CN108733985B - A method for identifying key environmental parameters that constrain the arsenic accumulation capacity of microalgae - Google Patents
A method for identifying key environmental parameters that constrain the arsenic accumulation capacity of microalgae Download PDFInfo
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- 229910052785 arsenic Inorganic materials 0.000 title claims abstract description 91
- RQNWIZPPADIBDY-UHFFFAOYSA-N arsenic atom Chemical compound [As] RQNWIZPPADIBDY-UHFFFAOYSA-N 0.000 title claims abstract description 90
- 230000007613 environmental effect Effects 0.000 title claims abstract description 53
- 238000009825 accumulation Methods 0.000 title claims abstract description 48
- 238000000034 method Methods 0.000 title claims abstract description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 20
- 238000004364 calculation method Methods 0.000 claims abstract description 14
- 238000004458 analytical method Methods 0.000 claims abstract description 7
- 238000012360 testing method Methods 0.000 claims description 25
- 241000192710 Microcystis aeruginosa Species 0.000 claims description 14
- 241000195493 Cryptophyta Species 0.000 claims description 10
- 238000013461 design Methods 0.000 claims description 9
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 8
- 238000010521 absorption reaction Methods 0.000 claims description 7
- 229910019142 PO4 Inorganic materials 0.000 claims description 5
- MMDJDBSEMBIJBB-UHFFFAOYSA-N [O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O.[NH6+3] Chemical compound [O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O.[NH6+3] MMDJDBSEMBIJBB-UHFFFAOYSA-N 0.000 claims description 5
- 239000010452 phosphate Substances 0.000 claims description 5
- 231100000693 bioaccumulation Toxicity 0.000 claims description 4
- 238000012258 culturing Methods 0.000 claims description 4
- 238000012163 sequencing technique Methods 0.000 claims description 4
- 235000007122 Scenedesmus obliquus Nutrition 0.000 claims description 3
- 241000195662 Tetradesmus obliquus Species 0.000 claims description 3
- 239000001963 growth medium Substances 0.000 claims description 3
- 238000005286 illumination Methods 0.000 claims description 3
- 238000005259 measurement Methods 0.000 claims description 3
- 239000002609 medium Substances 0.000 claims description 2
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- 230000009286 beneficial effect Effects 0.000 abstract description 5
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- 238000002474 experimental method Methods 0.000 abstract 1
- 229910052757 nitrogen Inorganic materials 0.000 description 14
- 229910052698 phosphorus Inorganic materials 0.000 description 11
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 10
- 238000005067 remediation Methods 0.000 description 5
- 229910052751 metal Inorganic materials 0.000 description 3
- 239000002184 metal Substances 0.000 description 3
- 241000224474 Nannochloropsis Species 0.000 description 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 2
- 238000000540 analysis of variance Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000001095 inductively coupled plasma mass spectrometry Methods 0.000 description 2
- 239000011574 phosphorus Substances 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 238000003723 Smelting Methods 0.000 description 1
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- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 230000009920 chelation Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 238000009713 electroplating Methods 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 150000002739 metals Chemical class 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
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- 231100000167 toxic agent Toxicity 0.000 description 1
- 231100001234 toxic pollutant Toxicity 0.000 description 1
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- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06F—ELECTRIC DIGITAL DATA PROCESSING
- G06F2219/00—Indexing scheme relating to application aspects of data processing equipment or methods
- G06F2219/10—Environmental application, e.g. waste reduction, pollution control, compliance with environmental legislation
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Abstract
本发明涉及一种确定制约微藻砷累积能力的关键环境参数的方法，可包括以下步骤：S1.获取与微藻砷累积能力有关的环境参数；S2.根据S1所获取的环境参数进行田口(Taguchi)试验设计；S3.根据S2得到的试验设计进行试验并分析计算试验结果获取微藻砷累积能力参数；S4.根据S3得到的微藻砷累积能力参数进行信噪比S/N、方差分析、因素百分比贡献度PC的统计计算，进而确定制约微藻砷累积能力的关键环境参数。本发明能够高效确定出制约微藻砷累积能力的关键环境参数，有利于水环境中砷污染的修复，能够为砷污染水体修复以及维护提供可靠的数据依据。
The present invention relates to a method for determining key environmental parameters that restrict the arsenic accumulation capacity of microalgae, which may include the following steps: S1. obtaining environmental parameters related to the arsenic accumulation capacity of microalgae; S2. carrying out Taguchi ( Taguchi) experimental design; S3. Carry out experiments according to the experimental design obtained in S2 and analyze and calculate the experimental results to obtain microalgae arsenic accumulation ability parameters; S4. According to the microalgae arsenic accumulation ability parameters obtained in S3, perform signal-to-noise ratio S/N and variance analysis , Statistical calculation of factor percentage contribution PC, and then determine the key environmental parameters that restrict the accumulation of arsenic in microalgae. The invention can efficiently determine the key environmental parameters that restrict the arsenic accumulation capacity of microalgae, is beneficial to the restoration of arsenic pollution in the water environment, and can provide reliable data basis for the restoration and maintenance of arsenic-contaminated water bodies.
