An anaerobic microbial combination with sulfonamide antibiotic degradation function and its application

Abstract

The present invention belongs to the technical field of antibiotic degradation treatment, and specifically relates to an anaerobic microorganism combination with sulfonamide antibiotic degradation function and its application. The present invention provides two anaerobic microorganisms for sulfonamide antibiotic treatment, namely ParaclostridiumbifermentansJ2 and Clostridium sp.J2. These two anaerobic microorganisms can degrade sulfonamide antibiotics, and the removal effect of sulfonamide antibiotics can be enhanced by combining the two. In addition, after ParaclostridiumbifermentansJ2 and / or Clostridium sp.J2 are made into microbial immobilized bacterial balls with sodium alginate and biochar, they can still be quickly started in a short time and sulfonamide antibiotics can be removed efficiently, which provides a new way for the degradation of sulfonamide antibiotics and the treatment of sulfonamide antibiotic pharmaceutical wastewater.

Description

Technical Field

[0001] The invention belongs to the technical field of antibiotic degradation treatment, and specifically relates to an anaerobic microbial combination with sulfonamide antibiotic degradation function and application thereof. Background Art

[0002] For a long time, antibiotics have been widely used as antibacterial drugs to prevent and treat human and animal diseases. At the same time, they are used as feed additives in animal breeding to promote animal growth. They are one of the most widely used drugs in the world. According to statistics, the production and sales of sulfonamide antibiotics are increasing year by year, among which sulfamethoxazole is the most common sulfonamide antibiotic. However, sulfamethoxazole cannot be completely absorbed by humans / animals. Up to 85% of antibiotics are discharged into the environment in the form of prototype or metabolites through the feces and urine of patients and livestock and poultry, causing pollution to soil and water bodies through different pathways. At the same time, due to the high solubility and fluidity of sulfonamide antibiotics, they are easy to migrate and distribute in the water environment. Due to their high stability and weak adsorption, they are often easier to persist in the environment for a long time than other antibiotics.

[0003] At present, the treatment methods for antibiotics in wastewater mainly include physical, chemical and biological methods. Among them, the traditional physical and chemical methods mainly include precipitation, ion exchange, adsorption, etc., but their application is limited due to their high cost. The microbial method has attracted widespread attention from scholars due to its advantages such as good effect and low cost. At present, aerobic microorganisms are often used in sewage treatment plants to treat wastewater, but this process will produce a large amount of residual sludge and the operating cost is too high. Anaerobic microorganisms have the advantages of low energy consumption and low residual sludge production, and have received widespread attention in wastewater treatment. Therefore, the use of anaerobic microorganisms to treat wastewater has broad application prospects.

[0004] In addition, the microbial fixation method is used to fix anaerobic microorganisms, and the pollutants in the environmental medium are adsorbed and biotransformed by the microorganisms' own metabolic process. This method has the outstanding advantages of fast startup, strong toxicity resistance and adaptability, easy solid-liquid separation and fast precipitation, small and lightweight treatment process equipment, and small footprint. In recent years, it has attracted widespread attention from scholars at home and abroad.

[0005] In summary, it can be seen that it is of great significance to explore anaerobic bacteria that can efficiently degrade sulfonamide antibiotics and their applications. Summary of the invention

[0006] In order to overcome the deficiencies of the above-mentioned prior art, the present invention provides two anaerobic microorganisms for the treatment of sulfonamide antibiotics - ParaclostridiumbifermentansJ2 and Clostridium sp.J2. These two strains or their combination have significant advantages such as high degradation efficiency and short treatment cycle.

[0007] In order to achieve the above object, the technical solution adopted by the present invention is:

[0008] The first aspect of the present invention provides the use of Paraclostridiumbifermentans J2 and / or Clostridiumsp. J2 in degrading sulfonamide antibiotics.

[0009] The second aspect of the present invention provides the use of Paraclostridiumbifermentans J2 and / or Clostridiumsp. J2 in treating pharmaceutical wastewater containing sulfonamide antibiotics.

