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Atexandrium tamarense culture liquid sterilizing process

A technology for culturing liquid and bacteria, which is applied to unicellular algae, sanitary equipment for toilets, water supply devices, etc. It can solve the problems of troublesome operation, high experimental experience in algae separation, and difficulty in balancing, and achieve the effect of short processing time.

Inactive Publication Date: 2007-01-03
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] The existing sterilization methods for algae culture fluid mainly include: 1. Repeated washing method: generally, a single algae cell is washed repeatedly with a sterile medium, and then cultivated in a culture fluid containing antibiotics; this method is relatively troublesome to operate , requires high experience in algae isolation experiments, and at the same time, it is difficult to cultivate a single algal cell
2. Chemical substance method: including SDS and other detergents. This method uses chemical substances to directly lyse cells to achieve the purpose of killing bacteria; A balance between killing all bacteria
However, considering that most microorganisms are uncultured, this test result is not reliable

Method used

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  • Atexandrium tamarense culture liquid sterilizing process
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  • Atexandrium tamarense culture liquid sterilizing process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Use the 2216E solid plate to isolate the bacteria in the algae culture solution, inoculate the isolated strains in 3mL 2216E liquid medium, and culture at 25°C, 180rpm for 20h to the late exponential stage. Take 100 μL of bacterial culture solution and evenly spread it on the 2216E plate, and place it upright to make it completely absorbed. Paste the drug-sensitive paper sheets containing antibiotics (filter paper sheets with a diameter of 6 mm) on the surface of the plate, and each drug-sensitive paper sheet contains 50 μg of gentamicin, 25 μg of neomycin, 25 μg of streptomycin, and 25 μg of kanamycin, Cephalosporin 10 μg and rifampicin 5 μg were incubated on an inverted plate at 25°C until the zone of inhibition was clear and stable, and the diameter of the zone of inhibition was recorded. Filter 250mL algae culture solution in the exponential growth phase with an 8μm filter membrane, and resuspend the algal cells on the filter membrane in 50mL sterile f / 2 culture sol...

Embodiment 2

[0051] Similar to Example 1, the difference is that the isolated strain was inoculated in 4 mL of 2216E liquid medium, and cultured at 30° C., 100 rpm for 24 hours until late exponential. Take 150 μL of bacterial culture solution and evenly spread it on the 2216E plate, and place it upright to make it completely absorbed. Paste the drug-sensitive paper sheet containing antibiotics (filter paper with a diameter of 7mm) on the surface of the plate, invert the plate and incubate at 30°C until the zone of inhibition is clear and stable, and record the diameter of the zone of inhibition. Filter 100 mL of algae culture solution in the exponential growth phase with a 6 μm filter membrane, and resuspend the algal cells on the filter membrane in 20 mL of sterile f / 2 culture solution. Centrifuge at 1000g for 10min, remove the supernatant, and resuspend the algae cells in 50mL sterile f / 2. Repeat centrifugation at 1000g for 10min, remove the supernatant, resuspend the algae cells twice ...

Embodiment 3

[0054] Similar to Example 1, the difference is that the isolated strain was inoculated into 5 mL of 2216E liquid medium, and cultured at 20° C. and 250 rpm for 18 hours until late exponential. Take 50 μL of bacterial culture solution and evenly spread it on the 2216E plate, and place it upright to make it completely absorbed. Paste a drug-sensitive paper sheet containing antibiotics (a filter paper sheet with a diameter of 6 mm) on the surface of the plate, invert the plate and incubate at 20°C until the zone of inhibition is clear and stable, and record the diameter of the zone of inhibition. Filter 20 mL of algae culture solution in the exponential growth phase with a 5 μm filter membrane, and resuspend the algal cells on the filter membrane in 80 mL of sterile f / 2 culture solution. Centrifuge at 2000g for 20min, remove the supernatant, and resuspend the algae cells in 20mL sterile f / 2. Repeat centrifugation at 2000g for 20min, remove the supernatant, resuspend the algae ce...

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Abstract

The present invention relates to sterilization, and is especially Atexadrium tamarense culture liquid sterilizing process. The effective reliable Atexadrium tamarense culture liquid sterilizing process includes the following steps: plate separating bacteria from the Atexadrium tamarense culture liquid, inoculating to culture medium and painting to plate; sticking antibiotic containing medicine sensitive paper to the surface of the plate, turning the plate for culturing to reaching stable inhibition zone, filtering culture liquid in exponential phase and suspending Atexadrium tamarense cell in the filter membrane in f / 2 culture liquid; centrifuging to eliminate supernatant, re-suspending in f / 2 culture liquid and setting under illumination; adding lysozyme, adding SDS, setting under illumination, eliminating lysozyme and SDS, re-suspending in f / 2 culture liquid and setting under illumination; at least thrice passage of the treating culture liquid, eliminating antibiotic and inspection.

Description

technical field [0001] The invention relates to a sterilization method, in particular to a sterilization method of Alexandrium tamarii culture fluid. Background technique [0002] The existing sterilization methods for algae culture fluid mainly include: 1. Repeated washing method: generally, a single algae cell is washed repeatedly with a sterile medium, and then cultivated in a culture fluid containing antibiotics; this method is relatively troublesome to operate , requires high experience in algae isolation experiments, and it is difficult to cultivate a single algal cell. 2. Chemical substance method: including SDS and other detergents. This method uses chemical substances to directly lyse cells to achieve the purpose of killing bacteria; There is a balance between killing all the bacteria. 3. Antibiotic method: It is the most convenient and widely used method. It not only uses antibiotics to kill bacteria specifically, but also generally has no effect on algae growth....

Claims

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Application Information

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IPC IPC(8): A61L2/00A61L2/16A61L2/08C12N1/12
Inventor 郑天凌苏建强杨小茹
Owner XIAMEN UNIV
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