Aspergillus polymorpha NXY1, mycelial pellets and their application in shale gas produced water treatment

Abstract

The invention relates to the technical field of biological sewage treatment, and in particular to Aspergillus polydorum NXY1, mycelial pellets and their application in shale gas produced water treatment. Aspergillus polydorum NXY1 provided by the invention is a salt-tolerant bacterium, and its deposit number is CGMCC No. 40215. The Aspergillus polydorum NXY1 can form mycelial pellets and degrade organic matter in shale gas produced water; at the same time, the mycelial pellets formed by the fungus can adsorb other microorganisms and activated sludge to form a composite microbial system that is easy to separate solid and liquid, and can efficiently remove organic matter and ammonia nitrogen from shale gas produced water in different salinity ranges (0-8wt.%). The problem that the activated sludge method requires a secondary sedimentation tank for mud and water separation is solved, and the construction of the secondary sedimentation tank can be reduced. At the same time, the problem that sludge is prone to floating when the activated sludge method treats shale gas produced water is solved, which is helpful to achieve green, economical and efficient treatment and disposal of shale gas produced water.

Description

Technical Field

[0001] The invention relates to the technical field of biological sewage treatment, and in particular to Aspergillus polymorpha NXY1, mycelial pellets and applications thereof in shale gas produced water treatment. Background Art

[0002] Shale gas is a clean, low-carbon, unconventional natural gas resource, with methane as its main component. Due to the small pores and low permeability of shale gas reservoirs, its extraction must be achieved through high-pressure hydraulic fracturing of rock formations. After hydraulic fracturing, the liquid gradually flows back to the surface, which is generally called fracturing flowback fluid in the early stage and produced water in the later stage, collectively referred to as shale gas produced water.

[0003] Shale gas produced water contains a large number of chemical additives from fracturing fluids, such as gelling agents, drag reducers, cross-linking agents, bactericides and surfactants, as well as metal ions from the formation and organic matter such as alkanes and aromatic hydrocarbons. In general, shale gas produced water has high and fluctuating total suspended solids (TSS), total organic carbon (TOC), ammonia nitrogen and salinity. Improper treatment of shale gas produced water can cause serious problems such as environmental pollution. Green, economical and efficient treatment of shale gas produced water is an important requirement in the context of large-scale shale gas exploitation.

[0004] As for the removal of organic matter and ammonia nitrogen from shale gas produced water, advanced oxidation methods, including electrocatalytic oxidation, Fenton oxidation and ozone oxidation, have problems such as high equipment and operating costs. In comparison, biological methods are low-cost and environmentally friendly, and have advantages in industrial wastewater treatment processes. However, the sludge settling of the activated sludge method in the biological method is slow, and a secondary sedimentation tank is required for mud and water separation, so the sewage treatment plant occupies a large area; and the activated sludge method is used for shale gas produced water, which has high salinity and large water quality fluctuations, and problems such as sludge floating will occur. Summary of the invention

[0005] In order to solve the above problems, the present invention provides Aspergillus polydorum NXY1, mycelial pellets and their use in shale gas produced water treatment. The Aspergillus polydorum NXY1 provided by the present invention can be used in the shale gas produced water treatment process, which can not only effectively remove organic matter and ammonia nitrogen in shale gas produced water, but also can adsorb activated sludge, reduce the construction of secondary sedimentation tanks, and solve the problem of sludge floating easily when treating shale gas produced water by activated sludge method.

[0006] In order to achieve the above object, the present invention provides the following technical solutions:

[0007] The invention provides a strain of Aspergillus sydowii NXY1 capable of forming a ball. The preservation number of the Aspergillus sydowii NXY1 is CGMCC No.40215.

[0008] The present invention also provides a mycelial pellet that can remove or assist in removing organic matter and / or ammonia nitrogen in water. The mycelial pellet is obtained by culturing the above-mentioned Aspergillus polymorpha NXY1.

[0009] The present invention also provides a method for preparing the mycelial pellets, comprising inoculating a spore suspension containing the Aspergillus polydorum NXY1 into a culture medium, culturing, and obtaining the mycelial pellets;

[0010] The components of the culture medium include: glucose 5-80 g / L, NH 4 Cl 0.5~8g / L, KH 2 PO 4 0.3~6g / L, MgSO 4 7H 2 O 0.25~4g / L and NaCl 0~80g / L.

[0011] Preferably, the culture conditions include: temperature 20-35°C, time 20-200h, and rotation speed 100-400rpm.

[0012] Preferably, the volume ratio of the spore suspension to the culture medium is 1:99; the spore concentration of the spore suspension is 10 4 ~10 9 spores / mL.

[0013] Preferably, the method for preparing the spore suspension comprises:

[0014] The Aspergillus polydophorus NXY1 is inoculated on a PDA medium, and after slant culture, the spores are washed into sterile physiological saline to obtain the spore suspension.

