Campylobacter and fermentation agent and their application in reducing refinery sludge

Abstract

The present invention relates to the technical field of refinery wastewater treatment, and specifically relates to Campylobacter and a fermentation agent, and their use in refinery sludge reduction. The latter is carried out according to the following method: Campylobacter is inoculated into LB liquid culture medium for cultivation, then transferred to sterilized refinery wastewater for two activations, then the bacterial liquid after two activations is transferred to a nutrient solution for further expansion and cultivation, and finally, the fermentation agent containing Campylobacter is added to the refinery wastewater for reduction treatment. The present invention directly reduces the wastewater at the source by directly applying Campylobacter to an A / O biochemical tank to treat the refinery wastewater, with a hydraulic retention time of more than 25 hours. The enriched Campylobacter can treat refinery wastewater with an initial COD of 800 mg / L to 1600 mg / L, and the residual activated sludge content in the discharged wastewater after Campylobacter treatment is no more than 50 mg / L, achieving a reduction of more than 90%, greatly reducing the cost of subsequent treatment of environmental pollution.

Description

Technical Field

[0001] The invention relates to the technical field of refinery wastewater treatment, in particular to Campylobacter and a fermentation agent and their application in reducing refinery sludge. Background Art

[0002] The biochemical treatment of refinery wastewater produces a large amount of excess activated sludge. This sludge is primarily composed of various bacteria, organic matter, pathogens, and viruses, and contains a high amount of water. With a water content of up to 99%, the excess activated sludge is bulky and fluffy, making it difficult to directly incinerate. If carelessly disposed of, it not only pollutes the air, water, and soil, but the toxic substances and pathogens it contains pose a threat to human health, causing cancer and birth defects. Therefore, the primary treatment for excess activated sludge is to reduce its volume.

[0003] Currently, excess activated sludge is treated primarily through centrifugal dehydration, high-temperature hydrolysis, ultrasound, microwaves, ozone, and alkaline electrochemical treatment. These methods require large equipment and high energy consumption to achieve sludge dehydration and reduction. Microorganisms are also used to reduce excess activated sludge.

[0004] Chinese patent, application number 202110583595.8, patent name is "Application of a Bacillus in Industrial Sludge Reduction and Detoxification". It is achieved by expanding a Bacillus and mixing it with dehydrated activated sludge and secondary powder for fermentation. The temperature in the fermentation tank is controlled at 55°C to 65°C. After 24 hours of reaction, it is ventilated and dried, which can achieve a 90% sludge reduction, and a 75% reduction in organic matter. This treatment of residual activated sludge using microorganisms is a subsequent process treatment and does not fundamentally achieve source reduction. At the same time, there is no essential difference between the reduction achieved through ventilation and dehydration and other dehydration reductions. It only reduces some energy consumption.

[0005] The Chinese patent, patent number 201811617269.9, is titled "A Thermophilic Microbial Strain for In-Situ Sludge Reduction and Its Application." The method for achieving sludge reduction is to add a thermophilic functional bacterium to an aerobic reaction tank. The thermophilic functional bacterium releases a hydrolytic extracellular enzyme to decompose macromolecular organic matter or dead microorganisms, and the decomposition products serve as a nutrient source for the microorganisms. This method only reduces the activated sludge content in the system by 40%, but still produces excess sludge. This part of the discharged excess sludge still needs to be subsequently treated, which still causes pollution to the environment.

[0006] It can be seen that the current methods for sludge reduction are mainly focused on the treatment of residual activated sludge, that is, the treatment of subsequent processes. Even if the reduction is achieved at the source, only a small reduction can be achieved, and the residual sludge discharged externally still exists. There is a lack of solutions that can truly achieve a significant reduction in sludge concentration at the source and reduce or even eliminate the discharge of residual sludge. Summary of the Invention

[0007] The present invention provides Campylobacter and a fermentation agent and their use in reducing refinery sludge, overcoming the shortcomings of the above-mentioned existing technologies. The residual activated sludge content in the discharged wastewater after Campylobacter treatment is no more than 50 mg / L, achieving a reduction of more than 90%.

[0008] One of the technical solutions of the present invention is achieved through the following measures: a Campylobacter sp., which was deposited in the General Microbiology Center of the China Culture Collection Administration on January 18, 2021, and its preservation number is CGMCC No. 21652.

[0009] The second technical solution of the present invention is achieved through the following measures: a fermentation agent containing Campylobacter.

[0010] The following is a further optimization and / or improvement of the second technical solution of the above invention:

[0011] The fermentation medium containing Campylobacter comprises Campylobacter and nutrient solution, and the concentration of Campylobacter in the fermentation solution is 1×10 8 CFU / mL or above.

[0012] The composition of the above-mentioned nutrient solution includes 20g / L to 24g / L of glucose, 2g / L to 5g / L of peptone, 1g / L to 2g / L of yeast powder, 2g / L to 3g / L of ammonium nitrate, 4g / L to 5g / L of sodium chloride, 0.5g / L to 1g / L of potassium dihydrogen phosphate, 0.3g / L to 0.4g / L of manganese sulfate, 1g / L to 2g / L of magnesium sulfate, 0.02g / L to 0.05g / L of ferrous sulfate and 1g / L to 2g / L of calcium carbonate, and the pH value of the nutrient solution is 7.0 to 7.2.

