Screening and application of efficient desulfurizing bacterium

Abstract

The invention belongs to the technical field of microorganisms, discloses screening and application of efficient desulfurizing bacteria, and particularly discloses acinetobacter which is characterized in that the preservation name of the acinetobacter is Acinetobacter sp.HSC-01, the acinetobacter is preserved in Guangdong Microbial Culture Collection Center, the preservation number is GDMCC No: 64605, and the preservation date is May 11, 2024. The invention provides Acinetobacter sp.HSC-01 with an efficient sulfur oxidation function, the strain can effectively remove sulfides in aquatic water, the action time of the strain is 8 h, and the removal rate of the sulfides can reach 96% or above. Meanwhile, the strain can also be used for oxidation of black mud in an aquaculture pond. The efficient desulfurization capacity has potential application value in aquaculture sewage and sludge treatment.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms, and in particular relates to the screening of a strain of efficient desulfurization bacteria and its application. Background Art

[0002] Sulfide in aquaculture water is seriously toxic to aquatic animals. After penetrating into the fish body, it can convert low-valent iron ions in hemoglobin into high-valent iron, causing hemoglobin to lose its oxygen-carrying capacity, resulting in tissue hypoxia and, in severe cases, death. Sulfide in aquaculture water includes soluble H2S, HS - , S 2- , as well as soluble sulfides and acid-soluble metal sulfides present in suspended matter. The sulfides in the water are easily converted into hydrogen sulfide (H2S), which produces a bad smell and is very toxic. Hydrogen sulfide can react with cytochromes, oxidases and disulfide bonds (-SS) in animal bodies, affecting the cell oxidation process, causing cell tissue hypoxia, thereby leading to tissue necrosis and even endangering life. The production of hydrogen sulfide in ponds is an intermediate product formed by incomplete decomposition of sulfur-containing organic matter under anaerobic conditions, or sulfate-rich water bodies are converted from sulfate to sulfide under the action of sulfate-reducing bacteria, and then regenerated into hydrogen sulfide. Under acidic conditions, most of the sulfides exist in the form of hydrogen sulfide. When the hydrogen ion concentration is high, the toxicity is enhanced. When the dissolved oxygen in the water increases, the hydrogen sulfide is immediately oxidized and disappears. The production of hydrogen sulfide is a very important and thorny problem that restricts the high yield of aquaculture. At present, in actual aquaculture production, the way to solve this problem mainly relies on the use of a large number of highly oxidizing chemicals, and the effects of these chemicals are very obvious. Although the large-scale use of chemical drugs may not have too much negative impact on aquaculture in the short term, in the long run there will inevitably be a large amount of drug residues in the aquaculture system, the pond bottom mud will become hardened, and the entire aquaculture will be affected to a relatively large extent.

[0003] In recent years, biological desulfurization technology has gradually attracted everyone's attention. Biological desulfurization technology mainly relies on a special type of microorganisms - sulfur-oxidizing bacteria. Sulfur-oxidizing bacteria are a type of bacteria that can completely oxidize low-valent reduced sulfides into sulfates (SO4 2– ) or partially oxidizes it to a higher valent sulfur state. In the process of oxidizing H2S, sulfur-oxidizing bacteria fix CO2 and synthesize organic matter. This pathway is a chemosynthetic process, and H2S provides the primary production energy. This process usually involves two enzymes, Sulfide:quinone reductase (Sqr) and Flavocytochrome csulfidedehydrogenase (Fcc). In the process of sulfide oxidation, Sqr converts the produced e –Through its cofactor FDA, it is transferred to the quinone pool; Fcc transfers e – In sulfur-oxidizing bacteria, Sqr and Fcc have high affinity for high and low concentrations of sulfide, respectively, so they play a role in these two situations. Summary of the invention

[0004] The purpose of the first aspect of the present invention is to provide a bacterium.

[0005] The second aspect of the present invention aims to provide a microbial agent.

[0006] The purpose of the third aspect of the present invention is to provide the use of the Acinetobacter of the first aspect of the present invention and / or the microbial agent of the second aspect of the present invention.

[0007] The fourth aspect of the present invention aims to provide a product.

[0008] The fifth aspect of the present invention aims to provide a sewage treatment system.

[0009] The sixth aspect of the present invention aims to provide a method for treating sewage.

