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Preparation method of avibacterium paragallinarum culture medium

A culture method and culture medium technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve the problems of easy pollution, low number of viable bacteria, slow proliferation, etc., and achieve easy scale production and culture methods Simple, short growth cycle results

Pending Publication Date: 2020-02-04
TIANJIN RINGPU BIO TECH +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The culture conditions of Avian bacilli paragallinarum are very harsh, and are very sensitive to changes in the temperature and physical and chemical conditions of the outside world. In the conventional culture process, there are often phenomena such as slow proliferation, low number of viable bacteria, and easy contamination. The existing chicken infectious rhinitis vaccine The effect of the culture medium is not ideal, and the number of viable bacteria cannot meet the production requirements
The cultivation of the existing Avulina paragallinarum generally adopts the method of preparing seed liquid step by step, and real-time monitoring is also carried out during each stage of preparation, which has many steps and takes a long time. Serotype, the time and conditions used by each serotype to reach the peak growth period are different. Therefore, it is particularly important to optimize the optimal ratio of each factor according to the influencing factors and provide high-quality medium formulations for large-scale production.

Method used

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  • Preparation method of avibacterium paragallinarum culture medium
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  • Preparation method of avibacterium paragallinarum culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1, the preparation of Avian bacillus paragallinarum bacterial classification

[0025] Dissolve A, B, and C freeze-dried strains of Avulina paragallinarum (provided by Tianjin Ruipu Biotechnology Co., Ltd. Airport Economic Zone Company) with PBS (PH=7.0), and then use an inoculation loop to dip Inoculate an appropriate amount on a culture medium plate and culture in a candle jar at 39°C for 20 hours; use a sterile pipette tip (10ul) to select a single typical colony, inoculate it into a 250ml Erlenmeyer flask (100ml liquid medium), put a rubber stopper on it and place it at 37 In a shaker at ℃, 180rpm / min, after 14 hours of cultivation, take it out and put it in a refrigerator at 4℃ for later use (storage should not exceed 2 days).

Embodiment 2

[0026] Embodiment 2, the influence of different concentrations of chicken serum on the growth of Avian bacillus paragallinarum

[0027] Take 6 bottles of experimental medium (100ml liquid medium in a 250ml Erlenmeyer flask), add 0.5g polypeptone, 0.5g casein peptone, 0.5g soaked yeast powder, 0.5g sodium chloride, 0.5g glutamic acid Sodium, 20ul coenzyme NAD aqueous solution (0.02%), 0.2g glucose (0.2%), then respectively add chicken serum 3%ml (3%), 6ml (6%), 9ml (9%) in each bottle culture medium, 12ml (12%), 15ml (15%), 18ml (18%), and then according to the inoculum size of 1%, add 1ml of the prepared seed solution to 6 bottles of medium respectively. Incubate in a constant temperature shaker at 37°C at 180rpm / min for 16h, take out every 1h to measure the absorbance OD 600 (taking the average value three times) and recording its change, as can be seen from the result, the best chicken serum concentration is 6%; detailed results are shown in Table 1:

Embodiment 3

[0028] Embodiment 3. the impact of different concentrations of glucose on the growth of Avian bacteria paragallinarum:

[0029] Take 6 bottles of experimental medium (100ml liquid medium in a 250ml Erlenmeyer flask), add 0.5g polypeptone, 0.5g casein peptone, 0.5g soaked yeast powder, 0.5g sodium chloride, 0.5g glutamic acid Sodium, 20ul coenzyme NAD aqueous solution (0.02%), chicken serum 6ml (6%), then respectively add glucose 0.1g (0.1%), 0.2g (0.2%), 0.3g (0.3%) in each bottle culture medium, 0.4 (0.4%), 0.5 (0.5%), 0.6 (0.6%), and then according to the inoculum amount of 1%, add 1 ml of the prepared seed solution into 6 bottles of medium respectively. Incubate in a constant temperature shaker at 37°C at 180rpm / min for 16h, take out every 1h to measure the absorbance OD 600 (take the average value three times) and record its change, as can be seen from the result, the optimal glucose concentration is 0.2%; Detailed results are shown in Table 2:

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Abstract

The present invention provides a formula of an avibacterium paragallinarum culture medium. The formula comprises the following raw materials: 0.5% of polypeptone, 0.5% of casein peptone, 0.5% of yeastextract powder, 0.5% of sodium chloride, 0.5% of sodium glutamate, 0.02% of a coenzyme NAD aqueous solution, 0.2% of glucose and 6% of chicken serum. The present invention also discloses a culture method established according to the formula. The method is simple, growth period is short, the number of viable bacteria of the cultured avibacterium paragallinarum is high, and moreover, the provided avibacterium paragallinarum culture medium is low in costs and easy to produce on a large scale.

Description

technical field [0001] The invention belongs to the technical field, and in particular relates to a preparation method of a culture medium of Avian bacteria paragallinarum. Background technique [0002] Avian bacillus paragallinarum is a spore-free Gram-negative bacterium. It is the pathogenic bacterium of chicken infectious rhinitis. It mainly causes acute respiratory diseases in chickens. Chickens of all ages are susceptible and can occur all year round, especially High incidence in cold and wet season. The disease can cause laying hens to drop eggs, grow poorly in bred chickens and broilers, increase the elimination rate, and bring serious harm to the chicken industry. At present, the vaccine of Avian bacilli paragallinarum is mainly used to control its occurrence and spread. [0003] The culture conditions of Avian bacilli paragallily are very harsh, and are very sensitive to changes in the temperature and physical and chemical conditions of the outside world. In the c...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/01
CPCC12N1/20
Inventor 李守军王冬雪唐荣宏郁宏伟范莉莉朱秀同
Owner TIANJIN RINGPU BIO TECH
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