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A method for in vitro preservation of pig intestinal chyme microbiome

A technology of microorganisms and chyme, which is applied in the field of biotechnology and animal husbandry, can solve the problems of poor reliability in the study of microorganisms in the colon of growing pigs, and achieve the effect of improving reliability

Active Publication Date: 2022-02-11
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current lack of systematic and validated microbial preservation studies leads to poor reliability in in vitro studies of colonic microorganisms in growing pigs

Method used

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  • A method for in vitro preservation of pig intestinal chyme microbiome
  • A method for in vitro preservation of pig intestinal chyme microbiome
  • A method for in vitro preservation 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 invention discloses an in vitro preservation method of pig intestinal chyme microbiome. The invention provides a method for cryopreserving colon microbes of growing pigs, which comprises the following steps: adding glycerin to cryopreservation of growing pig colon digesta or its inoculum, so as to realize cryopreservation of growing pig colon microbiomes. The microbiome with high similarity level preserved by this method can be directly used in fecal bacteria transplantation and dietary fiber fermentation test, which solves the problems of high cost, few donors and inconsistent standards when the same microbiome is used multiple times. It has played a positive role in promoting the production of pigs and the development of animal husbandry 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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Patent Type & Authority Patents(China)
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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