Heavy Metal Treatment Composite Microbial Agent in Water and Preparation Method Thereof

Abstract

The present invention discloses a heavy metal treatment composite microbial agent in water and a preparation method thereof, belonging to the field of heavy metal treatment. The microbial agent of the present invention is prepared from the following components in parts by weight: 20-30 parts of Pseudomonas, 15-30 parts of Bacillus, 5-15 parts of Staphylococcus, and 5-15 parts of Pichia pastoris. The microbial agent of the present invention can quickly and efficiently adsorb and remove heavy metal ions, and the removal efficiencies of the microbial agent of the present invention on the cadmium, copper, lead and chromium after 2 d reach 81.0%, 56.5%, 52.0% and 74.0% respectively, wherein the adsorption and removal effects on the cadmium and chromium are most obvious. In addition, the microbial agent of the present invention can effectively improve the removal efficiency of the pollutants in the sewage to be treated, can achieve 80% CODMn removal rate or more, 85% TN removal rate or more, 80% TP removal rate or more, and 80% NH.sub.4.sup.+-N removal rate or more with a small amount, meets the pollutant discharge standards of the sewage treatment plant, and has a good application prospect.

Claims

1. A heavy metal ion treatment method, wherein the method comprises adding the microbial agent to a sample to remove heavy metal ions in the sample, the microbial agent comprising the following components in parts by weight: 20-30 parts of Pseudomonas, 15-30 parts of Bacillus, 5-15 parts of Staphylococcus, and 5-15 parts of Pichia pastoris.

2. The method according to claim 1, wherein the Pseudomonas comprises one or more of Pseudomonas aeruginosa, Pseudomonas brenneri, Pseudomonas putida, and Pseudomonas stutzeri.

3. The method according to claim 1, wherein the Pichia pastoris is Pichia membranifaciens.

4. The method according to claim 1, the microbial agent comprising the following components in parts by weight: 25 parts of Pseudomonas, 20 parts of Bacillus, 15 parts of Staphylococcus and 10 parts of Pichia pastoris.

5. The method according to claim 2, the microbial agent comprising the following components in parts by weight: 25 parts of Pseudomonas, 20 parts of Bacillus, 15 parts of Staphylococcus and 10 parts of Pichia pastoris.

6. The method according to claim 3, the microbial agent comprising the following components in parts by weight: 25 parts of Pseudomonas, 20 parts of Bacillus, 15 parts of Staphylococcus and 10 parts of Pichia pastoris.

7. The method according to claim 1, the microbial agent comprising the following component in parts by weight: 5-20 parts of Fusarium.

8. The method according to claim 1, wherein the added amount of the microbial agent is not less than 0.2%.

9. The method according to claim 1, further comprises enriching the microbial agent onto a carrier, the carrier being a spongy cube carrier ACP or PM.

10. The method according to claim 9, wherein the content of the strain relative to the carrier is not less than 10 g/g.

11. The method according to claim 10, wherein the content of the strain relative to the carrier is 50-150 g/g.

12. A sewage treatment method, wherein the method comprises adding the microbial agent to the sewage to remove pollutants in the sewage, the microbial agent comprising the following components in parts by weight: 20-30 parts of Pseudomonas, 15-30 parts of Bacillus, 5-15 parts of Staphylococcus, and 5-15 parts of Pichia pastoris.

Description

DETAILED DESCRIPTION

[0029] The sewage of the present invention is taken from the river ecological sewage of a community in Wuxi City, Jiangsu Province, China: pH is 6.53, COD.sub.Mn is 54.61 mg/L, TN mass concentration is 35.15 mg/L, TP mass concentration is 3.14 mg/L, and NH.sub.4.sup.+-N mass concentration is 31.58 mg/L.

Example 1

[0030] Preparation of microbial agent: Pseudomonas aeruginosa CICC 10351 and Bacillus cereus CICC 21155 were respectively cultured in a nutrient broth agar medium to obtain P. aeruginosa CICC 10351 fermentation broth and B. cereus CICC 21155 fermentation broth; Staphylococcus CICC 10311 was cultured in a wort agar medium to obtain Staphylococcus CICC 10311 fermentation broth; Pichia membranifaciens CICC 33242 was cultured in a wort agar medium to obtain P. membranifaciens CICC 33242 fermentation broth;

[0031] According to parts by weight, 25 parts of the P. aeruginosa CICC 10351 fermentation broth, 20 parts of the B. cereus CICC 21155 fermentation broth, 15 parts of the Staphylococcus CICC 10311 fermentation broth, and 10 parts of the P. membranifaciens CICC 33242 fermentation broth were mixed to obtain a composite microbial agent.

