A kind of odor-removing water-purifying algae and its purification application

Abstract

The present invention belongs to the technical field of water treatment and recycling, and specifically relates to an odor-removing water-purifying algae species and its application. The odor-removing water-purifying algae species used in the present invention can achieve the combined removal of odor substances and environmental hormones in the tail water system while purifying nitrogen and phosphorus pollutants in the aquaculture system, and can solve the problem of odor substances and toxin accumulation in the tail water in an eco-friendly manner. The odor-removing water-purifying algae species and its preparation of the present invention are obtained by screening Oocystis ovale Oocystis sp. YTLJ‑NAPE‑RC3 was pre-cultured and expanded, and then vacuum-freeze-dried to obtain the corresponding algae powder, which was then mixed with an additive to prepare the product. Oocystis algae YTLJ‑NAPE‑RC3 was deposited in the General Microbiology Center of China Microbiological Culture Collection Administration, with the deposit number CGMCC No.40455.

Description

Technical Field

[0001] The invention belongs to the technical field of tail water deep treatment and resource utilization, and specifically relates to an odor-removing water-purifying algae and purification application thereof. Background Art

[0002] Problems such as water pollution and water resource scarcity restrict the rapid development of water-scarce areas. Currently, there are still relatively few biotechnologies that can be used for water treatment and reuse. Traditional biological water purification methods such as bacteria are mainly used to remove nitrogen and phosphorus nutrients. The effect is average and it is easy to cause the accumulation of toxins such as odor substances and environmental hormones, which greatly limits the promotion and application of such technologies.

[0003] In recent years, microalgae water purification technology has been developed in many industries, especially the aquaculture industry. This technology can not only achieve the rapid removal of ammonia nitrogen and nitrite in water and purify water quality, but also microalgae can be used as a high-protein bait for aquatic animals, thereby reducing feed costs. Microalgae have been used in tailwater treatment, and microalgae are often used in production to adjust water and increase oxygen, but there are still a series of problems. For example, in the process of eel farming, it is necessary to stimulate the growth of Microcystis, but once Microcystis is out of control, it will cause water blooms and accumulation of algal toxins. Chlorella and diatoms have the problem of easy algae collapse, and cannot remove odor substances, and diatoms are also prone to produce odor substances such as geosmin. Summary of the invention

[0004] The purpose of the present invention is to provide an odor-removing water-purifying algae and its purification application in view of the deficiencies in the prior art.

[0005] To achieve the above object, the technical solution adopted by the present invention is:

[0006] A deodorizing water purifying algae, the deodorizing water purifying algae is Oocystis Oocystis sp. YTLJ-NAPE-RC3;

[0007] Oocystis Oocystis sp. YTLJ-NAPE-RC3 was deposited in the General Microbiology Center of China Microbiological Culture Collection Administration on April 6, 2023, with the deposit address in Beijing, China, with the deposit number CGMCC No.40455, and the classification name is Oocystis Oocystis sp. .

[0008] An application of the odor-removing and water-purifying algae, the oocyst algae Oocystis sp. Application of YTLJ-NAPE-RC3 in water purification.

[0009] A deodorizing water purifying algae agent, the algae agent containing the oocyst algae Oocystis sp. YTLJ-NAPE-RC3.

[0010] The algae agent is a culture solution containing oocysts, a culture concentrate or a resuspension of a culture concentrate.

[0011] The algae agent is composed of the oocyst algae Oocystis sp. YTLJ-NAPE-RC3 and the additives are mixed in a mass ratio of 1:1-100. After mixing, 0.1% to 1.0% of YTLJ-NAPE-RC3 and 0.05% to 0.5% of NH4NO3 are added to the system, and the mixture is obtained. The total effective number of live algae in the algae agent is ≥1.0×10 9 Pieces / mL.

[0012] The oocyst algae YTLJ-NAPE-RC3 mixed with the auxiliary agent is a dried and crushed product of the oocyst algae after propagation;

[0013] The oocyst algae YTLJ-NAPE-RC3 added in the system is the oocyst algae culture solution after the oocyst algae is expanded.

[0014] The egg cysts are propagated by inoculating the egg cysts in an enrichment medium at a volume ratio of 2% to 10%, culturing at 20° C. and 2000 lx of light until the OD 680 is 1.0.