[0010] The present invention provides two sulfonamide antibiotic degrading bacteria, one of which is a paraclostridium strain, named Paraclostridiumbifermentans J2, and the other is a cereus strain, named Clostridium sp. J2. These two strains were obtained after long-term domestication of sulfate-reducing sludge collected from a sulfur-mediated bioreactor, and then screened, separated and purified multiple times. The preservation information of Paraclostridiumbifermentans J2 is: Preservation unit: Guangdong Microbiological Culture Collection Center (GDMCC), preservation number: GDMCC No: 62845, preservation address: Guangdong Institute of Microbiology, 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou City, Guangdong Province, preservation date: September 28, 2022. The preservation information of Clostridium sp. J2 is: Preservation unit: Guangdong Microbiological Culture Collection Center (GDMCC), preservation number: GDMCC No: 62934, preservation address: Guangdong Institute of Microbiology, 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou City, Guangdong Province, preservation date: October 28, 2022.

[0011] Preferably, the sulfonamide antibiotic comprises sulfamethoxazole.

[0012] More preferably, the concentration of sulfamethoxazole is no more than 10 mg / L.

[0013] The sulfamethoxazole removal experiment found that the removal rates of ParaclostridiumbifermentansJ2 for 1mg / L, 2mg / L and 5mg / L sulfamethoxazole within 72 hours were 53.0%, 52.6% and 44.25%, respectively; the removal rates of Clostridiumsp.J2 for 1mg / L, 2mg / L and 5mg / L sulfamethoxazole within 72 hours were 34.7%, 31.2% and 24.92%, respectively. When the two microorganisms were treated together (the total inoculation amount remained unchanged), the removal rates of 1mg / L, 2mg / L and 5mg / L sulfamethoxazole within 24 hours were 68.7%, 69.5% and 64.8%, respectively, which proved that ParaclostridiumbifermentansJ2 and Clostridium sp.J2 have good tolerance and degradability to sulfamethoxazole.

[0014] Preferably, the Paraclostridiumbifermentans J2 and / or Clostridium sp. J2 are made into immobilized bacterial pellets before use.

[0015] More preferably, rice straw biochar is added to ParaclostridiumbifermentansJ2 bacterial solution and / or Clostridium sp.J2 bacterial solution, and the biochar-microorganism solution is mixed with a sodium alginate solution after shaking culture, and the resulting mixture is dropped into a continuously stirred CaCl2 solution to form gel beads after cross-linking.

[0016] The sulfamethoxazole removal experiment found that the immobilized microbial pellets can start up quickly in a short time and efficiently remove sulfamethoxazole. The removal rate of sulfamethoxazole by sodium alginate-biochar-combined microbial pellets was as high as 26.61% in 24 hours, proving that the immobilized bacterial pellets have good tolerance and removal effect on sulfamethoxazole.

[0017] More preferably, the mass concentration of the sodium alginate solution is 3%, the mass concentration of the CaCl2 solution is 5%, and the concentration of the bacterial solution is OD 600 =1.5-2; the dosage ratio of rice straw biochar to bacterial solution is 0.4-0.6 g / 50 mL, the volume ratio of sodium alginate solution to bacterial solution is 1:1, and the volume ratio of sodium alginate solution to CaCl2 solution is 1:10.

[0018] More preferably, the shaking culture time is 1-3 h; and the cross-linking time does not exceed 1 h.

[0019] More preferably, the liquid culture medium used for culturing ParaclostridiumbifermentansJ2 bacterial solution and / or Clostridium sp.J2 bacterial solution includes Na2SO4, NH4Cl, CaCl2·6H2O, MgSO4·7H2O, KH2PO4, yeast extract, CH3COONa, FeSO4·7H2O, C3H8ClNO2S·H2O (L-cysteine ​​hydrochloride hydrate), and C6H7O6Na (ascorbic acid).

[0020] Further, the liquid culture medium used (per 1000 mL): 1000 mg Na2SO4, 1300 mg NH4Cl, 100 mg CaCl2·6H2O, 2000 mg MgSO4·7H2O, 550 mg KH2PO4, 1000 mg yeast extract, 1300 mg CH3COONa, 200 mg FeSO4·7H2O, 500 mg C3H8ClNO2S·H2O (L-cysteine ​​hydrochloride hydrate), 100 mg C6H7O6Na (ascorbic acid), and the rest is ultrapure water.