[0032] 1000 mL of sewage water sample was taken, and 0.2%, 0.25%, 0.3%, 0.5% and 1% by mass of microbial agents were respectively added for water degradation experiments. The culture was performed at a temperature of 30 C. for 72 h respectively. The removal effects on COD.sub.Mn, TN, TP and NH.sub.4.sup.+-N were determined. The specific removal rate results are shown in Table 1.

TABLE-US-00001 TABLE 1 Removal effects of different added amounts of microbial agent on COD.sub.Mn, TN, TP and NH.sub.4.sup.+N Added Amount of Microbial COD.sub.Mn TN TP NH.sub.4.sup.+N Agent (mg/L) (mg/L) (mg/L) (mg/L) 0 (not added) 54.61 35.15 3.14 31.58 0.2% 9.88 6.93 0.62 6.22 0.25% 7.35 5.75 0.49 4.96 0.3% 6.02 4.96 0.35 3.45 0.5% 5.64 4.20 0.22 2.89 .sup.1% 4.89 3.86 0.19 2.05

[0033] Detection methods: COD.sub.Mn was determined by an acidic permanganate oxidation method (GB 11892-1989); TN was determined by an alkaline potassium persulfate ultraviolet spectrophotometry (GB 11894-89); NH.sub.4.sup.+-N was determined by Nessler reagent colorimetry (GB 7479-87); and TP was determined by potassium persulfate oxidation-molybdenum antimony anti-spectrophotometry (GB11893-89).

[0034] It can be seen from Table 1 that the microbial agent can achieve 80% COD.sub.Mn removal rate or more, 85% TN removal rate or more, 80% TP removal rate or more, and 80% NH.sub.4.sup.+-N removal rate or more with a small amount, and meets the pollutant discharge standards of the sewage treatment plant.

Example 2

[0035] The microbial agent was prepared according to the formula shown in Example 1. 0.2% microbial agent was enriched in 20 mg spongy cube carrier ACP membrane at room temperature for 24 h; and 1000 mL of sewage water sample was taken, and the enriched carrier ACP membrane was added to the sewage for water degradation experiment. The culture was performed at a temperature of 30 C. for 72 h respectively. The removal effects on COD.sub.Mn, TN, TP and NH.sub.4.sup.+-N were determined. The removal rates were 89.8%, 86.5%, 90.5%, and 88.8% respectively.

Example 3

[0036] 4 parts of 200 mL sewage water sample was taken, and cadmium, copper, lead and chromium were respectively added in an amount of 0.04 mg to respectively obtain 4 samples, in which the metal ion concentration was 0.2 mg/kg;

[0037] The microbial agent in Example 1 was inoculated into the 4 samples in a dose of 0.3%, cultured at 30 C. in a dark shaker, and sampled on the second and third days, and the content of metal ions in the sample was determined by atomic absorption spectrophotometry, as shown in Table 2.

TABLE-US-00002 TABLE 2 Metal ion adsorption and removal effect of microbial agent Sampling Cadmium Copper Lead Chromium Time mg/kg mg/kg mg/kg mg/kg 0 0.2 0.2 0.2 0.2 48 h 0.038 0.087 0.096 0.052 72 h 0.021 0.075 0.087 0.033

[0038] It can be seen from Table 2 that the microbial agent of the present invention has good adsorption and removal effects on heavy metal ions such as cadmium, copper, lead and chromium, and the removal efficiencies on cadmium, copper, lead and chromium after 2 d respectively reach 81.0%, 56.5%, 52.0% and 74.0%, wherein the adsorption and removal effects on cadmium and chromium are the most obvious.

