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Preparation method and application of liquid complex microbial inoculant

A compound bacterial agent and liquid technology, applied in the field of environmental pollution control, can solve the problems of inability to maintain metabolic activities, additional regular adjustments, etc., and achieve the effects of low growth and degradation activity, large number of viable bacteria, and long storage time

Active Publication Date: 2015-03-04
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Generally, the environment required by strains to degrade organic pollutants is weakly acidic or neutral (pH6.5-7.5), but since strains will produce a certain amount of small molecule organic acids (such as Formic acid, acetic acid, propionic acid, succinic acid, etc.) make the culture system acidic, so it is necessary to artificially adjust the pH value of the system regularly during the metabolic process, otherwise normal metabolic activities cannot be maintained
This is a common drawback in current biodegradation

Method used

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  • Preparation method and application of liquid complex microbial inoculant
  • Preparation method and application of liquid complex microbial inoculant
  • Preparation method and application of liquid complex microbial inoculant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] See figure 1 , The preparation method of the compound bacterial agent of the present invention is as follows:

[0033] (1) Obtain the bacterial suspension

[0034] Pick the above-mentioned colonies from the preservation medium and inoculate them into the inorganic salt solution medium. After culturing for 48-64h with α-pinene, 1-chlorobenzene, dichloromethane, o-xylene and acetic acid as the sole carbon source, the different A bacterial suspension of the strain.

[0035] The said inorganic salt culture solution (g / L): KH 2 PO 4 0.376, K 2 HPO 4 ·3H 2 O 0.456, (NH 4 ) 2 SO 4 0.484, NaNO 3 0.68, Mg(NO 3 ) 2 0.256, CaCl 2 0.011, trace element (MnCl 2 ·H 2 O 0.06, ZnCl 2 0.088, KI 0.01, NaMoO 4 ·2H 2 O 0.1, H 3 BO 3 0.05), pH=7.2.

[0036] (2) Obtaining mixed bacterial seed liquid

[0037] After centrifuging the bacterial suspension of various bacteria under the condition of 12000g, the bacteria are collected, and washed with sterile water 3 times, and a certain volume of st...

Embodiment 2

[0041] Place the prepared liquid composite inoculum in a refrigerator (4°C) and store it for 1 week, 2 weeks, 4 weeks, and 8 weeks respectively. The number of active bacteria in the liquid inoculum was investigated by the plate counting method. The results are shown in Table 1. The number of effective viable bacteria contained in the freshly prepared liquid bacterial agent is 7.6×10 respectively 9 CFU / mL. The number of corresponding viable bacteria decreased slightly after storage for different time, and with the increase of storage time, the downward trend was obvious, but it remained at 10 8 Above CFU / mL. The attenuation of the number of viable bacteria in the composite liquid inoculum is small, which may be due to the low bacterial activity at low temperatures. At the same time, yeast powder and peptone can be used as carbon sources to maintain the activity of the strains at low temperatures, so that the activity of the inoculum is maximized and effectively stored The time ...

Embodiment 3

[0044] Place the prepared liquid composite inoculum in a refrigerator (4°C), and test the degradation activity of the inoculum after 1 week, 2 weeks, 4 weeks, and 8 weeks. The specific steps are as follows: After taking out the liquid compound bacterial agent, place it at room temperature, centrifuge at 12000g, collect the bacterial cells, wash them with sterile water 3 times, inoculate them into an inorganic salt liquid medium, and add mixed organic pollutants ( α-pinene, 1-chlorobenzene, methylene chloride, toluene), the concentration of each pollutant is 100mg / L. Place it on a shaker and shake culture at a temperature of 30℃. After culturing for 48 hours, analyze the concentration of each pollutant in the liquid. The results are as follows figure 2 Shown.

[0045] It can be seen that the degradation activity of the liquid bacterial agent is almost unchanged after being stored at 4°C for 1 week and 2 weeks, indicating that the degradation stability is better; after 4 weeks and...

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Abstract

1) taking PseudomonasveroniiZW, RalstoniapickettiiL2, PandoraeapnomenusaLX-1, ZoogloearesiniphilaHJ1 and StaphylococcuspasteuriLWX, inoculating an inorganic salt solution culture medium with the PseudomonasveroniiZW, RalstoniapickettiiL2, PandoraeapnomenusaLX-1, ZoogloearesiniphilaHJ1 and StaphylococcuspasteuriLWX, and culturing the PseudomonasveroniiZW, RalstoniapickettiiL2, PandoraeapnomenusaLX-1, ZoogloearesiniphilaHJ1 and StaphylococcuspasteuriLWX by adopting alpha-pinene, 1-chlorobenzene, dichloromethane, o-xylene and acetic acid as carbon resources only, so as to obtain bacterium suspensions; 2) performing centrifugal washing and diluting on the bacterium suspensions, and mixing the bacterium suspensions in an isovolumetric manner, so as to obtain an inoculation liquid; 3) preparing an inorganic salt culture solution with the pH of 6.8-7.5, putting the inoculation liquid into a mixture of the alpha-pinene, 1-chlorobenzene, dichloromethane and o-xylene, and culturing for 24-48 h at the temperature of 30 DEG C; 4) adding yeast powder and peptone, and preserving at the temperature of 4 DEG C. The preparation of the liquid complex microbial inoculant is simple, and the pH can be automatically adjusted.

Description

Technical field [0001] The invention belongs to the field of environmental pollution control, and relates to a preparation method and application of a liquid composite bacterial agent. Background technique [0002] In recent years, environmental pollution caused by volatile organic compounds (VOCs) has become increasingly serious. In particular, the multi-component VOCs emitted during industrial production have the characteristics of high toxicity and difficult degradation, which endanger the ecological environment and human health. Therefore, the prevention and control of VOCs pollution is of great significance. [0003] The current technologies for processing VOCs mainly include adsorption, combustion, absorption, etc., but these technologies all have disadvantages such as high investment costs, complex operation processes, and high energy consumption. Biological purification can effectively treat low-concentration atmospheric VOCs without secondary pollution problems, and has ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20B01D53/84B01D53/44C12R1/38C12R1/01C12R1/44
CPCB01D53/44B01D53/68B01D53/70B01D53/84B01D2257/206B01D2257/708C12N1/20Y02A50/20
Inventor 成卓韦陈建孟於建明
Owner ZHEJIANG UNIV OF TECH
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