CN116375214A - Method for treating aquaculture wastewater by combining polyhydroxy carbon nano-tube with algae-bacteria symbiota - Google Patents
Method for treating aquaculture wastewater by combining polyhydroxy carbon nano-tube with algae-bacteria symbiota Download PDFInfo
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Abstract
The invention discloses a method for treating aquaculture wastewater by combining polyhydroxy carbon nanotubes with algae and bacteria symbiota, which comprises the following steps: s1: culturing chlorella; s2: culturing ganoderma lucidum; s3: co-culturing chlorella-ganoderma lucidum; s4: transferring the obtained algae-bacteria symbiota into aquaculture wastewater, adding polyhydroxy carbon nanotubes with a certain concentration into the aquaculture wastewater, and then treating the aquaculture wastewater under a certain condition. The polyhydroxy carbon nanotube combined algae and fungus symbiont has obvious advantages in the treatment of aquaculture wastewater, and has great application potential.
Description
Technical Field
The invention belongs to the technical field of pollution ecology, and particularly relates to a method for treating aquaculture wastewater by combining polyhydroxy carbon nanotubes with algal and bacterial symbiota.
Background
In recent years, with the increase of demand of high-quality protein and the exhaustion of natural fishery resources, the aquaculture scale is increasingly enlarged, and the sustainable development of the aquaculture industry is seriously hindered due to the negative environmental problem caused by a large amount of wastewater generated in the aquaculture process. The microalgae-fungus symbiont has good application potential in the purification of aquaculture wastewater. However, the algae-bacteria symbiota takes longer to balling, the algae-bacteria balls grow slower, and the purification performance of the algae-bacteria symbiota has a certain improvement space.
Polyhydroxy carbon nanotubes (MWCNTs) are carbon nano materials with excellent performance and wide application, and the MWCNTs can help a plant photosynthetic reaction system to stimulate photosynthetic activity, improve stability of chloroplasts, and transfer absorbed energy to a photosynthetic device, so that photosynthetic capacity is improved. MWCNTs can help exogenous antioxidant or signal molecules to enter cells through cell walls, and improve the capability of plant cells to cope with stress. Through adding MWCNTs from an external source, the photosynthesis of microalgae and the growth of fungi are promoted, the balling speed of the algae is accelerated, the growth performance of the algae balls is optimized, the biological purification performance of the algae symbiotes is improved, and the maximization of the efficiency of the purification system is realized. However, MWCNTs have been relatively rarely used in microalgae and fungi.
Disclosure of Invention
The invention provides a method for treating aquaculture wastewater by combining polyhydroxy carbon nanotubes with algae and bacteria symbiota, which comprises the following steps:
(1) Amplifying and culturing chlorella;
(2) Culturing ganoderma lucidum: pre-culturing Ganoderma for 7d, homogenizing Ganoderma granule to obtain final Ganoderma strain;
(3) Co-culture of Chlorella-Ganoderma: washing, precipitating and re-suspending the chlorella expanded culture and the pre-cultured ganoderma lucidum, then mixing the ganoderma lucidum ball suspension and the chlorella suspension to obtain an algae-bacteria co-culture system, and culturing the algae-bacteria mixed system for 7d to obtain a domesticated algae-bacteria mixed system;
(4) Transferring the obtained algae-bacteria symbiota into aquaculture wastewater, adding polyhydroxy carbon nanotubes, and treating to purify the wastewater.
Further, in the step (1), the chlorella is amplified and cultivated, FACHB-8 variety of chlorella is selected, the chlorella is amplified and cultivated by utilizing a BG-11 culture medium, the amplified and cultivated illumination is provided by a cool white LED lamp, the illumination intensity is set to 200 mu mol m-2s-1, the light-dark ratio is 12h (day): 12h (night), the amplified and cultivated temperature is controlled to 23-27 ℃, and the amplified and cultivated time is 7 days; the chlorella is manually shake-bottled 3 times daily at regular intervals during the period of cultivating chlorella.
Further, the step (2) of preculture of the ganoderma lucidum is to place the ganoderma lucidum into a constant temperature shaking table, and culture the ganoderma lucidum at the rotation speed of 160rpm and the temperature of 24-26 ℃.
In the step (2), the ganoderma lucidum particles are homogenized into ganoderma lucidum particles which are obtained by washing and culturing with sterile deionized water, and the washed ganoderma lucidum particles are homogenized with the sterile deionized water.
Further, the washing, precipitation and re-suspension treatment in the step (3) is to wash, precipitate and re-suspend the chlorella expanded culture in the logarithmic phase and the pre-cultured ganoderma lucidum with a sterile BG-11 culture medium solution, and the washing, precipitation and re-suspension treatment is repeated for three times.
