Enterococcus faecalis and application thereof

A technology for Enterococcus faecalis and bacterial agent, which is applied in the field of microorganisms, can solve the problems of short fermentation period, secondary pollution of fermented soybean meal, breeding of miscellaneous bacteria, and unclear background of fermented soybean meal bacteria.

Pending Publication Date: 2021-08-06
ZHENGZHOU UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0005] At present, the background of the strains used for fermenting soybean meal is not clear enough, and the fermentation period is short. There are no reports on how to preserve the fermented soybean meal after opening, and whether there will be secondary pollution to breed miscellaneous bacteria. Animal husbandry is an important industry in the world. The demand for probiotics is large, and the feed probiotics market is still in its infancy, and the number of commercialized probiotic products is relatively small. Therefore, the present invention aims to comprehensively evaluate the effects of pig-derived lactic acid bacteria strains on females in vitro and in vivo. Both pigs and piglets have beneficial characteristics. Safe and good anti-ETEC K88 pig-derived lactic acid bacteria were isolated and screened. This strain can not only be used in soybean meal fermentation to improve soybean meal exposed to air during continuous fermentation for 30 days and after fermentation. At the same time, it has a good effect on breeding sows and piglets, so as to provide reference for the full utilization of fermented soybean meal and the development of new fermented feed, and provide probiotic resources for the development of complex microecological preparations for pigs

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  • Enterococcus faecalis and application thereof
  • Enterococcus faecalis and application thereof
  • Enterococcus faecalis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1. Isolation and Identification of Enterococcus faecalis PF95 CCTCC NO: M 2020852

[0042] 1. Isolation of Enterococcus faecalis PF95

[0043] Accurately weigh 20g of feces samples from weaned healthy piglets from a pig farm in Puyang City, Henan Province, China, put them into a triangular flask filled with 180mL of sterile water, seal it with a parafilm, and shake it in a shaker at 180r / min for 30 minutes , to fully disperse the microorganisms, let it stand for 20 seconds, and then 10 -1 diluent; pipette 100μL 10 -1 Add the diluted solution into a centrifuge tube filled with 900 μL sterile water, and mix well to obtain 10 -2 diluent; pipette 100μL 10 -2 Diluent, add to a centrifuge tube filled with 900 μL sterile water, mix well and serve as 10 -3 Diluent; use the same method to serially dilute to make 10 -4 、10 -5 Wait for a series of dilutions. Take the MRS medium that was sterilized at 121°C for 15 minutes, and use a marker pen to mark 10 -1 、10 -3 ...

Embodiment 2

[0058] Embodiment 2, Enterococcus faecalis (Enterococcus faecalis) PF95 inhibits the characteristics and physiological and biochemical characteristics of ETEC K88

[0059] 1. Determination of the ability of Enterococcus faecalis PF95 to inhibit ETEC K88 The antagonism of Enterococcus faecalis PF95 on ETEC K88 was determined by the agar block diffusion method. After culturing the target bacteria ETEC K88 overnight in Luria-Bertani (LB) liquid medium, take 100uL of the overnight culture, add K88 to the LB agar medium and cool to 50°C, mix well, and pour into the solidified LB agar plate Surface; after solidification, punch a hole in the center of the agar plate with a hole puncher with a diameter of 10.00 mm, and add 200 uL of PF95 bacterial solution cultured for 16 hours. Uninoculated blank MRS broth and penicillin were used as negative and positive controls, respectively. After diffusion at 4°C for 2 hours, the diameter of the inhibition zone was measured, and it was found th...

Embodiment 3

[0085] Embodiment 3, the bile salt resistance of Enterococcus faecalis (Enterococcus faecalis) PF95, simulated gastrointestinal tract and antibacterial spectrum determination

[0086] 1. Determination of Bile Salt Resistance of Enterococcus faecalis PF95

[0087] The seed liquid of the activated Enterococcus faecalis (Enterococcus faecalis) PF95 was mixed evenly with 0.2% bile salt solution in a ratio of 2%, and samples were taken at 0, 1, 2, 3 and 4 hours under room temperature conditions, and each sampling 20 μl was dropped on the MRS plate, incubated at 30°C for 48 hours, and the number of colonies in each sample was determined by the MRS plate colony counting method. The experiment was repeated 3 times, and the results were averaged.

[0088] Table 6. Bile salt tolerance of Enterococcus faecalis PF95

[0089] strain 0 hour bacteria count The number of viable bacteria in 1 hour 2 hours surviving bacteria count 3 hours surviving bacteria 4 hours survivi...

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Abstract

The invention relates to the field of microorganisms and fermented feed, in particular to enterococcus faecalis and application thereof. The enterococcus faecalis is specifically (Enterococcus faecalis) PF95, is preserved in the China Center for Type Culture Collection with the preservation number of CCTCC NO: M 2020852, has the stress resistance such as temperature resistance, salt resistance and acid and alkali resistance, and can effectively inhibit growth or generation of harmful infectious microbes; the content of lactic acid is increased, and meanwhile a large number of probiotics are generated; particularly, the quality of the feed can still be well improved in the unsealing stage when the feed is opened for feeding after fermentation; a lactobacillus agent is added in the perinatal period and the lactation period for feeding, so that the breeding performance and re-breeding performance of sows can be improved, the serum indexes of the sows are optimized, and meanwhile, the growth performance of suckling piglets can be improved; and by adding the mixed lactobacillus agent for feeding, the feed intake and the growth speed of piglets can be effectively increased, and the diarrhea rate and the case fatality rate of the piglets are reduced.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to a strain of Enterococcus faecalis and its application. Background technique [0002] Enterotoxic Escherichia coli (ETEC K88) expressing Acinetobacter K88 is the main microorganism that causes bacterial diarrhea in weaned piglets, and the economic loss caused by bacterial infectious diarrhea in piglets can reach as much as 26% in the pig industry. For a long time, antibiotics have been used to reduce diarrhea and improve growth performance in weaned piglets. Unfortunately, the indiscriminate use of antibiotics has caused many problems such as drug resistance, environmental pollution, and residues in human food. Since January 1, 2006, the European Union has banned the use of antibiotics as growth promoters in weaned pig diets (EC Regulation No. 1831 / 2003). The use of antibiotics in feed has been completely banned in my country since January 1, 2020. Some studies have shown that zinc ...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23K10/12A23K10/18A23K10/37A23K50/30A23K50/60A61K35/744A61P31/04C12R1/01
CPCA23K10/12A23K10/18A23K10/37A23K50/30A23K50/60A61K35/744A61P31/04Y02P60/87
Inventor 庞会利马皓王伟伟于豪杰秦广雍谈重芳王雁萍
Owner ZHENGZHOU UNIV
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