[0015] Preferably, the conditions for the slant culture include: temperature of 20-35°C and time of 3-15 days.

[0016] The present invention also provides the use of the above-mentioned Aspergillus polymorpha NXY1 or the above-mentioned mycelial pellet or the mycelial pellet prepared by the above-mentioned preparation method in removing or assisting in removing organic matter and / or ammonia nitrogen in liquid.

[0017] Preferably, the liquid includes liquid returned to the surface after shale gas hydraulic fracturing.

[0018] Preferably, the liquid includes fracturing flowback fluid and / or produced water.

[0019] Beneficial effects:

[0020] The present invention provides a ball-forming Aspergillus sydowii NXY1, and the deposit number of the Aspergillus sydowii NXY1 is CGMCC No. 40215. The Aspergillus sydowii NXY1 provided by the present invention is a salt-tolerant bacterium, which can form mycelial balls and degrade organic matter in shale gas produced water; at the same time, the mycelial balls formed by the fungus can adsorb other microorganisms and activated sludge to form a composite microbial system that is easy to separate solid and liquid, and can efficiently remove organic matter and ammonia nitrogen from shale gas produced water in different salinity ranges (0-8wt.%); the problem that the activated sludge method requires a secondary sedimentation tank for mud and water separation can be solved, and the construction of the secondary sedimentation tank can be reduced. At the same time, the problem that sludge is prone to floating when the activated sludge method treats shale gas produced water is solved, which is helpful to achieve green, economical and efficient treatment and disposal of shale gas produced water.

[0021] Biological Deposit Description

[0022] Aspergillus sydowii NXY1, Latin name Aspergillus sydowii, was deposited in the China General Microbiological Culture Collection on June 17, 2022. The deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, and the deposit number is CGMCC No.40215. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required to be used in the embodiments are briefly introduced below.

[0024] Figure 1 The colony morphology of NXY1 strain on the culture medium;

[0025] Figure 2 The morphological characteristics of NXY1 strain under a microscope;

[0026] Figure 3 is the phylogenetic tree of NXY1 strain;

[0027] Figure 4 The mycelial ball morphology prepared in Example 2;

[0028] Figure 5 The mycelial ball morphology prepared in Example 3;

[0029] Figure 6 The mycelial ball morphology prepared in Example 4;

[0030] Figure 7 The mycelial ball morphology prepared in Example 5;

[0031] Figure 8 The mycelial ball morphology prepared in Example 6;

[0032] Fig. 9 The mycelial ball morphology prepared in Example 7;

[0033] Fig.10 The mycelial ball morphology prepared in Comparative Example 1;

[0034] Fig.11 This is the morphology of the mycelium ball prepared in Comparative Example 2. DETAILED DESCRIPTION

[0035] The invention provides a spherical Aspergillus sydowii NXY1 strain, wherein the Aspergillus sydowii NXY1 has a deposit number of CGMCC No.40215.

[0036] The conidia of Aspergillus polydorum NXY1 of the present invention are spherical or nearly spherical, with obviously rough walls; the microconidial heads are loose columnar or scattered; the apices are small and elliptical, and almost the entire surface is fertile; and the spore-producing structure is double-layered.

[0037] The Aspergillus polydodecene NXY1 of the present invention has a round colony on a Czapek medium, a powdery texture, a white edge, a dark green interior and a small amount of dark brown, no exudate, and no radial grooves; the back of the culture medium is white, and no pigment diffusion; on a Czapek yeast culture medium, the colony is round, a powdery texture, a white edge, a light orange interior, radial grooves are formed, and a small amount of light orange exudate is present in the grooves; the back of the culture medium is dark brown-red; on a malt extract agar culture medium, the colony is round with irregular edges, a velvety texture, a white edge, a light bean green interior, and radial grooves; the back of the culture medium is dark purple-brown.

[0038] The Aspergillus polydophorus NXY1 of the present invention can tolerate a salinity of 0 to 8 wt.%, and can efficiently remove organic matter and ammonia nitrogen from shale gas produced water in different salinity ranges (0 to 8 wt.%).

[0039] In the present invention, the nucleotide sequence of the ITS rDNA of Aspergillus polymorpha NXY1 preferably includes the sequence shown in SEQ ID NO.1, which is as follows:

[0040] .

[0041] The Aspergillus polydophorus NXY1 of the present invention has the characteristic of being able to form balls, has a better separation effect during mud-water separation, requires a short sedimentation time, can save separation time, correspondingly increase reaction time and influent organic load, and improve treatment efficiency; at the same time, can reduce the construction of a secondary sedimentation tank, can be simply separated, is suitable for removing or assisting in removing pollutants in liquids, and in particular can remove or assist in removing organic matter and / or ammonia nitrogen in liquids, and is particularly suitable for shale gas produced water treatment processes.

