Microbial marker for bovine endometritis and application thereof
A technology for endometritis and microbiology, applied in the direction of microbiology, microbiological-based methods, microbiological measurement/testing, etc., can solve problems such as limitations of research methods
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Embodiment 1
[0030] Using EDTA-K 2 Anticoagulant vacuum blood collection tubes were used to collect 10 mL of tail vein blood from 8 cows and store them in a refrigerator at 4°C.
[0031] Use a fully automatic five-differentiation animal blood cell analyzer to measure the number of white blood cells, percentage of neutrophils, number of neutrophils, percentage of lymphocytes, percentage of lymphocytes, and percentage of monocytes in the blood of healthy and endometritis cows , monocyte count and other blood routine physiological indicators.
[0032] The relevant indicators of blood routine are as follows:
[0033]
[0034]
[0035] It can be seen from the above table that the percentage of neutrophils, the number of neutrophils and the percentage of lymphocytes in the endometritis group were significantly different from those in the healthy control group. The percentage of neutrophils and the number of neutrophils in the endometritis group were significantly higher than those in the...
Embodiment 2
[0037] Rinse the vulva of the cow with water before sampling, wipe it clean and disinfect it with 75% alcohol solution to avoid fecal contamination. Uterine mucus samples were collected using a sampler designed by Henan Academy of Agricultural Sciences, and the sampler was sterilized by ethylene oxide. Use the rectal grasping method to send the sampler into the vagina. After the sampler enters the uterus, pull the plastic catheter backward to expose the sampling head to the uterine mucus. After rotating 2-3 times, the sampling head returns to the plastic catheter to avoid The sampling head is in contact with the vagina. Immediately after taking it out, rinse the sampling head with normal saline, collect the washing liquid and put it into a 50mL sterile centrifuge tube, and quickly put it into the ice box. After all samples were collected, the samples were stored at -80°C.
[0038] Total bacterial DNA was extracted from cow uterine mucus samples using a DNA extraction kit. P...
Embodiment 3
[0041]The applicant used the Illumina HiSeq platform to perform high-throughput sequencing of the V4 region of the 16S rRNA gene in 8 vaginal mucus samples. After performing quality trimming and chimera inspection, 601,017 high-quality sequences were obtained. The effective length averaged 252bp. According to the above results, the ven diagram between the sick group and the healthy group is established, such as figure 1 shown.
[0042] figure 1 It shows that among the 2281 OTUs, 627 are shared by the two groups, and these 627 shared OTUs account for 80.07% and 41.85% of the total OTUs of cows in the healthy group and diseased group respectively, indicating that The levels of microbial diversity of the uterine flora were partially similar, sharing some bacterial species, but the diversity of the flora in the diseased group was more complex.
[0043] Draw the sparse curve and OUT Rank curve respectively according to the above results, such as figure 2 and image 3 shown. ...
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