[0021] More preferably, the culture environment of Paraclostridiumbifermentans J2 and / or Clostridium sp. J2 is: static culture at a constant temperature of 35° C. under anaerobic conditions, and the initial pH value is 7.0-7.5.

[0022] A third aspect of the present invention is a bacterial agent for degrading sulfonamide antibiotics, wherein the bacterial agent contains Paraclostridium bifermentans J2 and / or Clostridium sp. J2 as main active ingredients.

[0023] Preferably, the bacterial agent further comprises excipients acceptable in the field of bacterial agents, such as liquid culture medium and other fillers or carriers.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] The present invention performs long-term domestication on sulfate-reducing sludge collected from a sulfur-mediated bioreactor, and obtains two anaerobic bacteria after multiple screening and separation and purification, one of which is a paraclostridium strain, named ParaclostridiumbifermentansJ2, and the other is a cereus bacillus, named Clostridium sp.J2. These two anaerobic microorganisms can degrade sulfonamide antibiotics, and the removal effect of sulfonamide antibiotics can be enhanced by combining the two, and can be applied to the biological anaerobic method to treat pharmaceutical wastewater of sulfonamide antibiotics. In addition, after ParaclostridiumbifermentansJ2 and / or Clostridium sp.J2 are made into microbial immobilized bacterial balls with sodium alginate and biochar, they can still be quickly started in a short time and efficiently remove sulfonamide antibiotics. The present invention provides a new approach for the degradation of sulfonamide antibiotics and the treatment of sulfonamide antibiotic pharmaceutical wastewater. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 The removal effect of sulfamethoxazole antibiotics by different microbial treatment methods;

[0027] Figure 2 The removal effect of rice straw biochar on sulfamethoxazole antibiotics;

[0028] Figure 3 This is a physical picture of sodium alginate-biochar-microorganism immobilized bacterial balls;

[0029] Figure 4 Shown are the removal effects of different immobilized bacterial pellets on sulfamethoxazole antibiotics (a, b, c, and d are used to distinguish different experimental groups). DETAILED DESCRIPTION

[0030] The specific embodiments of the present invention are further described below. It should be noted that the description of these embodiments is used to help understand the present invention, but does not constitute a limitation of the present invention. In addition, the technical features involved in each embodiment of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0031] The experimental methods in the following examples are conventional methods unless otherwise specified, and the experimental materials used in the following examples are commercially available unless otherwise specified.

[0032] Example 1 Acquisition of anaerobic microorganisms

[0033] The components of the liquid culture medium used in this example are as follows:

[0034] The composition of each 1000 mL of culture medium is: 1000 mg Na2SO4, 1300 mg NH4Cl, 100 mg CaCl2·6H2O, 2000 mg MgSO4·7H2O, 550 mg KH2PO4, 1000 mg yeast extract, 1300 mg CH3COONa, 200 mg FeSO4·7H2O, 500 mg C3H8ClNO2S·H2O (L-cysteine ​​hydrochloride hydrate), 100 mg C6H7O6Na (ascorbic acid), and the rest is ultrapure water.

[0035] The method for screening and obtaining anaerobic microorganisms comprises the following steps:

[0036] (1) Screening and enrichment

[0037] Pour 100mL of sterile liquid culture medium with a sulfamethoxazole concentration of 10mg / L into the serum bottle, and take out 10mL of uniformly mixed sludge (sulfate-reducing sludge) from the sulfur-mediated bioreactor treating sulfamethoxazole, inoculate it into a 100mL serum bottle, and place it in a 35℃ constant temperature biological incubator after sufficient mixing and shaking, and let it stand and culture for 10 days to obtain the bacterial solution after the first screening and enrichment. Then take 10mL of supernatant from the bacterial solution enriched by the first screening and transfer it to a 100mL serum bottle, which is pre-filled with 100mL of sterile liquid culture medium with a sulfamethoxazole concentration of 10mg / L, and place it in a 35℃ constant temperature biological incubator after sufficient mixing and shaking, and let it stand and culture for 10 days to obtain the bacterial solution after the second screening and enrichment. Then, the concentration of sulfamethoxazole in the liquid culture medium was increased in a concentration gradient of 10 mg / L, and the above operation was repeated three times until the concentration of sulfamethoxazole in the culture medium reached 50 mg / L. The culture was statically cultured in a 35° C. incubator for 10 days to obtain the final mixed bacterial solution.

