Triton's bacillus mobilis and application thereof

A technology of Tritonella and movement, applied in application, bacteria, climate change adaptation, etc., can solve the problem of bacterial resistance to aquatic product quality decline, and achieve the effect of broad drug resistance and strong virulence

Pending Publication Date: 2022-05-27
THIRD INST OF OCEANOGRAPHY MINIST OF NATURAL RESOURCES
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The use of antibiotics in the farming industry can reduce the outbreak of Vibrio disease, b

Method used

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  • Triton's bacillus mobilis and application thereof
  • Triton's bacillus mobilis and application thereof
  • Triton's bacillus mobilis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Isolation, identification and physiological and biochemical characteristics of target strains:

[0029] The Philippine sea basin samples were coated on MA plates and incubated at 28°C for 48h. Pick a single colony from the plate for purification, and obtain pure bacteria and number R6-3G2. The DNA of strain R6-3G2 was extracted, and 27F and 1492R were used as primers to amplify the 16s rRNA gene to obtain a gene sequence fragment with a length of 1392bp. Tritonibacter mobilis.

[0030] R6-3G2 salinity, pH, temperature measurement:

[0031] Fresh R6-3G2 colonies were scraped from MA plates in MB medium. The corresponding pH gradient medium and the corresponding salt concentration medium were distributed on the 96-well plate, and then 5 μL of R6-3G2 bacterial suspension was added to the corresponding 96-well plate. OD600 was measured every 24h. Pipette 5 μL of R6-3G2 bacterial suspension and add it to the MA medium, incubate at 4°C, 10°C, 15°C, 20°C, 28°C, 37°C, 45°C...

Embodiment 2

[0043] Detection of antibacterial activity of strain R6-3G2 on VP-HL:

[0044] The strain R6-3G2 was fermented in 2216E medium at 120rpm and 28°C for 72h. After the pathogenic bacteria are prepared into a bacterial suspension, it is diluted to about OD600=0.4, and the diluted bacterial liquid is further diluted 100 times. Take 100 μL of the diluted bacterial solution on the MA plate, coat it with a glass coating rod, and let it stand for 20 to 30 minutes. After the surface is completely dry, use a hole punch with a diameter of 10 mm to punch 5 evenly on the plate. After marking the hole, use a pipette to take 100 μL of fermentation broth and add it to the punched hole. Wrap the plate with plastic wrap and place it in a 28°C incubator for 16-18h. Observe the size of the inhibition zone and take pictures to record. The result is as figure 1 As shown, it was found that the fermentation broth of R6-3G2 had a better inhibitory effect on VP-HL, the diameter of the inhibition zon...

Embodiment 3

[0046] Bacteriostatic effect of R6-3G2 on other pathogens:

[0047] From the above results, it can be found that R6-3G2 has a certain inhibitory effect on pathogenic bacteria, so R6-3G2 can be further scientifically researched. R6-3G2 was inoculated into MA liquid medium and YP30 liquid medium at 120rpm / min, and fermented at 28°C for 72h. The fresh pathogenic bacteria were prepared into a bacterial suspension with OD600=0.4. After the bacterial suspension was diluted 100 times, 100 μL was taken into MB medium, coated with a sterilized coating rod, and allowed to stand for a period of time. When there is no moisture on the surface of the plate, use a hole punch to punch holes. Take 100 μL of R6-3G2 fermentation broth into the hole of the plate and mark it on the plate. After sealing and fixing with plastic wrap, incubate at 28°C for 16-18h, observe the size of the inhibition zone, and make a record.

[0048] Table 4 Inhibition of pathogenic bacteria by two fermentation broth...

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Abstract

The invention relates to triton mobilis and application thereof, and belongs to the field of probiotic screening. The invention provides a strain of Tritonibacter mobilis R6-3G2, wherein the preservation number of the Tritonibacter mobilis R6-3G2 is GDMCC (China General Microbiological Culture Collection Center) No: 62115; the DNA gene sequence is as shown in SEQ ID NO. 1. The R6-3G2 can generate an antibacterial compound for inhibiting glass seedling pathogenic bacteria vibrio parahaemolyticus VP-HL. Bacteriostatic experiments prove that the fermentation liquor of R6-3G2 has an inhibition effect on vibrio parahaemolyticus VP-HL; it is proved through a resazurin color development method that intracellular and extracellular metabolic crude extracts of R6-3G2 can inhibit vibrio parahaemolyticus VP-HL; in R6-3G2 prawn culture experiments, the infection of the vibrio parahaemolyticus VP-HL to prawns can be reduced, the survival rate of young prawns is increased, the young prawns have no toxic or side effects, and the vibrio parahaemolyticus VP-HL can be used as probiotics to be added into a water body during culture; the feed can be used as an aquaculture water additive.

Description

technical field [0001] The invention belongs to the field of probiotic screening, in particular to a strain of Tritonella mobilis and its application in shrimp culture. Background technique [0002] my country is a big country in aquaculture, and its output ranks in the forefront of the world. But in the process of aquaculture, it is often faced with the outbreak of bacterial diseases, especially Vibrio (Wang Fengqing, Sun Yuzeng, Ren Lihua, et al. Research progress on the main pathogenic Vibrio species of aquatic animals in mariculture [J]. Quality and Standards of Chinese Fisheries, 2018, 8(02):49-56). There are more than 20 kinds of Vibrio pathogens in aquaculture, mainly including Vibrio parahaemolyticus, Vibrio vulnificus, Vibrio splendidus, Vibrio alginolyticus, etc. and drug susceptibility test analysis [J]. Science Fish Farming, 2020, 08: 51-52). "Glass seedling" disease affects 6-12-day-old larvae of Litopenaeus vannamei, and the mortality rate exceeds 90% within...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34A01K61/59A01K61/13C12R1/01C02F103/20
CPCC02F3/34A01K61/59A01K61/13C02F2103/20Y02A40/81
Inventor 赖其良邵宗泽刘天琪付欢
Owner THIRD INST OF OCEANOGRAPHY MINIST OF NATURAL RESOURCES
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