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Method for obtaining nitrate-reducing bacteria from rumen fluid

A technology of nitrate and rumen fluid, applied in the field of microbiological research, can solve the problems of black particles in the bacterial fluid, failure to obtain rumen nitrate-reducing bacteria, flocculent precipitation, etc., and achieve the effect of increasing the total bacterial content

Inactive Publication Date: 2017-05-10
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, rumen nitrate-reducing bacteria must reproduce under anaerobic conditions, and the use of Na 2 S, as a deoxygenating solvent, can easily lead to black particles in the bacterial solution, or flocculent precipitation after sterilization, which affects the purity and growth of bacteria, and the growth temperature is very different from that of soil and water.
At present, the culture method of soil or water body is still used, and rumen nitrate-reducing bacteria cannot be obtained

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach 1

[0013] (1) Preparation of rumen fluid

[0014] The rumen digesta was obtained through the rumen of an adult goat with a permanent rumen fistula, and the rumen fluid was obtained after filtering with 4 layers of sterile gauze.

[0015] (2) Preparation of rumen nitrate-reducing bacteria pure culture fermentation broth

[0016] First prepare the basic medium: add sodium chloride, magnesium chloride, calcium chloride, manganese chloride, ferrous sulfate, zinc chloride, cobalt chloride, yeast extract, resazurin to ultrapure water, and take 100ml of ultrapure Water, add 110mg sodium chloride, 10mg magnesium chloride, 8mg calcium chloride, 3mg manganese chloride, 2mg ferrous sulfate, 1mg zinc chloride, 0.3mg cobalt chloride, 19mg yeast extract, 19mg resazurin respectively, until complete After dissolving, add 1 ml 2.5% cysteine ​​hydrochloride solution, 180 μl acetic acid, 60 μl propionic acid, 15 μl each of butyric acid, valeric acid, isobutyric acid and isovaleric acid. The pH is...

Embodiment approach 2

[0023] (1) Preparation of rumen fluid

[0024] The rumen digesta was obtained through the rumen of an adult goat with a permanent rumen fistula, and the rumen fluid was obtained after filtering with 4 layers of sterile gauze.

[0025] (2) Preparation of rumen nitrate-reducing bacteria pure culture fermentation broth

[0026] First prepare the basic medium: add sodium chloride, magnesium chloride, calcium chloride, manganese chloride, ferrous sulfate, zinc chloride, cobalt chloride, yeast extract, resazurin to ultrapure water, and take 100ml of ultrapure The water contains 130mg of sodium chloride, 12mg of magnesium chloride, 10mg of calcium chloride, 5mg of manganese chloride, 4mg of ferrous sulfate, 2mg of zinc chloride, 0.5mg of cobalt chloride, 23mg of yeast extract, and 23mg of resazurin. After completely dissolving, add 2 ml of 2.5% cysteine ​​hydrochloride solution, 190 μl of acetic acid, 70 μl of propionic acid, 18 μl each of butyric acid, valeric acid, isobutyric acid...

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Abstract

The invention relates to a method for obtaining nitrate-reducing bacteria from a rumen fluid and belongs to the technical field of micro-organism studies. The method comprises steps of preparation of a pure culture fermentation broth of rumen nitrate-reducing bacteria, preparation of an enrichment culture solution and inoculation culture. The number of viable bacteria in each milliliter can be larger than or equal to 10<6> by use of a selective medium and a specific deoxidizer. According to the preparation method of the rumen nitrate-reducing bacteria, black precipitating particles possibly produced after use of sodium sulfide are avoided, meanwhile, enrichment and propagation of the bacteria are realized, the content of total bacteria is increased, and a material is provided for scientific research.

Description

technical field [0001] The invention relates to a method for obtaining nitrate-reducing bacteria from rumen fluid, and belongs to the technical field of microorganism research. Background technique [0002] Currently reported rumen anaerobic bacteria capable of reducing nitrate include Selenomonas ruminantium, Veillonella parvula, Wolinella succinogenes, Campylobacter ( Campylobacter fetus) and Mannheimia succiniciproducens. Among them, Crescentomonas ruminants was the dominant bacterium, which had the largest number in the rumen and was the most important rumen nitrate-reducing bacteria. Nitrate can provide nitrogen in the rumen and reduce methane emissions, and this process requires the participation of nitrate-reducing bacteria. Therefore, how to cultivate and obtain nitrate-reducing bacteria and conduct in-depth research is a key issue in this field that needs to be solved . [0003] At present, most of the research and reports on the cultivation of nitrate-reducing b...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02C12R1/01
CPCC12N1/20C12N1/02
Inventor 林淼赵国琦霍永久贡笑笑王梦琦张蒙
Owner YANGZHOU UNIV