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Citrobacter, application of citrobacter in treatment of sulphate-containing waste water, and separation and identification method of citrobacter

A Citrobacter and identification method technology, applied in the field of sulfate-containing wastewater treatment, can solve the problems of long cycle and cumbersome screening process, and achieve the effects of short cycle, simple screening process and easy cultivation

Active Publication Date: 2020-12-08
SOUTH CHINA UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional method of screening strains with sulfate reducing ability is mainly to enrich through selective medium, and then alternately screen solid-liquid medium to isolate and purify strains. This method is more practical for screening microorganisms, but the screening process is also cumbersome. , long cycle

Method used

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  • Citrobacter, application of citrobacter in treatment of sulphate-containing waste water, and separation and identification method of citrobacter
  • Citrobacter, application of citrobacter in treatment of sulphate-containing waste water, and separation and identification method of citrobacter
  • Citrobacter, application of citrobacter in treatment of sulphate-containing waste water, and separation and identification method of citrobacter

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Experimental program
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Effect test

Embodiment 1

[0043] Sediment samples collected from the Hengshi River Basin in Dabaoshan Mining Area, Guangdong Province were used for enrichment culture of sulfate-reducing bacteria under anaerobic conditions using Postgate B-type medium. In order to simulate the acidic environment of acid mine wastewater, the phosphate ratio was adjusted to make the pH of the culture medium 5.6. Dig 0.1-0.5 grams of sediment samples with a spoon and transfer them to a 25mL anaerobic tube in an ultra-clean bench, and add 20mL of postgate B-type medium that has been deaerated and sterilized by autoclaving. Isolated from outside air. The whole operation was carried out in an anaerobic operation box. After the inoculation, the cells were transferred to a constant temperature incubator at 25°C for 7 days of cultivation. During the cultivation process, it was judged whether the enrichment was successful by observing whether its color turned black. After repeated transfer 5 times, a stable sulfate-reducing ba...

Embodiment 2

[0046] Dilute the colony collected in the dialysis bag isolation group to 10 -3 、10 -4 、10 -5 In the anaerobic operation box, take each dilution of the bacterial suspension and spread it on the solid Postgate B-type medium respectively, put part of the plate upside down in the anaerobic tank, and place the other part in the aerobic incubator. The temperature for culturing under aerobic conditions and aerobic conditions was 30°C, and the culturing time for both anaerobic and aerobic conditions was 3 days until colonies grew out. Pick a single colony, inoculate it on a liquid medium, and culture it anaerobically in a shaker at 30°C and 150 rpm for 5-7 days, select an anaerobic tube that turns black, and then continue to streak the bacterial solution in the anaerobic tube with a plate. After 3 times of solid-liquid medium alternate screening, under anaerobic conditions, the 2+ and SO 4 2- On the solid medium, the target strain produced black colonies after one week; under ae...

Embodiment 3

[0050] Take the bacterial solution grown to the logarithmic phase (OD 600 is about 0.3), centrifuged at 4°C and 8000×g for 5 min, and discarded the supernatant. After adding 5mL of 2.5wt% pentanediol solution and fully shaking, place it in a refrigerator at 4°C for 2-4h. After washing with 10mmol / L phosphate buffer for 3 times, use ethanol gradient dehydration, followed by 30%, 50%, 70%, 85%, and 95% ethanol once each (gradient concentration of ethanol is prepared with phosphate buffer ), dehydrated twice with 100% ethanol, and finally washed twice with isoamyl acetate. Each step needs to be fully shaken and maintained for more than 20 minutes, and finally centrifuged at 8000×g for 5 minutes, discard the supernatant, and proceed to the next step. After the treated bacteria were freeze-dried, a small amount of bacteria was placed on a sample plate covered with conductive glue, sprayed with gold for 120 seconds, and observed with a field emission scanning electron microscope. ...

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Abstract

The invention discloses citrobacter, application of the citrobacter in treatment of sulphate-containing waste water, and a separation and identification method of the citrobacter. The citrobacter hasa sulphate reduction capability; the number of the citrobacter is Citrobacter sp.EPS8; the citrobacter is preserved in the Guangdong microbial strain preservation centre on July 13th, 2020; the preservation address is Floor 5, Building 59, Courtyard 100, Xianlie Middle Road, Guangzhou City, Guangdong Province; and the preservation number is GDMCC NO:61090. The strain can take lactic acid as a carbon source, and can reduce sulphate (the reduction rate is 100%) under an anaerobic condition; the strain has relatively strong sulphate reducing capacity within the initial pH value of 6-8; and the optimal initial pH value of the strain is 6.0. The citrobacter disclosed by the invention is facultative anaerobic bacteria, is rapid in aerobic growth, easy to culture and capable of anaerobically reducing sulphate, and has wide application in the aspect of treating wastewater containing SO4<2> (such as acid mine drainage).

Description

technical field [0001] The invention belongs to the field of sulfate-containing wastewater treatment, and in particular relates to a Citrobacter and its application in treating sulfate reduction and a method for separating and identifying Citrobacter. Background technique [0002] The development of mineral resources is often accompanied by the production of tailings. Under natural weathering and rain erosion, the exposed tailings will dissolve a large amount of heavy metals and sulfates into the water body, thus forming acid mine drainage (AMD). AMD has the characteristics of low pH value and high sulfate content, and often contains a large amount of heavy (class) metal ions and their cyanides, such as cadmium, arsenic, zinc, nickel, lead, iron, etc. and their cyanides. The discharge of AMD will pollute the surface water, groundwater and soil around the mining area, seriously threatening human health and the surrounding ecological environment. Sulfate-reducing bacteria (SR...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02C02F3/34C12R1/01C02F101/10C02F103/10
CPCC12N1/20C12N1/02C02F3/345C02F2101/101C02F2103/10C12R2001/01C12N1/205
Inventor 郭楚玲柯常栋姜梦戈张思宇曾宇飞卢桂宁党志
Owner SOUTH CHINA UNIV OF TECH
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