[0013] The third technical solution of the present invention is achieved through the following measures: an application of Campylobacter in reducing refinery sludge.

[0014] The following are further optimizations and / or improvements to the third technical solution of the above invention:

[0015] The application of the Campylobacter in the reduction of refinery sludge is carried out according to the following method:

[0016] In the first step, Campylobacter was inoculated into LB liquid culture medium and cultured on a shaking table to obtain a fermentation seed liquid. The culture temperature was 30°C to 35°C, the shaking speed was 100 rpm to 250 rpm, and the concentration of Campylobacter in the fermentation seed liquid was 1×10 8 CFU / mL or above;

[0017] The second step is to inoculate the fermentation seed liquid into the sterilized low-COD refinery wastewater for activation, wherein the activation conditions include a culture temperature of 30°C to 35°C, a pH of 8 to 10, a shaker speed of 100 rpm to 250 rpm, an activation time of 24 hours, an inoculation amount of the fermentation seed liquid of 3% to 6% by volume of the sterilized low-COD refinery wastewater, and a COD of 500 mg / L to 800 mg / L of the sterilized low-COD refinery wastewater;

[0018] The third step is to transfer the activated fermentation seed liquid to the sterilized high-COD refinery wastewater for reactivation, wherein the activation conditions include a culture temperature of 30°C to 35°C, a pH of 8 to 10, a shaker speed of 100 rpm to 250 rpm, an activation time of 24 hours, and an inoculation amount of the activated fermentation seed liquid of 3% to 6% by volume of the sterilized high-COD refinery wastewater, and the COD of the sterilized high-COD refinery wastewater is 800 mg / L to 1600 mg / L.

[0019] In the fourth step, the reactivated fermentation seed liquid is transferred to a fermentation tank containing a nutrient solution for continued fermentation and expansion culture to obtain a starter containing Campylobacter, wherein the culture temperature is 28°C to 38°C, the pH is 7.5 to 9.5, the rotation speed is 200 rpm, the pressure of the fermentation tank is 0.03 MPa to 0.06 MPa, the ventilation volume is 1.0 to 1.3:1, the fermentation time is 16 h to 20 h, the inoculation amount of the reactivated fermentation seed liquid is 3% to 6% of the volume percentage of the nutrient solution, and the concentration of Campylobacter in the starter containing Campylobacter is 1×10 8 CFU / ml or above;

[0020] In the fifth step, a fermentation agent containing Campylobacter is added to an A / O biochemical pool containing refinery wastewater and filled with fiber filler for reduction treatment, wherein the addition amount of the fermentation agent containing Campylobacter is 3% to 6% of the volume percentage of the refinery wastewater. During the reduction treatment, the temperature is 30°C to 35°C, the time is not less than 25 hours, the initial COD of the refinery wastewater is 800mg / L to 1600mg / L, the oil content is not higher than 20mg / L, the fiber filler is made of polypropylene, and the filling rate of the fiber filler in the A / O biochemical pool is 20% to 70%.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] The Campylobacter used in the present invention has excellent COD degradation and ammonia nitrogen removal capabilities for refinery wastewater. At the same time, it also has good resistance to sulfide shock and has strong shock resistance when the water quality of upstream water changes significantly. Generally, the COD of refinery wastewater in the regeneration treatment stage is mostly between 500mg / L and 800mg / L, while the COD tolerance of this strain is 1800mg / L. Moreover, after the sulfide shock of upstream water quality, this strain can quickly restore its original treatment capacity.

[0023] The present invention performs wastewater reduction treatment directly at the source, directly applying Campylobacter in an A / O biochemical pool to treat refinery wastewater with a hydraulic retention time of more than 25 hours. The enriched Campylobacter can treat refinery wastewater with an initial COD of 800 mg / L to 1600 mg / L and an oil content of no more than 20 mg / L. Moreover, the residual activated sludge content in the discharged wastewater after Campylobacter treatment is no more than 50 mg / L, achieving a wastewater reduction of more than 90%. This truly achieves a substantial reduction in sludge content at the source and greatly reduces the cost of subsequent treatment of environmental pollution. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is an electron micrograph of Campylobacter sp. DETAILED DESCRIPTION

[0025] The present invention is not limited to the following embodiments; specific implementation methods may be determined based on the technical solutions and actual conditions of the present invention. Unless otherwise specified, the various chemical reagents and chemicals mentioned in the present invention are all known and commonly used chemical reagents and chemicals in the prior art; and percentages in the present invention are all by mass unless otherwise specified.

[0026] The present invention will be further described below in conjunction with the embodiments:

[0027] Example 1: The Campylobacter sp. strain was deposited in the General Microbiology Center of the China Culture Collection Administration on January 18, 2021, and its deposit number is CGMCC No. 21652.

