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Method for screening probiotics

A technology of probiotics and bacterial strains, which is applied in the field of screening probiotics, can solve problems such as ineffective screening of bacterial strains, achieve high accuracy, promote animal health, and reduce diseases

Active Publication Date: 2013-05-08
江西华农恒青农牧有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, traditional phenotypic identification methods and 16S rDNA sequence homology analysis and identification methods are common methods for screening strains. These methods can better screen out strains with strong adhesion characteristics, high survival characteristics, and strong processability , but cannot effectively screen out strains that are beneficial to animal health, such as strains that can prevent animal colibacillosis

Method used

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  • Method for screening probiotics
  • Method for screening probiotics
  • Method for screening probiotics

Examples

Experimental program
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Embodiment 1

[0015] Dilute fresh pig manure with normal saline, streak culture on selective medium, and then purify and culture various bacteria, and select candidate probiotic strains (Lactobacillus, Streptococcus, Enterococcus genus) spare. The spleen of 21-day-old piglets was aseptically taken, cut into pieces, ground and homogenized, filtered through a 150-mesh copper mesh, and centrifuged to collect cells. Splenocytes can be obtained by processing red blood cell lysate and GKN buffer solution and centrifuging. Resuspend the cells with RPMI1640 nutrient solution, adjust the cell concentration to 1×10 6 pieces / ml. Inoculate 2×10 6 After cells / well were cultured in 6-well plate for 48 hours, 1×10 8 Escherichia coli K88 or candidate probiotic strains (Lactobacillus, Streptococcus, Enterococcus) were co-cultured for 2 hours, the cells were collected, and the expression levels of interleukin-4 and interleukin-8 in spleen cells were detected. The probiotic strains (Lactobacillus, Strept...

Embodiment 2

[0024] Dilute fresh chicken manure with normal saline, streak culture on selective medium, and then purify and culture various bacteria, and select candidate probiotic strains (Lactobacillus, Streptococcus, Enterococcus, etc.) genus) spare. Aseptically take the spleen of 6-day-old chickens, cut it into pieces, grind and homogenate, then filter through a 100-mesh mesh, collect the cell solution by centrifugation, add lymphocyte separation medium, and collect the middle white blood cell layer by centrifugation, resuspend the cells with RPMI1640 nutrient solution, and adjust the cell concentration 1×10 6 pieces / ml. Inoculate 2×10 6 After cells / well were cultured in 6-well plate for 48 hours, 1×10 8 Escherichia coli K88 or probiotics (Lactobacillus, Streptococcus, Enterococcus) were co-cultured for 2 hours, the cells were collected, and the expression levels of interleukin-4 and interleukin-8 in spleen cells were detected. The probiotic strains (Lactobacillus and Streptococcus...

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Abstract

The invention discloses a method for screening probiotics. The method comprises the following steps of: (1) carrying out a primary culture on spleen tissues of a pig or a domestic animal; (2) attacking toxicity of the tissues through colon bacillus K88, and testing variations of levels of an interleukin 8 and an interleukin 4; (3) respectively processing the tissues through candidate probiotic strains, and testing variations of levels of two interleukins; (4) comparing the variations to select probiotic strains which are similar in variation degrees of the two interleukins but opposite in effect due to treatment of the colon bacillus K88; and (5) through the colon bacillus K88 animal toxicity attacking test, observing protecting effects of primarily selected probiotic strains, and finally determining an efficient probiotic strain. The method for screening the probiotic disclosed by the invention has the remarkable advantages that: the method combining in vitro immunology experiment and in vivo toxicity attacking experiment is high in accuracy, strong in pertinency and time-saving; and the screened probiotics can effectively replace antibiotic, and greatly reduce diseases caused by colon bacillus so as to promote the health of the animal.

Description

technical field [0001] The invention belongs to the technical field of animal husbandry and veterinary medicine, and in particular relates to a method for screening probiotics. Background technique [0002] Probiotics refer to live bacterial preparations that play a beneficial role by improving the ecological balance of the host’s intestinal flora to improve the health level and health status of the host (human and animal). They can help maintain the normal function of the intestinal tract and promote nutrient absorption. , Stimulate the body's immune system and improve immunity. Probiotics are playing an increasingly important role in safeguarding human and animal health or preventing certain diseases. Screening probiotic strains from different sources and with special functions has become a research hotspot at home and abroad. At present, my country still lacks original and innovative key technologies and products of probiotics, and a double monopoly of foreign products a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/16C12Q1/02C12R1/225C12R1/46C12R1/10C12R1/125C12R1/01
Inventor 侯永清丁斌鹰易丹
Owner 江西华农恒青农牧有限公司
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