CN109224364B - Method for reducing alkalinity of Bayer process red mud by using penicillium oxalicum - Google Patents
Method for reducing alkalinity of Bayer process red mud by using penicillium oxalicum Download PDFInfo
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Abstract
本发明公开了一种利用草酸青霉降低拜耳法赤泥碱性的方法,属于微生物应用技术领域,包括:(1)将赤泥自然风干并过筛,将得到的赤泥分为第一部分和第二部分,然后将经预处理后的甘蔗渣和麸皮加入到第一部分赤泥中,并混合均匀,得到添加生物质的赤泥;(2)进行土柱装填,将石英砂装填于土柱底部,第二部分赤泥作为中间层,将添加生物质的赤泥装填于顶部,并将草酸青霉菌种接种到第二部分赤泥中;(3)向装填好的土柱中从下向上充水,保持含水率为60%~80%,在室温下进行培育,降低赤泥碱性。本发明利用生物质与草酸青霉相结合的方法,能将赤泥的pH值降低至6.66,本发明使用原料来源广泛,成本低廉,而且微生物修复无二次污染,效果显著,可应用于赤泥堆场。
The invention discloses a method for reducing the alkalinity of Bayer process red mud by using Penicillium oxalate, and belongs to the technical field of microorganism application. In the second part, the pretreated bagasse and bran are then added to the first part of the red mud, and mixed evenly to obtain the biomass-added red mud; (2) soil column filling is carried out, and the quartz sand is filled in the soil At the bottom of the column, the second part of the red mud is used as the middle layer, the red mud with biomass added is packed on the top, and the Penicillium oxalic acid species is inoculated into the second part of the red mud; (3) Into the packed soil column from the bottom Fill up with water, keep the water content at 60% to 80%, and cultivate at room temperature to reduce the alkalinity of red mud. The method of combining biomass and Penicillium oxalate can reduce the pH value of red mud to 6.66. The method uses a wide range of raw materials, has low cost, and has no secondary pollution in microbial restoration. The effect is remarkable and can be applied to red mud. mud yard.
Description
技术领域technical field
本发明属于微生物应用技术领域,具体涉及一种利用草酸青霉降低拜耳法赤泥碱性的方法。The invention belongs to the technical field of microorganism application, and in particular relates to a method for reducing the alkalinity of Bayer process red mud by using Penicillium oxalate.
背景技术Background technique
拜尔法赤泥是氧化铝工业生产过程中排放的高碱性固体废弃物,每生产1 t氧化铝产生1.5~2.0 t赤泥,综合利用难度大,外排赤泥以堆存为主,2017年全球积存的待处理赤泥约40亿t,并以每年1.7亿t的速度增长。赤泥堆场的环境安全问题日益严重,如何有效处置和利用赤泥问题已迫在眉睫。国内外已尝试开展对赤泥堆场的生态修复,但由于赤泥碱性强,盐分含量高,营养物质匮乏,植物难以生长。由中南大学环境生态工程团队薛生国教授提出赤泥土壤化的研究思路,拟通过物理-化学-生物等方法将赤泥转化为类似土壤的基质,使经过改良的赤泥具备植物生长的基本条件,实现赤泥的无害化处置。Bayer process red mud is a highly alkaline solid waste discharged in the alumina industrial production process. Every 1 ton of alumina produced produces 1.5-2.0 tons of red mud, which is difficult to comprehensively utilize. The discharged red mud is mainly stored. In 2017, the world's accumulated red mud to be treated was about 4 billion tons, and it is increasing at an annual rate of 170 million tons. The environmental safety problem of the red mud yard is becoming more and more serious, and the problem of how to effectively dispose and utilize the red mud is imminent. At home and abroad, attempts have been made to carry out ecological restoration of red mud yard, but due to the strong alkalinity of red mud, high salt content, and lack of nutrients, it is difficult for plants to grow. Professor Xue Shengguo from the Environmental and Ecological Engineering Team of Central South University proposed the research idea of red mud soilization. It is planned to convert red mud into a soil-like matrix through physical-chemical-biological methods, so that the improved red mud has the basic conditions for plant growth. Realize the harmless disposal of red mud.
微生物繁殖速度快,数量多,能分解有机质,释放出营养元素,改善土壤结构,增加土壤肥力;某些特殊的功能菌株能代谢产生有机酸,降低赤泥碱性,为植物提供适宜生长的条件,促进赤泥土壤化进展。近年来,已有研究发现Acidobacteriaceae,Nitrosomonadaceae 和 Caulobacteraceae 等能在未修复的赤泥环境中生长,为赤泥堆场生态修复提供了重要指标。Microorganisms reproduce quickly and in large numbers, can decompose organic matter, release nutrients, improve soil structure, and increase soil fertility; some special functional strains can metabolize to produce organic acids, reduce the alkalinity of red mud, and provide suitable growth conditions for plants , to promote the progress of red mud soilization. In recent years, studies have found that Acidobacteriaceae, Nitrosomonadaceae and Caulobacteraceae can grow in unrestored red mud environment, providing important indicators for the ecological restoration of red mud yard.
传统的赤泥基质改良方法有物理法和化学法,但这两种方法成本较高,赤泥堆场占地面积大,用化学试剂喷洒等耗费量大,因此这些方法在实际应用中困难重重。而微生物具有繁殖速度快、应用成本低、无二次污染等特点,应用前景广泛。但是有关利用生物方法对赤泥修复的研究却相对较少。因此,本领域迫切需要利用一种低成本的生物方法对赤泥进行实际修复,从而为赤泥土壤化的实施提供技术支撑。The traditional red mud matrix improvement methods include physical method and chemical method, but these two methods are expensive, the red mud yard covers a large area, and the consumption of chemical reagent spraying is large, so these methods are difficult to apply in practice. . Microorganisms have the characteristics of fast reproduction, low application cost, and no secondary pollution, and have broad application prospects. However, there are relatively few studies on red mud remediation using biological methods. Therefore, there is an urgent need in the field to use a low-cost biological method to actually remediate red mud, so as to provide technical support for the implementation of red mud soilization.