[0038] (2) Gradient dilution

[0039] Take 1 mL of the mixed bacterial solution obtained in step (1) and use 9 anaerobic tubes at 10 1 , 10 2 , 10 3 , 10 4 , 10 5 , 10 6 , 10 7 , 10 8 , 10 9 The gradient was diluted.

[0040] (3) Purification and enrichment

[0041] Three parallel samples of each of the nine gradient diluted mixed bacterial solutions obtained in step (2) were set up for separation and purification: 0.1 mL of the mixed bacterial solution was taken for plate spreading, so that the bacterial solution was evenly spread on a sterilized solid culture medium (1.5% agar was added to the liquid culture medium, the concentration of sulfamethoxazole was 20 mg / L, and the remaining components were the same as the aforementioned liquid culture medium), and inverted culture was carried out in a constant temperature anaerobic incubator at 35°C for 7 days.

[0042] Select the dilution gradient with moderate colony number and clear colonies (10 4 ) was purified and separated by selecting dominant colonies with obvious growth differences on the culture medium and inoculating them into 100 mL of sterile liquid culture medium with a sulfamethoxazole concentration of 20 mg / L. After being fully mixed and shaken evenly, the culture was placed in a 35°C constant temperature incubator for static culture. After 10 days of culture, black flocculent bacteria could be seen in the serum bottle.

[0043] (4) Further purification and enrichment

[0044] The bacterial solution obtained in step (3) was spread on a sterilized solid culture medium (1.8% agar was added, the concentration of sulfamethoxazole was 20 mg / L, and the other components were the same as the aforementioned liquid culture medium), and inverted culture was carried out in a constant temperature anaerobic incubator at 35°C for 7 days.

[0045] Then, the colonies with good growth on the plate culture medium were inoculated into 100 mL serum bottles for enrichment culture. The serum bottles were previously filled with 100 mL sterile culture medium (sulfamethoxazole concentration was 20 mg / L). After being fully mixed and shaken, they were placed in a 35°C constant temperature incubator for 7 days. The above operation was repeated three times until the growth morphology of the colonies on the plate was basically the same, and it was judged to be a pure bacterium that could tolerate sulfamethoxazole.

[0046] The two purified and isolated pure strains of sulfamethoxazole-degrading bacteria were subjected to 16S rRNA sequencing and matched with the NCBI database. The purified strains were highly similar to the strains in the table below.

[0047] Chinese name of fungus The most similar bacterial genus Similarity (%) Clostridium parasitica Paraclostridium bifermentans 99.66 Bacillus cereus Clostridium sp. 100

[0048] Finally, the two strains were named ParaclostridiumbifermentansJ2 and Clostridium J2, and sent to the strain collection center for preservation.

[0049] The deposit information of Clostridium parasitica J2 is as follows:

[0050] Genus name: Paraclostridium bifermentansJ2;

[0051] Deposit number: GDMCC No: 62845;

[0052] Deposit date: September 28, 2022;

[0053] Name of depository: Guangdong Microbiological Culture Collection Center;

[0054] Address of the preservation unit: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou.

[0055] The deposit information of Bacillus cereus J2 is as follows:

[0056] Genus name: Clostridium sp.J2;

[0057] Deposit number: GDMCC No: 62934;

[0058] Deposit date: October 28, 2022;

[0059] Name of depository: Guangdong Microbiological Culture Collection Center;

[0060] Address of the preservation unit: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou.