Further, the specific steps of mixing the ganoderma lucidum ball suspension in the step (3) with the chlorella suspension to obtain the algae-bacteria co-culture system are that the ganoderma lucidum ball suspension with the concentration of 75.88-88.38 mg L-1 after treatment and the chlorella suspension with the concentration of 70.67-81.89 mg L-1 are mixed according to the volume ratio of 1:20, and the initial concentration of the mixed algae-bacteria co-culture system is about 85.27 +/-7.34 mg L-1.
Further, the mixed system of algae and bacteria in the step (3) is cultivated for 7 days under the conditions that a constant-temperature shaking table is utilized, the rotation speed of the shaking table is 150rpm, and the temperature of the shaking table is 23-27 ℃.
Further, the diameter of the polyhydroxy carbon nano tube in the step (4) is 10-20nm, the purity is more than 95wt percent, and the length is 0.5-2 mu m.
Further, the treatment method in the step (4) is characterized in that the concentration of the polyhydroxy carbon nanotubes in the aquaculture wastewater is adjusted to 1mg L-1, and then the aquaculture wastewater is placed in an environment with the temperature of 25 ℃ and the illumination intensity of 200mol & m & lt-2 & gts & lt-1 & gt and the light-dark time ratio of 12h illumination/12 h darkness for 10 days.
Further, the BG-11 medium comprises the following components: naNO 3.5 g L-1, K2HPO4.3H2O 0.04g L-1, KH2PO4.3H2O 0.2g L-1, ethylenediamine tetraacetic acid 0.5mg L-1, ferric ammonium citrate 5mg L-1, citric acid 5mg L-1, na2CO3 0.025mg L-1 and trace metals solution 1mL L-1; the conditions of the LB solid medium are as follows: tryptophan: 10g/L, yeast extract: 5g/L, sodium chloride: 10g/L, agar powder: 15g/L, pH 7.0-7.2.
The invention has the following beneficial effects:
1. the polyhydroxy carbon nano tube is added from an external source, so that the photosynthesis of microalgae and the growth of fungi can be promoted, the balling speed of the algae is accelerated, the growth performance of the algae balls is optimized, the biological purification performance of the algae-bacteria symbiotes is improved, and the efficiency maximization of the purification system is realized.
2. The method for obtaining the algae bacteria community is simple and easy to operate, and the obtained biological community has higher activity and low cost, and can realize large-scale expansion and production.
3. The method provided by the invention has a good effect on the treatment of aquaculture wastewater, and opens up space for the application of polyhydroxy carbon nanotubes in the field of biotechnology and improves the commercial value of the polyhydroxy carbon nanotubes.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings can be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 shows the removal rate of COD in aquaculture wastewater after 10 days of reaction under different treatment modes;
FIG. 2 shows TN removal rate in aquaculture wastewater after 10 days of reaction under different treatment modes;
FIG. 3 shows the removal rate of TP from aquaculture wastewater after 10 days of reaction under different treatment modes.
Detailed Description
Various exemplary embodiments of the invention will now be described in detail, with reference to the examples using conventional methods, unless otherwise indicated, and with reference to reagents, either conventional commercial reagents or reagents configured using conventional methods. The detailed description is not to be taken as limiting, but is to be understood as a more detailed description of certain aspects, features, and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in this disclosure, it is understood that each intermediate value between the upper and lower limits of the ranges is also specifically disclosed. Every smaller range between any stated value or stated range, and any other stated value or intermediate value within the stated range, is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the invention described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from consideration of the specification of the present invention. The specification and examples of the present invention are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are open ended terms, meaning including, but not limited to.
Example 1
Firstly, washing, precipitating and re-suspending the chlorella expanded culture in logarithmic growth phase and the pre-cultured ganoderma lucidum with a sterile BG-11 culture medium solution (repeated for 3 times); then 5mL of the glossy ganoderma pellet suspension with the concentration of 82.13 + -6.52 mg L-1 after treatment is added to 100mL of the chlorella suspension with the concentration of 76.28+ -5.61 mg L-1, so that the initial concentration of the algae-bacteria co-culture system is about 85.27 + -7.34 mg L-1. Culturing the mixed system of algae and bacteria for 7d by using a constant temperature shaking table under the conditions that the rotation speed of the shaking table is 150rpm and the temperature of the shaking table is 25+/-2 ℃. The domesticated chlorella-fungus particles are used for the subsequent purification treatment of aquaculture wastewater.
