Acinetobacter strain as well as culture method and application thereof

Abstract

The invention discloses an acinetobacter strain as well as a culture method and application thereof, the acinetobacter strain is named as 450-8 and preserved in the China General Microbiological Culture Collection Center (CGMCC), the preservation number of the acinetobacter strain is CGMCC No.33205, the preservation date is December 25, 2024, and the 16S rDNA sequence of the acinetobacter strain 450-8 is shown as SEQ ID NO.1. The acinetobacter strain has the advantages that the acinetobacter strain can be used for culturing the acinetobacter strain; the acinetobacter strain (Acinetobacter sp.) 450-8 provided by the invention is an acinetobacter strain which is obtained from an activated sludge sample through directional screening, separation and purification screening and can improve the treatment efficiency of the salt-containing refined sewage, has a remarkable removal effect on COD (Chemical Oxygen Demand) in the salt-containing refined sewage, also has a relatively good removal effect on ammonia nitrogen in a water body, and can be used for treating the salt-containing refined sewage. Meanwhile, good salt tolerance, substrate broad-spectrum property and heavy metal resistance are achieved. The acinetobacter can remarkably reduce main pollution indexes of water and improve the treatment efficiency, shows huge application potential in the aspect of refining and chemical sewage treatment, and has a relatively good industrial application prospect.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, and in particular to an Acinetobacter strain and a culture method and application thereof. Background Art

[0002] A large amount of wastewater is generated during the oil refining process, which is divided into oily wastewater and saline wastewater. The saline wastewater has a complex composition and high salt characteristics. It usually contains various types of petroleum hydrocarbons, organic solvents, heavy metal ions and high concentrations of salts (such as sodium chloride, sulfate, nitrate, etc.). The high-salinity environment has a significant inhibitory effect on microorganisms in traditional sewage treatment. It is difficult for ordinary microorganisms to survive and perform the function of degrading pollutants under such harsh conditions. This greatly increases the difficulty of treating saline wastewater from refineries. Conventional biological treatment processes are often inefficient and it is difficult to achieve ideal pollutant removal effects and emission standards. Therefore, it is urgent to seek more effective treatment technologies.

[0003] Acinetobacter has shown great potential in the field of bioremediation due to its strong environmental adaptability and metabolic diversity. Acinetobacter can decompose a variety of organic pollutants, including petroleum hydrocarbons and polycyclic aromatic hydrocarbons, and plays an important role in improving the treatment of refinery wastewater. The decomposition ability of Acinetobacter makes it the key to solving the problem of wastewater treatment, and the salt tolerance of Acinetobacter makes it have potential application value in treating refinery saline wastewater.

[0004] Therefore, providing a strain of Acinetobacter that is highly salt-tolerant and efficient in treating refinery wastewater is crucial to promoting the development of the field of petroleum refinery wastewater treatment. Summary of the invention

[0005] The invention provides an Acinetobacter strain and a culture method and application thereof.

[0006] In order to achieve the above object, the technical solution of the present invention is:

[0007] In one aspect, the present invention provides an Acinetobacter sp. 450-8, which has been deposited in the General Microbiological Center of the National Microbiological Culture Collection Administration Committee, and its deposit number is CGMCC No. 33205. The deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing. The deposit date is December 25, 2024.

[0008] The surface of the colony of the Acinetobacter sp. 450-8 is moist, smooth and flat, with neat edges, opaque and milky white, and the colony is round and easy to pick.

[0009] The Acinetobacter sp. 450-8 was isolated from an activated sludge sample, which was taken from a biochemical pool of a refinery saline wastewater treatment plant.

[0010] On the other hand, the present invention provides a culture method for the Acinetobacter sp. 450-8. After the activated sludge sample is inoculated in a screening culture medium for screening, it is purified multiple times through a solid culture medium to obtain an Acinetobacter sp. strain. The purified strain is enriched and cultured using a liquid culture medium to obtain a bacterial suspension. Through experiments on the growth characteristics of the strain, the salt tolerance, substrate broad spectrum and heavy metal resistance of the strain are explored. The suitable culture conditions are pH 6.5-7.5 and temperature 26-30°C.