[0061] The 16S rRNA sequence of Paraclostridiumbifermentans J2 (SEQ ID NO.1) is as follows:

[0062]

[0063] The 16S rRNA sequence of Clostridium sp. J2 (SEQ ID NO. 2) is as follows:

[0064]

[0065] Example 2 Removal of sulfamethoxazole by Paraclostridiumbifermentans J2 and Clostridium sp. J2

[0066] In order to explore the removal effect of ParaclostridiumbifermentansJ2 and Clostridium sp.J2 on sulfamethoxazole (SMX), the following sulfamethoxazole removal experiment was carried out:

[0067] ParaclostridiumbifermentansJ2 and Clostridium sp.J2 were inoculated into 100 mL serum bottles for enrichment culture. The serum bottles were pre-filled with 100 mL of sterilized culture medium liquid (the composition was the same as the culture medium in Example 1, and the initial pH was 7.0-7.5). After being fully shaken and mixed, they were placed in a 35°C incubator for static culture for 1 day. It was observed that the bacterial liquid in the serum bottle was black and flocculent.

[0068] Take 10mL of the above two bacterial solutions (exponential growth phase) into 100mL anaerobic culture bottles (filled with nitrogen to maintain anaerobic state), and then put 100mL of sterilized liquid culture medium (v:v=10:100) into the anaerobic culture bottle, the culture medium contains different concentrations (1mg / L, 2mg / L, 5mg / L) of sulfamethoxazole. After fully shaking and mixing, place it in a 35℃ constant temperature incubator for 3 days, and finally use high performance liquid chromatography to determine the removal of sulfamethoxazole. The results showed that the removal rates of Paraclostridiumbifermentans J2 for 1mg / L, 2mg / L and 5mg / L sulfamethoxazole within 72 hours were 53.0%, 52.6% and 44.25% respectively; the removal rates of Clostridium sp. J2 for 1mg / L, 2mg / L and 5mg / L sulfamethoxazole were 34.7%, 31.2% and 24.92% respectively.

[0069] In addition, 5 mL of the above two bacterial solutions were taken into the same 100 mL anaerobic culture bottle, and then 100 mL of sterilized liquid culture medium (v:v=10:100) was added to the anaerobic culture bottle. The culture medium contained different concentrations (1 mg / L, 2 mg / L, 5 mg / L) of sulfamethoxazole. After thorough shaking and mixing, the mixture was placed in a 35°C constant temperature incubator for 3 days, and finally the removal of sulfamethoxazole was determined using a high performance liquid chromatograph. The results showed that ( Figure 1), the removal rates of the microbial combination of ParaclostridiumbifermentansJ2 and Clostridium sp.J2 for 1mg / L, 2mg / L and 5mg / L sulfamethoxazole were 68.7%, 69.5% and 64.8% within 72h, which proved that the two screened anaerobic microorganisms can degrade sulfamethoxazole, and the removal effect of sulfamethoxazole can be enhanced by the combination of the two.

[0070] Example 3 Immobilization of Paraclostridiumbifermentans J2 and Clostridium sp. J2 and Removal of Sulfamethoxazole by Immobilized Microorganisms

[0071] 1. Removal effect of rice straw biochar on sulfamethoxazole

[0072] (1) Obtaining rice straw biochar

[0073] The agricultural waste rice straw was placed in an oven at 80°C to dry out the moisture, then the material was crushed with a grinder, and then the raw material powder was compacted and loaded into a crucible with a lid, and then the crucible was transferred to a muffle furnace and heated at 700°C for 2 hours under constant temperature and oxygen-deficient conditions, and then cooled to room temperature, taken out, crushed and ground with a grinder, and passed through a 100-mesh sieve, and put into a white transparent wide-mouth bottle for later use.

[0074] (2) Sulfamethoxazole removal experiment

[0075] In order to explore the removal effect of biochar on sulfamethoxazole, the following sulfamethoxazole removal experiment was carried out:

[0076] 5 g of rice straw biochar was placed in a 1000 mL serum bottle, which was previously filled with artificial water from the bioreactor with sulfamethoxazole concentrations of 0.1, 0.5, 1, 3, and 5 mg / L, and fully shaken at 150 rpm and 30 °C for 24 h. The removal of sulfamethoxazole was then determined using a high performance liquid chromatograph. Figure 2 As shown in the figure, the removal rates of rice straw biochar for different concentrations of sulfamethoxazole within 24 hours were 77.94%, 56.33%, 34.75%, 23.47% and 16.63%, respectively. It can be seen that rice straw biochar can be used to treat wastewater containing sulfamethoxazole.