Description
Technical Field
The invention belongs to the field of water pollution treatment, and particularly relates to a method for determining key environmental parameters for restricting the arsenic accumulation capacity of microalgae.
Background
Arsenic (As), which is commonly present in the environment, is a highly mobile, highly toxic pollutant and is of high interest. A large amount of arsenic-containing wastewater is discharged into the environment in the production process of the industries of chemical industry, metal smelting, semiconductors, breeding, electroplating and the like. The method for repairing the water source polluted by the arsenic comprises a chemical ion exchange method, solvent extraction, chelation, filter membrane filtration, a reduction method, precipitation and the like, but the methods have high general operation cost and low economic efficiency, have no obvious effect on treating the water body polluted by the arsenic with lower solubility, are difficult to put into practical use, and are easy to generate precipitates and toxic compounds to cause secondary pollution. How to treat arsenic-containing polluted water efficiently and economically becomes one of the important problems in the field of water environment.
The microalgae are widely distributed in natural environment and have the characteristics of multiple types, large quantity and the like. As a common bioremediation agent, the microalgae can be used for removing heavy metals in a water environment, even can be used for treating and recovering metals in a water body, has the advantages of high efficiency, low consumption, environmental protection and the like, and has the following advantages when used for repairing arsenic-polluted water bodies: (1) the raw materials are rich in source, high in growth speed, low in price and easy to obtain; (2) high adsorption and absorption enrichment effects are simultaneously generated, and the removal rate is high; (3) has good selectivity; (4) arsenic absorbed by adsorption is easy to elute and discharge, and metal recovery is facilitated; (5) no secondary pollution is generated; (6) wide environmental adaptability and wide application range.
The environmental parameters of the water body polluted by arsenic in the natural environment are varied, and the method has great restriction influence on the accumulation capacity of the microalgae arsenic. By creating appropriate environmental parameters, the arsenic accumulation capacity of the microalgae can be improved, the arsenic removal performance of the microalgae can be promoted, and the bioremediation capacity of the microalgae on arsenic-polluted water bodies can be further improved. Therefore, how to determine the key environmental parameters which restrict the arsenic accumulation capacity of the microalgae becomes an important research problem.
Disclosure of Invention
The invention aims to provide a method for efficiently determining key environmental parameters restricting the arsenic accumulation capacity of microalgae. Therefore, the invention adopts the following specific technical scheme:
a method of determining key environmental parameters that restrict arsenic accumulation in microalgae can include the steps of:
s1, obtaining environmental parameters related to the arsenic accumulation capacity of microalgae;
s2, carrying out Taguchi test design according to the environmental parameters obtained in the step S1;
s3, testing according to the test design obtained in the step S2, analyzing and calculating a test result to obtain a microalgae arsenic accumulation capacity parameter;
and S4, performing signal-to-noise ratio S/N, variance analysis and statistical calculation of factor percentage contribution degree PC according to the microalgae arsenic accumulation capacity parameter obtained in the step S3, and further determining a key environmental parameter for restricting the microalgae arsenic accumulation capacity.
Further, the design of the Taguchi test specifically includes determining different influence levels of the environmental parameters according to the environmental parameters, and designing test conditions by using a Taguchi orthogonal array.
Further, the Takou orthogonal array is a four-factor three-level, wherein the four factors are four environmental parameters of nitrate nitrogen, phosphate, pentavalent arsenic and PH value, and the three levels are a high level, a middle level and a low level of the environmental parameters.