[0028] The Campylobacter of the present invention is obtained by separation and screening from activated sludge in a secondary sedimentation tank of a biochemical device of a petrochemical enterprise. The strain is characterized by a convex and moist colony morphology, an irregular shape, a rough surface, and a milky white color.

[0029] Example 2: The fermentation culture containing Campylobacter comprises Campylobacter and nutrient solution. The concentration of Campylobacter in the fermentation culture containing Campylobacter is 1×10 8CFU / mL or above, the composition of the nutrient solution also includes 20g / L to 24g / L glucose, 2g / L to 5g / L peptone, 1g / L to 2g / L yeast powder, 2g / L to 3g / L ammonium nitrate, 4g / L to 5g / L sodium chloride, 0.5g / L to 1g / L potassium dihydrogen phosphate, 0.3g / L to 0.4g / L manganese sulfate, 1g / L to 2g / L magnesium sulfate, 0.02g / L to 0.05g / L ferrous sulfate and 1g / L to 2g / L calcium carbonate, and the pH value of the nutrient solution is 7.0 to 7.2.

[0030] Example 3: Application of Campylobacter in reducing refinery sludge.

[0031] Example 4: Application of Campylobacter in Refinery Sludge Reduction is carried out according to the following method:

[0032] In the first step, Campylobacter was inoculated into LB liquid culture medium and cultured on a shaking table to obtain a fermentation seed liquid. The culture temperature was 30°C, the shaking speed was 100 rpm, and the concentration of Campylobacter in the fermentation seed liquid was 1×10 8 CFU / mL or above;

[0033] The second step is to inoculate the fermentation seed liquid into the sterilized low-COD refinery wastewater for activation. The activation conditions include a culture temperature of 30°C, a pH of 8, a shaker speed of 100 rpm, an activation time of 24 hours, and an inoculation amount of the fermentation seed liquid of 3% by volume of the sterilized low-COD refinery wastewater. The COD of the sterilized low-COD refinery wastewater is 500 mg / L.

[0034] In the third step, the activated fermentation seed liquid was transferred to the sterilized high-COD refinery wastewater for reactivation. The activation conditions included a culture temperature of 30°C, a pH of 8, a shaker speed of 100 rpm, an activation time of 24 hours, and an inoculum size of 3% of the volume of the sterilized high-COD refinery wastewater. The COD of the sterilized high-COD refinery wastewater was 800 mg / L.

[0035] In the fourth step, the reactivated fermentation seed liquid was transferred to a fermentation tank containing a nutrient solution to continue fermentation and expansion culture to obtain a starter containing Campylobacter, wherein the culture temperature was 28°C, the pH was 7.5, the rotation speed was 200 rpm, the pressure of the fermentation tank was 0.03 MPa, the ventilation volume was 1.0:1, the fermentation time was 16 h, the inoculation amount of the reactivated fermentation seed liquid was 3% of the volume percentage of the nutrient solution, and the concentration of Campylobacter in the starter containing Campylobacter was 1×10 8CFU / ml or above, the composition of the nutrient solution also includes 20g / L glucose, 2g / L peptone, 1g / L yeast powder, 2g / L ammonium nitrate, 4g / L sodium chloride, 0.5g / L potassium dihydrogen phosphate, 0.3g / L manganese sulfate, 1g / L magnesium sulfate, 0.02g / L ferrous sulfate and 1g / L calcium carbonate, and the pH value of the nutrient solution is 7.0;

[0036] In the fifth step, a fermentation agent containing Campylobacter is added to an A / O biochemical pool containing refinery wastewater and filled with fiber filler for reduction treatment, wherein the addition amount of the fermentation agent containing Campylobacter is 3% of the volume percentage of the refinery wastewater. During the reduction treatment, the temperature is 30°C, the time is 25 hours, the initial COD of the refinery wastewater is 800 mg / L, the oil content is 8 mg / L, the fiber filler is made of polypropylene, and the filling rate of the fiber filler in the A / O biochemical pool is 20%.

[0037] Example 5: Application of Campylobacter in Refinery Sludge Reduction is carried out according to the following method:

[0038] In the first step, Campylobacter was inoculated into LB liquid culture medium and cultured on a shaking table to obtain a fermentation seed liquid. The culture temperature was 35°C, the shaking speed was 250 rpm, and the concentration of Campylobacter in the fermentation seed liquid was 1×10 8 CFU / mL or above;

[0039] In the second step, the fermentation seed liquid was inoculated into the sterilized low-COD refinery wastewater for activation. The activation conditions included a culture temperature of 35°C, a pH of 10, a shaker speed of 250 rpm, an activation time of 24 hours, and an inoculation amount of the fermentation seed liquid of 6% by volume of the sterilized low-COD refinery wastewater. The COD of the sterilized low-COD refinery wastewater was 800 mg / L.

[0040] In the third step, the activated fermentation seed liquid was transferred to the sterilized high-COD refinery wastewater for reactivation. The activation conditions included a culture temperature of 35°C, a pH of 10, a shaker speed of 250 rpm, an activation time of 24 hours, and an inoculum size of 6% of the volume of the sterilized high-COD refinery wastewater. The COD of the sterilized high-COD refinery wastewater was 1600 mg / L.

