Method for evaluating activity of bio-feed
A technology of activity and feed, applied in the field of activity evaluation of biological feed, can solve the problems of evaluating active feed and lack of active feed evaluation system, and achieve the effects of improving growth performance, preventing sow constipation, and treating piglet diarrhea.
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Embodiment 1
[0062] Method Example 1 Establishment of a mouse model transplanted with pig-derived flora
[0063] Sterile C57BL / 6J mice (provided by the Experimental Animal Center of the Third Military Medical University) were kept in a sterile isolator in the Teaching and Research Office of Experimental Zoology, Basic Department, Third Military Medical University, and the breeding environment: 20-24°C, humidity 40%-70% , strict 12h light 12h dark. The feed was from the co60 irradiated rat growth and breeding feed sold by Beijing Keao Xieli Feed Co., Ltd. The litter was sterilized by irradiation with Co-60γ50kGy rays. Rat cages, drinking water and drinking bottles were all sterilized by high temperature and high pressure (121°C, 60min ). Thirty 4-week-old sterile C57BL / 6J mice were fed with 0.1 mL of pig feces suspension (from healthy young pigs, without gastrointestinal diseases, normal diet and living habits, and not fed 2 months before the start of the experiment) The first fresh feces...
Embodiment 2
[0064] Method Example 2 Ex vivo chemostat model operation
[0065] Inject 500ml of sterilized intestinal flora culture medium into the fermenter of the isolated chemostat model (assemble the model according to the instructions of the application (patent) No. 200710028346.2), seal it with wax, and make the whole system in a sterile, airtight environment . Turn on the peristaltic pump connected to the fresh medium for 35ml.h -1 Flow rate operation, conveying fresh medium to continuously replenish nutrients, and waste liquid is removed at the same flow rate to ensure a constant culture medium. Accurately weigh pig feces samples (collected from healthy young pigs, without gastrointestinal diseases, normal diet and living habits, and pigs that have not been fed any antibiotics and probiotics 2 months before the start of the experiment, the first time in the morning fresh feces) 5g, diluted 5 times with sterilized 0.85% normal saline, fully homogenized and filtered, got 3 equal sa...
Embodiment 316
[0066] Method Example 3 16S rDNA sequencing
[0067] Extract the total DNA of the sample, using 16S rDNA gene V 3 -V 4 The variable region is used as a target for amplification, universal primer sequence: 341F-806R Forward primer (5'-3'): ACTCCTACGGGRSGCAGCAG (341 F) (SEQ ID NO: 1), reverse primer (5'-3') : GGACTACVVGGGTATCTAATC (806 R) (SEQ ID NO: 2) (referring to Sun, W.et al.Mechanismand Effect of Temperature on Variations in Antibiotic Resistance Genes during Anaerobic Digestion of Dairy Manure.Sci.Rep.6,30237; doi:10.1038 / srep30237(2016).), the amplified PCR product was purified and then sequenced on the machine. The entire sequencing process was completed by Shanghai Ruiyi Biotechnology Co., Ltd., including DNA extraction and primer design, etc., using Illumina HiSeq PE250 for on-machine sequencing. By performing OUT (operational taxonomic unit) cluster analysis on the sequencing results, the species composition of the community, the evolutionary relationship between...
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