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Seawater selective chromogenic culture media for VP (Vibrio Parahaemolyticus) and authenticating and counting method thereof

A technology of chromogenic culture medium and Vibrio hemolyticus, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, and resistance to vector-borne diseases, etc. It can solve the problems of statistical analysis of inability and counting, and improve accuracy and efficiency, growth promotion, identification and enumeration cost-bottom effects

Inactive Publication Date: 2015-01-21
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are mainly traditional detection methods, molecular biology detection methods, chromatography, immunological detection methods, etc. in the identification of Vibrio parahaemolyticus, but these methods are not capable of counting statistical analysis
In addition, the identification and counting of Vibrio parahaemolyticus in the bath environment is a blank field

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Specificity experiment: Preparation of seawater selective chromogenic medium for Vibrio parahaemolyticus in marine environment: Each 1000mL medium contains 10g of beef extract, 5g of peptone, 15g of sodium thiosulfate, 3g of sodium citrate, 10g of choline, β - Glucoside 0.1g, sucrose 20g, agar 15g, seawater 1000mL, natural pH, heat and boil the culture medium to sterilize, cool to 50-60°C, pour plate, and set aside.

[0023] Inoculation: Mix Vibrio parahaemolyticus, Vibrio cholerae, Vibrio vulnificus, Enterobacter cloacae, Escherichia coli, Bacillus cereus, Staphylococcus aureus, and Salmonella, and then make gradient dilutions, and take 102-105 different concentrations of bacterial solutions to apply Spread on the plate prepared in step 1, and observe after incubating at a constant temperature of 28°C for 18 hours.

[0024] Results and analysis: Vibrio parahaemolyticus showed purple-red colonies in the medium, and other bacteria showed blue, green, white colonies or di...

Embodiment 2

[0026] 1. Preparation of seawater selective chromogenic medium of Vibrio parahaemolyticus in a marine environment: every 1000mL medium contains beef extract 10g, peptone 5g, sodium thiosulfate 15g, sodium citrate 3g, choline 10g, 0.1g of β-glucoside, 20g of sucrose, 15g of agar, 1000mL of seawater, natural pH; heat and boil the culture medium to sterilize, cool down to 50-60°C, pour it on a plate, and set aside.

[0027] 2. Membrane filtration of water samples: In August 2014, a sterile sampling bottle was used to collect 100mL of fresh seawater 0.5m below the water surface of Laohushi Bathing Beach, Beidaihe, Qinhuangdao, and a 0.22μm membrane was used to filter the water samples;

[0028] 3. Membrane culture: Use sterile tweezers to remove the filter membrane after filtering in step 2, and move it to the surface of the seawater selective color development plate prepared in step 1 so that the filter membrane and the plate are close to each other without air bubbles ;

[0029...

Embodiment 3

[0033] 1. Preparation of a seawater selective chromogenic medium for Vibrio parahaemolyticus in a marine environment: every 1000mL medium contains 5g of beef extract, 10g of peptone, 6g of sodium thiosulfate, 5g of sodium citrate, 5g of choline, and β - Glucoside 0.3g, sucrose 15g, agar 20g, seawater 1000mL, natural pH, heat and boil the medium to sterilize, cool to 50-60°C, pour plate, and set aside;

[0034] 2. Membrane filtration of water samples: In August 2014, 10mL of fresh seawater 0.5m below the water surface of Laohushi Bathing Beach, Beidaihe, Qinhuangdao was collected with a sterile sampling bottle, and the seawater samples were filtered with a 0.22μm membrane;

[0035] 3. Membrane culture: Use sterile tweezers to remove the filter membrane after filtering in step 2, and move it to the surface of the seawater selective color development plate prepared in step 1 so that the filter membrane and the plate are close to each other without air bubbles ;

[0036] 4. Place...

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Abstract

The invention discloses a method adopting seawater selective chromogenic culture media to perform quick separation, authentication, counting and analyzing on the VP (Vibrio Parahaemolyticus) in the marine environment. The method comprises the specific steps as follows: firstly, preparing a seawater selective chromogenic culture flat plate for the VP; secondly, taking a certain amount of seawater sample and filtering by a filter membrane with the filtration diameter of 0.22 mum; thirdly, horizontally adhering the filter membrane with sterilized tweezers to the seawater selective chromogenic culture flat plate obtained in the first step for culture at 28 DEG C; fourthly, counting the statistics of the bacterial colony which is claret-colored. According to the method provided by the invention, the blanks of quick authentication and counting of pathogenic bacteria VP in the present marine environment are filled, and enriched culture is not required, the testing time is short, and the result is accurate and reliable.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a seawater selective chromogenic culture medium for Vibrio parahaemolyticus and an identification and counting method thereof. Background technique [0002] Vibrio parahaemolyticus (Vibrio parahaemolyticus, VP) is a common pathogen. Specifically, it is a Gram-negative bacterium belonging to the genus Vibrio, which is widely found in marine environments such as estuaries, seawater, and seabed sediments, as well as seafood such as fish and shellfish, and can be infected by water and aquatic animals. . It not only seriously harms the mariculture industry, but also causes food poisoning diseases such as human gastroenteritis, and is an important pathogenic bacteria. In recent years, food poisoning incidents caused by accidentally eating seafood contaminated by Vibrio parahaemolyticus or improperly processed have emerged in an endless stream, and bacterial food poisonin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/06
CPCY02A50/30
Inventor 王中华
Owner TIANJIN UNIV
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