[0042] The present invention also provides a mycelial pellet that can remove or assist in removing organic matter and / or ammonia nitrogen in water. The mycelial pellet is obtained by culturing the Aspergillus polymorpha NXY1 described in the above scheme.

[0043] The mycelium ball of the present invention is spherical, ellipsoidal or mallet-shaped, and the diameter of the mycelium ball is 500-15000 μm; the sedimentation speed of the mycelium ball is 0.5-2 cm / s.

[0044] The mycelial balls described in the present invention have the characteristics of easy solid-liquid separation, fast sedimentation speed, and strong organic matter degradation and adsorption capacity. They can quickly degrade organic matter and ammonia nitrogen independently or by adsorbing activated sludge, solving the problem that the activated sludge method requires a secondary sedimentation tank for mud-water separation, and can reduce the construction of secondary sedimentation tanks. At the same time, it solves the problem that sludge floating is easy to occur when the activated sludge method is used to treat shale gas produced water. It has significant application potential in the field of shale gas produced water treatment.

[0045] The present invention also provides a method for preparing the mycelial pellets described in the above scheme, wherein a spore suspension containing the Aspergillus polymorpha NXY1 described in the above scheme is inoculated into a culture medium, and cultured to obtain the mycelial pellets.

[0046] In the present invention, the method for preparing the spore suspension preferably comprises:

[0047] 1) using an inoculating loop to streak the Aspergillus polydophorus NXY1 on a sterile potato dextrose agar (PDA) medium slant;

[0048] 2) placing the slope marked in step 1) in an incubator at 20-35° C. and culturing for 3-15 days;

[0049] 3) Wash the spores on the slope in step 2) into sterile physiological saline to prepare a spore suspension.

[0050] In the present invention, the culture temperature in step 2) is preferably 25-32°C, more preferably 30°C; the culture time is preferably 5-15d, more preferably 5-10d or 11-15d, more preferably 15d.

[0051] After obtaining the spore suspension, the present invention inoculates the spore suspension into a culture medium and cultures the culture medium to obtain the mycelium ball.

[0052] In the present invention, the volume ratio of the spore suspension to the culture medium is preferably 1:99; the spore concentration of the spore suspension is preferably 10 4 ~10 9 spores / mL, further preferably including 10 4 , 10 5 , 5×10 5 , 10 6 , 2×10 6 , 10 7 , 10 8 or 10 9 spores / mL, more preferably 10 7 The invention can cultivate mycelium balls with moderate volume, complete structure and fast sedimentation speed through appropriate inoculation amount.

[0053] The sterile culture medium of the present invention comprises 5 to 80 g / L of glucose, preferably 8 to 40 g / L, more preferably 9 to 20 g / L, and more preferably 10 g / L. In the present invention, the glucose preferably comprises anhydrous glucose.

[0054] The sterile culture medium of the present invention comprises NH 4 Cl 0.5 to 8 g / L, preferably 0.6 to 5 g / L, more preferably 0.8 to 3 g / L, and even more preferably 1 g / L.

[0055] The sterile culture medium of the present invention comprises KH 2 PO 4 0.3 to 6 g / L, preferably 0.4 to 3 g / L, more preferably 0.5 to 1 g / L, and even more preferably 0.72 g / L.

[0056] The sterile culture medium of the present invention comprises MgSO 4 7H 2 O 0.25 to 4 g / L, preferably 0.3 to 2 g / L, more preferably 0.4 to 1 g / L, and even more preferably 0.5 g / L.

[0057] The sterile culture medium of the present invention includes 0-80 g / L of NaCl; the concentration of NaCl in the culture medium is preferably determined according to the salinity of the wastewater to be treated, specifically including: the value before the percentage of the salinity of the wastewater to be treated is recorded as A, that is, when the salinity is 8%, A is 8, and the concentration of NaCl in the culture medium = A×10 g / L; wherein when A is less than 1, NaCl may not be added to the culture medium. The present invention determines the concentration of NaCl in the culture medium according to the salinity of the wastewater to be treated, so that the mycelial balls obtained by culture can directly play the role of degrading and adsorbing organic matter in saline wastewater, without the need for an adaptation process, thereby shortening the debugging time of the biochemical process.

[0058] In the present invention, the culture preferably includes constant temperature culture; the culture temperature is preferably 20-35°C, preferably 25-32°C, and further more preferably 30°C; the culture time is preferably 20-200h, further preferably 50-150h, and further more preferably 120h; the culture rotation speed is preferably 100-400rpm, further preferably 130-200rpm, and further more preferably 160rpm.

[0059] The present invention also provides the use of the above-mentioned Aspergillus polytoxin NXY1 or the above-mentioned mycelial pellet or the mycelial pellet prepared by the above-mentioned method in removing or assisting in removing organic matter and / or ammonia nitrogen in liquid.