[0015] The auxiliary agent is a mixture of polyglutamic acid, nisin-halloysite complex and azithromycin in a mass ratio of 0.01-0.1:1:0.01-0.001.

[0016] The nisin-halloysite complex is prepared by mixing nisin and halloysite in a ratio of 1:10-20, and then subjected to ultrasonic treatment and vacuum drying for later use.

[0017] An application of the deodorizing and water-purifying algae agent, and application of the deodorizing and water-purifying algae agent in purifying water.

[0018] The advantages of the present invention are:

[0019] The present invention can control emerging complex pollution such as odor substances and environmental hormones by adding odor-removing and water-purifying algae species while strengthening nitrogen and phosphorus removal, specifically:

[0020] 1) The water-purifying algae species used in the present invention can purify nitrogen and phosphorus pollutants in water, and can also remove odor substances and environmental hormones in the aquaculture system, and can solve the problem of accumulation of odor substances and environmental hormones in tail water in an eco-friendly manner. At the same time, the algae species can also be used as aquaculture bait algae, so as to achieve both water regulation and aquaculture.

[0021] 2) The growth cycle of the algae of the present invention is significantly longer than that of Chlorella, which overcomes the problem that Chlorella and diatoms are prone to premature aging and algae collapse, and ensures the simplicity and operability of in-situ treatment and purification of aquaculture water.

[0022] 3) The water purification algae species of the present invention replaces Microcystis or diatoms, which overcomes the disadvantages of the traditional Microcystis water conditioning method that easily causes water blooms and accumulation of algal toxins, and also overcomes the disadvantage of diatom water conditioning that easily produces odorous substances. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 A growth curve comparison diagram of the oocyst algae YTLJ-NAPE-RC3 provided in an embodiment of the present invention and commercial Chlorella vulgaris (purchased from Nanhua Qianmu). DETAILED DESCRIPTION

[0024] The present invention is further illustrated by the following examples, but the present invention is not limited to the following examples.

[0025] Example 1

[0026] Isolation of Oocystis YTLJ-NAPE-RC3:

[0027] The egg cyst algae YTLJ-NAPE-RC3 was isolated from the coastal environment. The specific isolation process was as follows: 10 g of Rongcheng offshore (Rongcheng Chengshantou Nature Reserve) water sample was placed in 50 mL of sterile water, and the mixture was shaken at a constant temperature of 25°C for 48 h. The bacterial solution was centrifuged at 8000 rpm for 10 min, and then the upper clear liquid was removed. The lower bacterial and algae solution was transferred to a sterilized enrichment medium, cultured at 30°C for 7 days, and then diluted and cultured in BG11 medium. Single algae were selected and repeatedly streaked on BG11 agar solid medium to obtain the purified egg cyst algae YTLJ-NAPE-RC3 ( Oocystis sp. ), and preserve it.

[0028] The enrichment medium used was a mass ratio of 1% NH4NO3, 0.05% NaNO2, 0.1% polyglutamic acid, 0.001% geosmin, 1% K2HPO4, 1% MgSO4, 2% NaCl, 0.01% FeSO4, 0.001% azithromycin, and 0.01% MnSO4.

[0029] The Oocystis sp. YTLJ-NAPE-RC3 ( Oocystis sp. ) is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, the deposit address is Beijing, China, the deposit date is April 6, 2023, and the deposit number is CGMCC No.40455.

[0030] By comparing the 18S rDNA gene sequence, it was confirmed that the oocysts YTLJ-NAPE-RC3 was a newly discovered algae species.

[0031] The Oocystis sp. YTLJ-NAPE-RC3 ( Oocystis sp.) is shown in SEQ ID NO: 1:

[0032] .

[0033] Example 2

[0034] Application of the oocyst algae YTLJ-NAPE-RC3 obtained in Example 1 in aquaculture tail water treatment:

[0035] The purified oocyst algae YTLJ-NAPE-RC3 was inoculated at a volume ratio of 2% in an enriched medium at 25°C, 2000 lx light intensity, and 12 h / day light frequency until OD 680 =1.0; the cultured algae liquid was inoculated into the simulated nitrogen-containing aquaculture tail water at an inoculation rate of 10wt%, and Chlorella vulgaris (purchased from Nanhua Qianmu) was used as the control algae, and the tail water was treated at 25°C (see Figure 1 );

[0036] The nitrogen-containing aquaculture tail water is prepared by adding ammonia nitrogen and nitrate to each liter of water, wherein the concentrations of ammonia nitrogen, nitrite, nitrate nitrogen and phosphate are 1, 0.5, 50 and 0.5 mg / L respectively, the concentration of estradiol is 0.1 mg / L, and the concentration of geosmin is 0.001 mg / L.

