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Haemophilus parasuis culture medium and preparation method thereof

A Haemophilus suis culture medium technology, applied in the field of Haemophilus parasuis culture medium and its preparation, can solve the problems of unable to carry out research work normally, difficult to separate and cultivate, unstable growth, etc., and achieve the goal of reducing the contamination of miscellaneous bacteria chance, increase the success rate of separation, and the effect of sufficient nutrition

Pending Publication Date: 2022-03-08
WUHAN CHOPPER BIOLOGY
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  • Description
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Problems solved by technology

[0003] Due to the biological characteristics of Haemophilus parasuis, it is very difficult to isolate and cultivate clinically: Taking several common microbial culture media as an example, Haemophilus parasuis cannot be cultured on the blood agar medium with high nutritional value. Growth, growth on the chocolate agar medium is also unstable, sometimes it cannot grow, and sometimes it needs to be cultivated for 72 hours to grow pinpoint-sized colonies, and the live bacteria are stored on the chocolate medium for a short period of time, and they will die if they exceed 72 hours. Post-purification, biochemical identification, drug susceptibility testing, strain preservation and other research work cannot be carried out normally

Method used

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  • Haemophilus parasuis culture medium and preparation method thereof
  • Haemophilus parasuis culture medium and preparation method thereof

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preparation example Construction

[0030] Based on above-mentioned embodiment, the present invention also proposes a kind of preparation method of Haemophilus parasuis culture medium, the preparation method of described Haemophilus parasuis culture medium comprises the following steps:

[0031] Step S10, dissolving coenzyme I in distilled water to prepare a coenzyme I solution;

[0032] Step S20, under a sterile environment, add the coenzyme I solution, defibrillated sheep blood and horse serum into the basal medium, and mix evenly to obtain a culture medium for Haemophilus parasuis.

[0033] It can be understood that, in the above steps, the addition amount of each component is as follows: 1000 parts of basal medium, 40-60 parts of coenzyme I solution, 80-100 parts of horse serum and 50-70 parts of defibrated sheep blood; The concentration of the coenzyme I solution is 1.8-2.2 g / ml, preferably 2 g / ml.

[0034] The culture medium proposed by the invention is simple to prepare and low in price.

[0035] During s...

Embodiment 1

[0043] Take 15g of tryptone, 5g of soybean peptone, 5g of sodium chloride, 18g of agar powder, and 25g of glucose, add distilled water to 1000ml, stir, heat and boil until completely dissolved, adjust the pH value to 7.1 with sodium hydroxide, pack in a triangular flask, 121°C Autoclave for 15 minutes. Take 1g of coenzyme I (nicotinamide adenine dinucleotide, NAD) fully dissolved in 50ml of distilled water, filter and sterilize; when the basal medium is cooled to 50-55°C, add sterile coenzyme I sequentially in a sterile environment 50ml of the solution, 50ml of defibrated sheep blood, and 100ml of horse serum were mixed thoroughly and distributed into sterile glass petri dishes in a sterile environment (shake constantly when dispensing), each dish was 15ml, after the dispensing was completed, put Haemophilus parasuis culture medium was obtained after the plate was left to stand overnight in a greenhouse at 37°C.

Embodiment 2

[0045] Take 16g of tryptone, 6g of soybean peptone, 5g of sodium chloride, 16g of agar powder, and 24g of glucose, add distilled water to 1000ml, stir, heat and boil until completely dissolved, adjust the pH value to 7.3 with sodium hydroxide, pack in a triangular flask, 121°C Autoclave for 15 minutes. Take 1.1g of coenzyme I (nicotinamide adenine dinucleotide, NAD) and fully dissolve it in 50ml of distilled water, filter and sterilize; when the basal medium is cooled to 50-55°C, add sterile coenzymes sequentially under a sterile environment 50ml of solution Ⅰ, 50ml of defibrated sheep blood, and 100ml of horse serum, mixed well and then packed into sterile glass plates in a sterile environment (shake constantly when filling), each plate is 15ml, after the filling is completed, Haemophilus parasuis culture medium was obtained after the plate was left to stand overnight in a greenhouse at 37°C.

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Abstract

The invention discloses a haemophilus parasuis culture medium and a preparation method thereof. The haemophilus parasuis culture medium comprises the following components: a basic culture medium, coenzyme I, horse serum and defibrated sheep blood. According to the method, coenzyme I, defibrated sheep blood and horse serum are added into a basic culture medium, so that the problem that staphylococcus aureus must be inoculated when haemophilus parasuis grows on a blood agar culture medium is solved, bacterial colonies are purer, and the probability of infectious microbe contamination in bacterial purification is effectively reduced; moreover, the culture medium provided by the invention has more sufficient nutrition, is more suitable for the growth characteristics of haemophilus parasuis, and is more stable in growth, larger in bacterial colony and longer in preservation time than a chocolate culture medium, and the cultured viable bacteria can be preserved for 30 days at the temperature of 2-8 DEG C. The haemophilus parasuis culture medium provided by the invention can be used for culturing, identifying and separating haemophilus parasuis, and effectively improves the separation success rate of the haemophilus parasuis.

Description

technical field [0001] The invention relates to the technical field of microorganisms of biological products, in particular to a culture medium of Haemophilus parasuis and a preparation method thereof. Background technique [0002] Haemophilus parasuis disease is polyserositis and arthritis of pigs caused by Haemophilus parasuis. The disease has no obvious seasonality. It is high in other viral infections, unsuitable feeding environment and poor feeding tube methods. Incidence rate (about 20%) and high fatality rate (up to 50%); diseased pigs and recessively infected pigs are the main source of infection of the disease, mainly transmitted through two ways of breathing and digestion; susceptible objects are within 4 months of age piglets, the younger the piglets are, the more harmful they are. Piglets in the weaning period often show acute attacks and die suddenly, while young pigs and big pigs often turn to chronic disease courses. 20d failure, death. Especially in pig far...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/21
CPCC12N1/20
Inventor 郑良益朱薇石宝兰漆世华谢红玲李晶梅王治江万佳秦红刚何珊
Owner WUHAN CHOPPER BIOLOGY
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