Composite culture medium capable of simultaneously enriching three food-borne pathogenic bacteria and preparation method

A technology for food-borne pathogenic bacteria and culture medium, which is applied in microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve problems such as increasing the workload of detection, and achieve rapid proliferation and good co-enrichment effect. Effect

Inactive Publication Date: 2021-01-01
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to overcome the problems in the prior art that different enrichment media are often used for cultivating different pathogenic bacteria, which increases the workload of detection, the present invention provides a method that can simultaneously cultivate Listeria monocytogenes and

Method used

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  • Composite culture medium capable of simultaneously enriching three food-borne pathogenic bacteria and preparation method
  • Composite culture medium capable of simultaneously enriching three food-borne pathogenic bacteria and preparation method
  • Composite culture medium capable of simultaneously enriching three food-borne pathogenic bacteria and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1: LES culture medium monoenrichment effect identification

[0032] Weigh 17.0 parts of tryptone, 3.0 parts of peptone, 6.0 parts of yeast extract, 5.0 parts of sodium chloride, 2.5 parts of dipotassium hydrogen phosphate, 9.6 parts of disodium hydrogen phosphate, 1.35 parts of potassium dihydrogen phosphate, 25.0 parts of peptone, 15.0 parts of beef powder, 14.0 parts of soybean peptone, 3.0 parts of glucose, 0.3 parts of cycloheximide, 3.0 parts of mannitol, 6.0 parts of sodium pyruvate, 0.003 parts of acridine yellow, 0.003125 parts of fosfomycin sodium, distilled water Set the volume to 1000 parts, stir and heat, boil until completely dissolved, after cooling to room temperature, adjust the pH to 7.2, autoclave at 121°C for 20min, and store at 4°C.

[0033] Take a certain amount of overnight cultured fresh bacteria liquids of Listeria monocytogenes (LM), Escherichia coli O157:H7 (O157:H7), and Salmonella (SE) and inoculate them into LES medium and their ...

Embodiment 2

[0035] Embodiment 2: LES culture medium co-enrichment bacterial effect identification

[0036]Add a certain amount of Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella bacterial liquid into the LES medium, so that the initial concentration of the three target bacteria is 10 CFU / mL. After inoculation, shake culture at 37°C and 200rpm, take 3h, 6h, 9h, 12h, 16h, and 20h cultures for plate counting, and count the culture medium for Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella They are PALCAM agar medium, Escherichia coli O157 chromogenic medium and bismuth sulfite agar medium respectively.

[0037] According to the colony counting results at each time point, the figure 2 In the growth curve shown, on the whole, 0-12h is the stage of rapid bacterial growth. After 12 hours of cultivation, the concentrations of the three bacteria basically reached or exceeded 10 7 CFU / mL (Listeria monocytogenes 0.87×10 7 CFU / mL, Escherichia coli O157:H7 7.5×10...

Embodiment 3

[0038] Embodiment 3: the inhibitory effect of LES medium to non-target bacteria

[0039] A certain amount of target bacteria and non-target bacteria were added to the LES medium, so that the initial concentration of non-target bacteria was 100 CFU / mL, and the initial concentration of target bacteria was 10 CFU / mL, and then cultured with shaking at 37 °C and 200 rpm. Take the cultures of 3h, 6h, 9h, 12h, 16h, and 20h to measure the McFarland turbidity value, and the results are shown in Table 4. By comparing the McFarland turbidity values ​​of target bacteria and non-target bacteria, it can be seen that the growth of non-target bacteria Bacillus subtilis, Yersinia enterocolitica, and Pseudomonas aeruginosa in LES medium was significantly inhibited.

[0040] Table 4 McFarland turbidity values ​​at different time points when target bacteria and non-target bacteria were cultured in LES medium

[0041]

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Abstract

The invention discloses a composite enrichment culture medium capable of simultaneously enriching three food-borne pathogenic bacteria, namely Listeria monocytogenes, Escherichia coli O157:H7 and salmonella, and a preparation method. The formula of the culture medium comprises the following components of, in parts by weight, 15.0-19.0 parts of tryptone, 2.7-3.3 parts of peptone, 5.5-6.5 parts of yeast extract, 4.5-5.5 parts of sodium chloride, 2.25-2.75 parts of dipotassium phosphate, 8.6-10.6 parts of disodium hydrogen phosphate, 1.22-1.48 parts of potassium dihydrogen phosphate, 24.0-30.0 parts of peptone, 14.0-18.0 parts of beef powder, 10.0-14.0 parts of soy peptone, 2.0-5.0 parts of glucose, 0.10-0.30 part of actinomycete ketone, 2.5-3.5 parts of mannitol, 4.5-6.5 parts of sodium pyruvate, 0.00125-0.00300 part of acridine yellow, 0.0007813-0.0032500 part of fosfomycin sodium and 1000 parts of distilled water, and the pH value of theculture medium is 6.8-7.4. The single enrichmenteffect of the composite enrichment culture medium is superior to that of respective selective enrichment liquid, growth of non-target bacteria can be well inhibited, under the condition that non-target bacteria exist, the culture medium can further achieve constant-speed and rapid proliferation of the three target bacteria.

Description

technical field [0001] The invention belongs to the technical field of pre-enrichment medium for pathogenic bacteria, and in particular relates to a composite medium capable of simultaneously enriching three kinds of food-borne pathogenic bacteria, that is, Listeria monocytogenes, Escherichia coli O157:H7 and Salmonella. Preparation. Background technique [0002] In recent years, foodborne diseases caused by eating food contaminated by foodborne pathogens such as Listeria monocytogenes, Escherichia coli O157:H7 and Salmonella have often occurred. The control and detection of foodborne pathogenic bacteria have been highly valued all over the world. [0003] Listeria monocytogenes, Escherichia coli O157:H7 and Salmonella are three common foodborne pathogens that must be detected in my country's national food safety standards. Usually, the number of foodborne pathogenic bacteria that may exist in food is often relatively small. In order to ensure the accuracy of the test resu...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/38C12R1/19C12R1/42C12R1/01
CPCC12N1/20C12N1/38
Inventor 孙春燕谢楠楠张晓光李爽郝莹刘子婕狄心瑜
Owner JILIN UNIV
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