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Mycoplasma anatis culture medium and separation and purification method for mycoplasma anatis

A technology of liquid medium and solid medium, which is applied in the field of separation and purification of mycoplasma anativa culture medium and mycoplasma anativa, can solve the problems of easy misdiagnosis, poor reproductive ability, and low titer of viable bacteria, so as to promote reproduction and prevent miscellaneous Bacterial growth, the effect of increasing the concentration

Active Publication Date: 2016-04-06
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The diagnosis of mycoplasma anativa is mainly polymerase chain reaction (PCR) and direct immunofluorescence (FA), both of which involve the isolation and cultivation of a pure culture of mycoplasma, that is, a single colony; and the artificial culture medium currently used to isolate mycoplasma anativa Generally, they are all improved from Frey or PPLO medium, but there are still problems such as low titer of viable bacteria and poor reproductive ability, which are easy to cause missed diagnosis; The method of Mycoplasma anatis, and then carry out the research on the treatment and prevention of Mycoplasma anatis infection

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Mycoplasma duck culture medium of the present invention is made up of liquid medium and solid medium, specifically:

[0029] The liquid medium formula is: 10% thallium acetate solution 4.8mL / L, PPLO broth 3g / L, glucose 5g / L, peptone 4.8g / L, tryptone 10g / L, 1% phenol red 2mL / L, CMRL-1066 medium containing L-glutamine 450mL / L, inactivated horse serum 180mL / L, yeast extract 45mL / L, penicillin 3 million units / L, L-cysteine ​​hydrochloride aqueous solution 0.2g / L, nicotinamide adenine dinucleotide aqueous solution 0.2g / L, the balance is deionized water.

[0030] The method for preparing 500ml of the liquid culture medium of the present invention is as follows: place a 1L conical flask on a magnetic stirrer, first measure 100mL of deionized water, slowly add 2.4mL of 10% thallium acetate solution dropwise, then weigh and add 1.5g of PPLO broth, 2.5g of glucose, 2.4g of peptone, 5g of tryptone, 2mL of 1% phenol red, mix well, adjust the pH to 7.8 with 20% NaOH by mass percen...

Embodiment 2

[0042] The difference between this embodiment and embodiment 1 is:

[0043] The liquid medium formula is: 10% thallium acetate solution 4.5mL / L, PPLO broth 5g / L, glucose 4g / L, peptone 4g / L, tryptone 9g / L, 1% phenol red 2.5mL / L, CMRL-1066 medium containing L-glutamine 500mL / L, inactivated horse serum 150mL / L, yeast extract 55mL / L, penicillin 2.5 million units / L, L-cysteine ​​hydrochloride aqueous solution 0.1g / L, nicotinamide adenine dinucleotide aqueous solution 0.1g / L, the balance is deionized water.

[0044] The solid medium formula is: 10% thallium acetate solution 4.5mL / L, PPLO broth 5g / L, glucose 4g / L, peptone 4g / L, tryptone 9g / L, agar powder 10g / L, containing L-glucose Aminoamide CMRL-1066 medium 500mL / L, inactivated horse serum 150mL / L, yeast extract 55mL / L, penicillin 2.5 million units / L, L-cysteine ​​hydrochloride aqueous solution 0.1g / L, smoke Amide adenine dinucleotide aqueous solution 0.1g / L, the balance is deionized water.

[0045] The rest are the same.

Embodiment 3

[0047] The difference between this embodiment and embodiment 1 is:

[0048] The liquid medium formula is: 10% thallium acetate solution 6.0mL / L, PPLO broth 4g / L, glucose 6g / L, peptone 6g / L, tryptone 8g / L, 1% phenol red 3mL / L, containing L-glutamine CMRL-1066 medium 480mL / L, inactivated horse serum 170mL / L, yeast extract 50mL / L, penicillin 2.8 million units / L, L-cysteine ​​hydrochloride aqueous solution 0.1g / L L, nicotinamide adenine dinucleotide aqueous solution 0.2g / L, the balance is deionized water.

[0049] The solid medium formula is: 10% thallium acetate solution 6.0mL / L, PPLO broth 4g / L, glucose 6g / L, peptone 6g / L, tryptone 8g / L, agar powder 11g / L, containing L-glucose Aminoamide CMRL-1066 medium 480mL / L, inactivated horse serum 170mL / L, yeast extract 50mL / L, penicillin 2.8 million units / L, L-cysteine ​​hydrochloride aqueous solution 0.1g / L, smoke Amide adenine dinucleotide aqueous solution 0.2g / L, the balance is deionized water.

[0050] The rest are the same.

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Abstract

The invention provides a mycoplasma anatis culture medium. The mycoplasma anatis culture medium is composed of a liquid culture medium and a solid culture medium. The liquid culture medium is prepared from deionized water, a 10% thallium acetate solution, PPLO broth, glucose, peptone, tryptone, 1% phenol red, a CMRL-1066 culture medium containing L-glutamine, inactivated horse serum, yeast leachate, penicillin, L-cysteine hydrochloride and nicotinamide adenine dinucleotide. Besides the situation that agar powder is added and phenol red is removed, other components of the solid culture medium are the same as those of the liquid culture medium. The culture medium is used for separating and purifying mycoplasma anatis, wherein a bacterial colony is separated from the solid culture medium, the separated bacterial colony is placed in the liquid culture medium to grow, then filtering, dilution and plate coating of liquid culture substances and purification of picked single colony multiplication are conducted, and the steps are repeated till purified mycoplasma anatis is obtained. The culture medium can promote growth of mycoplasma anatis, and the success rate of separation and purification of mycoplasma anatis is high.

Description

technical field [0001] The invention relates to a bacterial culture medium and a method for separation and purification, in particular to a culture medium for mycoplasma anatis and a method for separation and purification of mycoplasma anatis. Background technique [0002] Duck Mycoplasma ( Mycoplasma anatis ) mainly cause infectious sinusitis in ducks, clinically susceptible ducklings, leading to inflammation of the infraorbital sinus, nasal cavity, trachea and air sacs, among which the swelling of the infraorbital sinus is the most typical. When duck mycoplasma is mixed with other pathogens (such as influenza virus, coliform bacteria), it can cause a series of neurological symptoms such as dyspnea, soft feet, diarrhea, ataxia, torticollis, circling or backward movement in ducks, which seriously affects ducks. growth and development; histological lesions manifested as lymphocytic meningitis, fibrin exudative air sacculitis, interstitial pneumonia, and lymphofollicular trac...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02C12R1/35
CPCC12N1/02C12N1/20
Inventor 宫晓炜刘永生陈启伟郑福英储岳峰
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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