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.
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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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