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Bifidobacterium breve and method for detecting various antibiotic residues in milk and application

A technology for the detection of Bifidobacterium breve, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, and microorganism measurement/inspection. It can solve the problems of difficult detection of antibiotic residues, low sensitivity, and difficulty in simultaneous detection. Quantitative detection method is simple and easy, high sensitivity and precision, easy to store and operate

Active Publication Date: 2016-06-29
杭州老蚁科技有限公司
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AI Technical Summary

Problems solved by technology

There are 33 kinds of veterinary drugs in 5 categories, including sulfonamides, quinolones, β-lactams, tetracyclines, and chloramphenicol, which are commonly used in my country. Their chemical molecular structures and chemical properties are quite different, and it is difficult to detect them at the same time.
At the same time, it is difficult to detect multiple antibiotic residues in milk. At present, my country's food hygiene standard (GB / T4789.27-2008) stipulates the TTC method to detect antibiotic residues in milk. This method has simple equipment and low cost, but low sensitivity and Easy to be false positive, the sensitivity in detecting penicillin and chloramphenicol is roughly equal to that of the Bacillus subtilis disk method, but it is not very sensitive to streptomycin and neomycin, and the disinfectant can interfere with the test

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  • Bifidobacterium breve and method for detecting various antibiotic residues in milk and application
  • Bifidobacterium breve and method for detecting various antibiotic residues in milk and application
  • Bifidobacterium breve and method for detecting various antibiotic residues in milk and application

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Embodiment 1

[0039] The isolation and identification of embodiment 1 Bifidobacterium breve bacterial strain

[0040] Take the feces of healthy young men in Zhejiang Province as an isolated sample, 1g of feces is suspended and diluted with 10mL PBS, inoculated on TPY solid medium, and cultured anaerobically at 37°C for 48 hours to obtain Bifidobacterium breve DSQH-1 according to the present invention , identified as Bifidobacterium breve, was preserved on December 13, 2013 in the General Microbiology Center of China Committee for the Collection of Microbial Cultures (Address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Microbiology, Chinese Academy of Sciences Research Institute, Zip Code 100101), the deposit number is CGMCCNo.8579.

[0041] TPY agar medium and TPY liquid medium are products of Qingdao Hi-Tech Park Haibo Biotechnology Co., Ltd.

[0042] Bifidobacterium breve DSQH-1 strain of the present invention has the following microbiological characteristics:

[0043...

Embodiment 2

[0053] Example 2: Detection of antibiotic residues in milk samples

[0054] (1) Strain activation: inoculate the Bifidobacterium breve DSQH-1 strain on TPY solid medium, and culture at 37°C for 24-48h;

[0055] (2) Bacterial suspension preparation: Inoculate the Bifidobacterium breve DSQH-1 strain obtained in step (1) into TPY liquid, culture with shaking at 37°C and 200rpm for 4-5 hours, centrifuge at 3000rpm for 5 minutes, and adjust the bacterial solution with PBS The value of OD600 is 0.15-0.20, stored at 4°C;

[0056] (3) The obtained bacterial liquid was cultured anaerobically at 40°C for 12-16 hours; adjusted to zero with the culture medium, measured the OD600 of the negative control tube of the sample tube, and read the antibiotic concentration value (μg / L) from the standard curve.

[0057] (4) Preparation of screening medium: by weight, take peptone, 2.5g, yeast extract 3.0g, glucose 4.0g, soluble starch 1.0g, sodium chloride 1.0g, beef extract 1.0g, K 2 HPO 4 0.6g...

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Abstract

The invention belongs to the field of life sciences and food safety, and relates to bifidobacterium breve isolated from the intestinal tract of a healthy person and a method for detecting antibiotic residues in milk by means of the strain. The isolated strain is DSQH-1, and the preservation number is CGMCC No.8579; the DSQH-1 strain is sensitive to antibiotics such as penicillin, tetracycline and erythromycin. The method for detecting various antibiotic residues in milk by taking the strain as a working strain comprises the steps of strain activation, bacterial suspension preparation, screening culture medium preparation, quantitative detection culture medium preparation, standard tube preparation, standard curve drawing, sample preparation, sample screening and detecting, sample quantitative detection and the like. The method for detecting various antibiotic residues in milk is easy and convenient to operate, simple in equipment requirement, high in sensentivity, not prone to contamination and suitable for screening and quantitative detection of a large number of samples and can also be used for self inspection of producers.

Description

technical field [0001] The invention belongs to the fields of life science and food safety, and relates to a strain of Bifidobacterium breve and a method for rapidly detecting antibiotic residues in milk by using the strain. Background technique [0002] Milk safety is not only related to the health of the general public, but also the lifeblood of dairy enterprises. Antibiotic residues in milk are an important issue for milk safety. With the rapid development of animal husbandry, milk production has increased significantly. In order to prevent diseases and treat diseases such as mastitis, antibiotic veterinary drugs are increasingly widely used in animal husbandry. While the application of antibiotics to treat cow diseases is effective, the use of antibiotics is not standardized, such as increasing the dosage and prolonging the treatment period, and even giving the lactation period The long-term application of antibiotics to prevent diseases by dairy cows not only has anti...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12Q1/18C12R1/01
CPCC12Q1/18C12N1/205C12R2001/01G01N2415/00
Inventor 杜季梅刘佳明楼永良
Owner 杭州老蚁科技有限公司
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