Further, the S3 specifically includes the following steps:
s31, according to the designed test conditions, the microalgae are cultured in a suspension mode by adopting a proper culture medium under the illumination intensity of 30 mu mol m-2s-1Culturing for 96h at the temperature of 20 +/-3 ℃ and the light-dark ratio of 16: 8;
s32, collecting dynamic change data of microalgae cell density OD, microalgae growth rate mu, arsenic accumulated total amount As (algae) in algae and arsenic content As (water) in water under a test culture condition;
s33, calculating to obtain microalgae arsenic accumulation capacity parameters according to the collected data, wherein the microalgae arsenic accumulation capacity parameters comprise the maximum growth rate mu of the algaemaxThe total accumulated arsenic content and the arsenic absorption rate K of the algaeuAnd release rate KeBCF of arsenic bioaccumulation coefficient, whereinmax＝Ln(OD)/t,KuAnd KeAccording to the formula
The biological accumulation coefficient of arsenic, BCF ═ K, was obtainedu/Ke。
Further, the S4 specifically includes the following steps:
s41, calculating S/N ratio of S/N-10 × log (sigma)nyi 2N), i ═ 1, …, n, where n is the number of repeated determinations under the same test conditions, yiIs the reciprocal of the measurement;
s42, variance analysis and calculation SST、SSF、VErDOF, wherein SSTIs the sum of squared mean differences, SSFIs the sum of squares of factors, VErFor error variance, DOF is the degree of freedom;
s43, calculating the factor percentage contribution degree PC, wherein the PC is (SS)F-(DOF×VEr))/SST×100；
S44, analyzing the maximum signal-to-noise ratio of the corresponding parameters according to the signal-to-noise ratio calculation result, thereby obtaining the optimal environment restriction parameters; and further analyzing the percentage contribution degree of the factors, and sequencing the optimal environmental parameters so as to determine the key environmental parameters which restrict the arsenic accumulation capacity of the microalgae.
Furthermore, the microalgae is one or more than two of microcystis aeruginosa, scenedesmus obliquus and marine nannochloropsis.
By adopting the technical scheme, the invention has the beneficial effects that: the method can efficiently determine the key environmental parameters restricting the arsenic accumulation capacity of the microalgae, is beneficial to the remediation of arsenic pollution in the water environment, and can provide reliable data basis for the remediation and maintenance of arsenic polluted water bodies.
Drawings
FIG. 1 is a general flow diagram of the process of the present invention;
FIG. 2 shows the signal-to-noise ratio S/N of the maximum specific growth rate of microalgae under the influence of different environmental parameters;
FIG. 3(a) shows the arsenic absorption rate k under the influence of different environmental parametersuS/N;
FIG. 3(b) shows the arsenic release rate K under the influence of different environmental parameterseS/N;
FIG. 3(c) shows the signal-to-noise ratio S/N of the arsenic bio-accumulation factor BCF under the influence of different environmental parameters.
Detailed Description
To further illustrate the various embodiments, the invention provides the accompanying drawings. The accompanying drawings, which are incorporated in and constitute a part of this disclosure, illustrate embodiments of the invention and, together with the description, serve to explain the principles of the embodiments. Those skilled in the art will appreciate still other possible embodiments and advantages of the present invention with reference to these figures.
The invention will now be further described with reference to the accompanying drawings and detailed description.
FIG. 1 is a general flow diagram of the method of the present invention for determining key environmental parameters that dictate the arsenic accumulation capacity of microalgae. As shown in fig. 1, the method comprises the steps of:
s1, obtaining environmental parameters related to the arsenic accumulation capacity of the microalgae.
The microalgae refers to algae species with high arsenic absorption and enrichment capacity, such as one or more than two of microcystis aeruginosa, scenedesmus obliquus or marine nannochloropsis. That is, the microalgae may beA single variety, or a combination of several varieties. The description will be given by taking Microcystis aeruginosa as an example. Through reference to the literature, environmental parameters related to the arsenic accumulation capacity of microcystis aeruginosa, including nitrate Nitrogen (NO)3 —N, N), Phosphate (PO)4 3—P, P), pentavalent arsenic (As)V) And pH, etc.
S2, carrying out Taguchi experimental design according to the environmental parameters obtained in S1.
In this step, according to the environmental parameters (nitrate Nitrogen (NO)3 —N, N), Phosphate (PO)4 3—P, P), pentavalent arsenic (As)V) And pH), three different impact levels of high, medium and low were determined for the selection parameters (as shown in table 1), and four-factor three-level experimental condition design was performed using the Taguchi orthogonal array. The test conditions are shown in table 2 below.