发明内容SUMMARY OF THE INVENTION
针对现有技术的不足,本发明的目的在于提供一种操作简单、经济有效、环境友好的利用草酸青霉降低拜耳法赤泥碱性的方法,以解决大量拜耳法赤泥堆存的环境安全问题。In view of the deficiencies of the prior art, the object of the present invention is to provide a simple, cost-effective and environmentally friendly method for reducing the alkalinity of Bayer process red mud by utilizing Penicillium oxalate, so as to solve the environmental safety of a large amount of Bayer process red mud storage. question.
为了实现上述技术目的,本发明提供以下技术方案:In order to realize the above-mentioned technical purpose, the present invention provides the following technical solutions:
本发明提供一种利用草酸青霉降低拜耳法赤泥碱性的方法,包括以下步骤:The invention provides a method for reducing the alkalinity of Bayer process red mud by utilizing Penicillium oxalate, comprising the following steps:
(1) 将赤泥自然风干并过筛,将得到的赤泥分为第一部分和第二部分,然后将经预处理后的甘蔗渣和麸皮加入到第一部分赤泥中,并混合均匀,得到添加生物质的赤泥;(1) The red mud is naturally air-dried and sieved, and the obtained red mud is divided into a first part and a second part, and then the pretreated bagasse and bran are added to the first part of the red mud, and mixed evenly, obtain biomass-added red mud;
(2) 进行土柱装填,将石英砂装填于土柱底部,第二部分赤泥作为中间层,将步骤(1)所得添加生物质的赤泥装填于顶部,并将草酸青霉菌种接种到第一部分赤泥中,得到装填好的土柱;(2) carrying out soil column filling, filling quartz sand at the bottom of the soil column, the second part of the red mud as the middle layer, filling the red mud of the gained biomass in step (1) on the top, and inoculating the Penicillium oxalicum species into the In the first part of the red mud, a packed soil column is obtained;
(3) 向步骤(2)中装填好的土柱中从下向上充水,保持含水率为60%~80%,在室温下进行培育,降低赤泥碱性,增加赤泥中有机质含量,促进赤泥团聚体的形成。(3) filling the soil column filled in the step (2) with water from bottom to top, keeping the moisture content at 60% to 80%, and cultivating at room temperature, reducing the alkalinity of red mud, and increasing the content of organic matter in the red mud, Promote the formation of red mud aggregates.
作为优选,步骤(1)中,所述甘蔗渣、麸皮和第一部分赤泥的质量百分比为(6~18%):(5~15%):(67~89%)。Preferably, in step (1), the mass percentage of the bagasse, bran and the first part of red mud is (6~18%): (5~15%): (67~89%).
作为优选,步骤(1)中,所述预处理为热液处理、硫酸处理、氧化钙处理和双氧水处理中的一种。Preferably, in step (1), the pretreatment is one of hydrothermal treatment, sulfuric acid treatment, calcium oxide treatment and hydrogen peroxide treatment.
进一步,所述预处理采用热液处理,具体步骤为:Further, the pretreatment adopts hydrothermal treatment, and the specific steps are:
(a) 将甘蔗渣和麸皮烘干,然后用粉碎机将它们粉碎,过60~80目筛;(a) Dry the bagasse and bran, then pulverize them with a pulverizer and pass through a 60-80 mesh sieve;
(b) 将过筛后的甘蔗渣、麸皮和水加入到烧杯中,置于高压蒸汽灭菌锅,加热到150~180℃,保持压力为4.0~6.0MPa,处理5~10min,最后突降压力对生物质进行爆破,得到经预处理后的甘蔗渣和麸皮。(b) Add the sieved bagasse, bran and water into a beaker, place it in a high-pressure steam sterilizer, heat it to 150~180°C, keep the pressure at 4.0~6.0MPa, and treat it for 5~10min. The biomass is blasted by reducing the pressure to obtain pretreated bagasse and bran.
作为优选,步骤(2)中,土柱装填的具体步骤为:As preferably, in step (2), the concrete steps of soil column filling are:
1) 建立室内土柱装置模型,将透明的柱子固定在铁架上,土柱横截面内径为6.3cm,外径为7.0cm,在柱体上每隔10cm深度有带塞的小孔,土柱上方加有淋溶装置,模拟赤泥堆场雨水天气,土柱最下端尖端接一橡皮管,橡皮管接入锥形瓶中盛取渗滤液;1) Build an indoor soil column device model, fix the transparent column on the iron frame, the inner diameter of the cross section of the soil column is 6.3cm, and the outer diameter is 7.0cm. A leaching device is added above the column to simulate the rainy weather of the red mud yard. A rubber tube is connected to the tip of the bottom end of the soil column, and the rubber tube is connected to the conical flask to collect the leachate;
2) 进行土柱装填,在土柱底部(65cm深度以下)装填6cm厚的石英砂,在土柱25~65cm深度装填过第二部分赤泥,然后将步骤(1)所得添加生物质的赤泥装填在土柱0~25cm深度;2) Filling the soil column, filling the bottom of the soil column (below 65cm depth) with 6cm thick quartz sand, filling the second part of the red mud at the depth of the soil column 25~65cm, and then adding the biomass-added red mud obtained in step (1). The mud is filled in the soil column at a depth of 0~25cm;
3) 分别装填两根土柱,命名为E柱和F柱,在F柱中定期接种草酸青霉菌剂,E柱作为对照不接种菌剂。3) Fill two soil columns respectively, named as E column and F column, inoculate the F column with Penicillium oxalate regularly, and E column as a control without inoculating the bacterial agent.
