Bacillus stearothermophilus and method for antibiotic residue detection

A technology of stearothermophilic spore and detection method, which is applied in the field of microorganism detection of antibiotics, and achieves the effects of good dispersibility, strong diffusivity, and simple and feasible method.

Active Publication Date: 2010-06-09
河南花花牛乳业集团股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved in the present invention is to screen out Bacillus stearothermophilus strains that are not easily affected by the contamination of miscellaneous bacteria and are sensiti

Method used

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  • Bacillus stearothermophilus and method for antibiotic residue detection
  • Bacillus stearothermophilus and method for antibiotic residue detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Screening of strains

[0032] (1) Medium preparation

[0033] ①Nutrition agar medium peptone 10g, beef extract 3g, sodium chloride 5g, agar 20g, distilled water 1000mL, add the above ingredients to distilled water, mix well, and autoclave at 120°C for 15min.

[0034] ②Soybean tryptone agar medium tryptone 17.0g, vegetable peptone 3.0g, sodium chloride 10.0g, glucose 2.5g, agar 20g, distilled water 1000mL, add the above ingredients into distilled water, mix well, and autoclave at 120°C for 15min.

[0035] ③Manganese salt nutrient agar medium peptone 10g, beef extract 3g, sodium chloride 5g, agar 20g, distilled water 1000mL, add the above ingredients to distilled water, add 0.5% manganese sulfate solution 1mL, mix well, autoclave at 120°C for 15min .

[0036] ④Potassium bromide phenol purple glucose peptone medium peptone 10g, glucose 5g, 2% potassium bromide phenol purple ethanol solution 0.6mL, agar 4.0g, distilled water 1000mL, add peptone, glucose, agar to distilled ...

Embodiment 2

[0054] Example 2 Strain identification

[0055] The strain S8 isolated in Example 1 was further morphologically identified through observation of biological characteristics, and the results are as follows:

[0056] (1) Physiological identification

[0057] Cultivated on Tryptone Soy Agar Medium, only a needle-sized colony is formed. The bacterial body is rod-shaped and does not form filaments. It has flagella all over the body and can move. The spores are oval, sub-terminal or terminal, which usually enlarge the cysts , Can cause high-temperature sterilization milky acid spoilage and sweet curd and other deterioration; the minimum growth temperature is 28 ℃, the optimum growth temperature is 50 ~ 65 ℃, the maximum growth temperature is 70 ~ 77 ℃, at pH 6.8 ~ 7.2 It grows well in the medium and cannot grow when the pH is close to 5.

[0058] (2) Biochemical identification:

[0059] Cultivation at 65°C can grow rapidly: glucose produces acid;

[0060] Sabouraud dextrose broth, negative; ...

Embodiment 3

[0069] Embodiment 3 A method for detecting antibiotic residues includes the following steps:

[0070] (1) Strain activation

[0071] Bacillus stearothermophilus CGMCC NO.3287 strain was inoculated on the slant medium of tryptone soybean, adjusted the pH of the medium to 7.2, and optimized culture at 65℃, and planted once a week;

[0072] (2) Spore enrichment and suspension

[0073] Heating and centrifugation: culture the slant strain at 65°C for 4 days, wash the lawn with 10 mL of physiological saline, transfer it to a sterile Erlenmeyer flask with a small amount of glass beads, shake, filter sterile cotton into a sterile container, Prepare a suspension with a certain concentration, heat the suspension at 70~80℃ for 10min, let it stand overnight, centrifuge at 2000r / min for 15min, take the supernatant, centrifuge at 4000r / min for 15min, and repeat the centrifugation and washing for 4 to 5 times. Suspend the centrifugal sediment into a spore suspension with physiological saline and ad...

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Abstract

The invention relates to bacillus stearothermophilus and a method for carrying out antibiotic residue detection by utilizing the same. The preservation number of the screened strain is CGMCC NO.3287; and the antibiotic residue detection method which is established by taking the strain as indicator bacteria comprises the following steps of: rejuvenating and activating the strain; preparing spore suspending liquid in enrichment; preparing a purple culture solution; embedding bacillus; detecting a sample, and the like. The strain CGMCC NO.3287 is stable, easy to store and prepare, highly sensitive to B-lactam antibiotic, good in homogeneity and dispersibility in a culture medium, grows fast at 65 DEG C and has fast metabolism and acid production. Compared with a TIC method and a fermentation test method, the method has convenient operation, high sensitivity, 2-3h time-consumption which is shorter than 4h of national similar methods and is easy to judge the result; compared with a commercial kit, the bacillus stearothermophilus has low cost which is equal to 1/7 of the kit in price; and the method is simple and practicable and can be operated in a common laboratory.

Description

technical field [0001] The invention relates to the field of microbial detection of antibiotics, in particular to a Bacillus stearothermophilus and a method for detecting antibiotic residues using the bacterial strain as an indicator bacterium. Background technique [0002] Antibiotics are a class of metabolites or their derivatives synthesized by microorganisms and other organisms in the course of life activities, which can inhibit or interfere with the life activities of pathogenic bacteria and viruses at very low concentrations, such as penicillin, tetracycline, gold, etc. chloramphenicol, streptomycin, sulfonamides, etc. In medicine, antibiotics are widely used in the prevention and treatment of various microbial infectious diseases and certain cancers. At the source of the dairy industry, antibiotics are used frequently, especially for the treatment of bovine mastitis, and are often used repeatedly in large doses. According to statistics, about ten kinds of antibiotic...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12Q1/18C12R1/07
Inventor 李文献王少武李红李永军周霞冯利青马伟华刘燕张伟丽刘军王亚昆李爱华
Owner 河南花花牛乳业集团股份有限公司
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