[0010] The seventh aspect of the present invention aims to provide a method for oxidizing black mud in aquaculture ponds.

[0011] In order to achieve the above object, the technical solution adopted by the present invention is:

[0012] The first aspect of the present invention provides a kind of Acinetobacter, characterized in that the preservation name of the Acinetobacter is Acinetobacter sp.HCS-01, which is preserved in Guangdong Provincial Microbiological Culture Collection Center, with a preservation number of GDMCCNo:64605 and a preservation date of May 11, 2024.

[0013] The second aspect of the present invention provides a microbial agent, comprising at least one of the Acinetobacter of the first aspect of the present invention, a culture ... supernatant of the Acinetobacter of the first aspect of the present invention, or an extract of the Acinetobacter of the first aspect of the present invention.

[0014] The active ingredients of the above-mentioned bacterial agent may be the above-mentioned Acinetobacter sp. HCS-01, the culture of the above-mentioned Acinetobacter sp. HCS-01 (such as a fermentation product) and / or the fermentation supernatant of the above-mentioned Acinetobacter sp. HCS-01. The active ingredients of the above-mentioned bacterial agent may also contain other biological components or non-biological components. Those skilled in the art may determine the other active ingredients of the above-mentioned bacterial agent according to the effect of the bacterial agent.

[0015] The term "culture" refers to a liquid or solid product (all substances in the culture container, fermentation products) that has a microbial community after artificial inoculation and culture. That is, the product obtained by growing and / or amplifying microorganisms, which can be a biologically pure culture of microorganisms, or it can contain a certain amount of culture medium, metabolites or other components produced during the culture process.

[0016] In the above-mentioned microbial agent, in addition to the active ingredient, the microbial agent also contains a carrier. The carrier can be a commonly used and biologically inert carrier in the field (such as microorganisms, agriculture, and aquaculture). The carrier can be a solid carrier or a liquid carrier; the solid carrier can be a mineral material, a plant material, or a polymer compound; the mineral material can be at least one of clay, talc, kaolin, montmorillonite, white carbon, zeolite, silica, and diatomaceous earth; the plant material can be at least one of corn flour, soybean flour, and starch; the polymer compound can be polyvinyl alcohol and / or polyglycol; the liquid carrier can be an organic solvent or water; the organic solvent can be decane and / or dodecane.

[0017] The above-mentioned bacterial agents may be in various dosage forms, such as liquid, emulsion, suspension, powder, granule, wettable powder or water dispersible granule.

[0018] According to needs, a binder, a stabilizer (such as an antioxidant), a pH adjuster, etc. may be added to the bacterial agent.

[0019] In some embodiments of the present invention, the bacterial agent further comprises: nutrients. The nutrients include: at least one of proteins, carbohydrates, lipids, minerals, vitamins, plant extracts, amino acids, immunomodulators and milk substitutes.

[0020] The third aspect of the present invention provides the use of the Acinetobacter of the first aspect of the present invention and / or the microbial agent of the second aspect of the present invention in at least one of (1) to (6):

[0021] (1) Desulfurization;

[0022] (2) preparing desulfurized products;

[0023] (3) Wastewater treatment;

[0024] (4) Preparation of products for treating sewage;

[0025] (5) Oxidation of black mud in aquaculture ponds;

[0026] (6) Prepare products for oxidizing black mud in aquaculture ponds.

[0027] In some embodiments of the present invention, the sewage in (3) and (4) contains sulfide; preferably, it is aquaculture wastewater containing sulfide.

[0028] In some embodiments of the present invention, the black mud in the aquaculture ponds described in (5) and (6) contains sulfide.

[0029] The fourth aspect of the present invention provides a product comprising the Acinetobacter of the first aspect of the present invention and / or the microbial agent of the second aspect of the present invention.

[0030] In some embodiments of the present invention, the product further comprises a carrier, which may be a commonly used and biologically inert carrier in a field (such as microorganisms, agriculture, aquaculture). The carrier may be a solid carrier or a liquid carrier; the solid carrier may be a mineral material, a plant material or a polymer compound; the mineral material may be at least one of clay, talc, kaolin, montmorillonite, white carbon, zeolite, silica and diatomaceous earth; the plant material may be at least one of corn flour, soybean flour and starch; the polymer compound may be polyvinyl alcohol and / or polyglycol; the liquid carrier may be an organic solvent or water; the organic solvent may be decane and / or dodecane.