进一步,所述草酸青霉菌剂的制备过程为:Further, the preparation process of described Penicillium oxalate agent is:
a) 配制葡萄糖液体培养基,取100mL装于250mL锥形瓶中,进行高压蒸汽灭菌;a) Prepare glucose liquid medium, take 100mL and put it in a 250mL conical flask, and carry out high pressure steam sterilization;
b) 将草酸青霉接种至灭菌冷却后的培养基中,于25~30℃,160~180r/min培养72~96h;b) Inoculate Penicillium oxalate into the sterilized and cooled medium, and cultivate at 25~30℃, 160~180r/min for 72~96h;
c) 将培养好的草酸青霉菌剂接种至F柱中。c) Inoculate the cultured Penicillium oxalate agent into the F column.
在前期实验中,发明人团队从赤泥堆场上筛选出耐碱性能好,产酸性能高的微生物菌株草酸青霉,微生物序列号为:MF802280,其中草酸青霉在赤泥碱性调控方面作用显著,而且它作为一种重要的产纤维素酶微生物,可以降解纤维素,在碳素循环中有重要作用。In the previous experiments, the inventor team screened out the microbial strain Penicillium oxalicum with good alkali resistance and high acid production performance from the red mud yard. It has a significant effect, and as an important cellulase-producing microorganism, it can degrade cellulose and play an important role in the carbon cycle.
本发明的原理:拜耳法赤泥碱性强,营养物质匮乏,结构松散,植物在原生赤泥堆场上难以生长。本发明通过在拜耳法赤泥中添加经热液处理的甘蔗渣和麸皮,作为微生物生存所需的碳源和氮源,然后向赤泥中施加草酸青霉菌剂,其代谢产生大量的有机酸,能有效降低赤泥碱性;草酸青霉通过利用甘蔗渣和麸皮为碳氮源能分解生物质,同时草酸青霉也会分泌纤维素酶,增加赤泥中有机质含量;其次草酸青霉的菌丝物理缠绕作用,能进一步促进赤泥团聚体的形成。The principle of the invention: the Bayer process red mud has strong alkalinity, lack of nutrients and loose structure, and it is difficult for plants to grow on the original red mud yard. In the present invention, sugarcane bagasse and bran treated with hydrothermal treatment are added to the Bayer process red mud as the carbon source and nitrogen source required for the survival of microorganisms, and then the Penicillium oxalate agent is applied to the red mud, and its metabolism produces a large amount of organic Acid, can effectively reduce the alkalinity of red mud; Penicillium oxalate can decompose biomass by using bagasse and bran as carbon and nitrogen sources, and Penicillium oxalate will also secrete cellulase to increase the content of organic matter in red mud; The physical entanglement of mold mycelium can further promote the formation of red mud aggregates.
相对于现有技术,本发明具有以下有益技术效果:Compared with the prior art, the present invention has the following beneficial technical effects:
本发明提供一种利用草酸青霉降低拜耳法赤泥碱性的方法,利用生物质与草酸青霉相结合的方法,能将赤泥的pH值降低至6.66,且提高了赤泥中有机质的含量,促进赤泥中团聚体的形成。本发明使用原料来源广泛,成本低廉,而且微生物修复无二次污染,效果显著,可应用于赤泥堆场,且能有效解决赤泥堆场对环境的污染问题。The invention provides a method for reducing the alkalinity of Bayer process red mud by using Penicillium oxalate, and the method of combining biomass and Penicillium oxalate can reduce the pH value of the red mud to 6.66, and improve the organic matter in the red mud. content, promoting the formation of aggregates in red mud. The invention uses a wide range of raw materials, has low cost, and has no secondary pollution for microbial remediation, and has a remarkable effect, can be applied to a red mud yard, and can effectively solve the problem of environmental pollution caused by the red mud yard.
附图说明Description of drawings
图1为生物质的不同预处理方法对草酸青霉产酸效果的影响。Figure 1 shows the effect of different pretreatment methods of biomass on the acid production of Penicillium oxalicum.
图2为土柱模型图。(F:添加草酸青霉的赤泥柱;E:不添加草酸青霉的赤泥柱)Figure 2 shows the soil column model diagram. (F: Red mud column with Penicillium oxalate added; E: Red mud column without Penicillium oxalate)
图3为赤泥pH、EC(电导率)和总碱随时间的变化图。Figure 3 is a graph showing the changes in pH, EC (conductivity) and total alkali of red mud over time.
图4为赤泥中有机质含量随时间的变化图。Figure 4 is a graph showing the change of organic matter content in red mud with time.
图5为赤泥中脲酶、纤维素酶含量随时间的变化图。Figure 5 is a graph showing the changes of urease and cellulase contents in red mud with time.
图6为修复后第30天的赤泥碱性阳离子(Na+、Ca2+、K+、Mg2+)含量变化图。Figure 6 is a graph of changes in the content of alkaline cations (Na+, Ca2+, K+, Mg2+) in red mud on the 30th day after repair.
图7为赤泥的SEM图。Figure 7 is an SEM image of red mud.
具体实施方式Detailed ways
下面将对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅是本发明一部分实施例,而不是全部实施例,基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, ordinary skills in the art All other embodiments obtained by personnel without creative work fall within the protection scope of the present invention.
