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Indicator strain and application thereof

An indicator bacteria and indicator technology, applied in the field of microorganisms, can solve the problems of high price, high detection cost, increase the detection cost of milk enterprises, etc., and achieve the effect of short detection time and low cost

Active Publication Date: 2013-09-25
INST OF SOIL & FERTILIZER FUJIAN ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This technology uses different types of cells called indicators or microorganisms like these specialized ones which are able to react with specific substances found on them when they come into contact with certain compounds. These reactions help identify any harmful organism by measuring how much more drug remains there than usual. By doing this, we aimed at developing an effective way to measure the amount remaining within food samples containing many potentially dangerous drugs without requiring expensive equipment.

Problems solved by technology

This patented technology describes how certain types of bacilli called AnaeroBacillustrellium (AnA) may stop harmful substances like ampiciline that could cause illnesses such as tuberculosis when used against cow's diarrheal disease caused by Escherichia coli. However, these methods require specialized equipment and trained staff who work alongside them.

Method used

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  • Indicator strain and application thereof
  • Indicator strain and application thereof

Examples

Experimental program
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Embodiment 1

[0025] The standard for using the kit of the present invention to detect and determine that it is a certain antibiotic is: purchase milk powder that has been tested to contain no antibiotic residues, and restore it to milk with a concentration of 13%. Prepare milk containing different concentrations of antibiotics, such as milk containing 2 μg / L penicillin, when the kit detects milk containing 2 μg / L penicillin, it is positive, which means that the kit can detect penicillin sensitively.

[0026] Another example is to use a kit to detect the milk containing a certain concentration of antibiotics (milk containing 4 μg / L penicillin, milk containing μg / L ampicillin, milk containing 100 μg / L sulfonamide). Take 100 microliters of the above-prepared milk samples and add them to the microwells of the kit respectively, and take milk samples without antibiotic residues as a control. Seal the micropores with plastic film or scotch tape and keep warm in a water bath at 60-65°C for 2.5 hou...

Embodiment 2

[0035] Step 1: Dip a ring of indicator strains preserved on a slant ( Anoxybacillus sp. FHB009), inoculate it into liquid beef extract peptone medium to activate the strain, culture it at 50°C, 100rpm / min for 12h, and make seed liquid;

[0036] In terms of mass percentage, the formula of the beef extract-peptone medium is: 0.1% beef extract, 0.5% peptone, and 0.1% sodium chloride, and the pH is adjusted to 5; after stirring evenly, sterilize and cool in an autoclave to 50- 60 ℃ room temperature standby;

[0037] Step 2: Spread the above seed liquid evenly on the spore-producing solid medium at a volume ratio of 1:200, and cultivate it in an incubator at 50°C. After 3 days of cultivation, staining and microscopic examination, when the spore formation rate reaches more than 90% stop culturing and collect spores;

[0038] In terms of mass percentage, the formula of the spore-producing solid medium is: beef extract 0.1%-, soybean peptone 0.1%, NaCL 0.2%, MnSO 4 0.001%, pH5, ag...

Embodiment 3

[0047] Step 1: Dip a ring of indicator strains preserved on a slant ( Anoxybacillus sp. FHB009), inoculate it into liquid beef extract peptone medium to activate the bacteria, and cultivate it at 60°C and 100rpm for 12h to make a seed liquid; the formula of the beef extract peptone medium is: beef extract 1%, peptone 5 %, sodium chloride 0.1%, pH 7; after stirring evenly, sterilize in an autoclave and cool to room temperature of 60°C for later use.

[0048] The second step: inoculate the seed liquid into the spore-producing solid medium, and cultivate it in a high-temperature incubator at 50°C. After 3 days of cultivation, staining and microscopic examination, stop the cultivation when the spores reach more than 90%, and collect the spores and spore suspension. The formula of the spore-producing medium is: beef extract 1%, soybean peptone 1%, NaCL 0.2%, MnSO 4 0.01%, pH8, agar 2%. After stirring evenly, sterilize and cool to 60°C in an autoclave and pour it into a plate for...

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Abstract

The invention relates to an indicator strain with a classification name of anoxybacillussp. FHB009. The indicator strain is preserved at China Center for Type Culture Collection, and has a preservation number of CCTCC NO: M2013128. For prolonging the service life of a kit, the spores of the strain are adopted. A culture medium embedding the spores is a modified beef extract peptone medium with a pH value of 5-10. The indicator strain provided by the invention can sensitively detect various commonly used antibiotics such as sulfadiazine, sulfamethazine, oxytetracycline, tetracycline, chlortetracycline, gentamicin, doxycycline, penicillin, ampicillin, ampicillin, and the like, wherein a sensitivity limit satisfies an antibiotic highest residue standard of national standard (GB/T4789.27-2008).

Description

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Claims

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Application Information

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Owner INST OF SOIL & FERTILIZER FUJIAN ACADEMY OF AGRI SCI
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