CN101063096A

CN101063096A - Method for treating sewage water having heavy metallic ion Cd2+ and Cu2+ by using bacteria 575

Info

Publication number: CN101063096A
Application number: CN 200710051995
Authority: CN
Inventors: 祝香芝; 杨琳
Original assignee: Hubei University
Current assignee: Hubei University
Priority date: 2007-04-27
Filing date: 2007-04-27
Publication date: 2007-10-31
Anticipated expiration: 2027-04-27
Also published as: CN100535105C

Abstract

The invention discloses a method with effective enriched heavy metal ion Cd2+ and Cu2+ strain to treat waste water with heavy metal ion Cd2+ and Cu2+, which is characterized by the following: preserving the strain in Chinese typical culture preserved center with preserved number at CCTCC NO: M207050 and name as Ochrobium 575 (Ochrobactrum sp. 575); culturing Ochrobactrum sp. 575 single bacterial colony as OD not less than 0. 6 bacterial liquid; adding the bacterial liquid into waste water with Cd2+, Cu2+ heavy metal ion with 0. 5-1% proportion; utilizing the property of effective enriched heavy metal ion Cd2+ and Cu2+; decreasing the content of heavy metal ion; reaching the goal of treating waste water.

Description

Utilize bacterium 575 to handle and contain Cd 2+, Cu 2+The method of heavy metal ion sewage

Technical field

What the present invention relates to is the method that industrial sewage is handled, and particularly utilizes bacterium 575 to handle and contains Cd ²⁺, Cu ²⁺The method of heavy metal ion sewage.

Background technology

A large amount of dischargings of heavy industry waste water at present, the aggravation of heavy metal accumulation in soil and the water source, the pollution of heavy metal is also serious day by day.Because heavy metal is the biomagnification by food chain easily, constitute serious threat to biological and HUMAN HEALTH.How to administer heavy metal contamination effectively and become the human common problem of paying close attention to.And the pollution condition of heavy metal is also comparatively serious in the sewage in development area, and in recent years, Chinese scholars has been done a large amount of research to the governing problem of heavy metal contamination.Wherein, utilize biosorption process to handle the concern that heavy metal ion more and more is subjected to Chinese scholars.

At present many about the report of bacterium enriching heavy metal ion this respect, but the actually rare of using value really arranged.Faisal and Hasnain have reported that in 2004 the reddish brown Salmonella strain CrT-1 of its screening and CrT-13 can reach 17.3 ± 0.15 and 12.1 ± 0.18mg/g (dry cell weight) respectively to the maximal absorptive capacity that belongs to chromium.And domestic in this respect research is actually rare, and subject matter may be that its enriching quantity is low and culture condition is harsh, thereby using value is little.Therefore just extremely urgent to the exploitation and the research of the heavy dose of enriching heavy metal ionic novel bacterial of energy.

Summary of the invention

The present invention proposes effectively enriching heavy metal ion Cd of a kind of utilization ²⁺With Cu ²⁺Bacterial strain handle and be rich in heavy metal ion Cd ²⁺With Cu ²⁺The method of sewage.

We screen tool laccase activity and the very high reddish brown Bordetella gram negative bacillus of enriching quantity the perennial trees ' root soil near the oil field, this bacterial strain is deposited in Chinese typical culture collection center on April 20th, 2007, preserving number is CCTCC NO:M207050, and name is called Ochrobium 575 (Ochrobactrum sp.575).

Bacterium separates

Get the oil field and gather pedotheque 1g, soil sample is resuspended in 20ml M9 basis screening base, the vibration mixing, left standstill 10 minutes, and got the 2ml supernatant and be inoculated in the 100ml M9 basic medium 35-40 ℃, 250rpm cultivated 7 days, nutrient solution suitably is inoculated on the LB flat board after the dilution, and coating is even, 35-40 ℃ of incubated overnight.Single colony inoculation on the LB screening culture medium flat board is contained 0.2mM CuSO in 6ml ₄The LB substratum (adding 20 μ l concentration in every 100mlLB liquid is the CuSO of 1M ₄) in, 35-40 ℃, the 250rpm shaken overnight is cultivated.