Adding the chlorella-ganoderma lucidum fungus symbiont into aquaculture wastewater to ensure that the initial total biomass (dry weight) is about 100-120mg/L, adding MWCNT of 1mg L-1, taking the aquaculture wastewater from aquaculture (four-big-family fish aquaculture) enterprises in Nanjing city, and storing at 4 ℃ for later use. Before the purification treatment, in order to avoid the influence of microorganisms in the original aquaculture wastewater, a nylon microfilter with the diameter of 0.45 μm is used in advance for filtration and sterilization treatment, and the components after the treatment are as follows: COD (78.89 mg L-1), TN (31.73 mg L-1), TP (3.18 mg L-1), pH (7.4.+ -. 0.1). Adding the waste water into the chlorella-ganoderma lucidum symbiote and the MWCNT according to the conditions, setting the temperature to 25 ℃ in a sterile room with constant temperature and constant illumination, wherein the illumination intensity is 200 mu mol m & lt-2 & gts & lt-1 & gt, and the light-dark time ratio is 12h illumination/12 h darkness; culturing for 10 days. The removal rates of COD, TN and TP in the aquaculture wastewater are shown in Table 1 through water quality detection (national standard method). The removal rates of COD, TN and TP in the aquaculture wastewater are respectively as follows: 78.25%,81.06% and 80.31%.
Example 2
Adding the chlorella-ganoderma lucidum fungus symbiont obtained in the example 1 into aquaculture wastewater (the conditions are the same as those of the example 1), so that the initial total biomass (dry weight) of the symbiont is about 80-90mg/L, adding MWCNT of 1mg L-1, setting the temperature in a sterile room with constant temperature and constant illumination to 25 ℃, the illumination intensity to 200 mu mol m-2s-1, and the light-dark time ratio to 12h illumination/12 h darkness; culturing for 10 days. The removal rates of COD, TN and TP in the aquaculture wastewater are shown in Table 1 through water quality detection (national standard method). The removal rates of COD, TN and TP in the aquaculture wastewater are respectively as follows: 70.32%,74.68% and 76.24%.
Comparative example 1:
the chlorella (FACHB-8) is cultivated by utilizing a BG-11 culture medium, the cultivation-enlarging illumination is provided by a cool white LED lamp, the illumination intensity is set to 200 mu mol m-2s-1, the light-dark ratio is 12h (day): 12h (night), the cultivation-enlarging temperature is controlled to 25+/-2 ℃, and the cultivation-enlarging time is 7 days. The chlorella is manually shake-bottled 3 times daily at regular intervals during the period of cultivating chlorella. Adding the obtained chlorella into aquaculture wastewater (under the same conditions as in example 1) to ensure that the initial total biomass (dry weight) is about 100-120mg/L, adding MWCNT of 1mg L-1, setting the temperature to 25 ℃ in a sterile room with constant temperature and constant illumination, wherein the illumination intensity is 200 mu mol m-2s-1, and the light-dark time ratio is 12h illumination/12 h darkness; culturing for 10 days. The removal rates of COD, TN and TP in the aquaculture wastewater are shown in Table 1 through water quality detection (national standard method). The removal rates of COD, TN and TP in the aquaculture wastewater are respectively as follows: 58.13%,63.47% and 65.46%.
Comparative example 2:
culturing Ganoderma in a constant temperature shaker (rotation speed 160rpm and temperature 25+ -1deg.C) for 7 days. And then washing the ganoderma lucidum particles obtained by culture with sterile deionized water, and homogenizing the washed ganoderma lucidum particles with 100mL of sterile deionized water to obtain the final ganoderma lucidum strain. Adding the obtained ganoderma lucidum strain into aquaculture wastewater (the conditions are the same as those of example 1) to ensure that the initial total biomass (dry weight) is about 100-120mg/L, adding MWCNT of 1mg L-1, setting the temperature in a sterile room with constant temperature and constant illumination to 25 ℃, the illumination intensity to 200 mu mol m-2s-1, and the light-dark time ratio to 12h illumination/12 h darkness; culturing for 10 days. The removal rates of COD, TN and TP in the aquaculture wastewater are shown in Table 1 through water quality detection (national standard method). The removal rates of COD, TN and TP in the aquaculture wastewater are respectively as follows: 42.56%,51.19%,56.32%.
TABLE 1 effects of polyhydroxy carbon nanotube combined algae/microalgae treatment for removal of COD, TN and TP from aquaculture wastewater
As shown in Table 1, after adding 1mg L-1MWCNT to the aquaculture wastewater on the basis of the chlorella-ganoderma lucidum symbiote, the aquaculture wastewater is cultured for 10 days, the removal rate of COD, TN and TP in the aquaculture wastewater is above 70%, and the wastewater treatment effect is good, and the example 1 is optimal, and the example 2 is repeated, so that the wastewater treatment effect is best when the initial total biomass (dry weight) of the chlorella-ganoderma lucidum symbiote is about 100-120 mg/L. In comparative examples 1 and 2, after adding 1mg/L MWCNT to aquaculture wastewater, respectively, on the basis of adding chlorella and ganoderma lucidum, the removal rate of COD, TN and TP in the wastewater is low, and on the basis of adding ganoderma lucidum, the removal rate of COD, TN and TP after adding 1mg L-1MWCNT is below 56.32%. Therefore, the polyhydroxy carbon nanotube combined algae and bacteria symbiont has increased application potential in the treatment of aquaculture wastewater, and has the best effect of treating the aquaculture wastewater when the initial total biomass (dry weight) is about 100-120mg/L and the MWCNT concentration is 1mg/L, and has larger application potential.