[0077] The components of the artificial water distribution of the bioreactor used in this example are as follows:

[0078] The components per 1000mL are: 1232mg Na2SO4, 640.63mg CH3COONa, 26mg CaCl2, 38.95mg MgCl2·6H2O, 95.5mg NH4Cl, 0.72mg KH2PO4, 2.52mg K2HPO4, 0.2mg KI, 5mg FeCl3·6H2O, 0.5mg H3BO3, 0.5mg CuSO4, 0.63mg MnSO4·H2O, 0.83mg ZnSO4·7H2O, 0.5mg CoCl2·6H2O.

[0079] 2. Obtaining immobilized microbial pellets

[0080] In order to further improve the removal effect of sulfamethoxazole by Clostridium parasitica J2 and Bacillus cereus J2, the two anaerobic microorganisms were subjected to the following embedding and solidification treatment:

[0081] (1) Take 10 mL of bacterial suspension (OD 600 =1.5-2), each inoculated into a 100 mL serum bottle, which was previously filled with 100 mL of sterilized liquid culture medium (the composition is the same as the liquid culture medium in Example 1). After being fully mixed and shaken evenly, it was placed in a 35°C constant temperature incubator for static culture for 1 day, and the black flocculent bacteria in the serum bottle could be clearly observed.

[0082] (2) Preparation of sodium alginate-biochar-microorganism immobilized bacterial pellets: The rice straw biochar was sterilized by wet heat autoclave at 121°C for 20 min. After the biochar was cooled, 50 mL of Paraclostridium bifermentans J2 bacterial solution (OD 600 =1.5~2), Clostridium sp.J2 bacterial liquid (OD 600 =1.5~2), combined culture solution (OD 600 =1.5~2, the mixed volume ratio of the two bacterial solutions is 1:1) transferred to a 50mL serum bottle, and then 0.5g sterilized rice straw biochar was added, and the shaker was shaken at 35℃, 150rpm for 2h. Then the biochar-microorganism solution was mixed with 3% (w / v) sodium alginate solution in equal volumes, and the mixture was dripped into 500mL 5% (w / v) CaCl2 solution that was constantly stirred at a uniform rate, and cross-linked for 1h. After the gel beads were formed, the beads were rinsed with deionized water for at least three times, and placed in 0.75% NaCl solution at 4℃ for future use. It can be clearly observed that the appearance of the sodium alginate-biochar-microorganism ball is black spherical ( Figure 3 ).

[0083] (3) Preparation of sodium alginate-microorganism immobilized bacterial pellets: Take 50 mL of 3% (w / v) sodium alginate solution and 500 mL of 5% (w / v) CaCl2 solution, and take another 50 mL of the co-cultured bacterial solution (OD 600 =1.5~2) was transferred to a 50mL serum bottle and cultured on a shaker at 35°C and 120rpm for 2h. Then, the combined microbial culture and sodium alginate solution were mixed in equal volumes, and the mixture was uniformly dripped into the continuously stirred CaCl2 solution for cross-linking for 1h. After the gel beads were formed, the beads were rinsed with deionized water for at least three times and placed in a 0.75% NaCl solution at 4°C for future use.

[0084] 3. Removal effect of immobilized microbial pellets on sulfamethoxazole

[0085] The study on the removal effect of immobilized microbial pellets on sulfamethoxazole mainly includes the following steps:

[0086] (1) The microbial pellets were placed in 100 mL of sterilized liquid culture medium (the composition was the same as that of the liquid culture medium in Example 1) and cultured again at 35° C. for 24 h. Black flocculent cells were observed in the culture medium.