TABLE 1 test levels corresponding to environmental parameter factors
TABLE 2 Experimental design of orthogonal arrays of environmental parameter factors
And S3, testing according to the test design obtained in the step S2, analyzing and calculating the test result to obtain the microalgae arsenic accumulation capacity parameter. Specifically, S3 includes the steps of:
s31, culturing Microcystis aeruginosa in a BG-11 culture medium in a suspension manner under the illumination intensity of 30 mu mol m according to the designed orthogonal array test conditions-2s-1Culturing at 20 + -3 deg.C under light-dark ratio of 16:8 for 96 h. It should be noted that the above culture method is only exemplary and not limiting to the present invention.
S32, measuring the density (OD) of the microcystis aeruginosa by using a spectrophotometer, further collecting the growth rate of the microcystis aeruginosa under the experimental culture condition, and simultaneously collecting dynamic change data of the accumulated total amount (As (algae), mu g/g) and the content (AS (water), mu g/L) of arsenic in the microcystis aeruginosa by using inductively coupled plasma mass spectrometry (ICP-MS).
S33, for the selected microalgae arsenic accumulation capacity parameter: maximum growth rate of algae mumaxThe total accumulated arsenic content and the arsenic absorption rate K of the algaeuAnd release rate KeBCF of arsenic bioaccumulation coefficient, whereinmax＝Ln(OD)/t,KuAnd KeAccording to the formula
The biological accumulation coefficient of arsenic, BCF ═ K, was obtainedu/Ke. In particular, KuAnd KeThe data is obtained by iterative calculation through a plurality of data points.
And S4, performing signal-to-noise ratio S/N, variance analysis and statistical calculation of factor percentage contribution degree PC according to the microalgae arsenic accumulation capacity parameter obtained in the step S3, and further determining a key environmental parameter for restricting the microalgae arsenic accumulation capacity. Specifically, S3 includes the steps of:
s41, calculating the signal-to-noise ratio, wherein S/N is-10 × log (sigma)nyi 2N), i ═ 1, …, n, where n is the number of repeated determinations under the same test conditions, yiThe reciprocal of the measurement value. The calculation results are shown in FIGS. 2 and 3(a) to 3 (b). It should be noted that N, P, As in FIGS. 2, 3(a) -3(b) are nitrate Nitrogen (NO) respectively3 —N, N), Phosphate (PO)4 3—P, P), pentavalent arsenic (As)V) Is shown simplified.
S42, variance analysis and calculation SST、SSF、VErDOF. Wherein SSTIs the sum of the mean square differences; SSFIs the sum of the squares of the factors; vErIs the error variance; DOF is a degree of freedom.
S43, the factor percentage contribution degree (PC) is represented by the formula PC ═ SSF-(DOF×VEr))/SSTThe x 100 was calculated, and the calculation results are shown in tables 3 and 4.
TABLE 3 different environmental factors (N, P, pH, As) under ANOVAV) Contrast maximum specific growth rate (. mu.)max) Percentage Contribution (PC)
Note: type III SS is the sum of squares of Type III; MS is mean square; p is the probability; f is the statistic.
TABLE 4 different environmental factors (N, P, pH, As) under ANOVAV) For arsenic absorption rate kuArsenic Release Rate kePercentage contribution to arsenic biological accumulation factor BCF (PC)
Note: type III SS is the sum of squares of Type III; MS is mean square; p is the probability; f is the statistic.
S44, analyzing the maximum signal-to-noise ratio of the corresponding parameters according to the signal-to-noise ratio calculation result, thereby obtaining the optimal environment restriction parameters; and further analyzing the percentage contribution degree of the factors, and sequencing the optimal environmental parameters so as to determine the key environmental parameters which restrict the arsenic accumulation capacity of the microalgae. Sequencing is carried out according to the maximum signal-to-noise ratio calculation result and the larger factor percentage contribution degree, and the phosphorus is found to be the most key factor for restricting the arsenic accumulation capacity of the microcystis aeruginosa. Nitrogen is an important factor for the growth of microcystis aeruginosa and the release rate of microcystis aeruginosa arsenic. Pentavalent arsenic (As)V) Has little influence on the growth of algae. Therefore, the high level of nitrogen and the low concentration of phosphorus in alkaline conditions are beneficial to the accumulation of arsenic in the microcystis aeruginosa and reduce the release of arsenic, and the microcystis aeruginosa is the best condition for the pentavalent arsenic pollution remediation.
Therefore, according to the method for determining the key environmental parameter restricting the accumulation capacity of the arsenic in the microalgae provided by the embodiment of the invention, the key environmental parameter restricting the accumulation capacity of the arsenic in the microalgae determined in the embodiment is beneficial to the remediation of the arsenic pollution in the water environment, and a reliable data basis can be provided for the remediation and maintenance of the arsenic polluted water body.
