materials. The mass ratio of chicken manure to sawdust is 5-10:1, and the moisture content of compost raw materials is 55%-60. %, the high temperature-resistant bacterial agent is prepared as Cellulomonas flavogenes;

(2) High-temperature composting: control the turning frequency so that the composting temperature is between 45 and 65°C to achieve high-temperature fermentation;

(3) Medium-temperature composting: After high-temperature fermentation, the frequency of manual turning is controlled so that the temperature of the compost product is between 30 and 45°C to achieve medium-temperature fermentation.

In the method described above, the high-temperature-resistant bacterial agent is added in the form of liquid bacterial agent, and the dosage ratio is 1:10-1:25 (v/w), preferably 1:20 (v/w).

The method, wherein, the bacterial strain is Cellulomonas flavogenes, and the steps of domesticating and preparing the high-temperature-resistant strain are as follows:

Cellulomonas flavogenes stored on LB medium was activated, and Cellulomonas flavogenes was inoculated into selective medium at 2% and shaken continuously for 72 hours at a constant temperature of 60°C.

Selective medium: KH ₂ PO ₄ : 0.5g, NaCI : 0.5g, Peptone: 0.5g, MgCI ₂ : 0.5g, (NH ₄ )SO ₄ : 0.5g, NH ₄ NO ₃ : 1.0g, K ₂ HPO ₄ : 1.5g, distilled water 1000ml, pH is neutral.

LB medium: peptone: 10g; yeast extract: 5g; NaCI: 10g; distilled water 1000ml; PH: 7.0.

The properties of the inoculated production strain of the present invention are as follows: Gram-positive, its cells are spherical, capable of producing cellulase and protease, and in the selective medium, its cellulase activity is 20.79 u/ml, and the protease activity is 18.52 u/ml, the bacteria still maintained high activity at 55 ℃.

In the method, the chicken manure is fresh chicken manure, the mass ratio of it to sawdust is 6-8:1, and the C/N is kept at 20:1.

According to the method, the moisture content of the prepared compost raw materials is between 55% and 60%, and the temperature in the composting chamber is controlled at 25 to 30°C.

In the method, in the high-temperature composting step, the high-temperature stage is reached after 3-4 days of composting, and the temperature is controlled at 45-60° C. for 10-12 days to realize high-temperature fermentation.

In the method, in the medium-temperature composting step, the temperature of the composting product is between 30-45° C. for 10-14 days.

In the method, the fluoroquinolone antibiotics include one or more of norfloxacin, ciprofloxacin, sarafloxacin, lomefloxacin, and enrofloxacin.

The invention proposes a method for removing fluoroquinolone antibiotics in poultry manure by combining composting with exogenous bacterial agent addition. The method of the present invention makes full use of the degradation effect of heat-resistant bacteria and related microorganisms to remove fluoroquinolone antibiotics in poultry manure, reduces the fluoroquinolone antibiotics in organic fertilizers to a safe amount, and fills the gap of removing fluoroquinolone antibiotics in poultry manure. Technology gap. At the same time, it has the following advantages:

(1) Using poultry and livestock manure as raw material, no need to add other chemical reagents, and will not cause secondary pollution to the environment; (2) Adopting composting technology, no need to generate electricity and other energy consumption, and low economic cost; (3) Using manual turning Oxygen supply, easy to operate and popularize.

Detailed ways

The present invention will be further clarified through the detailed description of specific embodiments below, but it is not intended to limit the present invention, but only for illustration.

Example 1

The present invention removes the method for fluoroquinolone antibiotics in poultry manure, and its steps are as follows:

Step 1: Activate the Cellulomonas flavogenes stored on the LB medium, inoculate 2% of the Cellulomonas flavogenes into the selective medium, and shake continuously for 72 hours at a constant temperature of 60°C.

LB medium: peptone: 10g; yeast extract: 5g; NaCI: 10g; distilled water 1000ml; PH: 7.0.

Selective medium: KH ₂ PO ₄ : 0.5g, NaCI : 0.5g, Peptone: 0.5g, MgCI ₂ : 0.5g, (NH ₄ )SO ₄ : 0.5g, NH ₄ NO ₃ : 1.0g, K ₂ HPO ₄ : 1.5g, distilled water 1000ml, pH 6.8~7.2.

Step 2: Mix fresh chicken manure with a moisture content of about 65.0% and an organic carbon content of about 20% with sawdust and distilled water to control the C/N ratio at 20:1 and the moisture content between 55% and 60%. , the amount of bacteria agent is about 5% of the volume of the compost material.

Step 3: The initial material temperature of the pile is 25°C, the height of the pile is 1.0 m, and the room temperature is kept at 25~30°C. The oxygen is supplied by manual turning. After 2~3 days of fermentation, the temperature of the pile rises to 45 ℃, entering the high temperature period, the composting temperature rises rapidly to 60°C on the 3rd to 8th day of composting, and gradually drops from 60°C to 45°C on the 8th to 17th day. 12 days.