[0087] (2) Add the bacterial pellets to a 100 mL serum bottle (which has been filled with nitrogen in advance to expel air) at a volume ratio of 10% (v / v). The serum bottle contains 100 mL of a sterilized liquid culture medium containing 5 mg / L sulfamethoxazole (the composition of the culture medium is the same as that of the liquid culture medium described in Example 1). After sufficient shaking to mix the liquid evenly, place it in a 35°C constant temperature incubator for 1 day, and finally use a high performance liquid chromatograph to determine the removal of sulfamethoxazole by the immobilized bacterial pellets. Figure 4 As shown in the figure, within 24 hours, the immobilized microbial pellets can quickly remove sulfamethoxazole in the water, among which the sodium alginate-biochar-co-cultured pellets are the best, with a removal rate of up to 26.61%.

[0088] According to the results of Examples 2, 4 and 5, it can be concluded that the combined microorganisms and rice straw biochar have a high removal efficiency for sulfamethoxazole, and can be applied to the biological anaerobic method to treat pharmaceutical wastewater containing sulfonamide antibiotics, providing a practical method for the treatment of wastewater containing sulfonamide antibiotics.

[0089] The embodiments of the present invention are described in detail above, but the present invention is not limited to the described embodiments. For those skilled in the art, various changes, modifications, substitutions and variations of these embodiments are made without departing from the principles and spirit of the present invention, and still fall within the protection scope of the present invention.

Claims

1. Paraclostridium bifermentans J2 and / or Clostridium sp. J2 The application in the degradation of sulfonamide antibiotics is characterized in that: Paraclostridium bifermentans The deposit number of J2 is GDMCC No: 62845. Clostridium sp. J2 The deposit number is GDMCC No: 62934, and the application is for non-disease treatment purposes.

2. Paraclostridium bifermentans J2 and / or Clostridium sp. J2 The application in treating pharmaceutical wastewater containing sulfonamide antibiotics is characterized by: Paraclostridium bifermentans The deposit number of J2 is GDMCC No: 62845. Clostridium sp. J2 The deposit number is GDMCC No: 62934.

3. The use according to claim 1 or 2, characterized in that: The sulfonamide antibiotics include sulfamethoxazole.

4. The use according to claim 3, characterized in that: The concentration of sulfamethoxazole is no more than 10 mg / L.

5. The use according to claim 1 or 2, characterized in that: Said Paraclostridium bifermentans J2 and / or Clostridium sp. J2 The immobilized bacterial pellets were prepared before use.

6. The use according to claim 5, characterized in that: Past Paraclostridium bifermentans J2 bacterial solution and / or Clostridium sp. J2 Rice straw biochar is added to the bacterial solution, and after oscillation culture, the biochar-microorganism solution is mixed with the sodium alginate solution, and the resulting mixture is dropped into the continuously stirred CaCl2 solution to form gel beads after cross-linking.

7. The use according to claim 6, characterized in that: The mass concentration of the sodium alginate solution is 3%, the mass concentration of the CaCl2 solution is 5%, and the concentration of the bacterial solution is OD 600 =1.5-2; the dosage ratio of rice straw biochar to bacterial solution was 0.4-0.6 g / 50 mL, the volume ratio of sodium alginate solution to bacterial solution was 1:1, and the volume ratio of sodium alginate solution to CaCl2 solution was 1:

10.

8. The use according to claim 6, characterized in that: The shaking culture time is 1-3 h; the cross-linking time does not exceed 1 h.

9. The use according to claim 6, characterized in that: nourish Paraclostridium bifermentans J2 bacterial solution and / or Clostridium sp. J2 The liquid culture medium used for the bacterial liquid includes Na2SO4, NH4Cl, CaCl2·6H2O, MgSO4·7H2O, KH2PO4, yeast extract, CH3COONa, FeSO4·7H2O, L-cysteine ​​hydrochloride hydrate, and ascorbic acid.

10. A bacterial agent for degrading sulfonamide antibiotics, characterized in that: The bacterial agent Paraclostridium bifermentans J2 and / or Clostridium sp. J2 As the main active ingredient, Paraclostridium bifermentans The deposit number of J2 is GDMCC No: 62845. Clostridium sp. J2 The deposit number is GDMCC No: 62934.

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