[0041] In the fourth step, the reactivated fermentation seed liquid was transferred to a fermentation tank containing a nutrient solution to continue fermentation and expansion culture to obtain a starter containing Campylobacter, wherein the culture temperature was 38°C, the pH was 9.5, the rotation speed was 200 rpm, the pressure of the fermentation tank was 0.06 MPa, the ventilation volume was 1.3:1, the fermentation time was 20 h, the inoculation amount of the reactivated fermentation seed liquid was 6% of the volume percentage of the nutrient solution, and the concentration of Campylobacter in the starter containing Campylobacter was 1×10 8 CFU / ml or above, the composition of the nutrient solution also includes 24g / L glucose, 5g / L peptone, 2g / L yeast powder, 3g / L ammonium nitrate, 5g / L sodium chloride, 1g / L potassium dihydrogen phosphate, 0.4g / L manganese sulfate, 2g / L magnesium sulfate, 0.05g / L ferrous sulfate and 2g / L calcium carbonate, and the pH value of the nutrient solution is 7.2;

[0042] In the fifth step, a fermentation agent containing Campylobacter is added to an A / O biochemical pool containing refinery wastewater and filled with fiber filler for reduction treatment, wherein the addition amount of the fermentation agent containing Campylobacter is 6% of the volume percentage of the refinery wastewater. During the reduction treatment, the temperature is 35°C, the time is 25 hours, the fermentation time is 25 hours, the initial COD of the refinery wastewater is 1600 mg / L, the oil content is 20 mg / L, the fiber filler is made of polypropylene, and the filling rate of the fiber filler in the A / O biochemical pool is 70%.

[0043] Example 6: The application of Campylobacter in reducing refinery sludge is carried out according to the following method:

[0044] In the first step, Campylobacter was inoculated into LB liquid culture medium and cultured on a shaking table to obtain a fermentation seed liquid. The culture temperature was 32°C, the shaking speed was 200 rpm, and the concentration of Campylobacter in the fermentation seed liquid was 1×10 8 CFU / mL or above;

[0045] In the second step, the fermentation seed liquid was inoculated into the sterilized low-COD refinery wastewater for activation. The activation conditions included a culture temperature of 32°C, a pH of 9, a shaker speed of 200 rpm, an activation time of 24 hours, and an inoculation amount of the fermentation seed liquid of 4% by volume of the sterilized low-COD refinery wastewater. The COD of the sterilized low-COD refinery wastewater was 700 mg / L.

[0046] In the third step, the activated fermentation seed liquid was transferred to the sterilized high-COD refinery wastewater for reactivation. The activation conditions included a culture temperature of 32°C, a pH of 9, a shaker speed of 200 rpm, an activation time of 24 hours, and an inoculum size of 4% of the volume of the sterilized high-COD refinery wastewater. The COD of the sterilized high-COD refinery wastewater was 1000 mg / L.

[0047] In the fourth step, the reactivated fermentation seed liquid was transferred to a fermentation tank containing a nutrient solution to continue fermentation and expansion culture to obtain a starter containing Campylobacter. The culture temperature was 32°C, the pH was 8.5, the rotation speed was 200 rpm, the pressure of the fermentation tank was 0.04 MPa, the ventilation volume was 1.1:1, the fermentation time was 18 h, the inoculation amount of the reactivated fermentation seed liquid was 4% of the volume percentage of the nutrient solution, and the concentration of Campylobacter in the starter containing Campylobacter was 1×10 8 CFU / ml or above, the composition of the nutrient solution also includes 22g / L glucose, 4g / L peptone, 1.5g / L yeast powder, 2.5g / L ammonium nitrate, 4.5g / L sodium chloride, 0.8g / L potassium dihydrogen phosphate, 0.35g / L manganese sulfate, 1.5g / L magnesium sulfate, 0.04g / L ferrous sulfate and 1.5g / L calcium carbonate, and the pH value of the nutrient solution is 7.1;

[0048] In the fifth step, a fermentation agent containing Campylobacter was added to an A / O biochemical pool containing refinery wastewater and filled with fiber filler for reduction treatment. The amount of the fermentation agent containing Campylobacter added was 4% of the volume of the refinery wastewater. During the reduction treatment, the temperature was 32°C, the time was 25 hours, the initial COD of the refinery wastewater was 1000 mg / L, the oil content was 10 mg / L, the fiber filler was made of polypropylene, and the filling rate of the fiber filler in the A / O biochemical pool was 40%.

[0049] The A / O biochemical pools in Examples 4 to 6 of the present invention are suspended with fiber fillers, which are made of polypropylene. The filling rate of the fiber fillers in each stage of the reactor in the A / O biochemical pool is 20% to 70%, the take-up rate of the filler is 1:2500, its aspect ratio is ≥2500:1, the specific surface area is ≥20000m2 / kg, the main wire diameter is 80μm to 100μm, the side wire diameter is 10μm to 20μm, and the size is 1.2m×1.5m.