下面结合具体实施例和附图对本发明进行进一步说明:The present invention will be further described below in conjunction with specific embodiments and accompanying drawings:
实施例1Example 1
生物质预处理可以采用以下4种方法,通过对比不同生物质预处理方法,确定草酸青霉产酸的最优条件,最后选择最佳预处理方法:Biomass pretreatment can adopt the following four methods. By comparing different biomass pretreatment methods, the optimal conditions for acid production by Penicillium oxalicum are determined, and finally the optimal pretreatment method is selected:
(1) 热液处理(1) Hydrothermal treatment
利用粉碎机将甘蔗渣和麸皮粉碎,过60目筛(孔径0.25mm),然后将甘蔗渣、麸皮和水按固液比1:10放在烧杯中,置于高压蒸汽灭菌锅,加热到180℃,保持压力4.0MPa左右5min,最后突降压力对生物质进行爆破。Use a pulverizer to pulverize the bagasse and bran, pass through a 60-mesh sieve (pore size 0.25mm), then put the bagasse, bran and water in a beaker at a solid-liquid ratio of 1:10, and place it in a high-pressure steam sterilizer. Heating to 180 ℃, maintaining the pressure of about 4.0MPa for 5min, and finally depressurizing the biomass to blast.
(2) 硫酸处理(2) Sulfuric acid treatment
将甘蔗渣和麸皮粉碎,过60目筛(孔径0.25mm),称取适量甘蔗渣和麸皮,按固液比1:10加入1%(w/w)H2SO4溶液,搅拌均匀,高压蒸汽下121℃处理1h。Pulverize the bagasse and bran, pass through a 60-mesh sieve (pore size 0.25mm), weigh an appropriate amount of bagasse and bran, add 1% (w/w) H2SO4 solution at a solid-to-liquid ratio of 1:10, stir evenly, and steam under high pressure. Treat at 121°C for 1h.
(3) 氧化钙处理(3) Calcium oxide treatment
将石灰(主要成分为氧化钙)浆化(生成氢氧化钙),然后每克生物质配0.075g氢氧化钙和5g水,在120℃温度下加热4h。The lime (the main component is calcium oxide) is slurried (to generate calcium hydroxide), and then each gram of biomass is mixed with 0.075g of calcium hydroxide and 5g of water, and heated at 120 ° C for 4 hours.
(4) 双氧水处理(4) Hydrogen peroxide treatment
称取10.0g甘蔗渣和麸皮于250mL锥形瓶中,按固液比1:10加入5%(w/w)的H2O2溶液,搅拌均匀,置于水浴恒温震荡箱中30℃处理24h。Weigh 10.0g bagasse and bran into a 250mL conical flask, add 5% (w/w) H2O2 solution at a solid-to-liquid ratio of 1:10, stir evenly, and place it in a water bath constant temperature shaking box for 24h at 30°C.
将处理过后的生物质与蒸馏水以1:5的比例配制成培养液,再接种草酸青霉于液体培养基中,同时设置空白对照组,放置于 28℃,160r/min摇床培养9天后测定pH值。The treated biomass and distilled water were prepared into a culture solution at a ratio of 1:5, and then inoculated with Penicillium oxalate in the liquid medium. At the same time, a blank control group was set up, placed at 28°C, and cultured on a 160r/min shaker for 9 days. pH.
图1结果表明,热液处理和H2SO4处理效果均显著高于CaO处理和H2O2处理方法(P<0.05),且热液处理和H2SO4处理后无显著性差异,但两者相比,H2SO4处理需要添加化学试剂,若引用至赤泥修复中,会引入外源离子,因此采用热液处理法。The results in Figure 1 show that the effects of hydrothermal treatment and H2SO4 treatment are significantly higher than those of CaO treatment and H2O2 treatment (P<0.05), and there is no significant difference between hydrothermal treatment and H2SO4 treatment, but compared with the two, H2SO4 treatment requires Adding chemical reagents, if used in the restoration of red mud, will introduce exogenous ions, so the hydrothermal treatment method is adopted.
实施例2Example 2
本发明提供一种利用草酸青霉降低拜耳法赤泥碱性的方法,包括以下步骤:The invention provides a method for reducing the alkalinity of Bayer process red mud by utilizing Penicillium oxalate, comprising the following steps:
(1) 将赤泥自然风干并过筛,将得到的赤泥分为第一部分和第二部分,然后将经热液处理后的甘蔗渣和麸皮加入到第一部分赤泥中,甘蔗渣、麸皮和第一部分赤泥的质量百分比为6%:5%:89%,并混合均匀,得到添加生物质的赤泥;(1) The red mud is naturally air-dried and sieved, and the obtained red mud is divided into the first part and the second part, and then the bagasse and bran after the hydrothermal treatment are added to the first part of the red mud, bagasse, The mass percentage of the bran and the first part of the red mud is 6%: 5%: 89%, and mixed evenly to obtain the red mud added with biomass;
(2) 进行土柱装填,土柱模型如图2所示,E柱和F柱的0~25cm层为添加生物质的赤泥,25~65cm深度的为第二部分赤泥,柱子底部(65cm以下)装填6cm深度的石英砂,防止赤泥随渗滤液的流出而流失,E柱和F柱装填好后,不定期对其进行淋溶,模拟赤泥堆场的降雨天气,F柱中每隔3天接种一次草酸青霉菌剂;(2) Fill the soil column. The soil column model is shown in Figure 2. The 0-25cm layer of the E column and the F column is the red mud with biomass added, the 25-65cm depth is the second part of the red mud, and the bottom of the column ( 65cm or less) is filled with quartz sand with a depth of 6cm to prevent the loss of red mud with the outflow of leachate. After the E column and F column are filled, they are leached from time to time to simulate the rainy weather in the red mud yard. Inoculate Penicillium oxalate every 3 days;
草酸青霉菌剂的制备过程为:The preparation process of penicillium oxalate agent is as follows:
a) 配制葡萄糖液体培养基,取100mL装于250mL锥形瓶中,进行高压蒸汽灭菌(21℃,20min);a) Prepare a glucose liquid medium, take 100 mL of it and put it in a 250 mL conical flask, and sterilize it by autoclaving (21°C, 20min);
b) 将草酸青霉接种至灭菌冷却后的培养基中,于28℃,180r/min培养72h;b) Inoculate Penicillium oxalate into the sterilized and cooled medium, and cultivate at 28°C and 180r/min for 72h;
c) 将培养好的草酸青霉菌剂接种至F柱中;c) Inoculate the cultured Penicillium oxalate agent into the F column;
(3) 向装填好的土柱中从下向上充水,保持含水率为60%~80%,在室温下进行培育,降低赤泥碱性,增加赤泥中有机质含量,促进赤泥团聚体的形成。(3) Fill the packed soil column with water from bottom to top, keep the moisture content at 60%~80%, cultivate at room temperature, reduce the alkalinity of red mud, increase the content of organic matter in red mud, and promote red mud agglomeration Formation.