[0031] In some embodiments of the present invention, the product includes a reagent and a bacterial agent.

[0032] In some embodiments of the present invention, the product has the following functions: removal of sulfide; and / or, sewage treatment; and / or, oxidation of black mud in aquaculture ponds.

[0033] A fifth aspect of the present invention provides a sewage treatment system, comprising a regulating tank for stabilizing water quality and quantity, and a biochemical treatment device connected to the regulating tank for treating sewage stabilized by the regulating tank, wherein the biochemical treatment device uses the Acinetobacter of the first aspect of the present invention, the microbial agent of the second aspect of the present invention, or the product of the fourth aspect of the present invention to treat sewage.

[0034] The sixth aspect of the present invention provides a method for treating sewage, comprising the steps of treating sewage using the Acinetobacter of the first aspect of the present invention, the microbial agent of the second aspect of the present invention, or the product of the fourth aspect of the present invention; or the steps of treating sewage using the sewage treatment system of the fifth aspect of the present invention.

[0035] In some embodiments of the present invention, the sewage includes aquaculture wastewater containing sulfide.

[0036] In some embodiments of the present invention, the treatment time is 2 to 12 hours.

[0037] In some preferred embodiments of the present invention, the treatment time is 4 to 10 hours.

[0038] In some more preferred embodiments of the present invention, the treatment time is 6 to 9 hours.

[0039] The method provided by the present invention is suitable for desulfurization wastewater treatment, can effectively remove sulfur-containing compounds, reduce operating costs, and promote standard discharge; the microbial agent of the present invention is added to the wastewater treatment system in a certain amount to accelerate the degradation of pollutants by microorganisms to improve the biological treatment efficiency of the system and ensure the stable operation of the system; the strain of the present invention has high activity, large biomass, fast reproduction, good wastewater treatment effect, and the sulfide removal rate can reach more than 96%.

[0040] The seventh aspect of the present invention provides a method for oxidizing black mud in aquaculture ponds, comprising the step of treating the black mud in aquaculture ponds using the Acinetobacter of the first aspect of the present invention, the microbial agent of the second aspect of the present invention, or the product of the fourth aspect of the present invention.

[0041] In some embodiments of the present invention, the treatment method includes pouring, wetting, drenching, atomizing, drizzling, spraying, and atomizing the Acinetobacter, the microbial agent, or the product into the black mud of the culture pond.

[0042] In some embodiments of the present invention, the black mud in the aquaculture pond contains sulfide.

[0043] The beneficial effects of the present invention are:

[0044] The present invention provides an Acinetobacter sp. HCS-01 with a highly efficient sulfur oxidation function. The strain can effectively remove sulfide in aquaculture water. After 8 hours of action, the removal rate of sulfide can reach more than 96%. At the same time, the strain can also act on the oxidation of black mud in aquaculture ponds. The highly efficient desulfurization ability has potential application value in the treatment of aquaculture sewage and sludge. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] Figure 1 This is the morphological observation of strain Acinetobacter sp.HCS-01. The left picture shows the colony of strain Acinetobacter sp.HCS-01, and the right picture shows the Gram staining result.

[0046] Figure 2 This is the phylogenetic tree of strain Acinetobacter sp. HCS-01.

[0047] Figure 3 The left picture shows the results of the blood plate hemolysis test for Staphylococcus aureus, and the right picture shows the results of the blood plate hemolysis test for strain Acinetobacter sp. HCS-01.

[0048] Figure 4 This is the growth curve of strain Acinetobacter sp. HCS-01.

[0049] Figure 5 Agarose gel electrophoresis.

[0050] Figure 6 This is the sulfide removal effect of strain Acinetobacter sp. HCS-01.

[0051] Figure 7 This is the removal effect of sulfide in simulated aquaculture wastewater by strain Acinetobacter sp. HCS-01.

[0052] Figure 8 This is the oxidation effect of strain Acinetobacter sp. HCS-01 on the black mud in actual aquaculture ponds. DETAILED DESCRIPTION

[0053] The present invention is further described in detail below through specific examples.

[0054] It should be understood that these embodiments are only used to illustrate the present invention and are not used to limit the scope of the present invention.