[0011] Further, the components of the screening medium are: (NH 4 ) 2 SO 4 1.5g, KH 2 PO 4 1.3 g, MgSO 4 7H 2 O0.3g, CaCl 2 0.05 g, FeSO 4 7H 2 O 0.005g, refinery saline wastewater 1000mL, adjust the pH to 7.0 with phosphate buffer, and sterilize at 121℃ for 20min before use.

[0012] Furthermore, the components of the solid culture medium are: (NH 4 ) 2 SO 4 1.5g, KH 2 PO 4 1.3 g, MgSO 4 7H 2 O0.3g, CaCl 2 0.05 g, FeSO 4 7H 2 O 0.005g, agar 15g, refined saline wastewater 1000mL, adjust the pH to 6.5-7.5 with phosphate buffer, and sterilize at 121℃ for 20min before use.

[0013] Furthermore, the components of the liquid culture medium are: 2.5 g yeast powder, 5 g peptone, 5 g NaCl, 2 HPO 4 1g, MgSO 4 7H 2O 0.5g, refinery saline wastewater 1000ml, adjust the pH to 6.5-7.5 with phosphate buffer, sterilize at 121℃ for 20min before use.

[0014] The effects of the strains' tolerance to salinity, substrates and heavy metal resistance were determined by experiments on strain tolerance to different salinities, experiments on a broad spectrum of different substrates and experiments on resistance to different heavy metals. The setting of different salinity conditions was achieved by 1%, 1.5%, 2%, 2.5% and 3% NaCl. Different substrates including ethylene, n-hexane, toluene and hydroquinone were selected for the broad spectrum substrate experiment. The strain resistance to heavy metals experiment included verifying the effects of zinc sulfate, copper sulfate, nickel chloride and potassium dichromate on strain growth.

[0015] The above-mentioned suitable pH of 6.5-7.5 is determined by the growth of the strain after being cultured under different pH conditions, and the above-mentioned suitable temperature of 26-30° C. is determined by the growth of the strain after being cultured under different temperature conditions.

[0016] In another aspect, the present invention provides an application of Acinetobacter sp. 450-8 in treating high-salt wastewater from refineries.

[0017] Furthermore, the salinity of the refined high-salt wastewater is 1-3%.

[0018] In another aspect, the present invention provides the use of an Acinetobacter strain in treating refinery wastewater containing high concentrations of ethylene and n-hexane.

[0019] Furthermore, the concentrations of high-concentration ethylene and n-hexane in the refinery wastewater are both 5-80 mg / L.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] The present invention provides an Acinetobacter sp. 450-8, which is obtained from an activated sludge sample through directional screening, separation and purification, and is capable of improving the treatment efficiency of saline wastewater from refineries. The strain has a significant removal effect on COD in saline refinery wastewater, and also has a good removal effect on ammonia nitrogen in water bodies, and also has good salt tolerance, substrate broad spectrum and heavy metal resistance. The Acinetobacter sp. 450-8 is simple to culture, grows rapidly, has strong environmental adaptability, is used for refinery wastewater treatment, can significantly reduce the main pollution indicators of water bodies, improves treatment efficiency, and shows its huge application potential in refinery wastewater treatment, and has good industrial application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0023] Figure 1 This is a colony photo of Acinetobacter sp. 450-8;

[0024] Figure 2 is a growth curve graph of Acinetobacter sp. 450-8;

[0025] Figure 3 Optical microscope photo of Acinetobacter sp. 450-8;

[0026] Figure 4 This is an electron microscopic photograph of Acinetobacter sp. 450-8;

[0027] Figure 5 This is an experiment on the tolerance of Acinetobacter sp. 450-8 to different salinities;