The biological nature of this bacterium:

1, bacterial colony form and dyeing.

1) 575 colonial morphologies: be single bacterium colony circle, bacterium colony big (about 1.3mm), yellow, surface elevation.Neat in edge.

2) the gramstaining result of 575 bacterium: be gram negative bacillus.

3) the spore staining result of 575 bacterium: do not see gemma.

4) the capsule stain result of 575 bacterium: through the Anthony capsule stain, visible 575 thalline purples, the thalline edge is lilac or red-purple, and bacterium all has pod membrane.

5) flagellum puncture experiment: 575 bacterium, 37 ℃ of following stab culture in test tube are cultivated observations demonstration after 24 hours: Ochrobactrum sp.575 does not have flagellum by being cloud diffusion explanation around the puncture alignment.

6) the 16s DNA analysis identifies that this bacterium is Ochrobactrum.

Ochrobactrum sp.575 is to heavy metal Cd ²⁺, Cu ²⁺Enrichment detect

In the LB substratum, add heavy metal Cd respectively ²⁺, Cu ²⁺, make its final concentration be respectively 0,50mg/ml, 100mg/ml.Bacterium is connected to contains Cd ²⁺Or Cu ²⁺The LB substratum in, 5.7,37 ℃ of pH, 200r/min incubated overnight.Treat that the OD value of bacterium collects thalline between 0.8-1.2, and with rinsed with deionized water 4-5 time (each 10ml), with rinsing liquid and supernatant liquor merging.Use the heavy metal ion in aas determination supernatant liquor and the thalline respectively.The result is presented at two heavy metal species ionic content minimizing in the supernatant liquor, and the amount of this part minimizing is just in time coincide with metal content measured in the thalline.The result shows that Ochrobactrum sp.575 is to heavy metal Cd ²⁺, Cu ²⁺Enriching quantity can reach 16.5 ± 0.21mg/g (dry cell weight) and 13.9 ± 0.16mg/g (dry cell weight) respectively, for new bacterial classification has been opened up in the biomagnification of heavy metal ion.

Utilize bacterium 575 to handle and contain Cd ²⁺, Cu ²⁺The method of heavy metal ion sewage:

One) bacterium liquid preparation

1, the single bacterium colony of Ochrobactrum sp.575 is inserted shaking in the bottle of 50ml, substratum is the LB substratum, and 35-40 ℃, the 250rpm shaken overnight is cultivated.

2, when OD 〉=0.6, get above-mentioned bacterium liquid 1mL and cultivate in shaking in the bottle of 1L, substratum is the LB substratum, and 35-40 ℃, the 250rpm shaken overnight is cultivated.

3, when OD 〉=0.6, stop to cultivate, it is standby to collect bacterium liquid.

Two, sewage disposal:

1, will contain Cd ²⁺, Cu ²⁺Heavy metal ion sewage pumps in the treating water pond, in the ratio adding bacterium liquid of 0.5-1%, presses 1-5 rev/min and slowly stirs, and the 2-3 hour sudden and violent gas of logical oxygen was once placed 24 hours, and supernatant liquor is pumped into second handles in the pond,

2, repeat above-mentioned treating processes 3-5 time, treating water is qualified discharge after filtering,

3, after the sewage supernatant liquid pump of forebay arrived next pond, the precipitation of forebay was discharged from blow-off pipe, and with at the bottom of scraper plate and the water cleaning pond, so that receive the sewage in a last pond once more.

The present invention has tangible advantage:

1, this bacterium source the Nature is safe;

2, reddish brown Salmonella absorption biosorption process is handled the waste water that contains heavy metal, cost low (not needing to add extra nutritive ingredient), high efficiency, manageable, do not cause secondary pollution, help the improvement of ecotope to environment.

Description of drawings

Fig. 1 is the single pond sewage disposal synoptic diagram of the present invention.