The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the design spirit of the present invention.
Claims (10)
1. A method for treating aquaculture wastewater by combining polyhydroxy carbon nano-tubes with algae and bacteria symbiota, which is characterized by comprising the following steps:
(1) Amplifying and culturing chlorella;
(2) Culturing ganoderma lucidum: pre-culturing Ganoderma for 7d, homogenizing Ganoderma granule to obtain final Ganoderma strain;
(3) Co-culture of Chlorella-Ganoderma: washing, precipitating and re-suspending the chlorella expanded culture and the pre-cultured ganoderma lucidum, then mixing the ganoderma lucidum ball suspension and the chlorella suspension to obtain an algae-bacteria co-culture system, and culturing the algae-bacteria mixed system for 7d to obtain a domesticated algae-bacteria mixed system;
(4) Transferring the obtained algae-bacteria symbiota into aquaculture wastewater, adding polyhydroxy carbon nanotubes, and treating to purify the wastewater.
2. The wastewater treatment method according to claim 1, wherein the chlorella in step (1) is cultivated by amplification, FACHB-8 variety of chlorella is selected, the chlorella is cultivated by utilizing BG-11 medium, the cultivation light is provided by a cool white LED lamp, and the light intensity is set to 200 mu mol m -2 s -1 The light-dark ratio is 12 hours (day) and 12 hours (night), the culture expansion temperature is controlled at 23-27 ℃, and the culture expansion time is 7 days; the chlorella is manually shake-bottled 3 times daily at regular intervals during the period of cultivating chlorella.
3. The method for treating wastewater according to claim 1, wherein the step (2) is characterized in that the ganoderma lucidum is pre-cultured by placing the ganoderma lucidum in a constant temperature shaking table, and culturing at a rotation speed of 160rpm and a temperature of 24-26 ℃.
4. The method according to claim 1, wherein in the step (2), the ganoderma lucidum particles are homogenized by washing the ganoderma lucidum particles obtained by culture with sterile deionized water, and then homogenizing the washed ganoderma lucidum particles with sterile deionized water.
5. The method for treating wastewater according to claim 1, wherein the washing, precipitation and re-suspension treatment in the step (3) is repeated three times by washing, precipitation and re-suspension treatment of the chlorella expanded culture in the logarithmic growth phase and the precultured ganoderma lucidum with a sterile BG-11 medium solution.
6. The method for treating wastewater according to claim 1, wherein the step (3) of mixing the suspension of Ganoderma lucidum mycelia with the suspension of Chlorella to obtain an algae-bacteria co-culture system comprises the steps of subsequently treating the mixture to a concentration of 75.88-88.38 mg L -1 The concentration of the ganoderma lucidum fungus ball suspension is 70.67-81.89 mg L -1 Mixing chlorella suspension at a volume ratio of 1:20, and making the initial concentration of the mixed algae-bacteria co-culture system about 85.27 + -7.34 mg L -1 。
7. The method for treating wastewater according to claim 1, wherein the mixed system of algae and bacteria in the step (3) is cultured for 7 days by using a constant temperature shaking table at a rotation speed of 150rpm and a shaking table temperature of 23 to 27 ℃.
8. The method according to claim 1, wherein the polyhydroxycarbon nanotubes in the step (4) have a diameter of 10-20nm, a purity of > 95wt% and a length of 0.5-2. Mu.m.
9. The method for treating wastewater according to claim 1, wherein the concentration of the polyhydroxycarbon nanotubes in the aquaculture wastewater is adjusted to 1mg L -1 Then the aquaculture wastewater is placed at the temperature of 25 ℃ and the illumination intensity of 200 mu mol.m -2 ·s -1 In an environment with a light-dark time ratio of 12h light/12 h dark, the treatment time is 10 days.
10. The wastewater treatment method according to claim 1, wherein the BG-11 medium has a composition of: naNO 3 1.5g L -1 、K 2 HPO 4 ·3H 2 O 0.04g L -1 、KH 2 PO 4 ·3H 2 O 0.2g L -1 0.5mg L of ethylenediamine tetraacetic acid -1 Ferric ammonium citrate 5mg L -1 Citric acid 5mg L -1 、Na 2 CO 3 0.025mg L -1 And trace metalsSolution 1mL L -1 The method comprises the steps of carrying out a first treatment on the surface of the The conditions of the LB solid medium are as follows: tryptophan: 10g/L, yeast extract: 5g/L, sodium chloride: 10g/L, agar powder: 15g/L, pH 7.0-7.2.
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