While the invention has been particularly shown and described with reference to a preferred embodiment, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (5)
1. A method for determining key environmental parameters that restrict the arsenic accumulation capacity of microalgae, comprising the steps of:
s1, obtaining environmental parameters related to the arsenic accumulation capacity of microalgae;
s2, carrying out Taguchi test design according to the environmental parameters obtained in the step S1;
s3, testing according to the test design obtained in the step S2, analyzing and calculating the test result to obtain the microalgae arsenic accumulation capacity parameter, and specifically comprising the following steps:
s31, according to the designed test conditions, the microalgae are cultured in a suspension culture mode by adopting a proper culture medium under the illumination intensity of 30 mu mol-2s-1Culturing for 96h at the temperature of 20 +/-3 ℃ and the light-dark ratio of 16: 8;
s32, collecting dynamic change data of microalgae cell density OD, microalgae growth rate mu, arsenic accumulated total amount As (algae) in algae and arsenic content As (water) in water under a test culture condition;
s33, calculating to obtain microalgae arsenic accumulation capacity parameters according to the collected data, wherein the microalgae arsenic accumulation capacity parameters comprise the maximum growth rate mu of the algaemaxThe total amount of accumulated arsenic in algae, As (algae) and the arsenic absorption rate KuAnd release rate KeBCF of arsenic bioaccumulation coefficient, whereinmax＝Ln(OD)/t,KuAnd KeAccording to the formula
The biological accumulation coefficient of arsenic, BCF ═ K, was obtainedu/Ke；
And S4, performing signal-to-noise ratio S/N, variance analysis and statistical calculation of factor percentage contribution degree PC according to the microalgae arsenic accumulation capacity parameter obtained in the step S3, and further determining a key environmental parameter for restricting the microalgae arsenic accumulation capacity.
2. The method for determining key environmental parameters for restricting the arsenic accumulation capacity of microalgae according to claim 1, wherein the Taguchi test design is specifically that different influence levels of the environmental parameters are determined according to the environmental parameters, and the design of test conditions is carried out by utilizing a Taguchi orthogonal array.
3. The method of claim 2, wherein the Takokou orthogonal array is four-factor three levels, wherein four factors are nitrate nitrogen, phosphate, pentavalent arsenic and pH, and three levels are high, medium and low levels of the environmental parameters.
4. The method for determining key environmental parameters for restricting the arsenic accumulation capacity of microalgae according to claim 3, wherein the S4 specifically comprises the following steps:
s41, calculating S/N ratio of S/N-10 × log (sigma)nyi 2N), i ═ 1, …, n, where n is the number of repeated determinations under the same test conditions, yiIs the reciprocal of the measurement;
s42, variance analysis and calculation SST、SSF、VErDOF, wherein SSTIs the sum of squared mean differences, SSFIs the sum of squares of factors, VErFor error variance, DOF is the degree of freedom;
s43, calculating the factor percentage contribution degree PC, wherein the PC is (SS)F-(DOF×VEr))/SST×100；
S44, analyzing the maximum signal-to-noise ratio of the corresponding parameters according to the signal-to-noise ratio calculation result, thereby obtaining the optimal environment restriction parameters; and further analyzing the percentage contribution degree of the factors, and sequencing the optimal environmental parameters so as to determine the key environmental parameters which restrict the arsenic accumulation capacity of the microalgae.
5. The method of claim 1, wherein the microalgae is one or more of microcystis aeruginosa, scenedesmus obliquus, and parachlorococcus maritima.
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| CN102912000A (en) * | 2012-11-07 | 2013-02-06 | 中国科学院地球化学研究所 | Method for biologically dissolving limestone by quantitative microalgae |
| CN104370324A (en) * | 2014-03-25 | 2015-02-25 | 中国科学院海洋研究所 | Method for adsorbing heavy metal ions in environment by utilizing macroalgae |
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| CN102912000A (en) * | 2012-11-07 | 2013-02-06 | 中国科学院地球化学研究所 | Method for biologically dissolving limestone by quantitative microalgae |
| CN103173520A (en) * | 2012-11-07 | 2013-06-26 | 中国科学院地球化学研究所 | Bioerosion action method of quantitative microalgae to limestone |
| CN104370324A (en) * | 2014-03-25 | 2015-02-25 | 中国科学院海洋研究所 | Method for adsorbing heavy metal ions in environment by utilizing macroalgae |
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