CN107937299B - Thermophilic Pb mineralization bacteria and method for passivating Pb in sludge high-temperature compost by using same - Google Patents
Thermophilic Pb mineralization bacteria and method for passivating Pb in sludge high-temperature compost by using same Download PDFInfo
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- CN107937299B CN107937299B CN201711079841.6A CN201711079841A CN107937299B CN 107937299 B CN107937299 B CN 107937299B CN 201711079841 A CN201711079841 A CN 201711079841A CN 107937299 B CN107937299 B CN 107937299B
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Abstract
The invention discloses a thermophilic Pb mineralization strain Thermus thermophilus FAFU013 with a deposit number: CGMCC No. 14654. The thermophilic Pb mineralization bacteria FAFU013 can induce Pb (II) to be converted into the most stable and difficult-to-dissolve phosphorochloridite [ Pb [ ]5(PO4)3Cl，Ksp＝10‑84.4]The method not only can effectively passivate heavy metal lead in the sludge and reduce the bioavailability of Pb (II), but also can utilize the original nutrient substances of the sludge to the maximum extent. In addition, the Pb mineralizing bacteria FAFU013 provided by the invention can also utilize nutrient substances in sludge to grow and propagate at 70-80 ℃, has good thermal stability, is not inhibited by the high-temperature composting temperature in metabolic activity, is beneficial to improving and maintaining the temperature of a high-temperature composting pile, promotes the degradation of organic matters and the killing of pathogenic microorganisms, and shortens the fermentation period.
Description
Technical Field
The invention belongs to the technical field of environmental protection, and particularly relates to a thermophilic Pb mineralization bacterium and a method for synergistically passivating lead in sludge high-temperature compost.
Background
Sludge is a sediment inevitably generated in the urban domestic sewage treatment process. The annual sludge production of China is increased at a speed of 10-15% every year, and the annual wet sludge production exceeds four million tons by 2016. These increasingly large and highly concentrated sludges are long puzzling the healthy development of the sewage treatment industry in China, and also cause great potential safety hazards, environmental pressure and economic burden. On the other hand, the sludge contains rich nutrient elements such as N, P and organic matters, and is organic solid waste with high agricultural recycling value. The sludge can be used as organic fertilizer after composting treatment, so that the soil fertility is improved, and the yield of crops is increased. However, the existence of a large amount of heavy metals in the sludge causes concern about environmental risks and food safety which may be caused by sludge agriculture, and greatly limits the implementation and harmless disposal of sludge agricultural utilization. Lead (Pb) is one of the major heavy metal pollutants in sludge, and its content can reach hundreds of milligrams per kilogram of sludge, and can enter human body through the way of food chain enrichment, etc., and cause serious damage to heart, kidney, reproductive system and nervous system, especially to the development of children nervous system, and have the risk of teratogenesis, carcinogenesis and mutagenesis. Therefore, the heavy metal lead in the sludge is removed or passivated, the bioavailability of the heavy metal lead in the sludge is reduced, and the method has great significance for recycling and harmless disposal of the sludge.
The traditional method for removing heavy metals in sludge adopts a chemical leaching method and utilizes inorganic acid or complexing agent (H)2SO4、HNO3HCl, EDTA) and leaching heavy metals, which consumes a large amount of inorganic acid and complexing agent, has high treatment cost, is liable to cause secondary pollution, and may cause loss of nutrients in sludge, although it can remove most of heavy metals. In recent years, researchers report that the heavy metals in the sludge are removed by a microbial leaching method, which is to dissolve and remove the heavy metals in the sludge by using sulfur oxidizing bacteria and thiobacillus ferrooxidans to generate sulfuric acid by taking sulfur as an energy source (US 5217615, US5454948 and CN 02112924. X).
Researches show that microorganisms can promote heavy metal pollutants to be converted into thermodynamically stable and extremely insoluble minerals, so that the migration and the biological activity of the minerals are reduced, and the microorganisms are considered to be one of the most effective methods for in-situ remediation of heavy metal pollutants. The method not only can effectively passivate heavy metals in the sludge, but also can protect original nutrient substances of the sludge from losing to the maximum extent. At present, various microorganisms such as Bacillus cereus, Vibrio Hawaii, providencia alkalopenii, Burkholderia cepacia and the like have been reported to promote lead to form stable and refractory minerals. However, the lead mineralization bacteria reported at present are all normal temperature bacteria (the optimal growth temperature is less than 40 ℃), and can not grow and propagate in high temperature environment such as compost or ultrahigh temperature compost. So far, no research report about the synergism of the thermophilic lead mineralization bacteria and the mineralization action of thermophilic microorganisms to promote the Pb passivation in the sludge high-temperature compost is available.