[0055] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0056] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0057] Example 1 Isolation and identification of strain HCS-01

[0058] 1. Enrichment and Isolation of Strain HCS-01

[0059] 1) Enrichment culture: From the black carp breeding pond of Haida Group breeding base in Haiou Island, Panyu District, Guangzhou, 500g of black mud samples were collected from the bottom of the pond at 5-10cm with a mud sampler, and immediately stored in a 4℃ foam box and transported back to the laboratory for enrichment and separation of sulfur-oxidizing microorganisms. Weigh 5g of sample from the black mud, add it to 95mL of physiological saline containing glass beads, and shake it at 200rpm and 30℃ for 30min to mix it thoroughly. Take another 5mL of the mixed liquid and inoculate it into 95mL of cooled liquid enrichment culture medium (120℃, 20min), enrich it at 30℃, 200rpm for 3d, and then use the previous enrichment culture liquid as the inoculation liquid according to the 5% inoculation ratio, and enrich it 3 times in this way. Enrichment medium: Na2S2O3·5H2O, 8g / L; K2HPO4, 2g / L; NH4Cl, 0.4g / L; MgCl2·6H2O, 0.2g / L; NaHCO3, 3g / L; FeS04·7H20, 0.01g / L;

[0060] 2) Plate separation and purification: Add 1.5% agar to the liquid enrichment medium, and then sterilize at 120℃ for 20 minutes to make a solid enrichment medium plate. Then take 1mL of the above enrichment culture solution and dilute it into 10 -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 -7 , 10 -8 Different dilution gradients of bacterial suspension were prepared. 10 -6 , 10 -7 , 10 -8 100 μL of the bacterial suspension of three gradients was evenly spread on the enrichment medium plate, with 3 parallels for each gradient, and the plate was placed in a 30°C incubator for 48 hours. Finally, single colonies with obviously different characteristics such as colony morphology and size were selected from the three dilution gradient plates, and streaked on a new enrichment medium plate. Repeat streaking 4 to 5 times until a purified strain developed from a single microbial cell was obtained. Note: Do not scratch the surface of the culture medium when streaking.

[0061] 3) Screening and preservation: The 96-well plate rapid screening method and the shake flask culture rescreening method were combined to screen sulfur-oxidizing microorganisms. The isolated and purified strains were added to LB sterile liquid culture medium for activation, cultured at 180 rpm and 30°C for 14 to 18 hours, then centrifuged, the supernatant was discarded, and the culture was washed twice with sterile saline. The bacterial solution concentration was adjusted to a uniform OD 600=2; Take 0.2mL of bacterial liquid and 0.8mL of sodium sulfide solution (100mg / L) in a 2mL Ep tube, place it in a 30℃ incubator and incubate for 48h, then take 50μL of the incubated liquid in 950μL of ultrapure water (in a 96-well plate), add 80μL of color developing solution (weigh 1.6g N,N-dimethyl-p-phenylenediamine hydrochloride and 2.4g ferric chloride hexahydrate, dilute to 50mL with 18.5% hydrochloric acid, and mix 1:1 before use), shake well, let stand at room temperature for 20min, and measure the absorbance at 660nm. Select the corresponding strains with lighter color (significantly lower absorbance) than the blank control for shake flask culture rescreening to determine the strain with the best desulfurization effect. The results of the initial screening of strains are as follows, and the results are shown in Table 1.

[0062] Table 1 96-well plate initial screening results

[0063]

[0064]

[0065] Ten strains (with significantly lower absorbance than that of the CK group) were selected from the initial screening strains: HCS-01, HCS-02, HCS-07, HCS-10, HCS-12, HCS-15, HCS-18, HCS-20, HCS-22, and HCS-25 for shake flask rescreening. The 10 strains screened initially were activated with liquid LB medium, and then inoculated with 2% inoculation amount into GY degradation solution containing 100 mg / L sulfide substrate (shake flask culture), and cultured at 30°C, 120 rpm for 8 hours. After the end, 1 mL of culture solution was centrifuged at 5000 rpm to obtain the supernatant, and the sulfide concentration in the culture solution was determined by the national standard methylene blue spectrophotometry. The lower the sulfide concentration, the better the desulfurization effect of the strain. That is, according to the desulfurization rate, the above 10 strains were screened by shake flask rescreening, and finally the target strain HCS-01 with efficient sulfur oxidation function was screened by the desulfurization rate. The results are shown in Table 2. The composition of the LB medium is: peptone 10 g / L, yeast extract 5 g / L, NaCl 10 g / L. The formula of GY degradation solution is C4H4Na2O4 10g / L, KH2PO4 0.5g / L, MgCl2 0.2g / L, NH4Cl, 0.6g / L, NaHCO3 3g / L, NaCl 5g / L, Na2S·9H2O 0.8g / L, pH 9.5.