[0060] In the present invention, the liquid preferably includes liquid returned to the surface after shale gas hydraulic fracturing; the liquid returned to the surface after shale gas hydraulic fracturing preferably includes fracturing return fluid and / or produced water.

[0061] In the present invention, the mycelium pellet is preferably used as a carrier to adsorb activated sludge or other functional microorganisms, and degrade organic matter in the liquid or remove ammonia nitrogen in the liquid, and is particularly suitable for organic matter and ammonia nitrogen in shale gas production water.

[0062] In the present invention, the organic matter preferably includes one or more of polyacrylamide, guar gum, isopropanol, aromatic hydrocarbons and long-chain hydrocarbons.

[0063] In order to further illustrate the present invention, the Aspergillus polymorpha NXY1, mycelial pellets and their application in shale gas produced water treatment provided by the present invention are described in detail below in combination with the examples and drawings, but they should not be construed as limiting the scope of protection of the present invention.

[0064] The method for determining the average sedimentation velocity of the following examples or comparative examples is as follows: sodium chloride solution is placed in a 1000 mL measuring cylinder, 30 mycelial balls are randomly taken, and one mycelial ball is taken each time and released from the top of the liquid surface without applying an initial velocity, and the time it takes to settle to the bottom of the measuring cylinder is recorded, and the sedimentation velocity is calculated. The average value is taken to obtain the average sedimentation velocity of the mycelial balls in the sodium chloride solution.

[0065] Example 1

[0066] Isolation and Identification of NXY1 from Aspergillus sydowii

[0067] The Aspergillus polymorpha NXY1 described in the present invention is isolated from shale gas produced water in Xuyong County, Luzhou, Sichuan.

[0068] The details are as follows:

[0069] Yeast powder was added to the shale gas produced water, and the concentration of yeast powder in the shale gas produced water was 1g / L. After one day of enrichment culture, 10mL was transferred to 100mL of new shale gas produced water supplemented with yeast powder (yeast powder concentration was 1g / L). The enrichment was repeated three times, each time for one day, for a total of four enrichment cultures.

[0070] Agar was added to shale gas produced water (the mass volume ratio of agar to shale gas produced water was 20g:1L) to prepare shale gas produced water agar plates; the bacterial liquid in the shale gas produced water after enrichment for 4 times was dipped and separated by streaking on the shale gas produced water agar plates, and the fungal strain was obtained after multiple purifications, which was numbered as NXY1. Then, the spores were washed, diluted and spread to obtain a single fungal colony.

[0071] The fungal colonies were inoculated into Czapek's medium, Czapek's yeast medium and malt agar medium respectively, and cultured in an incubator at 30°C for 10 days. Figure 1 As shown, A is the front of Czapek's agar medium, B is the front of Czapek's yeast extract agar medium, and C is the front of malt extract agar medium; D is the back of Czapek's agar medium, E is the back of Czapek's yeast extract agar medium, and F is the back of malt extract agar medium.

[0072] Depend on Figure 1 It can be seen that on the Czapek medium, the colonies are round, powdery, white on the edges, dark green inside and a small amount of dark brown, no exudate, no radial grooves; the back of the medium is white, and there is no pigment diffusion. On the Czapek yeast medium, the colonies are round, powdery, white on the edges, light orange inside, forming radial grooves, and a small amount of light orange exudate in the grooves; the back of the medium is dark brown-red. On the malt agar medium, the colonies are round with irregular edges, velvety texture, white on the edges, light bean green inside, with radial grooves; the back of the medium is dark purple-brown.

[0073] The structure of the NXY1 strain under a microscope is shown below: Figure 2 As shown in the figure, the conidia are spherical or nearly spherical, with a rough wall; the microconidia are loosely cylindrical or scattered; the apices are small and elliptical, and almost the entire surface is fertile; the conidial structure is double-layered, and NXY1 was preliminarily identified as an Aspergillus fungus based on the colony morphology.

[0074] The genomic DNA of NXY1 strain was extracted using a fungal genomic DNA extraction kit and used as a template for ITS rDNA amplification. PCR amplification was performed using the fungal universal primers ITS1 (5'-TCCGTAGGTGAACCTGCGG-3', SEQ ID NO.2) and ITS4 (5'-TCCTCCGCTTATTGATATGC-3', SEQ ID NO.3). PCR reaction system (20 μL): 10× buffer 2 μL; dNTP Mix (2.5 mM) 1.6 μL; ITS1 and ITS4 0.8 μL each; DNA template 0.5 μL; Taq enzyme (5 U / μL) 0.2 μL; sterile water 14.1 μL. PCR reaction program: 95℃ pre-denaturation for 5 min; 95℃ denaturation for 30 s, 55℃ annealing for 30 s, 72℃ extension for 40 s, 35 cycles; 72℃ post-extension for 10 min.