[0037] Depend on Figure 1 It can be seen that the oocysts YTLJ-NAPE-RC3 was still in a rapid growth period after 42 days of growth, and was still in a stable period at 84 days. After 84 days of treatment at 25℃, the concentrations of ammonia nitrogen, nitrite, nitrate nitrogen, and phosphate dropped to 0.02, 0.01, 1, and 0.03 mg / L, respectively, estradiol was lower than 10ng / L, and odor substances (geosin) were lower than 10ng / L; while the control Chlorella was lower than 42 days due to OD 680 It continues to decline rapidly and then shows premature algae decay.

[0038] Example 3

[0039] YTLJ-NAPE-RC3 is used for water purification in an eel farming base in Fujian:

[0040] 1) Propagation of YTLJ-NAPE-RC3: The purified YTLJ-NAPE-RC3 was inoculated with 10% volume ratio in enriched medium at 25°C, 2000 lx light intensity, and 12 h / day light frequency until OD 680 is 1.0.

[0041] 2) Algae agent: the above-mentioned expanded oocyst algae YTLJ-NAPE-RC3 is dried and crushed to prepare algae powder, and the algae powder is then mixed with the auxiliary agent in a ratio of 1:1 (mass ratio), and after mixing, the expanded oocyst algae YTLJ-NAPE-RC3 and NH4NO3 are added to the system;

[0042] The total effective number of live algae in the algae agent is ≥ 1.0×10 9 / mL, the auxiliary agent is polyglutamic acid, nisin-halloysite complex, and azithromycin mixed in a mass ratio of 0.01:1:0.01; among them, the nisin-halloysite complex is a mixture of nisin (purchased from Meilun Bio, commercially available) and halloysite in a mass ratio of 1:10, and it is treated with 40 kHz ultrasound for 15 min and then vacuum dried for 24 h for use; after expansion, the oocyst algae YTLJ-NAPE-RC3 accounted for 0.1% (mass ratio) of the system, and NH4NO3 accounted for 0.05% (mass ratio) of the system.

[0043] 3) Water purification: The algae agent obtained above was added to the eel breeding pond at an addition amount of 1wt% (the concentrations of ammonia nitrogen, nitrite, nitrate and phosphate were 0.8, 0.2, 10.0 and 0.6 mg / L, respectively, the concentration of estradiol was 100 ng / L, and the concentration of geosmin was 100 ng / L). After one month of breeding at 20℃, 2000 lx light intensity and 8 h / day light frequency, the removal rates of ammonia nitrogen, nitrite, geosmin and estradiol in the breeding pond were more than 90%, and the removal rates of nitrate and phosphate in the breeding pond were more than 80%.

[0044] Example 4

[0045] Oocystis YTLJ-NAPE-RC3 is used in Yantai municipal tail water treatment:

[0046] 1) Algae agent: the expanded oocyst algae YTLJ-NAPE-RC3 obtained in the above example is dried and crushed to prepare algae powder, and the algae powder is then mixed with the auxiliary agent at a ratio of 1:10 (mass ratio), and after mixing, the expanded oocyst algae YTLJ-NAPE-RC3 and NH4NO3 are added to the system;

[0047] The total effective number of live algae in the algae agent is ≥ 1.0×10 9 / mL, the auxiliary agent is polyglutamic acid, nisin-halloysite complex, and azithromycin mixed in a mass ratio of 0.02:1:0.003; among them, the nisin-halloysite complex is a mixture of nisin and halloysite in a mass ratio of 1:12, and is vacuum dried after ultrasonic treatment for later use; after expansion, the oocyst algae YTLJ-NAPE-RC3 accounts for 0.2% (mass ratio) of the system, and NH4NO3 accounts for 0.06% (mass ratio) of the system.