[0066] Table 2 Rescreening results

[0067]

[0068] 2. Morphological characteristics of strain HCS-01

[0069] 1) Morphological characteristics of strains

[0070] The morphological characteristics of strain HCS-01 were analyzed, and the results were as follows Figure 1 As shown, the bacteria are Gram-negative, aerobic, short rod-shaped, and spore-free. The colonies on the LB medium plate are milky white, round, small, moist, with neat edges and slight protrusions, smooth and opaque surface, and 2 to 3 mm in diameter. The strain was preliminarily determined to be Bacillus.

[0071] 2) Taxonomic status of strain HCS-01

[0072] The purified strain HCS-01 was extracted with the TaKaRa MiniBEST Bacteria Genomic DNA Extraction Kit Ver.3.0 genome extraction kit. The universal primers 27F (5'-AGAGTTTGATCCTGGCTCAG-3', SEQ ID NO: 1) and 1492R (5'-GGTTACCTTGTTACGACTT-3, SEQ ID NO: 2) of the 16S rRNA sequence were used as PCR reaction primers, and PCR amplification was performed using the 2×TsingKE Master Mix system; the PCR reaction system was: genomic DNA 1μL, 2×Tsing KE Master Mix 25μL, 27F Primer (10μM) 1μL, 1492RPrimer (10μM) 1μL, dH2O 22μL. The PCR reaction conditions were: pre-denaturation at 94°C for 10 min, 30 cycles (94°C for 30 s, 55°C for 30 s, 72°C for 1.5 min), and extension at 72°C for 10 min. The PCR product was sent to Novogene Biotech Co., Ltd. for 16S rRNA sequencing, and the sequence is shown in SEQ ID NO:3.

[0073]

[0074] The above sequences were submitted to the NCBI website (https: / / www.ncbi.nlm.nih.gov / ) for BLAST comparison and phylogenetic tree construction. The results showed that the sequence similarity between strain HCS-01 and Acinetobacter sp.407 (GenBank: EU841481) was more than 99%. The phylogenetic tree is shown in Figure 2 shown.

[0075] Based on the morphological and molecular biological analysis of HCS-01, strain HCS-01 was identified as Acinetobacter sp. and named Acinetobacter sp. HCS-01. It was deposited in Guangdong Provincial Microbiological Culture Collection Center on May 11, 2024, with the deposit number GDMCC NO: 64605.

[0076] Example 2 Biosafety Assessment of Acinetobacter sp. HCS-01

[0077] The hemolytic activity of the strain Acinetobacter sp. HCS-01 was tested on the hemolytic plate, and the Staphylococcus aureus (positive control) and the strain Acinetobacter sp. HCS-01 were cultured for 48 hours.

[0078] The results are as follows Figure 3 As shown, it can be clearly seen that there is an obvious luminous hemolytic ring around the Staphylococcus aureus (left) colony, while there is no obvious luminous hemolytic ring around the strain Acinetobacter sp. HCS-01 (right). This result shows that the strain Acinetobacter sp. HCS-01 is non-pathogenic and has a higher safety.

[0079] Example 3 Growth curve drawing of strain Acinetobacter sp. HCS-01

[0080] The seed culture of strain Acinetobacter sp. HCS-01 was inoculated into the fermentation culture at a rate of 5%, the shaking speed was 200 rpm, the culture temperature was 30°C, the culture was carried out for 48 h, and the OD was measured every 2 hours. 600 , with bacterial concentration OD 600 The growth curve was drawn with the y-axis and the culture time as the x-axis. Figure 4 shown.