[0028] Figure 6 This is an experiment on the utilization of different substrates by Acinetobacter sp. 450-8;

[0029] Figure 7 It is an experiment on the resistance of Acinetobacter sp. 450-8 to different heavy metals;

[0030] Figure 8 It is the removal effect of COD by the control group and the experimental group in Example 6;

[0031] Fig. 9 The control group and the experimental group in Example 6 are NH 3 -N removal effect;

[0032] Fig.10 is the removal effect of TN by the control group and the experimental group in Example 6;

[0033] Fig.11 It is the removal effect of COD for the control group and the experimental group in Example 7;

[0034] Fig.12 The control group and the experimental group in Example 7 are3 -N removal effect;

[0035] Fig.13 It is the removal effect of TN by the control group and the experimental group in Example 7. DETAILED DESCRIPTION

[0036] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0037] Example 1: Screening and cultivation of strains

[0038] Acinetobacter sp. was isolated and screened from activated sludge samples from a refinery saline wastewater treatment plant, as follows:

[0039] Take 1 ml of sludge sample and add it to sterile saline solution, shake it and make a sludge-water mixture. Use gradient dilution method to dilute the mixture into 10 -1 -10 -8 Take 0.1 mL of each gradient dilution and spread it evenly on a solid culture medium plate, and place the plate in a 30°C constant temperature incubator and invert it for 48 hours.

[0040] Use an inoculation loop to pick up a single colony and inoculate it into the screening medium. After 48 hours of culture, measure the chemical oxygen demand (COD) and ammonia nitrogen (NH) in the medium. 3 -N removal effect, select COD and NH 3 -N removal effect is good. Take an appropriate amount of bacterial solution and use an inoculation loop to streak the plate on the solid culture medium, and then invert and culture for 48 hours. The components of the above screening culture medium are: (NH 4 ) 2 SO 4 1.5g, KH 2 PO 4 1.3 g, MgSO 4 7H 2 O 0.3g, CaCl 2 0.05 g, FeSO 4 7H 2 O 0.005g, refine salty sewage 1000mL, adjust pH to 6.5-7.5 with phosphate buffer, sterilize at 121℃ for 20min before use. The components of the solid culture medium for purifying Acinetobacter are: (NH4 ) 2 SO 4 1.5g, KH 2 PO 4 1.3 g, MgSO 4 7H 2 O 0.3g, CaCl 2 0.05 g, FeSO 4 7H 2 O 0.005g, agar 15g, refined saline wastewater 1000mL, adjust the pH to 6.5-7.5 with phosphate buffer, and sterilize at 121℃ for 20min before use.

[0041] Repeat the above steps several times until COD and NH 3 -N removal effect is good and stable strains, the obtained strain 450-8, the surface of its colony is moist, smooth and flat, the edge is neat, opaque, milky white, the colony is round, easy to pick, such as Figure 1 shown.

[0042] The purified strain was inoculated into the liquid culture medium, and the inoculated culture medium was placed in a constant temperature shaking incubator and cultured at 30°C and 180 rpm. Appropriate amounts of bacterial liquid were taken out from the culture system at 0h, 2h, 4h, 6h, 8h, 10h, 12h, 18h, and 24h after culture, and the OD of the bacterial liquid was measured at a wavelength of 600nm using a spectrophotometer. 600 The growth curve of strain 450-8 is as follows Figure 2 The components of the liquid culture medium are as follows: 2.5 g yeast powder, 5 g peptone, 5 g NaCl, 2 HPO 4 1g, MgSO 4 7H 2 O0.5g, refine saline wastewater 1000ml, adjust the pH to 6.5-7.5 with phosphate buffer, and sterilize at 121℃ for 20min before use.