[0048] 2) Water purification: The algae agent obtained above was added into the Yantai municipal tailwater pond at a dosage of 1wt% (the concentrations of ammonia nitrogen, nitrite, nitrate and phosphate were 4.5, 0.8, 18.2 and 0.9 mg / L respectively, the concentration of estradiol was 60 ng / L and the concentration of geosmin was 50 ng / L). After one week of breeding at 25℃, 3000 lx light intensity and 12 h / day light frequency, the removal rates of ammonia nitrogen, nitrite, geosmin and estradiol in the tailwater pond were more than 95%, and the removal rates of nitrate and phosphate were more than 85%.

[0049] Example 5

[0050] Deep treatment and recycling of aquaculture water in a crucian carp farming process in coastal areas:

[0051] 1) Algae agent: the expanded oocyst algae YTLJ-NAPE-RC3 obtained in the above example is dried and crushed to prepare algae powder, and the algae powder is then mixed with the auxiliary agent at a ratio of 1:50 (mass ratio), and after mixing, the expanded oocyst algae YTLJ-NAPE-RC3 and NH4NO3 are added to the system;

[0052] The total effective number of live algae in the algae agent is ≥ 1.0×10 9 / mL, the auxiliary agent is polyglutamic acid, nisin-halloysite complex, and azithromycin mixed in a mass ratio of 0.05:1:0.005; wherein, the nisin-halloysite complex is a mixture of nisin (commercially available) and halloysite in a mass ratio of 1:15, and is vacuum dried after ultrasonic treatment for later use; after expansion, the oocyst algae YTLJ-NAPE-RC3 accounts for 0.2% (mass ratio) of the system, and NH4NO3 accounts for 0.2% (mass ratio) of the system.

[0053] 2) Water purification: The algae agent obtained above was added at a dosage of 0.5wt% into the aquaculture water of a crucian carp aquaculture process in the coastal area (the concentrations of ammonia nitrogen, nitrite, nitrate and phosphate were 1.2, 0.2, 4.9 and 0.8 mg / L, respectively, and the concentration of geosmin was 60 ng / L). After one month of breeding at 20℃, 2000 lx light intensity and 10 h / day light frequency, the removal rates of ammonia nitrogen, nitrite and geosmin in the aquaculture water were more than 85%, and the removal rates of nitrate and phosphate were more than 80%.

[0054] 3) Recycling: The treated aquaculture water is reused in the crucian carp aquaculture pond. Compared with the control aquaculture group (no tailwater reuse), the crucian carp growth rate increased by 20%, the mortality rate decreased by 10%, and the feed coefficient decreased by 0.1.

[0055] Example 6

[0056] YTLJ-NAPE-RC3 is used for water purification in shrimp breeding bases:

[0057] 1) Algae agent: the expanded oocyst algae YTLJ-NAPE-RC3 obtained in the above example is dried and crushed to prepare algae powder, and the algae powder is then mixed with the auxiliary agent at a ratio of 1:60 (mass ratio), and after mixing, the expanded oocyst algae YTLJ-NAPE-RC3 and NH4NO3 are added to the system;

[0058] The total effective number of live algae in the algae agent is ≥ 1.0×10 9 / mL, the auxiliary agent is polyglutamic acid, nisin-halloysite complex, and azithromycin mixed in a mass ratio of 0.06:1:0.006; among them, the nisin-halloysite complex is a mixture of nisin and halloysite in a mass ratio of 1:18, and is vacuum dried after ultrasonic treatment for later use; after expansion, the oocyst algae YTLJ-NAPE-RC3 accounts for 0.6% (mass ratio) of the system, and NH4NO3 accounts for 0.4% (mass ratio) of the system.

[0059] 2) Water purification: The algae agent obtained above was added to the shrimp culture pond at an addition amount of 0.5wt% (the concentrations of ammonia nitrogen, nitrite, nitrate, and phosphate were 0.8, 0.1, 11.8, and 1.4 mg / L, respectively, and the concentration of geosmin was 50 ng / L). After two weeks of breeding at 28℃, 3000 lx light intensity, and 8 h / day light frequency, the removal rates of ammonia nitrogen, nitrite, and geosmin in the culture pond were more than 75%, and the removal rates of nitrate and phosphate were more than 75%. Compared with before the addition of the algae agent, the shrimp feed coefficient decreased by 0.2.