[0075] The PCR product obtained by amplification was recovered by 1% agarose electrophoresis, and purified and sequenced by Beijing Liuhe BGI Technology Co., Ltd. The ITS rDNA sequencing result of the NXY1 strain is shown in SEQ ID NO.1. The obtained sequence was compared with the NCBI GenBank DNA sequence database for homology analysis, and the results showed that the obtained strain had a high homology with the sequence of Aspergillus sydowii SCAU241 (OK275112.1). The phylogenetic tree was constructed by the neighbor-joining method, as shown in Figure 3 As shown. Based on homology comparison and phylogenetic analysis, the strain was Aspergillus sydowii and was named Aspergillus sydowii NXY1. The strain was deposited in the General Microbiology Center (CGMCC) of the China Microbiological Collection Administration Committee in Chaoyang District, Beijing on June 17, 2022, with the deposit number CGMCC No.40215.

[0076] Example 2

[0077] A mycelium ball capable of removing or assisting in removing organic matter and / or ammonia nitrogen in water is prepared by the following method:

[0078] 1) taking out the glycerol tube containing the Aspergillus polydophorus NXY1 strain described in Example 1 from a -80°C refrigerator, and streaking on a slant of a sterile potato dextrose agar (PDA) medium using an inoculating loop;

[0079] 2) Place the marked slope in an incubator at 30°C for 15 days;

[0080] 3) Wash the spores on the slope into sterile saline in a clean bench to prepare a spore suspension, and count the number of spores per milliliter of the spore suspension under a microscope using a hemocytometer; adjust the spore concentration of the spore suspension to 10 with sterile saline. 7 spores / mL;

[0081] 4) Inoculate 1 mL of spore suspension into a shake flask containing 99 mL of sterile culture medium; the sterile culture medium consists of 10 g of anhydrous glucose, 1 g of NH 4 Cl, 0.72 g KH 2 PO 4 、0.5g MgSO 4 7H 2 O and 30g NaCl, add deionized water to 1L, and then sterilize;

[0082] 5) The shake flask inoculated with the spore suspension was placed in a shaker and cultured at 30° C. and 160 rpm for 120 h to obtain mycelial balls.

[0083] The mycelium ball is white and spherical. Figure 4 As shown, the average diameter is 1680 μm and the average sedimentation velocity in 30 g / L NaCl solution is 1.15 cm / s.

[0084] Example 3

[0085] A mycelium ball capable of removing or assisting in removing organic matter and / or ammonia nitrogen in water is prepared by the following method:

[0086] Steps 1) to 5) are similar to those in Example 2, except that the spore concentration of the spore suspension in step 3) is 10 6 spores / mL.

[0087] The mycelium ball is white and spherical. Figure 5 As shown, the average diameter is 2850 μm and the average sedimentation velocity in 30 g / L NaCl solution is 1.30 cm / s.

[0088] Example 4

[0089] A mycelium ball capable of removing or assisting in removing organic matter and / or ammonia nitrogen in water is prepared by the following method:

[0090] Steps 1) to 5) are similar to those in Example 2, except that the spore concentration of the spore suspension in step 4) is 10 5 spores / mL.

[0091] The mycelium ball is white and spherical. Figure 6 As shown, the average diameter is 3050 μm and the average sedimentation velocity in 30 g / L NaCl solution is 1.05 cm / s.

[0092] Example 5

[0093] A mycelium ball capable of removing or assisting in removing organic matter and / or ammonia nitrogen in water is prepared by the following method:

[0094] Steps 1) to 5) are similar to those in Example 2, except that the spore concentration of the spore suspension in step 4) is 10 4 spores / mL.

[0095] The mycelium ball is white and spherical. Figure 7 As shown, the average diameter is 3150 μm and the average sedimentation velocity in 30 g / L NaCl solution is 1.08 cm / s.

[0096] Example 6

[0097] A mycelium ball capable of removing or assisting in removing organic matter and / or ammonia nitrogen in water is prepared by the following method:

[0098] Steps 1) to 5) are similar to those in Example 2, except that the sterile culture medium in step 4) does not contain NaCl;

[0099] The mycelium ball is white and spherical. Figure 8 As shown, the average diameter is 1850 μm, the average sedimentation velocity in 30 g / L NaCl solution is 0.35 cm / s, and the average sedimentation velocity in 8 g / L NaCl solution is 0.85 cm / s.

[0100] Example 7

[0101] A mycelium ball capable of removing or assisting in removing organic matter and / or ammonia nitrogen in water is prepared by the following method:

[0102] Steps 1) to 5) are similar to those in Example 2, except that the sterile culture medium in step 4) consists of 10 g anhydrous glucose, 1 g NH 4 Cl, 0.72 g KH 2 PO 4 、0.5g MgSO 4 7H 2 O and 80g NaCl, add deionized water to 1L, and then sterilize;

[0103] The mycelium ball is white and spherical. Fig. 9 As shown, the average diameter is 1350 μm and the average sedimentation velocity in 80 g / L NaCl solution is 0.82 cm / s.