[0043] Example 2: Identification of strains

[0044] Gram staining of strain 450-8 obtained in Example 1 showed that Acinetobacter was red and was a Gram-negative bacterium. Figure 3 As shown in Figure 2, under an optical microscope, they are club-shaped or rod-shaped, mostly existing alone, but sometimes in pairs or short chains. Using an electron microscope, we can observe more subtle cell structures, such as Figure 4 As shown, it can be more accurately observed that the fungus 450-8 does not have special structures such as pili.

[0045] The 16S rDNA sequence of the 450-8 strain was extracted and determined, and its 16S rDNA gene sequence is shown in the sequence as shown in SEQ ID NO.1. The strain is Acinetobacter sp., named Acinetobacter 450-8, and the Acinetobacter was deposited in the General Microbiology Center (CGMCC) of the China Microbiological Culture Collection Administration on December 25, 2024, with a deposit number of CGMCC No. 33205, and the address is No. 3, Yard 1, Beichen West Road, Beijing.

[0046] Example 3: Tolerance experiment of Acinetobacter sp. 450-8 at different salinities

[0047] Weigh different masses of sodium chloride (NaCl) and add them to the prepared liquid culture medium (the composition is consistent with the liquid culture medium in Example 1) to prepare 5 kinds of culture medium with final NaCl concentrations of 1%, 1.5%, 2%, 2.5%, and 3%, respectively. The prepared culture medium is divided into several 250 mL conical bottles, 50 mL per bottle, and then sealed with sealing film.

[0048] The conical flask containing the culture medium was sterilized in a high pressure steam autoclave, sterilized at 121°C for 20 minutes, and then taken out and cooled to room temperature. The strain was inoculated into 50 mL conical flasks of liquid culture medium with the above 5 different salinities (1%, 1.5%, 2%, 2.5%, 3% NaCl). The inoculated conical flask was placed in a constant temperature shaking incubator, set the temperature to 37°C, the shaking speed to 180 rpm, and culture was started.

[0049] At 0h, 12h, 24h, 36h, and 48h after the start of culture, the uninoculated culture medium of the corresponding salinity was used as a blank control. The blank control and the bacterial solution sample were poured into a cuvette, placed in a UV-visible spectrophotometer, and the OD of the bacterial solution was measured at a wavelength of 600nm. 600 Before each measurement, the spectrophotometer was calibrated to zero using distilled water.

[0050] like Figure 5 As shown, under the five salinity conditions of 1%, 1.5%, 2%, 2.5%, and 3%, the OD of Acinetobacter strain 450-8 600 The values ​​increased with time. This shows that the strain can adapt and grow within the salinity range set in the experiment. In the 0-24h period, the OD 600 The value grows relatively fast and is in the logarithmic growth period; during the 24-48h period, the growth rate slows down and gradually enters the stable period.

[0051] Under 1% NaCl conditions, the growth rate of the strain was significantly faster than that of other salinities. At 48h, its OD 600 The maximum value reached 0.546. This shows that the cell metabolism of this strain is active in a low-salinity environment, the nutrient utilization efficiency is high, and it is more conducive to its growth and reproduction. As the NaCl concentration increases, at the same time point, OD 600 At 24h, the OD 600 The value was 0.366, while that under 3% NaCl was 0.247. This indicates that increased salinity will inhibit the growth of the strain, and the higher the salinity, the more obvious the inhibitory effect.

[0052] Example 4: Experiment on the utilization of different substrates by Acinetobacter sp. 450-8

[0053] Liquid culture medium (the components are consistent with those of the liquid culture medium in Example 1) was prepared, and different concentrations (5 mg / L, 10 mg / L, 20 mg / L, 40 mg / L, 80 mg / L) of ethylene, n-hexane, toluene, and hydroquinone were added as the sole carbon source, and several parallel groups were set up for each substrate-concentration combination.

[0054] Under a sterile environment, Acinetobacter sp. 450-8 was inoculated into the above culture medium containing different substrates and concentrations at the same inoculation amount (1% by volume). The inoculated conical flask was placed in a constant temperature shaking incubator, the temperature was set at 30° C., the shaking speed was set at 180 rpm, and the culture was carried out.