In this step, the compost material becomes weakly alkaline, the pH is stable between 7.3 and 8.1, and the moisture content is stable between 55% and 65%.

Step 4: The compost material fermented by the above-mentioned high temperature, the room temperature is 25~30°C, and the oxygen is supplied by manual turning, and it enters the medium temperature period. The temperature of the compost material lasts for 10 to 14 days between 30°C and 45°C.

In this step, the compost material is weakly alkaline, the pH is stable between 7.6 and 8.1, and the moisture content is stable between 50% and 55%.

By the end of the middle temperature fermentation stage, the seed germination index (GI) has reached more than 85%.

After three parallel experiments, the degradation effects of fluoroquinolone antibiotics are shown in Table 1.

Table 1 Norfloxacin Ciprofloxacin Sarafloxacin Lomefloxacin Enrofloxacin Initial concentration (mg/kg) 12.11 11.79 12.34 15.35 14.88 Removal rate 1 (high temperature section, %) 58.31 59.62 55.28 43.19 65.32 Removal rate 2 (medium temperature section, %) 37.43 15.97 23.62 33.08 17.12 Total removal rate (%) 73.91 66.06 65.84 61.99 71.26

Example 2

According to the proportioning of materials and the method of operation of Example 1, the experiment was carried out again. The difference between Example 1 and this Example 2 is: the initial concentration of fluoroquinolone antibiotics in the chicken manure is different. Through three parallel tests, the fluoroquinolone antibiotics The degradation effect is shown in Table 2.

Table 2 Norfloxacin Ciprofloxacin Sarafloxacin Lomefloxacin Enrofloxacin Initial concentration (mg/kg) 21.72 22.19 22.39 27.73 24.71 Removal rate 1 (high temperature section, %) 48.25 52.07 63.39 35.89 54.40 Removal rate 2 (medium temperature section, %) 66.81 56.18 35.87 39.31 27.93 Total removal rate (%) 82.82 79.00 76.52 61.09 67.13

Show from above two examples, through adopting the present invention to process fluoroquinolone antibiotics in chicken manure, make the fluoroquinolone antibiotics of 61.09～82.82% in chicken manure be removed, reduce the harm of chicken manure as organic fertilizer, make wherein fluorine The content of quinolone antibiotics reached the safe limit.

Claims (9)

1. A kind of 1. method for removing fluoroquinolone antibiotics in fowl and animal excrement, which is characterized in that include the following steps：

   (1) composting material is configured：Composting material, wherein chicken manure is made in chicken manure, sawdust, sterile distilled water and high temperature resistant microbial inoculum Mass ratio with sawdust is 5 ~ 10:1, for composting material moisture content between 55% ~ 60%, high temperature resistant microbial inoculum is made as production yellowish fiber unit cell Bacterium；The high temperature resistant microbial inoculum is added in the form of liquid bacterial agent, and the ratio that adds is 1：10~1：25（v/w）；

   (2) During High-Temperature Composting：Control turn pile frequency is so that compost temperature between 45 ~ 65 DEG C, realizes hot fermentation；

   (3) medium temperature compost：After hot fermentation, artificial turn pile frequency is controlled, makes composting production temperature between 30 ~ 45 DEG C, it is real Existing mesophilic digestion.
2. 2. according to the method described in claim 1, it is characterized in that, the high temperature resistant microbial inoculum is added in the form of liquid bacterial agent, throw It is 1 to add ratio：20（v/w）.
3. 3. according to the method described in claim 1, it is characterized in that, the preparation process of high temperature resistant microbial inoculum is as follows：Activation production is yellow fine Monad is tieed up, yellowish fiber monad is seeded to by 2% in selective medium, it is small continuously to shake 72 under the conditions of 60 DEG C of constant temperature When.
4. 4. according to the method described in claim 1, it is characterized in that, the chicken manure be fresh chicken manures, the mass ratio with sawdust It is 6 ~ 8:1, and C/N is maintained at 20：1.
5. 5. according to the method described in claim 1, it is characterized in that, the composting material moisture content of configuration is between 55% ~ 60%.
6. 6. according to the method described in claim 1, it is characterized in that, the control of compost indoor temperature is at 25 ~ 30 DEG C.
7. 7. according to the method described in claim 1, it is characterized in that, the During High-Temperature Composting step, compost reach a high temperature after 3 ~ 4 days Stage, temperature control continue 10 ~ 12 days between 45 ~ 60 DEG C, realize hot fermentation.
8. 8. according to the method described in claim 1, it is characterized in that, the medium temperature composting step, composting production temperature is 30 ~ 45 Between DEG C, continue 10 ~ 14 days.
9. 9. according to claim 1 to 8 any one of them method, which is characterized in that the fluoroquinolone antibiotics include promise It is one or more in Flucloxacillin, Ciprofloxacin, sarafloxacin, Lomefloxacin, Enrofloxacin.