Disclosure of Invention
The invention aims to provide a thermophilic Pb mineralization bacterium and a method for synergistically passivating Pb in sludge high-temperature compost by using the mineralization effect of a thermophilic bacterium Thermus thermophilus (thermophilic Thermus thermophilus), and the Pb in the sludge high-temperature compost is synergistically promoted to be converted into thermodynamically stable and extremely insoluble minerals, so that the migration and the biological activity of the Pb are reduced.
The technical scheme adopted by the invention is as follows:
a thermophilic Pb mineralization strain Thermus thermophilus FAFU013 with a deposit number: CGMCC No. 14654.
Application of Thermus thermophilus FAFU013 strain in sludge high-temperature composting.
A method for synergistically inactivating Pb in sludge high-temperature compost by using the strain of claim 1, comprising the steps of,
(1) preparation of a fermented seed solution of Thermus thermophilus FAFU013 Strain: selecting colonies, inoculating the colonies into a DSMZ Medium 74 liquid culture Medium, and performing activation culture; inoculating 2% of the activated bacteria liquid into a seed tank filled with DSMZ Medium 74 culture Medium, and performing fermentation culture;
(2) preparing a Pb passivation microbial inoculum: inoculating the fermented seed liquid into a sludge fermentation culture medium according to the inoculation amount of 2%, and performing fermentation culture; after the fermentation is finished, adding a conditioner, uniformly stirring, and adjusting the water content;
(3) mixing Pb passivating agent and materials in the sludge high-temperature compost according to the weight ratio of 1-4%.
Preferably, the sludge fermentation medium is: taking the concentrated sludge of a sewage treatment plant, adjusting the sludge to a solid content of 4%, adding 0.2% of natural sodium chloride, 1.5g/L of yeast powder, 0.5g/L of sodium chloride and 0.5g/L of cane sugar, adjusting the pH to 7.0, and uniformly stirring.
Preferably, the conditioning agent is at least one of corn flour, wheat bran and bean cake flour.
Preferably, the content of the conditioning agent is 3-5%.
Preferably, when the Pb passivation microbial inoculum is prepared, the water content is adjusted to be 45-55%.
Preferably, the conditions of the fermentation culture are: the fermentation temperature is 75 deg.C, and the ventilation amount is 20-100m3The stirring speed is 180 DEG and the fermentation time is 48 hours at 220 rpm.
Preferably, the Thermus thermophilus FAFU013 is capable of converting available lead to pnictogen.
The invention has the beneficial effects that: the thermophilic Pb mineralizing bacteria FAFU013 can induce Pb (II) to be converted into the phosphorochloridite (Pb)5(PO4)3Cl) which is the most stable and difficult-to-dissolve lead-containing mineral (Ksp ═ 10)-84.4) The method not only can effectively passivate heavy metal lead in the sludge and reduce the bioavailability of Pb (II), but also can utilize the original nutrient substances of the sludge to the maximum extent. In addition, the Pb mineralizing bacteria FAFU013 provided by the invention can also utilize nutrient substances in sludge to grow and propagate at 70-80 ℃, has good thermal stability, is not inhibited by the high-temperature composting temperature in metabolic activity, is beneficial to improving and maintaining the temperature of a high-temperature composting pile, promotes the degradation of organic matters and the killing of pathogenic microorganisms, and shortens the fermentation period.
Drawings
FIG. 1 Effect of different lead ion concentrations on growth of Thermus thermophilus FAFU 013.
FIG. 2 XRD analysis of adsorbed lead ions of Thermus thermophilus FAFU 013.
FIG. 3 shows the measurement of the lead content adsorbed by the cells of FAFU013 by Atomic Absorption Spectroscopy (AAS), and the measurement of the conversion of the lead-containing mineral mediated by the cells of FAFU013 by XRD.
Detailed Description
Example 1 screening and identification of FAFU013 Strain
(1) Screening of FAFU013 Strain
Weighing 5g of a sludge compost sample in a high temperature period (<70 ℃) in a triangular flask containing 15 glass beads and 50mL of sterile water, placing the mixture on a shaker for 1 hour under 180rpm to fully disperse the sample, then placing the mixture at 75 ℃ and standing the mixture for 3 hours.
And (3) sucking 500 mu L of high-temperature enrichment solution and low-power dilution solution, and uniformly coating the mixture in an improved YTPG solid culture medium. The evenly coated flat plate is placed into a sealing bag in an inverted mode, a culture dish filled with distilled water is placed at the same time, and the flat plate is sealed and then placed into an incubator at 75 ℃ for culture. After the colonies grow out, the characteristics of the colonies are carefully observed to determine different types of strains, and the strains are separated and purified by streaking on a good YTPG solid plate culture medium until FAFU013 pure colonies are obtained. The FAFU013 is transferred to a culture medium slant, cultured to be thick and preserved at 4 ℃ for later use.
