Culture medium for separating enterococcus and streptococcus from milk sample and preparation method of culture medium

A technology for streptococci and enterococci, applied in the field of culture medium for isolating bacteria from milk samples, to achieve the effect of improving sorting efficiency, obvious effect, and simplifying the process of downstream analysis and identification

Inactive Publication Date: 2015-07-22
GANSU AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is currently no general-purpose medium that can separate enterococ

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0029] Example 2-Simultaneous sorting of Enterococcus and Streptococcus in milk-like samples:

[0030] Each 1000g medium contains: tryptone 15g, yeast extract / yeast powder 5g, beef heart extract / beef extract 3g, glucose 1.5g, sodium chloride 2.5g, disodium hydrogen phosphate 0.3g, sodium carbonate 2.0g, Nalidixic acid 18mg, polymyxin B 1.8mg, crystal violet 0.15mg, esculin 4g, ferric citrate 0.3g, agar 12g, of which glucose and polymyxin B need to be pre-prepared, filtered and sterilized, Store in aliquots at -20°C, and take it for use as needed; add water, heat and stir to dissolve the rest of the ingredients, boil for 3 minutes, and sterilize at 115°C for 12 minutes. When the temperature drops to 50°C—65°C, add the Mix the prepared glucose and polymyxin, fill it in a petri dish, and use it when the temperature drops below 37°C (to room temperature), or perform aseptic operation. Use a sealing film or sterile newspaper for this plate Wait for sealing, temporarily store at 2-8℃ ...

Example Embodiment

[0032] Example 3-For simultaneous sorting of Enterococcus and Streptococcus in the milk sample:

[0033] Each 1000g medium contains: 25g monthly peptone, 2g yeast extract / yeast powder, 2g beef extract, 1g glucose, 2g sodium chloride, disodium hydrogen phosphate 0.5g, sodium carbonate 2.6g, nalidixic acid 12mg, Polymyxin B 2mg, crystal violet 0.3mg, esculin 2g, ferric citrate 0.8g, and agar 18g. The preparation and storage methods are the same as in Example 1.

Example Embodiment

[0034] Example 4-For simultaneous sorting of Enterococcus and Streptococcus in milk-like samples:

[0035] Each 1000g medium contains: peptone 15g, yeast extract / yeast powder 5g, beef extract 5g, glucose 3g, sodium chloride 2.5g, disodium hydrogen phosphate 0.4g, sodium carbonate 3g, nalidixic acid 20mg, polyviscous Bacteriocin B is 1.2 mg, crystal violet 0.1 mg, esculin 5 g, ferric citrate 0.5 g, and agar 10 g. The preparation and preservation methods are the same as in Example 1.

[0036] Example 5-Separate and simultaneous separation of Enterococcus in the milk sample: The difference from Example 1 is that the sodium chloride is 65 g, and the sorting culture temperature is 45°C.

[0037] Example 6-Separate and simultaneous separation of Enterococcus in the milk sample: The difference from Example 1 is that the sodium chloride is 70 g, and the sorting culture temperature is 47°C.

[0038] Example 5-Separate and simultaneous separation of Enterococcus in the milk sample: The differe...

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PUM

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Abstract

The invention discloses a culture medium for separating enterococcus and streptococcus from a milk sample and a preparation method of the culture medium. The culture medium is used for separating enterococcus and streptococcus simultaneously and comprises components as follows: 15-25 g of bacterial peptone, 2-5 g of yeast, 2-5 g of beef heart extract or beef extract, 1-3 g of glucose, 1.8-2.5 g of sodium chloride, 0.3-0.5 g of disodium hydrogen phosphate, 2.0-3 g of sodium carbonate, 12-20 mg of nalidixic acid, 1.2-2 mg of polymyxin B, 0.1-0.3 mg of crystal violet, 2-5 g of aesculin, 0.3-0.8 g of ferric citrate and 10-18 g of agar, and the strain culture temperature is in a range from 35 DEG C to 37 DEG C. The culture medium has the advantages that the overall separation process is simple, visual and fast, the culture medium can separate enterococcus and streptococcus from the milk sample simultaneously, pathogen species are directly positioned to specific species or genuses, and the downstream analysis and identification process is simplified greatly. Besides, the invention further provides a culture medium component for independently separating streptococcus from the milk sample.

Description

technical field [0001] The invention belongs to the biological field, in particular to a culture medium for isolating bacteria from milk samples. Background technique [0002] The currently available systems for the isolation and identification of Enterococcus and Streptococcus in dairy products include the VITEK bacterial identification system and the API20E bacterial identification system. The principle is to integrate a variety of biochemical identification tests for Gram-positive bacteria on a standardized carrier (such as 96 Orifice plate), by inoculating bacteria, machine, or human-assisted machine reading and comparison with the result judgment library (table), so as to distinguish different bacterial species. Efficient preliminary sorting of target bacteria that need to be detected and monitored is the basis for rapid and accurate downstream identification. Enterococci widely exist in the environment, the digestive tract of animals and humans. With the natural evol...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/46C12R1/01
CPCC12N1/20
Inventor 赵兴绪武小虎候书宝马友记张勇
Owner GANSU AGRI UNIV
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