In vitro preservation method of pig intestinal chyme microbiome

A technology of microorganisms and chyme, applied in the fields of biotechnology and animal husbandry, can solve problems such as poor reliability of research on colonic microorganisms of growing pigs, and achieve the effects of high cost, improved reliability and credible test results.

Active Publication Date: 2020-01-31
CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The current lack of systematic and validated microbial preservation studies leads

Method used

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  • In vitro preservation method of pig intestinal chyme microbiome
  • In vitro preservation method of pig intestinal chyme microbiome
  • In vitro preservation method of pig intestinal chyme microbiome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1. Establishment and selection of short-term cryopreservation method for colonic microbiome

[0056] 1. Obtaining fresh digesta and inoculum from porcine colon

[0057] 1. Obtaining fresh chyme from porcine colon

[0058] Three 30-50kg Du long three-way hybrid sows and three castrated boars were used. The animals were fasted for 8 hours, and then fed freely for 12 hours. After neck bleeding, the abdominal cavity was quickly opened, and the colon was ligated and placed in an ice box and transported to the laboratory immediately. The surface of the intestinal segment was disinfected with alcohol and placed in a constant temperature anaerobic operation box at 37°C. The same amount of chyme from the same position in the colon of each pig was mixed evenly to obtain fresh colonic mixed chyme of growing pigs.

[0059] 2. Preparation of inoculum

[0060] In the constant temperature anaerobic operation box, the fresh colon mixed chyme of the growing pig obtained in t...

Embodiment 2

[0110] Example 2, establishment and selection of long-term (greater than 48 hours) cryopreservation methods in the colonic microbiome

[0111] 1. Inoculation fermentation and result detection of methods B1, B2 and C1, C2

[0112] Method B1 Add a certain amount of glycerol to the fresh colonic mixed chyme of growing pigs obtained in 1-1 of Example 1 in a constant temperature anaerobic operation box at 37°C, and the volume ratio of chyme to glycerol is 10:1. Mix evenly and store in a refrigerator at -80°C for 18 weeks. After 18 weeks, quickly thaw and recover under anaerobic conditions at 37°C. The thawed digesta is prepared according to method B in Example 1 for inoculation and fermentation.

[0113] Method B2 Add a certain amount of glycerol to the fresh colonic mixed digesta of growing pigs obtained in Part 1 of Example 1 in a constant temperature anaerobic operation box at 37°C, and the volume ratio of digesta to glycerol is 5:1. Mix evenly and store in a refrigerator at -8...

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Abstract

The present invention discloses an in vitro preservation method of pig intestinal chyme microbiome and provides a method for freezing and storing colon microorganisms of growing pigs. The method comprises the following steps: glycerol is added into growing pig colon chyme or an inoculation solution thereof for freezing and storing, so that the freezing and storing of the colon microorganisms of the growing pigs are realized. The preserved obtained microbiome with high similarity level can be directly used in fecal microorganism transplantation and dietary grain fiber fermentation experiments,solves problems of high cost, few donors, inconsistent standards, etc. when the same microbiome is used multiple times, and actively promotes development of pig production and livestock industry.

Description

technical field [0001] The invention relates to the fields of biotechnology and animal husbandry, in particular to an in vitro preservation method of pig intestinal chyme microbiome. Background technique [0002] There are a variety of microbial flora distributed on the surface of the animal body, but more microorganisms exist in the hindgut of the animal body, and the number of microorganisms in the hindgut even exceeds the total number of cells in the body. In fact, the microbiome of the hindgut can almost be regarded as an organ with complex physiological functions, which is of great significance to the health of the body. The microecological balance in the intestinal tract is a key factor in determining the health of the intestinal tract. As people's extensive use of antibiotics breaks the balance of intestinal microecology, the imbalance of intestinal microecology leads to a large number of intestinal diseases, Clostridium difficile infection (CDI) is one of the importa...

Claims

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Application Information

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IPC IPC(8): C12N1/04
CPCC12N1/04
Inventor 王军军白宇张帅黄世猛杨光新何贝贝李娜
Owner CHINA AGRI UNIV
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