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Culture medium and fermentation culture method used for high-density culture of mycoplasma capricolumsubsp. capripneumonia

A culture medium and mycoplasma technology, applied in the biological field, can solve problems such as hindering isolation culture, difficulty in in vitro culture, nutritional requirements and harsh culture conditions

Inactive Publication Date: 2020-05-19
北京龙科方舟生物工程技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it is a long and difficult process to isolate the subspecies of Mycoplasma capricosum pneumonia, and it is also difficult to culture in vitro. It has strict nutritional requirements and culture conditions, which hinders its isolation and culture to a large extent.
In addition, the genome of Mycoplasma capricosum subspecies goat pneumonia is very small, and its biosynthetic ability is low. It needs exogenous cholesterol, nucleic acid precursors, vitamins, etc. for growth. The composition of the medium requires a lot of work to try and screen

Method used

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  • Culture medium and fermentation culture method used for high-density culture of mycoplasma capricolumsubsp. capripneumonia
  • Culture medium and fermentation culture method used for high-density culture of mycoplasma capricolumsubsp. capripneumonia
  • Culture medium and fermentation culture method used for high-density culture of mycoplasma capricolumsubsp. capripneumonia

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0088] The preparation method of the 25% (mass fraction) yeast infusion is as follows: Weigh 250 g of dry yeast powder, add 750 mL of sterile water, and mix well to obtain a 25% yeast infusion. Yeast dry powder is a product of Angel Yeast Co., Ltd., the item number is 81002166.

[0089] The preparation method of 10% (mass fraction) thallium acetate is as follows: Weigh 10 g of thallium acetate, add sterile water to make up to 100 mL, and mix well to obtain a 10% thallium acetate solution. Thallium acetate is a product of Beijing Yongze Haojia Biotechnology Development Center, with a CAS number of 563-58-8.

[0090] The experimental instruments and equipment used in the following examples are as follows: LRH-150 biochemical incubator; HZC-280 constant temperature shaking incubator; DL-CJ-2DN clean bench; 722-type visible spectrophotometer; 20L automatic fermentation tank; High-speed refrigerated centrifuge; inverted microscope; electronic scale with an accuracy of 0.01; 20μL-5...

Embodiment 1

[0091] Example 1. Isolation, identification and preservation of bacterial strain ZFY17

[0092] 1. Isolation and cultivation of strain ZFY17

[0093] Collect the respiratory secretions of goats that have been clinically diagnosed as suffering from mycoplasma pneumonia in the Inner Mongolia experimental sheep farm, add a small amount of normal saline to mix evenly, divide the tissue fluid into 1.5mL EP tubes, centrifuge at 2000r / min for 2-3min, and take 0.3mL of the supernatant for inoculation In mycoplasma solid medium (the solvent is sterile water, the solute and its concentration are as follows: inactivated horse serum 20%, 25% yeast extract 10%, 10% thallium acetate 0.1%, penicillin 200 μg / mL, phenol red 0.4%, 2.1% PPLO broth, 0.2% sodium pyruvate, 0.1% glucose, 2% agar powder) (37°C), cultured for 6-7 days, after the color of the medium became lighter, a single colony was taken with a scalpel, and inserted into Mycoplasma liquid medium (the solvent is sterile water, the s...

Embodiment 2

[0099] Embodiment 2, the cultivation of Mycoplasma capricum goat pneumonia subspecies

[0100] 1. The culture medium optimization experiment of Mycoplasma capricosum subsp. goat pneumonia under shake flask conditions

[0101] 1. Culture medium design

[0102] In the present invention, a relatively typical case in the currently published reports at home and abroad is used as the medium formula of the control group; glucose, fructose, sucrose, hydrolyzed milk protein, and casein are used to replace part of the horse serum respectively to optimize the medium, and each experimental group and The medium formula of the control group is shown in Table 1.

[0103] Table 1, the solute formula of each experimental group and control group medium

[0104] name Experimental group 1 Experimental group 2 Experimental group 3 Experimental group 4 Experimental group 5 Experimental group 6 control group inactivated horse serum 10％ 10％ 10％ 10％ 10％ 10％ 20％ ...

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Abstract

The invention discloses a culture medium and fermentation culture method used for high-density culture of mycoplasma capricolumsubsp. capripneumonia. The mycoplasma capricolumsubsp. capripneumonia isisolated from respiratory secretions of a goat clinically diagnosed with mycoplasmal pneumonia; the mycoplasma capricolumsubsp. capripneumonia has been collected in China General Microbiological Culture Collection Center on 4 December, 2019, and the collection number is CGMCC No.18878; according to the fermentation culture method, liquid fermentation culture is carried out on the resuscitated-passaging mycoplasma capricolumsubsp. capripneumonia ZFY17 CGMCC No.18878 in a specific fermentation culture medium under specific conditions; and the specific fermentation culture medium replaces part ofhorse serum with a combination of glucose, hydrolyzed yeast protein, a hydrolyzed yeast culture, lactoalbumin hydrolysate and casein, thus greatly reducing strain fermentation cost.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a culture medium for high-density cultivation of Mycoplasma capricum subsp. goat pneumonia and a fermentation cultivation method thereof. Background technique [0002] Goat infectious pleuropneumonia is a common acute or chronic highly contagious respiratory infectious disease in goat breeding. It is mainly characterized by fibrinous pleurisy. The infected flock has no age and gender differences, and the morbidity and mortality are high. It is now widely prevalent in sheep-raising countries in Africa and Asia, and is extremely harmful. It is listed as one of the animal infectious diseases that must be reported by the World Organization for Animal Health (OIE). It was first discovered in Algeria in 1873. So far, it has been reported in more than 40 countries around the world. There have been reports of the disease occurring in my country and other regions. Our country notif...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/35
CPCC12N1/20C12R2001/35C12N1/205
Inventor 宋青龙薛新升
Owner 北京龙科方舟生物工程技术有限公司
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