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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

Inactive Publication Date: 2020-12-15
ANHUI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The research on the uterine flora of cows suffering from endometritis has attracted the attention of many scholars, but due to the limitation of research methods, the current research on the flora of endometritis is still limited

Method used

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  • Microbial marker for bovine endometritis and application thereof
  • Microbial marker for bovine endometritis and application thereof
  • Microbial marker for bovine endometritis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses a microbial marker for bovine endometritis. The microbial marker comprises the following microorganisms with remarkably increased abundance: Bacteriodetes, Elusimicrobia, Verrucomicrobia, Spirochaetae and Lentisphaerae. The microbial marker also comprises the following microorganisms with remarkably increased abundance: Akkermansia, biogas bacillus, CF231 and Oscillosporia.The microbial marker also comprises the following microorganisms with remarkably decreased abundance: Ureaplasma urealyticum, Butyrivibrio, Doria, Sphingomonas sp, Rothia and Streptococcus. The invention discloses an application of the microbial marker for bovine endometritis in preparation of a tool for early screening or prediction of bovine endometritis. The applicant performs 16S rRNA gene sequencing on microbial DNA of a uterine mucus sample, compares structural differences of uterine florae between healthy and sick dairy cows, and finds differences in microbial community composition ofthe uterine florae between the healthy and sick dairy cows. The result provides a theoretical basis for better prevention and treatment of clinical endometritis.

Description

technical field [0001] The invention relates to the technical field of microbial markers, in particular to a microbial marker of bovine endometritis and its application. Background technique [0002] After the cow gives birth, the cervix opens, and various microorganisms "settle" in the uterus. Studies have shown that there are also microbial flora in the uterus of healthy cows, which contain not only normal microbial flora, but also pathogenic bacteria and opportunistic pathogenic bacteria. When host immunosuppression, external stress response and other conditions occur, the immune response of the cow is difficult to successfully resist the pathogenic effect of pathogenic bacteria, which will cause endometritis in the cow. [0003] Studies have found that 40% of dairy cows suffer from metritis and endometritis through bacterial infection within 3 weeks after delivery, and 15-20% of cows will develop clinical endometritis within 3 weeks after delivery. Clinical endometriti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12Q1/689G16B20/30G16B5/00C12R1/01C12R1/145
CPCC12Q1/6883C12Q1/689C12Q2600/124G16B5/00G16B20/30
Inventor 刘洪瑜刘亚欧阳少芬徐德志陈敬梦庄静仪刘善斋仝笑李开发鲁敏
Owner ANHUI AGRICULTURAL UNIVERSITY
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