[0055] 48 h after the start of culture, the OD of the bacterial solution was measured at a wavelength of 600 nm using the uninoculated corresponding substrate-concentration culture medium as a blank control. 600 Before each measurement, the spectrophotometer was zeroed and calibrated with distilled water.

[0056] The results are as follows Figure 6 As shown in Figure 2, the strain has obvious differences in its ability to utilize different substrates, and has better ability to utilize ethylene and n-hexane. The corresponding OD values ​​at each concentration are 600 The value is higher; the utilization capacity of toluene and hydroquinone is weaker. At the same concentration, OD 600 The lower value is related to the relevant metabolic enzyme system and substrate toxicity.

[0057] The substrate concentration had a significant effect on the growth of the strain. As the substrate concentration increased from 5 mg / L to 80 mg / L, the OD of the strain under each substrate condition increased. 600The values ​​decreased overall, and high concentrations of substrates could affect osmotic pressure and produce toxicity. Different substrates were affected by concentrations to different degrees, with relatively strong tolerance to ethylene and n-hexane, and more sensitive to toluene and hydroquinone.

[0058] Example 5: Experiment on the resistance of Acinetobacter sp. 450-8 to different heavy metals

[0059] Liquid culture medium was prepared by adding different concentrations (10 mg / L, 20 mg / L, 40 mg / L, 80 mg / L, 120 mg / L) of zinc sulfate, copper sulfate, nickel chloride, and potassium dichromate as culture medium containing different heavy metals. Parallel groups were set up for each heavy metal-concentration combination and the culture medium was sterilized with high-pressure steam for later use.

[0060] Under a sterile environment, the same inoculation amount (1% by volume) of Acinetobacter sp. 450-8 in the logarithmic growth phase was inoculated into the above-mentioned culture medium containing different heavy metals and concentrations. The inoculated culture medium was placed in a constant temperature shaking incubator, the temperature was set at 30° C., the shaking speed was 180 rpm, and the culture was carried out.

[0061] After 48 h of culture, samples were taken and the OD of the bacterial solution was measured using a spectrophotometer at a wavelength of 600 nm. 600 Before each measurement, the spectrophotometer was zeroed and calibrated with distilled water, and the OD of the strain under each heavy metal-concentration combination was recorded. 600 value.

[0062] The results are as follows Figure 7 As shown, Acinetobacter sp. 450-8 showed relatively good tolerance to zinc sulfate. At each concentration, its growth corresponding to OD 600 The value is relatively high. The tolerance to copper sulfate is second. At lower concentrations (10mg / L, 20mg / L), the strain can still maintain a certain growth, but as the concentration increases, the growth is significantly inhibited. Copper ions will bind to biological macromolecules such as proteins and nucleic acids in the cells, affecting their normal functions and causing the growth of the strain to slow down. The strain is also quite tolerant to nickel chloride. At low concentrations, the strain can grow, but as the concentration of nickel chloride increases, the growth is gradually inhibited; the tolerance to potassium dichromate is poor. At each concentration, the corresponding OD 600 The values ​​were all low. At 10 mg / L, OD 600 The value was about 0.05, and at 120 mg / L, almost no obvious growth was detected.

[0063] Example 6: Treatment of saline wastewater from a refinery wastewater treatment plant by Acinetobacter sp. 450-8

[0064] The saline wastewater from a refinery wastewater treatment plant that had been pretreated (grid degreasing) was used as experimental water, and its initial water quality parameters (COD 854 mg / L, NH 3 -N 16.8mg / L, total nitrogen TN 72.5mg / L, salinity 1.2), the water COD is relatively high, and the main pollutants leading to high COD are alkanes and aromatic hydrocarbons. An experimental group and a control group were set up. The experimental group was added with activated sludge and Acinetobacter sp. 450-8, and the control group was added with activated sludge without adding strains. The same amount of experimental water and the same amount of activated sludge were placed in 250mL conical flasks of the experimental group and the control group, and the pH was adjusted to about 7.0. For the experimental group, the strain was added to the conical flask at a 2% inoculation rate, and cultured at 180rpm in a constant temperature shaker at 26-30℃ for 72 hours; the control group was cultured under the same conditions, but no strains were added. During the treatment process, samples were taken from the experimental group and the control group every 6 hours to measure the water quality parameters of the samples (COD, NH 3 -N, TN).