Fig. 2 is the many ponds of the present invention series connection sewage disposal synoptic diagram.

1-water intaking valve wherein, the 2-treatment tank, the sudden and violent tracheae of 3-, the 4-swabbing pig, the 5-wash water valve, 6-bacterium liquid adds inlet, 7-outlet valve, 8-water pump, 9-standby pool.

Embodiment

Embodiment 1-handles in single pond

1, will contain Cd ²⁺, Cu ²⁺Heavy metal ion sewage pumps in the treating water pond (2), in the ratio adding bacterium liquid of 0.5-1%, presses 1-5 rev/min and slowly stirs, and the 2-3 hour sudden and violent gas of logical oxygen was once placed 24 hours, and supernatant liquor is pumped in the standby pond (9);

2, will precipitate from wash water valve (5) discharge, and with at the bottom of swabbing pig (4) and the water cleaning pond;

3, the sump pump in standby pond (9) is delivered in the processing pond (2), in the ratio adding bacterium liquid of 0.5-1%, pressed 1-5 rev/min and slowly stir, the 2-3 hour sudden and violent gas of logical oxygen was once placed 24 hours, and supernatant liquor is pumped in the standby pond (9);

4, repeat above-mentioned treating processes 3-5 time, treating water is qualified discharge after filtering,

Embodiment 2-many ponds series connection is handled

1, will contain Cd ²⁺, Cu ²⁺Heavy metal ion sewage pumps into 1# and handles in the pond, in the ratio adding bacterium liquid of 0.5-1%, presses 1-5 rev/min and slowly stirs, and the 2-3 hour sudden and violent gas of logical oxygen was once placed 24 hours, supernatant liquor is pumped into 2# handles in the pond;

2,1# is handled in the pond precipitation and discharge from wash water valve (5), and at the bottom of clearing up the pond with swabbing pig (4) and water, and accept new sewage;

3, in 2# handles the pond, in the ratio adding bacterium liquid of 0.5-1%, to press 1-5 rev/min and slowly stir, the 2-3 hour sudden and violent gas of logical oxygen was once placed 24 hours, supernatant liquor is pumped into 3# handles in the pond;

4, in like manner, sewage repeats above-mentioned processing in 4# handles the pond after, treating water is qualified discharge after filtering.

5, said process carries out continuously, can reach the purpose of disposing of sewage.

Claims

1, a kind of energy enrichment Cd ²⁺, Cu ²⁺The Ochrobium 575 of heavy metal ion, preserving number are CCTCC NO:M207050.

2, a kind ofly utilize bacterium 575 to handle to contain Cd ²⁺, Cu ²⁺The method of heavy metal ion sewage is characterized in that:

One, bacterium liquid preparation

1), the single bacterium colony of Ochrobactrum sp.575 is inserted shaking in the bottle of 50ml, substratum is the LB substratum, and 35-40 ℃, the 250rpm shaken overnight is cultivated;

2), when OD 〉=0.6, get above-mentioned bacterium liquid 1mL and cultivate in shaking in the bottle of 1L, substratum is the LB substratum, 35-40 ℃, the 250rpm shaken overnight is cultivated;

3), when OD 〉=0.6, stop to cultivate, it is standby to collect bacterium liquid;

Two, sewage disposal:

1), will contain Cd ²⁺, Cu ²⁺Heavy metal ion sewage pumps in the treating water pond, in the ratio adding bacterium liquid of 0.5-1%, presses 1-5 rev/min and slowly stirs, and the 2-3 hour sudden and violent gas of logical oxygen was once placed 24 hours, supernatant liquor is pumped into second handles in the pond;

2), repeat above-mentioned treating processes 3-5 time, treating water is qualified discharge after filtering;

3), when the sewage supernatant liquid pump of forebay behind next pond, the precipitation of forebay is discharged from blow-off pipe, and with at the bottom of scraper plate and the water cleaning pond, so that the sewage in the last pond of reception once more.