(2) Identification of FAFU013 Strain
The FAFU013 strain of the present invention was inoculated into the modified YTPG medium, cultured at 75 ℃ for 24 hours, gram-stained, and observed for its characteristics. The bacterium is gram-negative and aerobic bacterium, does not form spores, and has a rod shape.
Extracting the total DNA of the strain FAFU013, and performing PCR amplification on the 16S rRNA sequence by using the total DNA as a template. And (5) sequencing the amplified product after detecting the purity of the amplified product by electrophoresis. The obtained 16S rDNA sequences were aligned with the EzTaxon database, and only the strain with homology of more than 97% was Thermus thermophilus HB8(T) (similarity 99.71%), so FAFU013 was identified as Thermus thermophilus.
The strain provided by the invention is named as Thermus thermophilus FAFU 013. And is preserved in China general microbiological culture Collection center (CGMCC for short, the address: No. 3 of West Lu No.1 of Beijing Kogyo, Chaozhou, China academy of sciences, postal code: 100101) in 2017, 9 months and 21 days, with the preservation number: CGMCC No. 14654.
The strain can utilize the nutrient components in the organic solid wastes such as sludge, livestock and poultry manure and the like to carry out growth and propagation, and actively grows at 70-80 ℃; can induce Pb (II) to convert into the lead-phosphorus-chloride ore under the condition of 30-85 ℃.
Example 2 tolerance of the FAFU013 strain to lead and its mediation to mineralize lead
(1) Colonies were picked from a preserved normal Thermus thermophilus FAFU013 plate Medium, inoculated into DSMZ Medium 74 liquid Medium, and activated and cultured at 75 ℃ and 180rpm for 16 hours. Activated Thermus thermophilus FAFU013 at 1% ratio respectively containing Pb (II) (Pb (NO) at 50mg/L, 100mg/L, 200mg/L, 300mg/L, 400mg/L3)2) The Medium was cultured in DSMZ Medium 74 liquid Medium at 75 ℃ at 180 rpm. The influence of different lead ion concentrations on the growth of Thermus thermophilus FAFU013 is determined by detecting the OD600 value of the bacterial liquid. The results are shown in FIG. 1, and show that 400mg/L Pb (II) grew on Thermus thermophilus FAFU013Has no influence.
(2) After 48 hours of incubation, the cells were centrifuged at 12000rpm for 10 minutes, and the supernatant was taken to measure the residual lead content. The cells obtained after centrifugation were washed three times with distilled water, then dried at 60 ℃ and analyzed for lead-containing minerals on the cells by XRD. As a result, as shown in FIG. 2, Thermus thermophilus FAFU013 was able to adsorb lead ions and convert them into pnotide (Pb)5(PO4)3Cl)。
Example 3FAFU013 mediated mineralization of lead ions in lead-containing wastewater
(1) Taking lead-containing wastewater of a certain enterprise in Fujian (the basic properties of a water sample are shown in Table 1), and adjusting the pH of the wastewater to 5.5-6.5.
TABLE 1 basic Properties of lead-containing wastewater to be tested
(2) Activated Thermus thermophilus FAFU013 was inoculated into DSMZ Medium 74 liquid Medium at 1%, cultured at 75 ℃ for 48 hours at 180rpm, and centrifuged to collect the cells.
(3) After the culture, the cells of FAFU013 were added to the lead-containing wastewater at an amount of 10g/L (wet weight), and the mixture was stirred at 180 ℃ for 220 rpm for 24 hours at 70 ℃ to measure the lead content adsorbed by the cells of FAFU013 by Atomic Absorption Spectroscopy (AAS), and the conversion of the lead-containing mineral mediated by the cells of FAFU013 by XRD.
The results showed that Thermus thermophilus FAFU013 was able to adsorb lead ions in lead-containing wastewater and convert them to pnictogen (Pb) in 24 hours5(PO4)3Cl)。
Example 4 synergistic Pb inactivation of FAFU013 Strain in sludge high temperature composting
(1) Preparation of a fermented seed solution of Thermus thermophilus FAFU013 Strain: colonies were picked from a preserved normal Thermus thermophilus FAFU013 plate Medium, inoculated into DSMZ Medium 74 liquid Medium, and activated and cultured at 75 ℃ and 180rpm for 16 hours. Inoculating 2% of the activated bacteria solution into a seed tank containing DSMZ Medium 74 culture Medium, ventilating at 75 deg.CThe amount is 20-100m3The stirring speed is 180 DEG and the fermentation time is 48 hours at 220 rpm.
