Screening and application of enterococcus faecium G12

A technology of Enterococcus faecium and G12, applied in the field of microbial additives for aquatic products, can solve the problems of increased overall drug resistance and unfavorable treatment of animal diseases, and achieve the effect of sustainable development, significant probiotics, and high resistance to stress.

Inactive Publication Date: 2019-11-05
TIANJIN AGRICULTURE COLLEGE
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  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

On the other hand, the resistance of bacteria to antibiotics has a horizontal transmission effect, and it is likely to transmit the level of drug resistance to

Method used

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  • Screening and application of enterococcus faecium G12
  • Screening and application of enterococcus faecium G12
  • Screening and application of enterococcus faecium G12

Examples

Experimental program
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Example Embodiment

[0027] Example 1: Screening, isolation and identification of strains

[0028] Screening and isolation: A strain of Enterococcus faecium was selected from the hepatopancreas of shellfish, especially the hepatopancreas of Philippine clam. The specific operation of the screening is as follows: the body surface of the clam clams is sterilized with 75% medical alcohol and then dissected, the liver and pancreas are taken, and then 1 mL of pre-chilled sterile normal saline is added, homogenized thoroughly, centrifuged, and the supernatant is taken 10 times series dilution. Take stock solution, 10 -1 , 10 -2 The samples of the three dilutions were each 100uL, and they were coated on the MRS plate. Each dilution gradient was set to 3 parallel, and the samples were incubated at 37°C for 24h. Pick a single colony and purify the bacteria by streaking multiple times. The purified strain was made into a bacterial suspension with 40% glycerol and MRS liquid medium 1:1, and stored at -80°C for...

Example Embodiment

[0035] Example 2: Bacteriostatic test

[0036] Aeromonas victorini, Pseudomonas aeruginosa and Vibrio vulnificus (the above-mentioned strains are provided by the Aquatic Animal Disease and Immunology Laboratory of the Fisheries College of Tianjin Agricultural College) as indicator bacteria. Methods Refer to the literature (Evidence for the competitive exclusion of Aeromonas salmonicida from fish with a tressinducible furunculosis by a fluorescent pseudomonad. Smith P and Dabey S, "Journal of Fish Diseases", 1993 Issue 16), and the activated indicator bacteria and the test Strains were cultured in LB liquid medium for 24h, diluted with saline to 1×10 7 cfu / mL, take 0.2mL of indicator strain and spread it on fresh LB solid medium, and then take 2μL of test strain to spot on the above plate, and observe whether there is obvious inhibition zone around the spot seeding area for 24h.

[0037] The Oxford cup method (using the Oxford cup method to determine the antibacterial activity of pr...

Example Embodiment

[0040] Example 3: Safety test

[0041] Hemolysis test: Streak the tested strain on LB solid medium for 24h, scrape a single colony with an inoculation loop and spot it on the blood plate, incubate at 37℃ for 24h, observe whether a transparent circle is formed around the colony, and judge the production of hemolysin . The experimental results showed that: no hemolysis was observed on the blood plate, indicating that the strain did not produce hemolysin and had no potential pathogenicity.

[0042] Animal test: refer to the literature, select the immersion test of Penaeus monodon juveniles, select 1×10 7 , 1×10 5 The concentration of cfu / mL bacterial solution was soaked, the positive control was selected as Vibrio vulnificus, and the negative control group was selected as normal saline. After being soaked in the bacterial suspension for four hours, the juvenile shrimp were transferred to violent water. Each treatment group made three parallel observations for 7 days, and recorded th...

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Abstract

The invention provides enterococcus faecium G12. The enterococcus faecium G12 is isolated from hepato-pancreas of shellfish, is safe and has probiotic characteristics. A strain enterococcus faecium isnamed the enterococcus faecium G12, and is deposited in the China General Microbiological Culture Collection Center, the deposit number is CGMCC NO.15230, and the date of deposit is January 17, 2018.The enterococcus faecium G12 has high heat resistance, high resistance to stress and remarkable probiotic properties, has an obvious inhibitory effect on a common aquatic pathogenic bacterium aeromonas veronii, is sensitive to most broad-spectrum antibiotics, can tolerate a small intestine high-bile-salt environment and is colonized in the intestine, and is suitable for being used as a feed additive to be widely applied to aquaculture, thereby being capable of greatly reducing the pollution to the environmental and the adverse effects on human health caused by the addition of traditional antibiotics in the aquaculture, and achieving sustainable development of the aquaculture.

Description

technical field [0001] The invention relates to the technical field of microbial additives for aquatic products, in particular to the isolation, identification and safety of Enterococcus faecium (Enterococcus faecium), which has obvious inhibitory effect on the common aquatic pathogen Aeromonas vietori and is sensitive to most spectrum antibiotics. Performance test and identification and application of stress resistance performance. Background technique [0002] Enterococcus faecium (Enterococcus faecium) is a kind of anaerobic, non-spore-forming Gram-positive bacteria. The bacteria are round or oval, arranged in chains, and do not produce spores. Off-white, opaque, smooth-surfaced, circular colonies with a diameter of 0.5-1 mm can be formed on the blood plate. According to the "Catalogue of Feed Additives" issued by the Ministry of Agriculture of China, the types of feed microbial additives are clearly stipulated. Enterococcus faecium is among them, and it has become a com...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01K61/13A23K10/18C12R1/01
CPCA23K10/18A01K61/13C12N1/205C12R2001/46Y02A40/81
Inventor 毛晴孙敬锋谢云丹张燕玉王轶凡
Owner TIANJIN AGRICULTURE COLLEGE
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