实施例3Example 3
本发明提供一种利用草酸青霉降低拜耳法赤泥碱性的方法,包括以下步骤:The invention provides a method for reducing the alkalinity of Bayer process red mud by utilizing Penicillium oxalate, comprising the following steps:
(1) 将赤泥自然风干并过筛,将得到的赤泥分为第一部分和第二部分,然后将经热液处理后的甘蔗渣和麸皮加入到第一部分赤泥中,甘蔗渣、麸皮和第一部分赤泥的质量百分比为18%:15%:67%,并混合均匀,得到添加生物质的赤泥;(1) The red mud is naturally air-dried and sieved, and the obtained red mud is divided into the first part and the second part, and then the bagasse and bran after the hydrothermal treatment are added to the first part of the red mud, bagasse, The mass percentage of the bran and the first part of the red mud is 18%: 15%: 67%, and mixed evenly to obtain the red mud added with biomass;
(2) 进行土柱装填,土柱模型如图2所示,E柱和F柱的0~25cm层为添加生物质的赤泥,25~65cm深度的为第二部分赤泥,柱子底部(65cm以下)装填6cm深度的石英砂,防止赤泥随渗滤液的流出而流失,E柱和F柱装填好后,不定期对其进行淋溶,模拟赤泥堆场的降雨天气,F柱中每隔3天接种一次草酸青霉菌剂;(2) Fill the soil column. The soil column model is shown in Figure 2. The 0-25cm layer of the E column and the F column is the red mud with biomass added, the 25-65cm depth is the second part of the red mud, and the bottom of the column ( 65cm or less) is filled with quartz sand with a depth of 6cm to prevent the loss of red mud with the outflow of leachate. After the E column and F column are filled, they are leached from time to time to simulate the rainy weather in the red mud yard. Inoculate Penicillium oxalate every 3 days;
(3) 向装填好的土柱中从下向上充水,保持含水率为60%~80%,在室温下进行培育,降低赤泥碱性,增加赤泥中有机质含量,促进赤泥团聚体的形成。(3) Fill the packed soil column with water from bottom to top, keep the moisture content at 60%~80%, cultivate at room temperature, reduce the alkalinity of red mud, increase the content of organic matter in red mud, and promote red mud agglomeration Formation.
实施例4Example 4
分析实施例2所得赤泥中pH、EC、总碱(Total alkalinity)的变化:Analyze the variation of pH, EC, total alkali (Total alkalinity) in the red mud gained in Example 2:
分别在第6、12、18、24、30天对E柱和F柱的0、5cm、15cm、25cm、35cm和45cm深度处的赤泥进行取样,样品风干至恒重后,分别称取5.0g赤泥于离心管中,然后加入25mL去离子水(1:5),摇匀,静置30min后,测定上清液pH和EC。吸取10mL土水比为1:5的赤泥浸出液,放入100mL锥形瓶中,用溴酚蓝指示剂滴定法测定样品中CO32-(HCO3-)含量,向上清液加入溴酚蓝指示剂,用H2SO4标准溶液滴定至无色(pH 4.5),通过计算得到CO32-含量和HCO3-含量。赤泥中总碱是CO32-、HCO3-、Al(OH)4- 和OH-含量的总和,其中Al(OH)4-含量通过电感耦合等离子体原子发射光谱法(ICP-AES,Optima 5300DV,美国 Perkin Elmer 公司)测定上清液中Al含量并计算而得。On the 6th, 12th, 18th, 24th, and 30th days, the red mud at the depths of 0, 5cm, 15cm, 25cm, 35cm and 45cm of the E column and the F column was sampled. After the samples were air-dried to constant weight, 5.0 g red mud was placed in a centrifuge tube, then 25 mL of deionized water (1:5) was added, shaken well, and after standing for 30 min, the pH and EC of the supernatant were measured. Draw 10mL of red mud leachate with a soil-water ratio of 1:5, put it into a 100mL conical flask, measure the CO32- (HCO3-) content in the sample by bromophenol blue indicator titration, and add bromophenol blue indicator to the supernatant , titrated with H2SO4 standard solution to colorless (pH 4.5), and obtained CO32- content and HCO3- content by calculation. The total alkalinity in red mud is the sum of CO32-, HCO3-, Al(OH)4- and OH- content, where Al(OH)4- content was determined by inductively coupled plasma atomic emission spectrometry (ICP-AES, Optima 5300DV, American Perkin Elmer Company) determined the Al content in the supernatant and calculated it.