[0081] Example 4 Detection of sulfur oxidation functional gene SoxB of strain Acinetobacter sp. HCS-01

[0082] By analyzing the genome information of Acinetobacter sp. HCS-01, we found that the strain contained the SOX pathway in the sulfur oxidation pathway, including the key sulfur oxidation functional gene SoxB. Using the genomic DNA of the strain as a template, PCR amplification was performed using conventional primers SoxB-710F / SoxB-1184R of the SoxB gene, and the obtained products were analyzed by agarose gel electrophoresis, among which SoxB-710F: 5'-ATCGGYCAGGCYTTYCCSTA-3' (SEQ ID NO: 4), 5'-MAVGTGCCGTTGAARTTGC-3' (SEQ ID NO: 5).

[0083] The results are as follows Figure 5 As shown, a target band of 511 bp can be seen; in addition, the PCR amplification product was sent to Novogene Biotechnology Co., Ltd. for sequence sequencing, and the sequencing results were submitted to Genbank for homology comparison analysis. The analysis results showed that the amplified sequence was the sequence of the sulfur oxidation functional gene SoxB.

[0084] Example 5 Comparison of sulfur oxidation function between strain Acinetobacter sp. HCS-01 and conventional bacteria

[0085] Prepare sodium succinate culture medium with the formula of C4H4Na2O4 10g·L -1 、KH2PO4 0.5g·L -1 、MgCl2 0.2g·L -1 、NH4Cl 0.6g·L -1 , NaHCO3 3g·L -1 、NaCl 5g·L -1 、Na2S·9H2O 0.8g·L -1 , pH 9. Among them, S 2-The concentration was designed to be 100 mg / L. Acinetobacter sp. HCS-01, Acinetobacter pittii, Acinetobacter baumanii, Bacillus tropicalus, Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus licheniformis, and Bacillus pumilus were activated and cultured with LB liquid medium for 18 hours, respectively, until the concentration of the bacteria reached 1.0×10^10 cfu / mL, and then 5 mL of the bacterial solution was centrifuged, and the bacteria were rinsed with physiological saline, and then added to a 250 mL conical flask containing 100 mL of sodium sulfide with a concentration of 100 mg / L (i.e., sodium succinate medium), and then placed in a shaker at 30°C and 120 rpm for culture. After 8 hours, take a sample to determine the concentration of sulfide ions (see: "Determination of sulfide in water quality - Methylene blue spectrophotometry" (HJ 1226-2021)).

[0086] The results are as follows Figure 6 As shown, it can be seen that the sulfide removal effect of strain Acinetobacter sp. HCS-01 is significantly better than that of other conventional strains.

[0087] Example 6 Application of strain Acinetobacter sp. HCS-01 in removing sulfide from simulated aquaculture wastewater

[0088] Prepare sodium succinate culture medium with the formula of C4H4Na2O4 10g·L -1 、KH2PO4 0.5g·L -1 、MgCl2 0.2g·L -1 、NH4Cl 0.6g·L -1 , NaHCO3 3g·L -1 、NaCl 5g·L -1 、Na2S·9H2O 0.8g·L -1 , pH 9. Among them, S 2- The concentration was designed to be 100 mg / L. The strain Acinetobacter sp. HCS-01 was activated and cultured with LB liquid medium for 18 hours, then 5 mL of the bacterial solution was centrifuged, the bacteria were rinsed with physiological saline, and the bacteria were added to a 250 mL conical flask containing 100 mL of sodium sulfide with a concentration of 100 mg / L, and then placed in a shaker, cultured at 30°C and 120 rpm, and samples were taken at 0h, 2h, 4h, 6h, and 8h to determine the concentration of sulfide ions.

[0089] The results are as follows Figure 7 As shown, it can be seen that the concentration of sulfur ions is 3.80 mg / L after 8 hours, and the sulfide removal rate reaches 96.06%. The sulfide removal effect is very obvious.

[0090] Example 7 Application of strain Acinetobacter sp. HCS-01 in removing sulfide from actual aquaculture wastewater

[0091] 10L of aquaculture sewage was collected from the sewage outlet of the raw fish aquaculture pond, and the silt and other impurities visible to the naked eye were filtered out with gauze, and immediately brought back to the laboratory for sulfide determination. The aquaculture sewage was then divided into 250mL conical flasks, with 100mL in each flask. The strain Acinetobacter sp.HCS-01 was activated and cultured with LB liquid culture medium for 18 hours, and the concentration of the bacteria reached 1.0×10^10cfu / mL. Then 5mL of the bacterial solution was centrifuged, and the bacteria were rinsed with physiological saline, and then added to a 250mL conical flask containing 100mL of aquaculture sewage, and then placed in a shaker, 30℃, 120rpm for culture. After 8h, samples were taken to determine the concentration of sulfide. The treatment method of the blank control: 100mL of aquaculture sewage was placed in a shaker without any treatment, and then placed in a shaker for culture with the experimental group.

