Method for separating mycoplasma ovipneumoniae

Abstract

The invention provides a method for separating mycoplasma ovipneumoniae, which comprises the following steps: (1) collecting a sample and treating the sample to obtain a sample inoculation solution; (2) inoculating chick embryos; (3) separating the mycoplasma ovipneumoniae: collecting dead chick embryos after inoculation and / or non-dead chick embryos after 9 days, taking chick embryo allantoic fluid to carry out viable bacteria detection on the mycoplasma ovipneumoniae, and if the chick embryo allantoic fluid is collected to be positive, storing the material, if the chick embryo allantoic fluid of the first generation is negative, the collected chick embryo allantoic fluid is blindly transmitted to the chick embryo by the same method, and if the chick embryo allantoic fluid is still negative after three generations of blind transmission, determining the sample to be negative for mycoplasma ovipneumoniae, and discharging the material. The method disclosed by the invention is simple to operate, does not need to prepare a mycoplasma ovipneumoniae culture medium with complex components, and avoids the problem that in the prior art, high-pressure sterilization or a high-capacity filteris often needed in preparation of an isolation culture medium; and according to the method, the mycoplasma ovipneumoniae without animal serum can be obtained.

Description

technical field [0001] The invention belongs to the technical field of veterinary biotechnology research, and in particular relates to a method for isolating ovine mycoplasma pneumoniae. Background technique [0002] Mycoplasma ovis pneumoniae is a microorganism that can cause respiratory disease in sheep. Mycoplasma ovis can infect both sheep and goats. The pathogen can also be isolated from the nasal cavity of healthy sheep. The pathogen has a global distribution. The pathogen is currently prevalent in my country and poses a serious threat to the sheep industry. [0003] Mycoplasma ovis pneumoniae is relatively delicate, and its isolation and culture require special media. The improved Thiaucourt's medium is a commonly used isolation medium for Mycoplasma ovis pneumoniae. The effect of culturing Mycoplasma ovis pneumoniae is higher than that of other media such as the improved KM2 medium, and the effect is better. In the prior art, the composition of the culture mediu...

AI Technical Summary

Problems solved by technology

In the prior art, the composition of the culture medium for Mycoplasma pneumoniae in ovis is complicated, and in addition to containing many nutritional components such as carbon source, nitrogen source, protein, inorganic salt, etc., it generally needs to add animal serum ranging from 10% to 20%

The preparation steps of the prior art culture medium are cumbersome, and often need to prepare as many as 8 to 9 kinds of reagents. In addition, it is often necessary to prepare reagents such as fresh yeast extract, 1% NaOH, and 1% phenol red. The prepared culture medium must be sterilized or filtered. Filtration and other steps

There are still problems such as poor reproductive ability in the cultivation of Mycoplasma pneumoniae in the prior art

In addition, there are many problems in the use of serum in the culture medium of the prior art, such as differences in serum batches may easily lead to large differences in culture medium batches, high cost, biological safety, and easy increase in immune side effects, etc.

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