[0065] from Figure 8 It can be clearly seen that with the passage of time, the COD removal rates of both the control group and the experimental group gradually increased. At around 36h, the COD removal rate of the experimental group began to be significantly higher than that of the control group, and reached a peak of 89.5% at 48h. The highest COD removal rate of the control group was 70.2%. From 48h to 72h, the COD removal rate of the experimental group decreased slightly, but remained at around 80%, while the COD removal rate of the control group remained at around 70%. Overall, the COD removal rates of the experimental groups were higher than those of the control group, indicating that the addition of strains in the experimental group can improve the COD removal effect, and Acinetobacter sp. 450-8 has a better COD removal effect.

[0066] Fig. 9 The experimental data showed that the removal rate of the experimental group was higher than that of the control group within 72 hours. This shows that the experimental group is more effective in removing ammonia nitrogen. At 48 hours, the removal rate of the experimental group was close to 90%, while that of the control group was only about 70%. The experimental group was able to remove ammonia nitrogen more efficiently in the same time.

[0067] Fig.10The experimental data showed that the TN removal rates of both the control group and the experimental group gradually increased over time in the initial stage. At around 24 hours, the TN removal rate of the experimental group began to be significantly higher than that of the control group. The TN removal rate of the experimental group reached a peak of 74.6% at 48 hours, then decreased slightly, but still remained above 70%, while the TN removal rate of the control group reached a peak of only 55.5% at 60 hours.

[0068] The experimental results showed that Acinetobacter sp. 450-8 showed good removal efficiency in treating refinery saline wastewater, especially in treating COD and NH 3 -N aspects.

[0069] Example 7: Treatment of saline wastewater from a refinery wastewater plant by Acinetobacter sp. 450-8

[0070] The saline wastewater from a refinery wastewater treatment plant that had been pretreated (flotation oil removal) was used as experimental water, and its initial water quality parameters (COD 538 mg / L, NH 3 -N 19.4mg / L, TN 51.3mg / L, salinity 1.5). An experimental group and a control group were set up. The experimental group was added with activated sludge and Acinetobacter sp. 450-8, and the control group was added with activated sludge without adding the strain. The same amount of experimental water and the same amount of activated sludge were placed in 250mL conical flasks of the experimental group and the control group, and the pH was adjusted to about 7.0. For the experimental group, the strain was added to the conical flask at a 2% inoculation rate, and cultured at 26-30℃ constant temperature shaker at 180-240rpm for 72h; the control group was placed under the same conditions, but no strain was added. During the treatment process, samples were taken from the experimental group and the control group every 6h to measure the water quality parameters of the samples (COD, NH 3 -N, TN).

[0071] from Fig.11 It can be clearly observed that as time goes on, at around 36h, the COD removal rate of the experimental group begins to be significantly higher than that of the control group. The COD removal efficiency of the experimental group reaches a maximum of 89.3% at 54h, while the COD removal rate of the control group is the highest at 42h, not exceeding 65%. From 48h to 72h, the COD removal rate of the experimental group decreases slightly, but remains at around 80%, while the COD removal rate of the control group remains at around 55%.

[0072] Fig.12 It shows that the NH 3-N removal efficiency exceeded 80% at 48h. From 48h to 72h, the removal rate of the experimental group remained at around 80%, which was relatively stable. 3 The removal rate of -N was around 30%, with a maximum of 59.6% at 48 h, but then decreased slightly and remained between 52% and 56%.