[0092] The results are shown in Table 3. It can be seen that the strain Acinetobacter sp. HCS-01 has an obvious removal effect on sulfide in aquaculture wastewater.

[0093] Table 3 Sulfide removal effect in aquaculture wastewater

[0094]

[0095] Example 8 Oxidation effect of strain Acinetobacter sp. HCS-01 in black mud of actual aquaculture ponds

[0096] Collect black and smelly sludge from the bottom of the fish pond and bring it back to the laboratory. Mix the sludge and water in a ratio of 1:4 to make muddy water. Then take 200mL of muddy water and put it into a 300mL transparent plastic bottle and let it stand. The strain Acinetobacter sp.HCS-01 was activated and cultured with LB liquid for 18 hours. The concentration of the bacteria reached 1.0×10^10 cfu / mL. Then take 50mL of bacterial solution and add it directly into a 300mL transparent plastic bottle. The system in the bottle was 250mL in total. Then it was placed in a 30℃ incubator for 96 hours. Then the height of the oxidized sludge was measured with a vernier caliper, and photos were taken. Blank control: Take 200mL of muddy water and put it into a 300mL transparent plastic bottle without any treatment. Then, after 96 hours, the oxidation height of the sludge was measured with a vernier caliper.

[0097] from Figure 8 From the data in Table 4, it can be seen that the strain Acinetobacter sp. HCS-01 has a significant oxidation effect on pond black mud.

[0098] Table 4 The height of sediment oxidation

[0099] Treatment Group 0h(mm) 96h(mm) Blank control 0 10.01 Experimental Group 0 25.09

[0100] The embodiments of the present invention are described in detail above in conjunction with the accompanying drawings, but the present invention is not limited to the above embodiments. Various changes can be made within the knowledge of ordinary technicians in the relevant technical field without departing from the purpose of the present invention. In addition, the embodiments of the present invention and the features in the embodiments can be combined with each other without conflict.

Claims

1. An Acinetobacter, characterized in that The deposited name of the Acinetobacter is Acinetobacter sp. HCS-01, which is deposited in Guangdong Provincial Microbiological Culture Collection Center. The deposit number is GDMCC No: 64605, and the deposit date is May 11, 2024.

2. A microbial agent, comprising at least one of the Acinetobacter according to claim 1, the culture of Acinetobacter according to claim 1, the culture supernatant of Acinetobacter according to claim 1, or the extract of Acinetobacter according to claim 1.

3. Use of the Acinetobacter according to claim 1 and / or the microbial agent according to claim 2 in at least one of (1) to (6): (1) Desulfurization; (2) preparing desulfurized products; (3) Wastewater treatment; (4) Preparation of products for treating sewage; (5) Oxidation of black mud in aquaculture ponds; (6) Prepare products for oxidizing black mud in aquaculture ponds.

4. The use according to claim 3, characterized in that: The wastewater described in (3) and (4) contains sulfide.

5. The use according to claim 3, characterized in that: The black mud in the aquaculture ponds described in (5) and (6) contains sulfide.

6. A product comprising the Acinetobacter according to claim 1 and / or the microbial agent according to claim 2.

7. A sewage treatment system, comprising a regulating tank for stabilizing water quality and quantity, and a biochemical treatment device connected to the regulating tank for treating sewage stabilized by the regulating tank, wherein the biochemical treatment device uses the Acinetobacter described in claim 1, the microbial agent described in claim 2, or the product described in claim 6 to treat sewage.

8. A method for treating sewage, comprising the step of treating sewage using the Acinetobacter described in claim 1, the microbial agent described in claim 2, or the product described in claim 6; or the step of treating sewage using the sewage treatment system described in claim 7.

9. The method according to claim 8, characterized in that The sewage includes aquaculture water containing sulfide; preferably, the treatment time is 2 to 12 hours.

10. A method for oxidizing black mud in aquaculture ponds, comprising the step of treating the black mud in aquaculture ponds with the Acinetobacter described in claim 1, the microbial agent described in claim 2, or the product described in claim 6.

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