Membrane forming bacteria agent for biologic membrane and method for structuring and analyzing membrane

A technology of biofilm and film-forming bacteria, applied in biochemical equipment and methods, chemical instruments and methods, microorganisms, etc., can solve the problems of complex operation, poor real-time performance, and damage to the integrity of biofilm, and achieve simple and convenient operation, real-time good sex effect

Inactive Publication Date: 2005-07-27
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods have the following disadvantages: the integrity of the biofilm

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The green fluorescent protein gene carrier pEGFP was treated with CaCl 2 Escherichia coli Escherichia coli JM109 strain is marked by transformation method, and the obtained marker strain is used as a model bacterium to inoculate 50 mL / bottle of LB medium containing 50 μg / mL ampicillin with an inoculum of 5 rings / bottle, and mix with volcanic rock particles (5 g / bottle ) to co-culture the hanging membrane (37°C, 120r / min, 1 day). A laser confocal scanning microscope was used to capture image stacks of different layers of the volcanic rock filler biofilm area of ​​250×250 μm, and the obtained image stacks were processed by the COMSTAT program to obtain relevant quantitative parameters: the average thickness of the biofilm in one day was 0.120844 μm, and the largest biofilm The thickness is 10.5 μm, and the biofilm volume per unit area is 0.136986 μm 3 / μm 2 , biofilm surface area 21338.1μm 2 , the biofilm specific surface area is 3.36854μm 2 / μm 3 .

Embodiment 2

[0023] The green fluorescent protein gene carrier pEGFP was transformed by electroporation to mark Escherichia coli JM109 strain, and the obtained marked strain was used as a model bacterium to inoculate 50 mL / bottle of LB medium containing 150 μg / mL ampicillin with an inoculation amount of 5 rings / bottle, and shaken Mixed with porous carbon particles (5g / bottle) in the bottle to cultivate the hanging film (37°C, 120r / min, 1 day). A laser confocal scanning microscope was used to capture and obtain image stacks of different layers of the porous carbon filler biofilm area of ​​250×250 μm. The obtained image stacks were processed by the COMSTAT program to obtain relevant quantitative parameters: the average thickness of the biofilm in one day was 0.110685 μm, and the biofilm The maximum thickness is 9.1 μm, and the biofilm volume per unit area is 0.116928 μm 3 / μm 2 , biofilm surface area 18982.3μm 2 , the biofilm specific surface area is 3.05456μm 2 / μm 3 .

Embodiment 3

[0025] The green fluorescent protein gene carrier pEGFP was treated with CaCl 2 The transformation method marked Nitrobacter sp.N-20 strain, and the obtained marker strain was used as a model bacterium to inoculate 50 mL / bottle of LB medium containing 50 μg / mL ampicillin with an inoculum of 5 rings / bottle, and mixed with volcanic rock particles ( 5g / bottle) to culture the hanging film by blending (30°C, 120r / min, 7 days). A laser confocal scanning microscope was used to capture image stacks of different layers of the volcanic filler biofilm area of ​​250×250 μm. The obtained image stacks were processed by the COMSTAT program to obtain relevant quantitative parameters. The average thickness of the biofilm in 7 days was 0.121085 μm, and the biofilm The maximum thickness is 11.2 μm, and the biofilm volume per unit area is 0.198646 μm 3 / μm 2 , biofilm surface area 21338.1μm 2 , the biofilm specific surface area is 4.36854μm 2 / μm 3 .

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Abstract

This invention relates to a bacterial membrane agent and methods therefore. The membrane bacterial strain is labeled by color fluorescent protein gene. The said agent and methods verify in-situ analysis, i.e., analyzing under a condition of non-destructing biologic membrane; easy operation, i.e., no need of microbe culture and counting or extraction of cell nucleic acid for cross-breed or electrophoretic determination; and good real time online analysis.

Description

(1) Technical field [0001] The invention relates to a film-forming bacteria agent for constructing biofilm and methods for constructing and analyzing biofilm. (2) Background technology [0002] Biofilm is one of the important methods for microbial control of environmental pollution. It can be applied to biofilters used in sewage treatment plants of various industrial enterprises and municipal sewage treatment plants, and treatment towers or treatment pools for various industrial or domestic waste gas. [0003] The use of biofilms to control waste gas or sewage pollution is a research hotspot and a development direction at home and abroad. So far, the main problem in this field of research is the online rapid detection of biofilm flora composition and spatial structure, that is, the analysis of biofilms. Various process research and optimization are carried out in the "dark box" mode, so it is difficult to reveal the relationship between the composition of the flora and the e...

Claims

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Application Information

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IPC IPC(8): C02F3/10C02F3/34C12N1/00C12N1/20C12N15/65
CPCY02W10/10
Inventor 钟卫鸿陈建孟叶海仁汪琨宋艳绒路争
Owner ZHEJIANG UNIV OF TECH
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