[0060] Example 7

[0061] YTLJ-NAPE-RC3 is used in livestock and poultry breeding tail water treatment:

[0062] 1) Algae agent: the expanded oocyst algae YTLJ-NAPE-RC3 obtained in the above example is dried and crushed to prepare algae powder, and the algae powder is then mixed with the auxiliary agent at a ratio of 1:100 (mass ratio), and after mixing, the expanded oocyst algae YTLJ-NAPE-RC3 and NH4NO3 are added to the system;

[0063] The total effective number of live algae in the algae agent is ≥ 1.0×10 9 / mL, the auxiliary agent is polyglutamic acid, nisin-halloysite complex, and azithromycin mixed in a mass ratio of 0.1:1:0.001; among them, the nisin-halloysite complex is a mixture of nisin and halloysite in a mass ratio of 1:20, and is vacuum dried after ultrasonic treatment for later use; after expansion, the oocyst algae YTLJ-NAPE-RC3 accounts for 1.0% (mass ratio) of the system, and NH4NO3 accounts for 0.5% (mass ratio) of the system.

[0064] 2) Water purification: The algae agent obtained above was added to the tail water of livestock and poultry breeding at an addition amount of 0.5wt% (the concentrations of ammonia nitrogen, nitrite, nitrate and phosphate were 15, 2.0, 20.5 and 2.5 mg / L respectively, the concentration of estradiol was 300 ng / L, and the concentration of geosmin was 250 ng / L). One month after the algae agent was started for breeding at 25℃, 4000 lx light intensity and 10 h / day light frequency, the removal rates of nitrite, geosmin and estradiol in the tail water were more than 70%, and the removal rates of ammonia nitrogen, nitrate and phosphate were more than 70%.

Claims

1. A deodorizing water purifying algae, characterized in that: The deodorizing and water purifying algae is Oocystis ( Oocystis sp. )YTLJ-NAPE-RC3, deposited in the General Microbiological Center of China Microbiological Culture Collection Administration on April 6, 2023, with the deposit number CGMCC No.40455, and the classification name is Oocystis ， The storage address is Beijing, China.

2. An application of the odor-removing water-purifying algae according to claim 1, characterized in that: The application of the oocyst algae YTLJ-NAPE-RC3 in purifying water bodies.

3. A deodorizing algae purifying agent, characterized in that: The algae agent contains the oocyst algae YTLJ-NAPE-RC3 described in claim 1.

4. The deodorizing and water purifying algae agent according to claim 3, characterized in that: The algae agent is a culture solution containing oocysts, a culture concentrate or a resuspension of a culture concentrate.

5. The deodorizing and water purifying algae agent according to claim 3 or 4, characterized in that: The algae agent is prepared by mixing the dried and crushed material of the expanded egg cyst algae YTLJ-NAPE-RC3 according to claim 1 with an auxiliary agent in a mass ratio of 1:1-100, adding 0.1% to 1.0% by weight of the egg cyst algae culture solution of the expanded egg cyst algae YTLJ-NAPE-RC3 and 0.05% to 0.5% by weight of NH4NO3 to the system, and mixing them to obtain the algae agent; the total effective number of live algae in the algae agent is ≥1.0×10 9 Pieces / mL; The auxiliary agents are polyglutamic acid, nisin-halloysite complex and azithromycin.

6. The deodorizing and water purifying algae agent according to claim 5, characterized in that: The egg cysts are propagated by inoculating the egg cysts in an enrichment medium at a volume ratio of 2% to 10%, culturing at 20° C. and 2000 lx of light until the OD 680 is 1.

0.

7. The deodorizing and water purifying algae agent according to claim 5, characterized in that: The mass ratio of the polyglutamic acid, the nisin-halloysite complex and the azithromycin is 0.01-0.1:1:0.01-0.

001.

8. The deodorizing and water purifying algae agent according to claim 7, characterized in that: The nisin-halloysite complex is prepared by mixing nisin and halloysite in a ratio of 1:10-20, and then subjected to ultrasonic treatment and vacuum drying for later use.

9. An application of the deodorizing algae purifying agent according to claim 3, characterized in that: The application of the deodorizing and water purifying algae agent in purifying water bodies.

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