[0050] In the actual refinery wastewater reduction treatment process of this invention, five A / O biochemical tanks are connected in series. The A / O biochemical tanks have two to four A tank stages. Each A / O biochemical tank uses two to four anaerobic reactors in combination with a single aerobic reactor. The reactors are all of the same size. Fiber filler is suspended in each reactor, with a filling rate of 40%. Campylobacter bacteria adhere to the fiber filler in each reactor. The hydraulic retention time of the device is 25 hours.

[0051] The fiber filler used in this invention has a high microbial capacity, three to five times that of conventional fillers, providing an optimal environment for microbial growth and reproduction. The high dissolved oxygen utilization rate also significantly reduces operating costs. The continuous multi-stage filler packing also provides excellent filtration and shock absorption.

[0052] Example 7: The application of Campylobacter in reducing refinery sludge is carried out according to the following method:

[0053] In the first step, Campylobacter was inoculated into LB liquid culture medium and cultured on a shaking table to obtain a fermentation seed liquid. The culture temperature was 35°C, the shaking speed was 200 rpm, and the concentration of Campylobacter in the fermentation seed liquid was 1×10 8 CFU / mL or above;

[0054] The second step is to inoculate the fermentation seed liquid into the sterilized low-COD refinery wastewater for activation. The activation conditions include a culture temperature of 35°C, a pH of 8, a shaker speed of 200 rpm, an activation time of 24 hours, and an inoculation amount of the fermentation seed liquid of 4% by volume of the sterilized low-COD refinery wastewater. The COD of the sterilized low-COD refinery wastewater is 500 mg / L.

[0055] In the third step, the activated fermentation seed liquid was transferred to the sterilized high-COD refinery wastewater for reactivation. The activation conditions included a culture temperature of 35°C, a pH of 8, a shaker speed of 200 rpm, an activation time of 24 hours, and an inoculum size of 4% of the volume of the sterilized high-COD refinery wastewater. The COD of the sterilized high-COD refinery wastewater was 500 mg / L.

[0056] In the fourth step, the reactivated fermentation seed liquid was transferred to a fermentation tank containing a nutrient solution to continue fermentation and expansion culture to obtain a starter containing Campylobacter, wherein the culture temperature was 35°C, the pH was 8, the rotation speed was 200 rpm, the pressure of the fermentation tank was 0.04 MPa, the ventilation volume was 1.1:1, the fermentation time was 20 h, the inoculation amount of the reactivated fermentation seed liquid was 4% of the volume percentage of the nutrient solution, and the concentration of Campylobacter in the starter containing Campylobacter was 1×10 8CFU / ml or above, the composition of the nutrient solution also includes 22g / L glucose, 4g / L peptone, 1.5g / L yeast powder, 2.5g / L ammonium nitrate, 4.5g / L sodium chloride, 0.8g / L potassium dihydrogen phosphate, 0.35g / L manganese sulfate, 1.5g / L magnesium sulfate, 0.04g / L ferrous sulfate and 1.5g / L calcium carbonate, and the pH value of the nutrient solution is 7.1;

[0057] In the fifth step, a fermentation agent containing Campylobacter was added to an A / O biochemical pool containing refinery wastewater and filled with fiber filler for reduction treatment. The amount of the fermentation agent containing Campylobacter added was 4% of the volume of the refinery wastewater. During the reduction treatment, the temperature was 35°C, the time was 35 hours, the initial COD of the refinery wastewater was 812 mg / L, the oil content was 20 mg / L, the fiber filler was made of polypropylene, and the filling rate of the fiber filler in the A / O biochemical pool was 40%.

[0058] In Example 7 of the present invention, the A / O biochemical tank employed a four-stage A-tank anaerobic reactor coupled with a single-stage aerobic reactor. Each reactor had identical dimensions. Fiber filler was suspended within each reactor, with a filling rate of 40%. Campylobacter adhered to the fiber filler. The hydraulic retention time of the device was 35 hours.

[0059] Example 8: The application of Campylobacter in reducing refinery sludge is carried out according to the following method:

[0060] In the first step, Campylobacter was inoculated into LB liquid culture medium and cultured on a shaking table to obtain a fermentation seed liquid. The culture temperature was 35°C, the shaking speed was 200 rpm, and the concentration of Campylobacter in the fermentation seed liquid was 1×10 8 CFU / mL or above;

[0061] The second step is to inoculate the fermentation seed liquid into the sterilized low-COD refinery wastewater for activation. The activation conditions include a culture temperature of 35°C, a pH of 8, a shaker speed of 200 rpm, an activation time of 24 hours, and an inoculation amount of the fermentation seed liquid of 4% by volume of the sterilized low-COD refinery wastewater. The COD of the sterilized low-COD refinery wastewater is 800 mg / L.