结果如图3所示,在F柱中,赤泥pH随着时间的变化而变化,在第18天时,F柱的pH略微上升,可能是因为草酸青霉代谢的酸性物质中和了大量的自由碱,导致赤泥中结合碱的溶出。第24天后,F柱的pH维持在7左右,且第30天的pH与第6天的pH存在显著差异(P<0.05)。而在E柱中,赤泥的pH基本不变,其pH值维持在9左右,30天内未有显著差异。在第30天时,F柱的pH显著低于E柱的pH(P<0.05),说明草酸青霉能有效降低赤泥的pH,修复效果较好。在E柱中,EC随时间的增加而降低,可能是因为淋溶作用导致柱中碱性离子的迁移。而F柱中,EC随时间的增加而增加,可能是因为草酸青霉产生的酸性物质将赤泥中碱性离子溶出。两柱中第24天的EC显著高于第30天,可能主要由淋溶作用导致。F柱的总碱要显著低于E柱的总碱(P<0.05),说明草酸青霉产生的酸性物质对赤泥的碱性有很大影响。The results are shown in Figure 3. In the F column, the pH of the red mud changed with time. On the 18th day, the pH of the F column increased slightly, probably because the acidic substances metabolized by Penicillium oxalicum neutralized a large amount of Free base, leading to dissolution of bound base in red mud. After the 24th day, the pH of the F column was maintained at around 7, and the pH on the 30th day was significantly different from that on the 6th day (P<0.05). In the E column, the pH of the red mud was basically unchanged, and its pH value was maintained at around 9, and there was no significant difference within 30 days. On the 30th day, the pH of column F was significantly lower than that of column E (P<0.05), indicating that Penicillium oxalicum could effectively reduce the pH of red mud, and the repair effect was better. In the E column, the EC decreased with time, probably due to the migration of basic ions in the column due to leaching. In the F column, the EC increased with time, probably because the acidic substances produced by Penicillium oxalicum dissolved the alkaline ions in the red mud. The EC on day 24 was significantly higher than that on
F柱中,0~5cm层的赤泥pH显著低于E柱(P<0.05),说明草酸青霉在表层深度的活性较大,作用效果较明显。而F柱中赤泥EC值在0~5cm层显著高于E柱,说明草酸青霉能溶解出赤泥中大量的结合碱,使得碱性离子含量增加。在35~45cm层中,F柱的赤泥pH显著低于E柱(P<0.05),说明草酸青霉代谢的酸性物质在淋溶的作用下能迁移到下层与赤泥中的碱性物质反应。在F柱中,赤泥总碱含量均低于E柱,且F柱中未经改良的35~45cm层的赤泥总碱含量低于E柱,说明草酸青霉代谢的有机酸能明显降低赤泥的碱性,对赤泥碱性具有很好的调控作用。In the F column, the pH of the red mud in the 0-5cm layer was significantly lower than that in the E column (P<0.05), indicating that Penicillium oxalicum had greater activity at the surface depth and had a more obvious effect. The EC value of red mud in the F column was significantly higher than that in the E column in the 0-5 cm layer, indicating that Penicillium oxalicum can dissolve a large amount of bound alkali in the red mud, which increases the content of alkaline ions. In the 35-45cm layer, the pH of the red mud in the F column was significantly lower than that in the E column (P<0.05), indicating that the acidic substances metabolized by Penicillium oxalicum could migrate to the lower layer and the alkaline substances in the red mud under the action of leaching. reaction. In the F column, the total alkali content of red mud is lower than that of the E column, and the total alkali content of the red mud in the unmodified 35-45 cm layer in the F column is lower than that in the E column, indicating that the organic acids metabolized by Penicillium oxalicum can be significantly reduced. The alkalinity of red mud has a good control effect on the alkalinity of red mud.
实施例5Example 5
分析实施例2所得赤泥中有机质、脲酶、纤维素酶的变化:Changes in organic matter, urease and cellulase in the red mud obtained in Example 2 were analyzed:
(1) 有机质含量的测定(1) Determination of organic matter content
分别在第6、12、18、24、30天对E柱和F柱的0、5cm、15cm、25cm、35cm和45cm深度处的赤泥进行取样,样品风干至恒重后研磨,准确称取过100目筛的赤泥1.000g,放入50mL烧杯中,加入3mL水,充分摇散,加入10mL 1mol/L(1/6 K2Cr2O7)溶液,然后加入10mL浓硫酸不断摇动,放置20min,加水10mL,摇匀,静置过夜。吸取上清液3mL于10mL比色管中,加水至刻度摇匀,在590nm波长处测定,根据标准曲线计算出有机质含量。On the 6th, 12th, 18th, 24th and 30th days, the red mud at the depths of 0, 5cm, 15cm, 25cm, 35cm and 45cm of the E column and the F column was sampled, and the samples were air-dried to constant weight and then ground, and accurately weighed. Put 1.000g of red mud that has passed through a 100-mesh sieve into a 50mL beaker, add 3mL of water, shake well, add 10mL of 1mol/L (1/6 K2Cr2O7) solution, then add 10mL of concentrated sulfuric acid, shake continuously, leave for 20min, add 10mL of water , shake well and let stand overnight. Pipette 3mL of the supernatant into a 10mL colorimetric tube, add water to the mark, shake well, measure at a wavelength of 590nm, and calculate the organic matter content according to the standard curve.