Example 4

[0039] Preparation of microbial agent: P. aeruginosa CICC 10351, Pseudomonas stutzeri CICC 23621 and B. cereus CICC 21155 were respectively cultured in a nutrient broth agar medium to obtain P. aeruginosa CICC 10351 fermentation broth, P. stutzeri CICC 23621 fermentation broth and B. cereus CICC 21155 fermentation broth; Staphylococcus CICC 10311 was cultured in a wort agar medium to obtain Staphylococcus CICC 10311 fermentation broth; P. membranifaciens CICC 33242 was cultured in a wort agar medium to obtain P. membranifaciens CICC 33242 fermentation broth;

[0040] According to parts by weight, 10 parts of the P. aeruginosa CICC 10351 fermentation broth, 10 parts of the P. stutzeri CICC 23621 fermentation broth, 20 parts of the B. cereus CICC 21155 fermentation broth, 5 parts of the Staphylococcus CICC 10311 fermentation broth, and 5 parts of the P. membranifaciens CICC 33242 fermentation broth were mixed to obtain a composite microbial agent.

Example 5

[0041] Preparation of microbial agent: P. aeruginosa CICC 10351, P. stutzeri CICC 23621, P. brenneri CICC 10271 and B. cereus CICC 21155 were respectively cultured in a nutrient broth agar medium to obtain P. aeruginosa CICC 10351 fermentation broth, P. stutzeri CICC 23621 fermentation broth, P. brenneri CICC 10271 fermentation broth and B. cereus CICC 21155 fermentation broth; Staphylococcus CICC 10311 was cultured in a wort agar medium to obtain Staphylococcus CICC 10311 fermentation broth; P. membranifaciens CICC 33242 was cultured in a wort agar medium to obtain P. membranifaciens CICC 33242 fermentation broth;

[0042] According to parts by weight, 10 parts of the P. aeruginosa CICC 10351 fermentation broth, 10 parts of the P. stutzeri CICC 23621 fermentation broth, 10 parts of P. brenneri CICC 10271 fermentation broth, 15 parts of the B. cereus CICC 21155 fermentation broth, 10 parts of the Staphylococcus CICC 10311 fermentation broth, and 10 parts of the P. membranifaciens CICC 33242 fermentation broth were mixed to obtain a composite microbial agent.

Example 6

[0043] Preparation of microbial agent: P. stutzeri CICC 23621 and B. cereus CICC 21155 were respectively cultured in a nutrient broth agar medium to obtain P. aeruginosa CICC 10351 fermentation broth, P. stutzeri CICC 23621 fermentation broth and B. cereus CICC 21155 fermentation broth; Staphylococcus CICC 10311 was cultured in a wort agar medium to obtain Staphylococcus CICC 10311 fermentation broth; P. membranifaciens CICC 33242 was cultured in a wort agar medium to obtain P. membranifaciens CICC 33242 fermentation broth; Fusarium fujikuroi CICC 2489 was cultured in a potato agar medium to obtain F. fujikuroi CICC 2489 fermentation broth;

[0044] According to parts by weight, 25 parts of the P. stutzeri CICC 23621 fermentation broth, 30 parts of the B. cereus CICC 21155 fermentation broth, 5 parts of the Staphylococcus CICC 10311 fermentation broth, 5 parts of the P. membranifaciens CICC 33242 fermentation broth and 10 parts of F. fujikuroi CICC 2489 fermentation broth were mixed to obtain a composite microbial agent.

[0045] 1000 mL of sewage water sample was taken, and 0.2% by mass of microbial agents obtained in Examples 4-6 were respectively added for water degradation experiments. The culture was performed at a temperature of 30 C. for 72 h respectively. The removal effects on COD.sub.Mn, TN, TP and NH.sub.4.sup.+-N were determined. The specific removal rate results are shown in Table 3.

TABLE-US-00003 TABLE 3 Removal effects of microbial agents obtained in Examples 4-6 on COD.sub.Mn, TN, TP and NH.sub.4.sup.+N Microbial COD.sub.Mn TN TP NH.sub.4.sup.+N Agent (mg/L) (mg/L) (mg/L) (mg/L) Example 4 9.16 6.08 0.55 5.94 Example 5 8.95 5.41 0.53 5.50 Example 6 7.46 5.12 0.48 4.29

[0046] Referring to the test method of Example 4, the heavy metal removal capacity of the composite microbial agents obtained in Examples 4-6 was respectively tested. The composite microbial agent was inoculated into the sample in a dose of 0.3%, cultured at 30 C. in a dark shaker, and sampled on the second day. The content of metal ions in the sample was determined by atomic absorption spectrophotometry. The results are shown in Table 4.