(2) Preparing a sludge fermentation culture medium: taking concentrated sludge (the basic properties of the sludge are shown in table 2) of a certain sewage treatment plant of Fujian province, adjusting the sludge to solid content of 4%, adding 0.2% of natural pond molide, 1.5g/L of yeast powder, 0.5g/L of sodium chloride and 0.5g/L of cane sugar, adjusting the pH to 7.0, uniformly stirring, sterilizing at 100 ℃ for 45 minutes, and cooling to 70-80 ℃ for later use.
TABLE 2 basic Properties of the sludge tested
(3) Preparation of Pb (II) inactivating inoculum: inoculating 2% seed liquid of Thermus thermophilus FAFU013 into sludge fermentation culture medium at 75 deg.C and with aeration rate of 20-100m3The stirring speed is 180 DEG and the fermentation time is 220 rpm for 48 hours. After fermentation is finished, 4% of corn flour is added, and after uniform stirring, the corn flour is subjected to plate-and-frame filter pressing to ensure that the water content is 45% -55%.
(4) Pb (II) method for synergistically passivating Pb by using passivating microbial inoculum in sludge high-temperature composting: mixing the Pb (II) passivating agent and the materials in the sludge high-temperature compost according to the weight ratio of 1%. After municipal sludge, sawdust and corn crushed stalks are taken according to the weight ratio of 2:1:1 and are uniformly mixed, a Pb (II) passivating microbial inoculum is added according to 1 percent of the total material, and the mixture is piled and fermented. The test was carried out without inoculation, but with conventional treatment. Wherein, the conventional treatment is carried out, and the pile turning is carried out every 5 days or at the temperature of more than 65 ℃; inoculating treatment, and turning the pile every 5 days. And judging the composting progress through the composting temperature. Taking the composted sample, and detecting the content of the effective lead in the composted sample according to the national standard of the people's republic of China, namely determination of effective lead and cadmium in soil quality (GB/T23739-2009) (see table 3).
TABLE 3 change in available lead content in sludge before and after composting
| Treatment of | Total lead (mg/kg) | Active state lead (mg/kg) |
| Sludge sample before composting | 179.45 | 98.70 |
| Samples after conventional composting | 178.65 | 85.96 |
| Composting sample after adding lead passivation microbial inoculum | 181.10 | 34.33 |
The result shows that compared with sludge samples before composting and after composting by the conventional method, the total amount of lead in the sludge is basically unchanged after the Pb (II) passivating agent is added, but the content of the effective lead in the sludge is greatly reduced.
Claims (6)
1. A thermophilic Pb mineralization strain is characterized in that: the thermophilic Pb mineralization bacterium is thermophilic Thermus thermophilus (Thermus thermophilus) FAFU013 with a deposit number: CGMCC No. 14654.
2. The strain of claim 1 for use in sludge high temperature composting.
3. A method for synergistically inactivating Pb in sludge high-temperature compost by using the strain of claim 1, comprising the steps of,
(1) preparation of Thermus thermophilus (Thermus thermophilus) FAFU013 Strain fermented seed liquid: selecting colonies, inoculating the colonies into a DSMZ Medium 74 culture Medium, and performing activation culture; inoculating the activated bacterium liquid into a seed tank filled with a DSMZ Medium 74 culture Medium according to the inoculation amount of 2 percent, and performing fermentation culture;
(2) preparing a Pb passivation microbial inoculum: inoculating the fermented seed liquid into a sludge fermentation culture medium according to the inoculation amount of 2%, and performing fermentation culture;
after the fermentation is finished, adding a conditioner, uniformly stirring, and adjusting the water content;
(3) mixing a Pb passivating agent and materials in the sludge high-temperature compost according to the weight ratio of 1-4%;
wherein the sludge fermentation medium is as follows: taking the concentrated sludge of a sewage treatment plant, adjusting the sludge to a solid content of 4%, adding 0.2% of natural sodium chloride, 1.5g/L of yeast powder, 0.5g/L of sodium chloride and 0.5g/L of cane sugar, adjusting the pH to 7.0, and uniformly stirring.
4. The method for the synergistic passivation of Pb in sludge high-temperature compost as claimed in claim 3, wherein the conditioner is at least one of corn flour, wheat bran and bean cake flour.
5. The method for the synergistic passivation of Pb in the sludge high-temperature compost as claimed in claim 3, wherein the mass ratio of the conditioner to the sludge fermentation medium is 3% -5%.
6. The method for the synergistic passivation of Pb in sludge high-temperature compost according to claim 3, characterized in that the conditions of fermentation culture are as follows: the fermentation temperature is 75 deg.C, and the ventilation amount is 20-100m3The stirring speed is 180 DEG and the fermentation time is 48 hours at 220 rpm.
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