[0073] Fig.13 The experimental data showed that the highest TN removal rate in the experimental group was 90.2% at 54 h, while the highest TN removal rate in the control group was 63.3%, which further confirmed that the Acinetobacter strains in the experimental group played a role in the decomposition and transformation of nitrogen-containing pollutants, significantly enhancing the removal efficiency of TN.

[0074] In the treatment of high-salinity refinery wastewater, Acinetobacter sp. 450-8 is added to utilize its salt tolerance to adapt to the high-salinity environment of refinery wastewater, and to exert the biodegradation effect of Acinetobacter sp. on petroleum alkanes and nitrogen-containing substances, thereby significantly improving the removal effect of pollutants.

[0075] In summary, the present invention provides a special strain of Acinetobacter, which has a high efficiency in treating refinery saline wastewater, and has salt tolerance, a broad spectrum of substrates and heavy metal resistance. At the same time, the present invention also relates to a culture method and application technology of the strain, aiming to reduce treatment costs and environmental pollution through this efficient microbial technology, and provide new solutions for the sustainable development of the petrochemical industry.

[0076] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. An Acinetobacter strain, characterized in that The Acinetobacter strain (Acinetobacter sp.) is named 450-8 and is deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration. Its preservation number is CGMCC No. 33205, and the preservation date is December 25, 2024. The 16S rDNA sequence of the Acinetobacter strain 450-8 is shown in SEQ ID NO.

1.

2. The method for culturing the Acinetobacter strain according to claim 1, characterized in that: The Acinetobacter strain is derived from an activated sludge sample from a refinery saline wastewater treatment plant. The activated sludge sample is inoculated in a screening medium for screening, and then purified multiple times through a solid medium to obtain an Acinetobacter strain. The purified strain is enriched and cultured using a liquid medium to obtain a bacterial suspension. The culture conditions are a suitable pH of 6.5-7.5 and a suitable temperature of 26-30°C.

3. The culture method according to claim 2, characterized in that The screening medium comprises: (NH4)2SO4 1.5 g, KH2PO4 1.3 g, MgSO4·7H2O 0.3 g, CaCl2 0.05 g, FeSO4·7H2O 0.005 g, 1000 mL of refinery saline wastewater, pH adjusted to 7.0 with phosphate buffer, and sterilized at 121°C for 20 min before use.

4. The culture method according to claim 2, characterized in that The solid culture medium comprises: (NH4)2SO4 1.5g, KH2PO4 1.3g, MgSO4·7H2O 0.3g, CaCl2 0.05g, FeSO4·7H2O 0.005g, agar 15g, and 1000mL of refined saline wastewater. The pH value is adjusted to 6.5-7.5 with phosphate buffer and sterilized at 121°C for 20min before use.

5. The culture method according to claim 2, characterized in that: The liquid culture medium for enriching and culturing Acinetobacter comprises the following components: 2.5 g yeast powder, 5 g peptone, 5 g NaCl, 1 g K2HPO4, 0.5 g MgSO4·7H2O, 1000 ml refined saline wastewater, the pH value of which is adjusted to 6.5-7.5 with phosphate buffer, and sterilized at 121° C. for 20 min before use.

6. Use of the Acinetobacter strain described in claim 1 in treating high-salt wastewater from refineries.

7. The use of the Acinetobacter strain according to claim 6 in treating high-salt refinery wastewater, characterized in that: The salinity of the refining high-salt wastewater is 1-3%.

8. Use of the Acinetobacter strain according to claim 1 in treating refinery wastewater containing high concentrations of ethylene and n-hexane.

9. The use of the Acinetobacter strain according to claim 8 in treating high-salt refinery wastewater, characterized in that: The concentrations of high-concentration ethylene and normal hexane in the refinery wastewater are both 5-80 mg / L.

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