[0062] In the third step, the activated fermentation seed liquid was transferred to the sterilized high-COD refinery wastewater for reactivation. The activation conditions included a culture temperature of 35°C, a pH of 8, a shaker speed of 200 rpm, an activation time of 24 hours, and an inoculum size of 4% of the volume of the sterilized high-COD refinery wastewater. The COD of the sterilized high-COD refinery wastewater was 800 mg / L.

[0063] In the fourth step, the reactivated fermentation seed liquid was transferred to a fermentation tank containing a nutrient solution to continue fermentation and expansion culture to obtain a starter containing Campylobacter, wherein the culture temperature was 35°C, the pH was 8, the rotation speed was 200 rpm, the pressure of the fermentation tank was 0.04 MPa, the ventilation volume was 1.1:1, the fermentation time was 20 h, the inoculation amount of the reactivated fermentation seed liquid was 4% of the volume percentage of the nutrient solution, and the concentration of Campylobacter in the starter containing Campylobacter was 1×10 8 CFU / ml or above, the composition of the nutrient solution also includes 22g / L glucose, 4g / L peptone, 1.5g / L yeast powder, 2.5g / L ammonium nitrate, 4.5g / L sodium chloride, 0.8g / L potassium dihydrogen phosphate, 0.35g / L manganese sulfate, 1.5g / L magnesium sulfate, 0.04g / L ferrous sulfate and 1.5g / L calcium carbonate, and the pH value of the nutrient solution is 7.1;

[0064] In the fifth step, a fermentation agent containing Campylobacter is added to an A / O biochemical pool containing refinery wastewater and filled with fiber filler for reduction treatment, wherein the addition amount of the fermentation agent containing Campylobacter is 4% of the volume percentage of the refinery wastewater. During the reduction treatment, the temperature is 35°C, the time is 40 hours, the COD of the refinery wastewater is 1503 mg / L, the oil content is 16 mg / L, the fiber filler is made of polypropylene, and the filling rate of the fiber filler in the A / O biochemical pool is 40%.

[0065] In Example 8 of the present invention, the A / O biochemical tank employed a four-stage A-tank anaerobic reactor coupled with a single-stage aerobic reactor. Each reactor had identical dimensions. Fiber filler was suspended within each reactor, with a filling rate of 40%. Campylobacter adhered to the fiber filler. The hydraulic retention time of the device was 40 hours.

[0066] Using the treatment methods of Examples 4 to 8 of the present invention, enriched Campylobacter bacteria were added to an A / O biochemical tank containing refinery wastewater and filled with fiber fillers for volume reduction treatment. This treatment was able to treat refinery wastewater with an initial COD of 800 mg / L to 1600 mg / L and an oil content of no more than 20 mg / L. The residual activated sludge content in the discharged wastewater after Campylobacter treatment was no more than 50 mg / L, achieving a volume reduction of over 90%. The following describes the screening and identification of strains and an evaluation of the effectiveness of using Campylobacter bacteria in reducing refinery wastewater in Examples 7 and 8 of the present invention.

[0067] Experiment 1: Screening and identification of strains

[0068] Test method:

[0069] Step 1: Take 50 mL of activated sludge from the secondary sedimentation tank of the biochemical treatment unit in the industrial water workshop of a petrochemical enterprise and place it in a 250 mL Erlenmeyer flask. Incubate the flask in a shaker at 30°C and 150 rpm for 24 hours. Then, perform a graded dilution and spread the solution on LB plates. Incubate the plate at 30°C for 48 hours, and select individual colonies of different morphologies.

[0070] Step 2: The obtained single colonies of different morphologies were inoculated into LB medium for enrichment, and the culture time was 24 hours to obtain bacterial suspensions of different strains;

[0071] Step 3: sterilize the wastewater from the biochemical section of the industrial water unit of the refinery with high-pressure steam at a sterilization temperature of 120°C, a sterilization pressure of 0.1 MPa, and a sterilization time of 30 minutes;

[0072] Step 4: After the sterilized refinery wastewater is cooled to room temperature, 5 ml of bacterial suspension of different strains is inoculated into 100 mL of sterilized refinery wastewater for reduction treatment. After inoculation, the solution is placed in a shaker at a temperature of 30°C and a speed of 120 rpm for evaluation. The optimal strain is screened by the COD value and ammonia nitrogen degradation rate of the sterilized refinery wastewater after 48 hours of reduction treatment of the bacterial suspension of different strains. At the same time, the optimal strain is morphologically identified.

[0073] Test results: As shown in Table 1, Table 1 shows the COD value and ammonia nitrogen degradation rate of the sterilized refinery wastewater treated by different strains. As shown in Table 1, strain No. 7 (Campylobacter) has the best treatment effect on the COD value and ammonia nitrogen degradation rate of the sterilized refinery wastewater.

[0074] Morphological identification results of strain No. 7: Figure 1 As shown, the strain is characterized by a raised, moist, irregularly shaped colony with a rough surface and a milky white color. Morphological, physiological, biochemical, and molecular biological identification confirmed strain 7 as Campylobacter sp. It was deposited with the General Microbiology Center of the China Culture Collection of Microorganisms on January 18, 2021, under the CGMCC No. 21652. The deposit address is Building 3, No. 1 Beichen West Road, Chaoyang District, Beijing.

