Vacuum freeze-drying protective agent for vibrio mazei and its preservation method

A technology of vacuum freeze-drying and Vibrio maxima, applied in the field of preparation of freeze-drying protective agent for Vibrio maxima, can solve the problems of decreased cell activity, accelerated cell metabolism, cell damage, etc., so as to reduce the loss of bacteria and promote repair. , the effect of stable nature

Active Publication Date: 2021-06-01
INSPECTION & QUARANTINE TECH CENT SHANDONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, in the freeze-drying process, the freezing and drying stress caused by the severe environment will cause different degrees of damage to the cells and cause them to die or lose their activity.
Freezing damage is mainly caused by the formation of ice crystals, too fast a freezing rate can cause mechanical damage, and too slow a freezing rate can cause solute damage
Sublimation drying can also cause cell damage. Excessive drying will lead to cell death due to dehydration. Ending drying early will lead to excessive water, thereby accelerating cell metabolism and reducing cell activity.

Method used

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  • Vacuum freeze-drying protective agent for vibrio mazei and its preservation method
  • Vacuum freeze-drying protective agent for vibrio mazei and its preservation method
  • Vacuum freeze-drying protective agent for vibrio mazei and its preservation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] (1) Strain F5-1 was isolated from the abdomen of American lobster (Homarus americanus) imported from the United States. Aseptically take 25g of the abdomen of American lobster, put it in a sterile homogeneous cup filled with 225mL alkaline peptone water, incubate at a constant temperature of 36±1°C for 18h, inoculate it on a TCBS agar plate, incubate at 36±1°C for 24h, observe For colony morphology, obtain a dominant bacterial strain with basically the same colony morphology, pick a single colony of this strain, and transfer it to a new TCBS agar medium for streak culture. Repeat the above steps 2-3 times until a pure culture of the bacteria is obtained. The strain was identified as Vibrio metschnikovii (Vibrio metschnikovii) using an automatic microbial identification instrument and a Gram-negative bacteria identification card.

[0023] (2) Inoculate sterile alkaline peptone water with the purified and cultured Vibrio maslovakia, cultivate for 16-20 hours at 36±1°C, a...

Embodiment 2

[0046] (1) Strain F14-11 was isolated from frozen arctic sweet shrimp imported from Estonia. Aseptically take 25g of frozen arctic sweet shrimp, place it in a sterile homogeneous cup filled with 225mL alkaline peptone water, incubate at a constant temperature of 36±1°C for 18h, inoculate it on a TCBS agar plate, and incubate at 36±1°C for 24h. Observe the colony shape, obtain a dominant bacterial strain with basically the same colony shape, pick a single colony of this strain, and transfer it to a new TCBS agar medium for streak culture. Repeat the above steps 2-3 times until a pure culture of the bacteria is obtained. The strain was identified as Vibrio metschnikovii (Vibrio metschnikovii) using an automatic microbial identification instrument and a Gram-negative bacteria identification card.

[0047] (2) Inoculate sterile alkaline peptone water with the purified and cultured Vibrio maslovakia, cultivate for 16-20 hours at 36±1°C, and then enrich and cultivate to the station...

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Abstract

The invention discloses a vacuum freeze-drying protective agent for Vibrio marzini and a preservation method thereof. The protective agent is characterized in that the composition is: sodium alginate, sodium glutamate, sodium pyruvate, skimmed milk powder, chlorine NaCl and sterile distilled water. The preparation method of the freeze-dried powder of Vibrio mazei comprises the following steps: (1) The suspension of Vibrio mazei is uniformly dispersed in a freeze-drying protective agent to form a suspension of Vibrio mazei, which is divided into vials. Pre-cool at 4°C for 30min‑60min; prefreeze at ‑70°C for 8‑10h. (2) Transfer the above-mentioned pre-frozen Vibrio mazei suspension to a lyophilizer, and vacuum freeze-dry to obtain a lyophilized Vibrio mazei powder. The present invention adopts the method of pre-cooling first and then pre-freezing to carry out adaptive rapid pre-freezing of Vibrio martensilii, which reduces the loss of bacteria in the pre-freezing process, optimizes the vacuum drying parameters, can effectively reduce freezing damage and drying stress damage, and greatly improves The rejuvenation survival rate of the Vibrio mazei freeze-dried powder, the obtained freeze-dried powder of the Vibrio mazei has stable properties and is convenient for long-term storage and scientific research at room temperature.

Description

technical field [0001] The invention relates to a freeze-drying protective agent and a freeze-drying method for preparing Vibrio mazei, belonging to the field of strain preservation in the aspect of microbiological detection. Background technique [0002] Pathogenic Vibrio is widely distributed in seawater and seafood, especially in coastal estuaries and estuaries. Humans ingest or contact contaminated water and food to cause infection. Among them, Vibriometschnikovii (Vibriometschnikovii) is one of the 11 kinds of pathogenic Vibrio recognized internationally, and it is also a pathogenic bacterium that causes infectious diarrhea, mainly causing traumatic infection and sepsis in humans. Vibrio cholerae has the same cultural characteristics as Vibrio cholerae, can grow at pH 10 and 40°C, and is positive in serological tests for detecting Vibrio cholerae, so it is easy to be misdiagnosed as Vibrio cholerae. In addition, the oxidase-negative Vibrio mazei is easily mistaken for ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/04C12R1/63
CPCC12N1/04C12N1/20
Inventor 赵丽青贾俊涛姜英辉黄小华吴振兴厉艳张晓良方佩佩王勇
Owner INSPECTION & QUARANTINE TECH CENT SHANDONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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