[0104] Comparative Example 1

[0105] A fungal mycelium ball is prepared by the following method:

[0106] Steps 1) to 5) are similar to those in Example 2, except that the spore concentration of the spore suspension in step 4) is 10 2 spores / mL, a large number of loose hyphae and a small number of mycelial balls were obtained after cultivation (see Fig.10 ).

[0107] Depend on Figure 4 and Fig.10 It can be seen that if the inoculation amount is too low, the effect of the prepared mycelium balls will be poor.

[0108] Comparative Example 2

[0109] Steps 1) to 5) are similar to those in Example 2, except that the shaking speed in step 4) is 10 rpm; after cultivation, a large number of long, loose mycelial masses are obtained (see Fig.11 ).

[0110] Depend on Figure 4 and Fig.11 It can be seen that the shaking speed is too low and dense mycelium balls cannot be prepared.

[0111] Comparative application example 1

[0112] Activated sludge (taken from the activated sludge pool of the sewage treatment plant of East China University of Science and Technology on November 20, 2021, the same below) was added to the shale gas produced water of Changning, Sichuan (salinity is 3wt.%, COD concentration is 820mg / L, ammonia nitrogen concentration is 65mg / L), the dosage is 2.5g / L, and the sequencing batch reactor (reactor height 101cm, inner diameter 6.5cm, effective volume 2.2L) is operated for 6h per cycle, water exchange ratio is 50%, pH is maintained at 7.0-8.5, and temperature is maintained at 25℃-30℃. The organic load in the system is 1.64kgCOD / (m 3 ·d). In the first three days, a large amount of sludge floated up and flowed away. On the fourth day, the amount of sludge was only 0.7g / L. On the ninth day, the amount of sludge was 1.2g / L, the organic matter removal rate was only 32%, and the ammonia nitrogen removal rate was only 11%. The removal effect of organic matter and ammonia nitrogen in wastewater was not good.

[0113] Application Example 1

[0114] In a clean bench, the mycelium pellets prepared in Example 2 were inoculated with 2.5 g / L of biomass into a sterilized shake flask filled with simulated shale gas pressure return liquid, and cultured at 30°C and 160 rpm for 72 h. The simulated shale gas produced water consisted of 200 mg polyacrylamide, 220 mg isopropanol, 100 mg guar gum, 25 mg ethylene glycol, 40 mg n-hexadecane, 20 mg phenol, 30 mg aniline, and NH 4 Cl 153mg / L, CaCl 2 40mg, MgSO 4 7H 2 O 37mg, FeSO 4 7H 2 O 37mg and NaCl 30g, add deionized water to 1L, and then sterilize to prepare; the salinity of the simulated shale gas pressure return liquid is 3wt.%, the COD concentration is 850mg / L, and the ammonia nitrogen concentration is 40mg / L.

[0115] After 72 hours, the organic matter removal rate can reach 45.9%, and the ammonia nitrogen removal rate can reach 25.6%. The organic matter removal load per unit biomass is 0.052kg COD / (kg MLSS·d), and the ammonia nitrogen removal load per unit biomass is 0.0014kg NH 4 +-N / (kg MLSS·d).

[0116] Application Example 2

[0117] The mycelium pellets prepared in Example 2 were added to shale gas production water (salinity of 3wt.%, COD concentration of 820mg / L, ammonia nitrogen concentration of 65mg / L) in Changning, Sichuan, with a dosage of 2.5g / L. The reactor (reactor height 101cm, inner diameter 6.5cm, effective volume 2.2L) was operated in a sequencing batch reactor with a cycle of 6h, a water exchange ratio of 50%, a pH maintained at 7.0-8.5, and a temperature maintained at 25°C-30°C. The organic load in the system was 1.64kg COD / (m 3 ·d). In the first 9 days, the removal rates of organic matter and ammonia nitrogen gradually increased. After the 9th day, the organic matter removal rate could reach 69%, and the ammonia nitrogen removal rate could reach 30%. At this time, the mycelium balls acted as carriers to absorb the functional microorganisms to jointly treat the salt-containing organic wastewater. The total biomass reached 3.6g / L, and the removal effect was more efficient and stable than that of comparative application example 1.