[0075] The DNA nucleic acid sequence of strain No. 7 is shown in the sequence listing.

[0076] Strain No. 7 was found to be a conventional facultative anaerobic bacterium after puncture culture.

[0077] Experiment 2: Evaluating the effectiveness of Campylobacter in reducing refinery wastewater

[0078] Test method: According to the treatment methods of Examples 7 and 8 of the present invention, the enriched Campylobacter was used to reduce the amount of refinery wastewater, and the water quality after the reduction treatment was tested. The water quality indicators included chemical oxygen demand (COD), ammonia nitrogen, sulfide, total phosphorus, oil content, and external drainage sludge concentration. The chemical oxygen demand (COD) was determined according to the method specified in GB / T 11914-2015 "Water quality - Determination of chemical oxygen demand - Potassium dichromate method", the ammonia nitrogen was determined according to the method specified in HJ536-2015 "Water quality - Determination of ammonia nitrogen - Nessler's reagent spectrophotometric method", the sulfide was determined according to the method specified in GB / T 16489-2015 "Water quality - Determination of sulfide - Methylene blue spectrophotometric method", the total phosphorus was determined according to the method specified in GB / T 11893-2015 "Water quality - Determination of total phosphorus - Ammonium molybdate spectrophotometric method", and the oil content was determined according to the method specified in HJ 637-2015 "Water quality - Determination of petroleum and animal oils - Infrared spectrophotometry"

[0079] Test results: The water quality indicators of the refinery wastewater before and after the reduction treatment in Example 7 of the present invention are shown in Table 2. As can be seen from Table 2, the indicators of the refinery wastewater before the reduction treatment in Example 7 of the present invention are: COD value of 812 mg / L, oil content of 20 mg / L, sulfide content of 18 mg / L, ammonia nitrogen content of 31 mg / L. The water quality indicators of the refinery wastewater after the Campylobacter reduction treatment are: COD value of 81 mg / L, oil content of 1.5 mg / L, sulfide content of 2.1 mg / L, ammonia nitrogen content of 0.1 mg / L.

[0080] The water quality indicators of the refinery wastewater before and after the reduction treatment in Example 8 of the present invention are shown in Table 3. As can be seen from Table 3, the indicators of the refinery wastewater before the reduction treatment in Example 8 of the present invention are: COD value of 1503 mg / L, oil content of 16 mg / L, sulfide content of 68 mg / L, and ammonia nitrogen content of 27 mg / L. The water quality indicators of the refinery wastewater after the Campylobacter reduction treatment are: COD value of 121 mg / L, oil content of 1.5 mg / L, sulfide content of 6.1 mg / L, and ammonia nitrogen content of 3.1 mg / L.

[0081] It shows that Campylobacter can treat refinery wastewater with an initial COD of 800 mg / L to 1600 mg / L and an oil content not higher than 20 mg / L, and the residual activated sludge content in the discharged wastewater after Campylobacter treatment is no higher than 50 mg / L.

[0082] In summary, the present invention directly reduces wastewater at the source by directly applying Campylobacter to the A / O biochemical pool to treat refinery wastewater with a hydraulic retention time of more than 25 hours. The enriched Campylobacter can treat refinery wastewater with an initial COD of 800 mg / L to 1600 mg / L and an oil content of no more than 20 mg / L. Moreover, the residual activated sludge content in the discharged wastewater after Campylobacter treatment is no more than 50 mg / L, achieving a reduction of more than 90%. This truly achieves a significant reduction in sludge content at the source and greatly reduces the cost of subsequent treatment of environmental pollution.

[0083] The above technical features constitute the embodiments of the present invention, which have strong adaptability and implementation effect. Non-essential technical features can be added or removed according to actual needs to meet the requirements of different situations.

[0084] Sequence Listing <110> PetroChina Karamay Petrochemical Co., Ltd., China National Petroleum Corporation <120> Campylobacter and fermentation agent and their application in reducing refinery sludge <130> 2021 <160> 1 <170> PatentIn version 3.5 <210> 1 <211> 1000 <212> DNA <213> Campylobacter sp. <400> 1 aagtgatccc cttcgacggc tccttccaca aggggttaggc caccggcttc gggtgttacc 60 gactttcatg acttgacggg cggtgtgtac aaggcccggg aacgtattca ccgcagcgtt 120 gctgatctgc gattactagc gactccgact tcatgaggtc gagttgcaga cctcaatccg 180 aactgagacc ggctttttgg gattcgctcc accttacggt atcgcagccc tttgtaccgg 240 ccattgtagc atgcgtgaag cccaagacat aaggggcatg atgatttgac gtcatcccca 300 ccttcctccg agttgacccc ggcagtctcc tatgagtccc cggcataacc cgctggcaac 360 atagaacgag ggttgcgctc gttgcgggac ttaacccaac atctcacgac acgagctgac 420 gacaaccatg caccacctgt acaccgacca caagggggcg accatctctg gccgtttccg 480 gtgtatgtca agccttggta aggttcttcg cgttgcatcg aattaatccg catgctccgc 540 cgcttgtgcg ggcccccgtc aattcctttg agttttagcc ttgcggccgt actccccagg 600 cggggcgctt aatgcgttag ctacgacaca gaaaccgtgg aaaggtccct acatctagcg 660 cccaacgttt acggcatgga ctaccagggt atctaatcct gttcgctccc catgctttcg 720 ctcctcagcg tcagttacgg cccagagatc tgccttcgcc atcggtgttc ctcctgatat 780 ctgcgcattc caccgctaca ccaggaattc caatctcccc taccgcactc tagtctgccc 840 gtacccactg caagcccgag gttgagcctc gggatttcac agcagacgcg acaaaccgcc 900 tacgagctct ttacgcccaa taattccgga caacgcttgc accctacgta ttaccgcggc 960 tgctggcacg tagttagccg gtgcttttct gcaggtaccg 1000