(2) 脲酶(Urease)含量的测定(2) Determination of urease (Urease) content
分别在第6、12、18、24、30天对E柱和F柱的0、5cm、15cm、25cm、35cm和45cm深度处的赤泥进行取样,样品烘干至恒重,称取5.0g赤泥样于50mL锥形瓶中,加1mL甲苯,振荡均匀,15min后加10mL 10%尿素溶液和20mL pH 6.7的柠檬酸盐缓冲溶液,摇匀后在37℃恒温箱培养24小时。培养结束后过滤,过滤后取1mL滤液加入50mL容量瓶中,再加4mL苯酚钠溶液和3mL次氯酸钠溶液,边加边摇匀。20min后显色,定容。1h内在分光光度计于578nm波长处比色。(靛酚的蓝色在1h内保持稳定)On the 6th, 12th, 18th, 24th and 30th days, the red mud at the depths of 0, 5cm, 15cm, 25cm, 35cm and 45cm of the E column and the F column was sampled. The samples were dried to constant weight and weighed 5.0g. The red mud sample was placed in a 50 mL conical flask, 1 mL of toluene was added, and the mixture was shaken evenly. After 15 minutes, 10 mL of 10% urea solution and 20 mL of pH 6.7 citrate buffer solution were added. After culturing, filter, add 1 mL of filtrate into a 50 mL volumetric flask, add 4 mL of sodium phenolate solution and 3 mL of sodium hypochlorite solution, and shake up while adding. After 20 minutes, the color develops and the volume is fixed. Within 1h, the spectrophotometer was colorimetric at 578nm wavelength. (The blue color of indophenol remains stable within 1h)
(3) 纤维素酶(Cellulolytic enzyme)含量的测定(3) Determination of Cellulolytic enzyme content
分别在第6、12、18、24、30天对E柱和F柱的0、5cm、15cm、25cm、35cm和45cm深度处的赤泥进行取样,样品烘干至恒重,称10.0g赤泥置于50mL锥形瓶中,加入1.5ml甲苯,摇匀后放置15min,再加5ml 1%羧甲基纤维素溶液和5ml pH5.5醋酸盐缓冲液,将三角瓶放在37℃恒温箱中培养72h。培养结束后,过滤并取1mL滤液,然后按绘制标准曲线显色法比色测定。(为了消除土壤中原有的蔗糖、葡萄糖而引起的误差,每一土样需做无基质对照,整个试验需做无土壤对照;如果样品吸光值超过标曲的最大值,则应该增加分取倍数或减少培养的土样。)On the 6th, 12th, 18th, 24th and 30th days, the red mud at the depths of 0, 5cm, 15cm, 25cm, 35cm and 45cm of the E column and the F column was sampled, and the samples were dried to constant weight and weighed 10.0g red mud. Put the mud into a 50mL conical flask, add 1.5ml toluene, shake well and leave for 15min, add 5ml 1% carboxymethyl cellulose solution and 5ml pH5.5 acetate buffer, and place the flask at 37°C at a constant temperature Incubate for 72h. After the incubation, filter and take 1 mL of filtrate, and then draw a standard curve colorimetric method for colorimetric determination. (In order to eliminate the errors caused by the original sucrose and glucose in the soil, each soil sample needs to be a matrix-free control, and the whole experiment needs to be a soil-free control; if the sample absorbance value exceeds the maximum value of the standard curve, the fractionation multiple should be increased. Or reduce the soil samples for cultivation.)
实验结果如图4所示,随着时间的增加,两柱中的有机质含量略微增加。第12天时,柱中有机质含量最高,随后有所降低。两柱中0~25cm层的有机质含量显著高于35~45cm层(P<0.01)。从图5可以看出,E、F两柱中脲酶活性随时间的增加而增加,而E柱中的脲酶活性与F柱并没有明显差异,说明草酸青霉对脲酶活性影响不大。E、F两柱中0~25cm层的脲酶活性显著高于35~45cm层(P<0.01),说明添加了甘蔗渣和麸皮能明显改善赤泥中的酶活,提高氮的循环。E柱中纤维素酶的活性低于F柱,说明添加草酸青霉能改善赤泥中纤维素酶的活性,从而提高碳的循环。F柱中0~5cm层中的纤维素酶活性显著高于15~25cm层,说明草酸青霉在表层作用较大。在F柱中,第30天的酶活较高,说明草酸青霉在第30天时存活状态和数量达到最高。The experimental results are shown in Figure 4. With the increase of time, the content of organic matter in the two columns increased slightly. On the 12th day, the content of organic matter in the column was the highest and then decreased. The organic matter content of the 0-25cm layer in the two columns was significantly higher than that of the 35-45cm layer (P<0.01). It can be seen from Figure 5 that the urease activity in the E and F columns increased with time, while the urease activity in the E column was not significantly different from the F column, indicating that Penicillium oxalicum had little effect on the urease activity. The urease activity in the 0-25cm layer in the two columns E and F was significantly higher than that in the 35-45cm layer (P<0.01), indicating that the addition of bagasse and bran could significantly improve the enzyme activity in red mud and improve the nitrogen cycle. The activity of cellulase in column E was lower than that in column F, indicating that adding Penicillium oxalicum could improve the activity of cellulase in red mud, thereby increasing the carbon cycle. The cellulase activity in the 0-5 cm layer in the F column was significantly higher than that in the 15-25 cm layer, indicating that Penicillium oxalicum played a greater role in the surface layer. In the F column, the enzyme activity was higher on the 30th day, indicating that the survival state and number of Penicillium oxalicum reached the highest on the 30th day.
