Simple method for screening aquatic source aeromonas

A single-cell bacteria and aquatic technology, applied in the field of microbiology, can solve the problems of expensive, cumbersome, low separation rate and accuracy

Inactive Publication Date: 2013-07-10
PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the technologies for screening Aeromonas include: 1. After cultivating and purifying a single colony through common culture medium, use the rapid automatic identification system for bacteria to screen. This method requires expensive instruments and equipment and skilled operators; 2. Through common culture medium After cultivating and purifying a single colony, nucleic acid detection technology, including DNA and PCR technology (polymerase chain reaction technology), is fast, concise and sensitive, but this method requires expensive equipment and skilled operators; 3. The classic physiological and biochemical identification method is not only cumbersome and time-consuming, but also requires operators to have good professional quality
4. At present, there are some commercial Aeromonas culture medium, such as RS medium, AHM medium and APM medium, etc., but the separation rate and accuracy rate are not high (Ling Hongli, et al., Aeromonas hydrophila selective Comparison of Media Differentiation Effects, 1998)

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Screen and identify the identified bacteria isolated and preserved in our laboratory.

[0019] Experimental steps:

[0020] 1. Open the freeze-dried storage tube of the strain, inoculate it into 3ml of salt-free alkaline peptone water for enrichment, culture with shaking at 28°C for 18-20 hours, and obtain the enrichment solution.

[0021] 2. Dip the enrichment solution with an inoculation loop, inoculate it on an RS medium plate, and incubate at 28°C for 18-20 hours. If the colony is yellow and smooth on the RS medium plate, it is initially judged as suspected Aeromonas bacteria.

[0022] 3. Pick the suspected Aeromonas colony on the RS medium plate, inoculate it on the tryptone soybean agar medium plate for purification and culture, and culture it at 28°C for 18-20 hours, and then mark the grown colony on the oxidase Test strip, observe within 1min, if the line is purple or purple, it is positive for oxidase, otherwise it is negative for oxidase.

[0023] 4. Result...

Embodiment 2

[0033] Example application:

[0034] The sediment, water and fish tissues of Sanshui, Zhuhai and Shunde aquaculture ponds were randomly collected to isolate Aeromonas of aquatic origin.

[0035] A: Take about 3g of the bottom sludge, add 2mL of sterile water, mix well and let it stand for 10min, then take 100μL of the upper liquid;

[0036] B: Water sample (20-30cm below the pond surface) 100μL;

[0037] C: Take a mung bean-sized gill tissue sample with tweezers or scissors;

[0038] 1. Inoculate the above three types of samples into 3ml of salt-free alkaline peptone water for enrichment culture, shake culture at 28°C for 18-20h, and obtain the enrichment solution.

[0039] 2. Dip the enrichment solution with an inoculation loop, inoculate it on an RS medium plate, and incubate at 28°C for 18-20 hours. If the colony is yellow and smooth on the RS medium plate, it is initially judged as suspected Aeromonas bacteria;

[0040] 3. Pick the suspected Aeromonas colony on the RS ...

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PUM

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Abstract

The present invention discloses a simple method for screening aquatic source aeromonas. The method includes that: a sample to be detected is subjected to enrichment culture in salt free alkali peptone water, wherein the culture condition is a conventional culture condition of the aquatic source aeromonas; enrichment broth is inoculated to an RS medium flat plate for culturing, wherein the culture condition is the conventional culture condition of the aquatic source aeromonas, if yellow and smooth bacterial colonies appear on the RS medium flat plate, the sample is suspected bacteria of aeromonas; the suspected bacteria of aeromonas is inoculated to a tryptone soybean agar medium flat plate for culturing, wherein the culture condition is the conventional culture condition of the aquatic source aeromonas, oxidase activity of the cultured bacterial colony is detected; if the bacterial colony can grow well in the salt free alkali peptone water, the bacterial colony appears yellow and smooth on the RS medium flat plate, and the oxidase test is positive, then the bacterial colony is determined to be the aquatic source aeromonas, and the sample to be detected contains the aquatic source aeromonas.

Description

Technical field: [0001] The invention belongs to the field of microorganisms, and in particular relates to a simple and convenient method for screening Aeromonas from aquatic products. Background technique: [0002] Aeromonas belongs to Aeromonas family, widely distributed in water environment, and often detected in human body, aquatic animals and food. Aquatic animal-derived Aeromonas mainly include Aeromonas hydrophila, Aeromonas temperatus, Aeromonas victoria, Aeromonas caviae, etc., and are also common pathogenic bacteria in humans and fish, which can cause human food Poisoning, diarrhea, sepsis, etc. Aeromonas has a wide range of pathogenicity. In the high temperature season, it is easy to infect a variety of aquatic animals including fish, shrimp, shellfish, reptiles and amphibians alone or in combination. It is a highly harmful pathogen in aquaculture. It is easy to cause the outbreak of various bacterial diseases of aquatic animals, and the degree of harm is higher ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02C12Q1/04C12R1/01
Inventor 邓玉婷吴雅丽谭爱萍姜兰罗理王伟利赵飞梁爱玲
Owner PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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