Claims

1. A Campylobacter sp., characterized in that The strain was deposited at the General Microbiology Center of China Culture Collection Administration on January 18, 2021, and its deposit number is CGMCC No.21652.

2. A fermentation culture containing the Campylobacter according to claim 1.

3. The fermentation culture containing Campylobacter according to claim 2, characterized in that The fermentation culture containing Campylobacter comprises Campylobacter and nutrient solution. The concentration of Campylobacter in the fermentation culture containing Campylobacter is 1×10 8 CFU / mL or above.

4. The fermentation culture containing Campylobacter according to claim 3, characterized in that The composition of the nutrient solution includes 20g / L to 24g / L of glucose, 2g / L to 5g / L of peptone, 1g / L to 2g / L of yeast powder, 2g / L to 3g / L of ammonium nitrate, 4g / L to 5g / L of sodium chloride, 0.5g / L to 1g / L of potassium dihydrogen phosphate, 0.3g / L to 0.4g / L of manganese sulfate, 1g / L to 2g / L of magnesium sulfate, 0.02g / L to 0.05g / L of ferrous sulfate and 1g / L to 2g / L of calcium carbonate, and the pH value of the nutrient solution is 7.0 to 7.

2.

5. Use of the fermentation agent containing Campylobacter according to claim 2, 3 or 4 in reducing refinery sludge.

6. Use of the fermentation agent containing Campylobacter according to claim 5 in reducing refinery sludge, characterized in that Follow these steps: In the first step, Campylobacter was inoculated into LB liquid culture medium and cultured on a shaking table to obtain a fermentation seed liquid. The culture temperature was 30°C to 35°C, the shaking speed was 100 rpm to 250 rpm, and the concentration of Campylobacter in the fermentation seed liquid was 1×10 8 CFU / mL or above; The second step is to inoculate the fermentation seed liquid into the sterilized low-COD refinery wastewater for activation, wherein the activation conditions include a culture temperature of 30°C to 35°C, a pH of 8 to 10, a shaker speed of 100 rpm to 250 rpm, an activation time of 24 hours, an inoculation amount of the fermentation seed liquid of 3% to 6% by volume of the sterilized low-COD refinery wastewater, and a COD of 500 mg / L to 800 mg / L of the sterilized low-COD refinery wastewater; The third step is to transfer the activated fermentation seed liquid to the sterilized high-COD refinery wastewater for reactivation, wherein the activation conditions include a culture temperature of 30°C to 35°C, a pH of 8 to 10, a shaker speed of 100 rpm to 250 rpm, an activation time of 24 hours, and an inoculation amount of the activated fermentation seed liquid of 3% to 6% by volume of the sterilized high-COD refinery wastewater, and the COD of the sterilized high-COD refinery wastewater is 800 mg / L to 1600 mg / L. In the fourth step, the reactivated fermentation seed liquid is transferred to a fermentation tank containing a nutrient solution for continued fermentation and expansion culture to obtain a starter containing Campylobacter, wherein the culture temperature is 28°C to 38°C, the pH is 7.5 to 9.5, the rotation speed is 200 rpm, the pressure of the fermentation tank is 0.03 MPa to 0.06 MPa, the ventilation volume is 1.0 to 1.3:1, the fermentation time is 16 h to 20 h, the inoculation amount of the reactivated fermentation seed liquid is 3% to 6% of the volume percentage of the nutrient solution, and the concentration of Campylobacter in the starter containing Campylobacter is 1×10 8 CFU / ml or above; In the fifth step, a fermentation agent containing Campylobacter is added to an A / O biochemical pool containing refinery wastewater and filled with fiber filler for reduction treatment, wherein the addition amount of the fermentation agent containing Campylobacter is 3% to 6% of the volume percentage of the refinery wastewater. During the reduction treatment, the temperature is 30°C to 35°C, the time is not less than 25 hours, the initial COD of the refinery wastewater is 800mg / L to 1600mg / L, the oil content is not higher than 20mg / L, the fiber filler is made of polypropylene, and the filling rate of the fiber filler in the A / O biochemical pool is 20% to 70%.

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