[0118] Application Example 3

[0119] The mycelium pellets prepared in Example 2 were added to shale gas production water (salinity of 3wt.%, COD concentration of 820mg / L, ammonia nitrogen concentration of 65mg / L) in Changning, Sichuan, with a dosage of 2.5g / L. 2.5g / L of activated sludge was added at the same time. The reactor (reactor height 101cm, inner diameter 6.5cm, effective volume 2.2L) was operated in a sequencing batch reactor with a cycle of 6h, a water exchange ratio of 50%, a pH maintained at 7.0-8.5, and a temperature maintained at 25°C-30°C. The organic load in the system was 1.64kg COD / (m 3 ·d). In the first three days, the sludge was adsorbed by the mycelium balls in large quantities, and there was no obvious floating or loss phenomenon. The removal rates of organic matter and ammonia nitrogen gradually increased. After the 9th day, the organic matter removal rate could reach 76%, and the ammonia nitrogen removal rate could reach 55%. At this time, the mycelium balls acted as carriers to adsorb the activated sludge to jointly treat the salt-containing organic wastewater, and the total biomass reached 5.4g / L; the removal effect was more efficient and stable than that of comparative application example 1.

[0120] Application Example 4

[0121] The mycelium pellets prepared in Example 3 were added to shale gas production water (salinity of 3wt.%, COD concentration of 820mg / L, ammonia nitrogen concentration of 65mg / L) in Changning, Sichuan, with a dosage of 2.5g / L. The reactor (reactor height 101cm, inner diameter 6.5cm, effective volume 2.2L) was operated in a sequencing batch reactor with a cycle of 6h, a water exchange ratio of 50%, a pH maintained at 7.0-8.5, and a temperature maintained at 25°C-30°C. The organic load in the system was 1.64kg COD / (m 3·d). In the first 13 days, the removal rates of organic matter and ammonia nitrogen gradually increased; after the 13th day, the organic matter removal rate could reach 69%, and the ammonia nitrogen removal rate could reach 30%. At this time, the mycelium balls acted as carriers to absorb the functional microorganisms to jointly treat the salt-containing organic wastewater, and the total biomass reached 3.2g / L. The removal effect was more efficient and stable than that of comparative application example 1.

[0122] Application Example 5

[0123] The mycelium pellets prepared in Example 4 were added to shale gas production water (salinity of 3wt.%, COD concentration of 820mg / L, ammonia nitrogen concentration of 65mg / L) in Changning, Sichuan, with a dosage of 2.5g / L. The reactor (reactor height 101cm, inner diameter 6.5cm, effective volume 2.2L) was operated in a sequencing batch reactor with a cycle of 6h, a water exchange ratio of 50%, a pH maintained at 7.0-8.5, and a temperature maintained at 25°C-30°C. The organic load in the system was 1.64kg COD / (m 3 ·d). In the first 16 days, the removal rates of organic matter and ammonia nitrogen gradually increased; after the 16th day, the organic matter removal rate could reach 69%, and the ammonia nitrogen removal rate could reach 29%. At this time, the mycelium balls acted as carriers to absorb the functional microorganisms to jointly treat the salt-containing organic wastewater, and the total biomass reached 3.0g / L. The removal effect was more efficient and stable than that of comparative application example 1.

[0124] Application Example 6

[0125] The mycelium pellets prepared in Example 5 were added to shale gas production water (salinity of 3wt.%, COD concentration of 820mg / L, ammonia nitrogen concentration of 65mg / L) in Changning, Sichuan, with a dosage of 2.5g / L. The reactor (reactor height 101cm, inner diameter 6.5cm, effective volume 2.2L) was operated in a sequencing batch reactor with a cycle of 6h, a water exchange ratio of 50%, a pH maintained at 7.0-8.5, and a temperature maintained at 25°C-30°C. The organic load in the system was 1.64kg COD / (m 3 ·d). In the first 16 days, the removal rates of organic matter and ammonia nitrogen gradually increased; after the 17th day, the organic matter removal rate could reach 69%, and the ammonia nitrogen removal rate could reach 28%. At this time, the mycelium balls acted as carriers to absorb the functional microorganisms to jointly treat the salt-containing organic wastewater, and the total biomass reached 2.8g / L. The removal effect was more efficient and stable than that of comparative application example 1.

[0126] Application Example 7

[0127] The mycelium pellets prepared in Example 6 were added to shale gas production water (salinity of 3wt.%, COD concentration of 820mg / L, ammonia nitrogen concentration of 65mg / L) in Changning, Sichuan, with a dosage of 2.5g / L. The reactor (reactor height 101cm, inner diameter 6.5cm, effective volume 2.2L) was operated in a sequencing batch reactor with a cycle of 6h, a water exchange ratio of 50%, a pH maintained at 7.0-8.5, and a temperature maintained at 25°C-30°C. The organic load in the system was 1.64kg COD / (m 3 ·d). On the first day, the mycelial balls were difficult to settle, and floated and lost seriously; on the third day, the total biomass was only 0.5g / L, the organic matter removal rate was only 8%, and the ammonia nitrogen removal rate was only 4%. The reason may be that the mycelial balls have not been cultured at a suitable salinity, and no salt ions have accumulated in the cells or a large amount of extracellular polymers have been released outside the cells. When directly used in shale gas produced water with a salinity of 3wt.%, it is difficult to resist the high salt environment, resulting in floating and being discharged with the effluent. The biomass is too low, and the organic matter and ammonia nitrogen removal rates are also correspondingly poor.

