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Method for screening live pig intestinal micro-ecological preparation with high adhesive capacity

A micro-ecological preparation and high-adhesion technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as drug residues, affecting meat quality, flavor and food safety, and achieve simple methods , highly targeted effect

Inactive Publication Date: 2014-11-26
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the process of livestock breeding, the occurrence of animal diseases inevitably leads to the extensive use of antibiotics, resulting in extremely serious drug residues, seriously affecting the quality, flavor and food safety of meat

Method used

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  • Method for screening live pig intestinal micro-ecological preparation with high adhesive capacity
  • Method for screening live pig intestinal micro-ecological preparation with high adhesive capacity
  • Method for screening live pig intestinal micro-ecological preparation with high adhesive capacity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Screening and identification of strongly adherent bacteria in pig feces:

[0020] 1. Collection and preservation of samples: collect feces and intestinal wall mucosal layer substances in the cecum of healthy pigs, transfer them to anaerobic environment, add glycerin, whose weight is 10% of the total weight of feces and glycerin, mix well with a vortex shaker, - Store at 8O ℃;

[0021] 2. Assembly and use of the chemostat: connect the storage tank, fermentation tank, waste liquid tank, rubber plug, silicone tube, glass bottle containing NaOH and ventilation filter in order, place them on the workbench, connect and mix Gas, peristaltic pump, pH regulator, magnetic stirrer, exhaust gas removal device;

[0022] 3. Preparation of culture medium in the chemostat: add the following ingredients according to the mass percentage of the added ingredients in the final product: 5‰ soluble starch, 3‰ bovine casein, 3‰ peptone, 0.6‰ pectin, 0.6‰ xylan, 0.6 ‰ Arabinogalactan, 0.6‰ Pu...

Embodiment 2

[0024] Embodiment 2: the identification of screening bacterial strain

[0025] Scrape the material on the wall of the chemostat bottle, use the viable count method to isolate the microorganism and send it to the sequencing company for identification. The identification results are shown in Table 1. A total of 3 microorganisms were isolated, which are Lactobacillus plantarum WCFS1Lactobacillus plantarum, and the gene sequence number It is NR_076229.1; Lactobacillus reuteri, whose gene sequence number is EF421935.1; Enterococcus faecalis, whose gene sequence number is AY735444.1. The above three kinds of probiotics can be obtained from public institutions such as, and their gene sequences can be obtained from the public database NCBI.

Embodiment 3

[0026] Example 3: Evaluation of probiotic properties of screening strains

[0027] 1. Determination of adhesion strength: HT-29 cells were cultured in 1640 medium with 10% fetal bovine serum, and after confluence, the three kinds of bacteria selected were added Lactobacillus plantarum, Lactobacillus reuteri, Enterococcus faecalis , calculate the number of bacteria per 100 cells after culturing for 2 hours;

[0028] 3. Determination of acid-base tolerance: prepare MRS medium, set the pH to five gradients of 1.5, 2.5, 3.5, 4.5, and 7.0, inoculate 1% of the three bacterial solutions respectively, and count the viable bacteria after overnight culture;

[0029] 4. Bile salt tolerance test: Prepare MRS medium, set the bile salt concentration to four gradients of 0, 0.10%, 0.30%, and 0.50%, inoculate 1% of the three bacterial solutions respectively, and count the viable bacteria after overnight culture.

[0030] 5. The results of the determination of intestinal microbial adhesion ...

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Abstract

The invention discloses a method for screening a live pig intestinal micro-ecological preparation with high adhesive capacity. The method comprises the following steps: culturing pig manure by adopting a chemostat, picking microorganisms adhered to a bottle wall of the chemostat, performing identification, and then evaluating the influences of the microorganisms on the adhesive capacity of intestinal epithelial cells HT-29 and the influences of different factors (temperature, cholate, PH values and the like) on the growth of screened bacteria. The method disclosed by the invention has the advantages as follows: the method directly adopts the adhesive capacity on the bottle wall of the chemostat as a screening indicator, and has strong pertinence. Because the adhesive capacity is a key indicator for probiotics to play roles, the method can be used for screening micro-ecological potential strains of animal husbandry and human beings.

Description

technical field [0001] The invention relates to the screening of strongly adherent bacteria in pig feces and the research on their growth characteristics, which is closely related to the pig breeding industry and can be used for the development of breeding industry feed and the treatment of animal diseases. Background technique [0002] With the relatively sufficient supply of meat products in the domestic market, the contradiction between the rapid growth of meat pigs and the quality of meat pigs has become prominent. Effectively improving meat production efficiency and improving pork quality is an urgent problem to be solved. Probiotics are a type of healthy bacteria that are safe and have no toxic side effects. They can resist the invasion of foreign pathogens, improve the immunity and nutrient utilization of pigs, and can be used in feed fermentation to replace the use of antibiotics and drugs that catalyze the growth of pigs. production of safe and high-quality meat pr...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/25C12R1/225C12R1/01
Inventor 陈廷涛王鑫辛洪波
Owner NANCHANG UNIV
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