实施例6Example 6
分析实施例2所得赤泥中碱性阳离子的变化:Changes of basic cations in the red mud obtained in Example 2 were analyzed:
在第30天对E柱和F柱的0、5cm、15cm、25cm、35cm和45cm深度处的赤泥进行取样,样品风干至恒重后,称取5.0g,加入25mL 去离子水,摇匀,取上清液,采用电感耦合等离子体原子发射光谱法(ICP-AES)测定赤泥中的碱性阳离子Na+、K+、Ca2+、Mg2+。On the 30th day, the red mud at the depths of 0, 5cm, 15cm, 25cm, 35cm and 45cm of the E column and the F column was sampled. After the samples were air-dried to a constant weight, 5.0 g was weighed, 25 mL of deionized water was added, and the samples were shaken well. , the supernatant was taken, and the basic cations Na+, K+, Ca2+, Mg2+ in red mud were determined by inductively coupled plasma atomic emission spectrometry (ICP-AES).
由于第30天赤泥柱的碱性最低,故对第30天的水溶性碱性阳离子进一步分析。发现F柱的阳离子含量均显著高于E柱,尽管F柱的pH低于E柱,但其Na+含量高于E柱,说明草酸青霉代谢的酸性物质能促进Na+的溶出。在F柱和E柱中Na+含量分别占总碱性阳离子的86%和90%,说明Na+在土柱赤泥中的基本阳离子中占主导地位。F柱表层的Na+含量最高为5069.1mg/kg,且深度越深,离子含量越低,在45cm层Na+含量为998.25mg/kg;E柱表层的Na+含量最高为103.26mg/kg,且越到土柱的下部,离子含量越低,在45cm层Na+含量为65.26mg/kg。在F柱中Ca2+含量从表层的263.66mg/kg降低到土柱45cm层的72.28mg/kg;E柱中Ca2+含量较低,表层为31.24mg/kg,在35~45cm层基本检测不到。F柱中,K+在0~25cm处含量高于35~45cm层,尤其是表层含量达到了316.01mg/kg,而E柱中K+含量在0~25cm处和35~45cm处差异并不明显;在F柱中, Mg2+含量从表层的35.87mg/kg降低到土柱45cm层的2.61mg/kg,而E柱中Mg2+含量很低,在35cm~45cm层基本检测不到。说明草酸青霉的添加能显著提高赤泥中的营养元素,对赤泥基质改良作用效果显著。Since the red mud column had the lowest alkalinity on the 30th day, the water-soluble alkaline cations on the 30th day were further analyzed. It was found that the cation content of the F column was significantly higher than that of the E column. Although the pH of the F column was lower than that of the E column, its Na+ content was higher than that of the E column, indicating that the acidic substances metabolized by Penicillium oxalicum could promote the dissolution of Na+. The Na+ content in the F column and E column accounts for 86% and 90% of the total basic cations, respectively, indicating that Na+ dominates the basic cations in the red mud of the soil column. The Na+ content in the surface layer of the F-column is up to 5069.1 mg/kg, and the deeper the depth, the lower the ion content. The Na+ content in the 45cm layer is 998.25 mg/kg; the Na+ content in the E-column surface is up to 103.26 mg/kg, and the more In the lower part of the soil column, the lower the ion content, the Na+ content in the 45cm layer is 65.26mg/kg. The Ca2+ content in column F decreased from 263.66 mg/kg in the surface layer to 72.28 mg/kg in the 45 cm layer of the soil column; the content of Ca2+ in the E column was lower, 31.24 mg/kg in the surface layer, which was basically undetectable in the 35-45 cm layer. In column F, the content of K+ at 0-25 cm is higher than that at 35-45 cm, especially the surface layer content reaches 316.01 mg/kg, while the content of K+ in column E is not significantly different between 0-25 cm and 35-45 cm; In the F column, the Mg2+ content decreased from 35.87 mg/kg in the surface layer to 2.61 mg/kg in the 45 cm layer of the soil column, while the Mg2+ content in the E column was very low and could not be detected in the 35 cm-45 cm layer. It shows that the addition of Penicillium oxalate can significantly increase the nutrient elements in red mud, and has a significant effect on the improvement of red mud matrix.
实施例7Example 7
分析实施例2中土柱中赤泥的颗粒微观形貌变化图:Analysis of the change diagram of the microscopic morphology of the red mud particles in the soil column in Example 2:
在第30天对E柱和F柱的表层(0cm)深度处的赤泥进行取样,样品自然风干后轻微研磨,过300目筛,利用电镜扫描(SEM),测定两柱中赤泥细微颗粒的变化情况。On the 30th day, the red mud at the surface (0cm) depth of the E column and the F column was sampled. The samples were air-dried and ground slightly, passed through a 300-mesh sieve, and the fine particles of red mud in the two columns were determined by scanning electron microscope (SEM). changes.
在未添加草酸青霉的E柱的赤泥中,较多0.1~0.5μm细小颗粒分布在2~5μm团聚体上,结晶度差,颗粒分布相对分散、无序。而F柱中的赤泥相比E柱中的赤泥,细小颗粒明显减少,结晶度有一定的改善,大颗粒增加明显,团聚结构较好。F柱与E柱相比,其Na离子含量由1.47%降低至0.24%,Ca离子含量由6.65%升高至24.21%,进一步说明草酸青霉能促进赤泥碱性降低,并改善赤泥团聚结构。In the red mud of the E-column without Penicillium oxalate added, many fine particles of 0.1-0.5 μm are distributed on the aggregates of 2-5 μm, the crystallinity is poor, and the particle distribution is relatively scattered and disordered. Compared with the red mud in the E column, the fine particles of the red mud in the F column are significantly reduced, the crystallinity is improved to a certain extent, the large particles are increased significantly, and the agglomeration structure is better. Compared with the E column, the Na ion content of the F column decreased from 1.47% to 0.24%, and the Ca ion content increased from 6.65% to 24.21%, which further indicated that Penicillium oxalicum could promote the reduction of red mud alkalinity and improve red mud agglomeration. structure.
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