[0128] Application Example 8

[0129] The mycelium pellets prepared in Example 6 were added to the shale gas pressure return liquid in Chongqing (salinity of 0.8wt.%, COD concentration of 1050mg / L, ammonia nitrogen concentration of 102mg / L), the dosage was 2.5g / L, and the reactor was operated in a sequencing batch mode (reactor height 101cm, inner diameter 6.5cm, effective volume 2.2L), a cycle of 6h, water exchange ratio 50%, pH maintained at 7.0-8.5, and temperature maintained at 25℃-30℃. The organic load in the system was 2.1kg COD / (m 3 ·d). On the first day, the mycelial pellets settled slowly, but could be completely retained in the reactor; in the first 7 days, the pellets gradually settled faster, and the organic matter and ammonia nitrogen removal rates gradually increased. After the 7th day, the organic matter removal rate could reach 73%, and the ammonia nitrogen removal rate could reach 42%. At this time, the mycelial pellets acted as carriers to absorb the functional microorganisms to jointly treat the salt-containing organic wastewater, and the total biomass reached 3.4g / L. The removal effect was more efficient and stable than that of Application Example 7.

[0130] Application Example 9

[0131] In a clean bench, the mycelium pellets prepared in Example 7 were inoculated with a biomass of 2.5 g / L into a sterilized shake flask containing simulated shale gas pressure return liquid, and cultured at 30°C and 160 rpm for 72 h. The simulated shale gas produced water consisted of 200 mg polyacrylamide, 220 mg isopropanol, 100 mg guar gum, 25 mg ethylene glycol, 40 mg n-hexadecane, 20 mg phenol, 30 mg aniline, and NH 4 Cl 153mg / L, CaCl 2 40mg, MgSO4 7H 2 O 37mg, FeSO 4 7H 2 O 37mg and NaCl 80g, add deionized water to 1L, and then sterilize to prepare; the salinity of the simulated shale gas pressure return liquid is 8wt.%, the COD concentration is 850mg / L, and the ammonia nitrogen concentration is 40mg / L.

[0132] After 72 hours, the organic matter removal rate can reach 41.2%, and the ammonia nitrogen removal rate can reach 23.6%. The organic matter removal load per unit biomass is 0.047kg COD / (kgMLSS·d), and the ammonia nitrogen removal load per unit biomass is 0.0013kgNH 4 + -N / (kgMLSS·d).

[0133] In summary, the Aspergillus polymorpha NXY1 provided by the present invention can be used in the shale gas produced water treatment process. It can not only effectively remove organic matter and ammonia nitrogen in shale gas produced water, but also adsorb activated sludge, reduce the construction of secondary sedimentation tanks, and solve the problem of sludge floating easily when treating shale gas produced water by activated sludge method.

[0134] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A strain of Aspergillus polymorpha that can form a ball Aspergillus sydowii )NXY1, characterized in that The deposit number of the Aspergillus polymorpha NXY1 is CGMCC No.40215.

2. A mycelial pellet capable of removing or assisting in the removal of organic matter and / or ammonia nitrogen in water, characterized in that: The mycelial pellet is obtained by culturing the Aspergillus polymorpha NXY1 described in claim 1.

3. The method for preparing mycelial pellets according to claim 2, characterized in that: Inoculating a spore suspension containing the Aspergillus polydorum NXY1 of claim 1 into a culture medium, culturing, and obtaining the mycelial pellet; The components of the culture medium include: 5-80 g / L glucose, 0.5-8 g / L NH4Cl, 0.3-6 g / L KH2PO4, 0.25-4 g / L MgSO4·7H2O and 0-80 g / L NaCl.

4. The preparation method according to claim 3, characterized in that: The culture conditions include: temperature 20-35° C., time 20-200 h, and rotation speed 100-400 rpm.

5. The preparation method according to claim 3 or 4, characterized in that: The volume ratio of the spore suspension to the culture medium is 1:99; the spore concentration of the spore suspension is 10 4 ~10 9 spores / mL.

6. The preparation method according to claim 3 or 4, characterized in that: The preparation method of the spore suspension comprises: The Aspergillus polydorum NXY1 of claim 1 is inoculated on a PDA medium, and after slant culture, the spores are washed into sterile physiological saline to obtain the spore suspension.

7. The preparation method according to claim 6, characterized in that: The conditions of the slant culture include: temperature 20-35° C., time 3-15 days.

8. Use of the Aspergillus polymorpha NXY1 described in claim 1 or the mycelial pellets described in claim 2, or the mycelial pellets prepared by the preparation method described in any one of claims 3 to 7 in removing or assisting in the removal of organic matter and / or ammonia nitrogen in liquids; the liquid is fracturing flowback fluid and / or produced water.

Images