TABLE-US-00004 TABLE 4 Adsorption and removal effects of microbial agents obtained in Examples 4-6 on metal ions Microbial Cadmium Copper Lead Chromium Agent mg/kg mg/kg mg/kg mg/kg Example 4 0.033 0.081 0.084 0.045 Example 5 0.029 0.078 0.089 0.048 Example 6 0.022 0.065 0.071 0.039

Comparative Example

[0047] The formula of the microbial agent was replaced by the microbial agent 1, the microbial agent 2, the microbial agent 3, and the microbial agent 4, wherein the components of the microbial agents 1, 2, 3, and 4 are as follows:

[0048] Microbial agent 1: No Pseudomonas was added, and other conditions were kept unchanged with reference to the preparation method of the microbial agent in Example 1.

[0049] Microbial agent 2: P. pastoris was replaced with Saccharomyces cerevisiae, other conditions are kept unchanged with reference to the preparation method of the microbial agent in Example 1; (Saccharomyces cerevisiae ACCC21144, see document Dai Youzhi, Xu Caixia. Adsorption of Cr (VI) in Water by Saccharomyces cerevisiae [J]. Natural Science Journal of Xiangtan University, 2007, 29(3), 79-83.).

[0050] Microbial agent 3: No P. pastoris was added, and other conditions were kept unchanged with reference to the preparation method of the microbial agent in Example 1.

[0051] Microbial agent 4: No Staphylococcus was added, and other conditions were kept unchanged with reference to the preparation method of the microbial agent in Example 1.

[0052] Microbial agent 5: The parts by weight of the Staphylococcus in Example 1 were replaced with 2 parts, and other conditions were kept unchanged with reference to the preparation method of the microbial agent in Example 1.

[0053] Microbial agent 6: The parts by weight of the Staphylococcus in Example 1 were replaced with 25 parts, and other conditions were kept unchanged with reference to the preparation method of the microbial agent in Example 1.

[0054] The sewage treatment was carried out in accordance with the method of Example 1, and the test results of the treated sewage are as shown in Table 5.

TABLE-US-00005 TABLE 5 Index results of treated sewage (0.2% dose) Microbial COD.sub.Mn TN TP NH.sub.4.sup.+N Agent (mg/L) (mg/L) (mg/L) (mg/L) 0 (not added) 54.61 35.15 3.14 31.58 Microbial 38.35 28.79 2.75 28.91 agent 1 Microbial 36.18 25.45 2.56 25.33 agent 2 Microbial 42.12 30.30 2.33 24.85 agent 3 Microbial 26.80 22.24 1.94 18.17 agent 4 Microbial 20.21 17.46 2.03 15.94 agent 5 Microbial 35.37 26.60 2.42 27.11 agent 6

[0055] The heavy metal ions were treated in accordance with the method of Example 3, and the contents of the heavy metal ions after the treatment are as shown in Table 6.

TABLE-US-00006 Microbial Cadmium Copper Lead Chromium Agent mg/kg mg/kg mg/kg mg/kg Microbial 0.135 0.155 0.145 0.120 agent 1 Microbial 0.128 0.146 0.148 0.153 agent 2 Microbial 0.153 0.166 0.170 0.161 agent 3 Microbial 0.165 0.184 0.187 0.152 agent 4 Microbial 0.088 0.112 0.125 0.092 agent 5 Microbial 0.146 0.150 0.161 0.045 agent 6

[0056] Referring to Table 1 to Table 4, it can be seen that the interaction between the various strains in the microbial agent of the present invention exists, and the various strains can be well fermented and symbiotic. It can be seen from the microbial agent 1 that the decontamination performance of the system without Pseudomonas is significantly decreased, and the adsorption capacity is also not good. It can be seen from the microbial agent 2 that the S. cerevisiae has poor symbiotic effect with other strains in the microbial agent system of the present invention, and the corresponding decontamination effect and metal ion adsorption capacity are poor. In addition, Staphylococcus has a very important influence on the heavy metal ion adsorption performance of the microbial agent, and the microbial agent without Staphylococcus (microbial agent 4) has a certain nitrogen and phosphorus removal effect, but the metal ion adsorption performance is very poor. At the same time, according to the microbial agents 5 and 6, the added amount of the Staphylococcus has a great influence on the decontamination and metal ion removal effects, and too little or too much additive will obviously inhibit the effect of the composite microbial agent.

[0057] Although the present invention has been disclosed in the above preferred examples, the present invention is not limited thereto. Any modifications and variations can be made without departing from the spirit and scope of the present invention, and therefore, the scope of the present invention should be determined by the scope of the appended claims.