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Method for detecting total number of bacterial colonies in activated lactobacillus drink

A technology of active lactic acid bacteria and total number of colonies, applied in the field of microbial detection, can solve problems such as insufficient detection sensitivity, influence of detection accuracy, and reduction of result accuracy.

Inactive Publication Date: 2012-04-25
GUANGDONG HUANKAI MICROBIAL SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there is no unified method for the determination of the total number of colonies of active lactic acid bacteria beverages in my country. There are ISO 13559: 2002 (E) "Butter, fermented milk products and fresh cheese. Counting of polluting microorganisms. Colony counting method at 30 °C" and Japan "Determination of Total Colony Count of Lactic Acid Bacteria Powder Refreshing Beverages"
ISO13559: 2002 (E) due to the coating method, the inspection accuracy is affected to a certain extent, and the inspection time is long; Japan "Determination of the Total Colony of Lactic Acid Bacteria Powder Refreshing Drinks", using glucose agar medium with penicillin G potassium and 4% chloride The difference of the number of colonies grown on the sodium BCP plate counting agar medium is used as the total number of colonies, which is cumbersome to operate, because the selective inhibitory components added to the medium have complex effects on the detection, reducing the accuracy of the results, and there is also insufficient detection sensitivity. The problem

Method used

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  • Method for detecting total number of bacterial colonies in activated lactobacillus drink
  • Method for detecting total number of bacterial colonies in activated lactobacillus drink

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Embodiment 1: TTC nutrient agar preparation

[0013] Weigh 33g of nutrient agar, add 1 liter of distilled water or deionized water, divide into Erlenmeyer flasks, stir, heat and boil to dissolve, and autoclave at 121°C for 15min. After sterilization, when the culture medium is cooled to about 45°C, add sterile TTC to make the concentration of TTC reach 2.5mg / 100ml, mix well, and pour onto the plate.

Embodiment 2

[0014] Embodiment 2: Detection of the total number of bacterial colonies in artificially polluted samples

[0015] Take fresh cultures of Escherichia coli, Staphylococcus aureus, and Bacillus cereus, dilute them with sterile saline to a concentration of 0.5 McFarland turbidimetric tubes, and then serially dilute them 10 times. Add the solution to sterile samples A, B, C, and D respectively, so that the concentration of each contaminating bacteria in the sample reaches about 1000cfu / ml. Take 1ml of the bacterium-added sample and mix it with 9ml sterile normal saline to make a 10-fold dilution, take 1ml of the diluted solution and mix it with an appropriate amount (25ml) of sterile normal saline, filter it through a 0.45μm filter membrane, and then use 25ml Rinse the filter membrane with physiological saline, and stick it on the TTC nutrient agar plate described in Example 1 with the filter side up after rinsing. 37 ℃, 48 ± 2h inverted culture. Its count is as follows:

[001...

Embodiment 3

[0018] Embodiment 3: Comparison between the present invention and ISO 13559:2002 (E) "Butter, fermented milk products and fresh cheese. Enumeration of contaminating microorganisms. Colony counting method at 30°C"

[0019] 30 kinds of yoghurt samples were purchased in the market, and the method of the present invention and ISO 13559: 2002 (E) "Butter, fermented milk products and fresh cheese. Counting of contaminating microorganisms. Colony counting method at 30°C" was adopted respectively to check the total number of colonies .

[0020] The present invention detects that 2 samples are positive for the total number of colonies, and the ISO method detects 1 sample. In the present invention, at 10 times dilution concentration, 28 samples have no colony growth on the filter membrane, and sample 1 grows 3 colonies. After identification, none of them are lactic acid bacteria, which can be included in the total number of colonies. The total number of colonies in sample 1 is 30cfu / ml....

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Abstract

The invention relates to a method for detecting the total number of bacterial colonies in an activated lactobacillus drink, relating to a microorganism detecting method. The method comprises the following steps of: filtering diluent of the activated lactobacillus drink with a filter membrane, flushing the filter membrane with physiological saline, pasting the flushed filter membrane on a TTC nutrient agaragar tablet with the filtering face facing upwards, converting the nutrient agaragar tablet and culturing at 37 DEG C, and counting after culturing for 48+ / -2h. By adopting the method, the total number of pollution bacterial colonies in the activated lactobacillus drink can be accurately, simply and quickly detected so as to provide an effective method for detecting the sanitation qualityof the of the activated lactobacillus drink.

Description

【Technical field】 [0001] The invention relates to a method for detecting microorganisms. 【Background technique】 [0002] As consumer groups pay more and more attention to food safety, the state's monitoring and control of food safety and requirements are becoming more and more stringent, and various food manufacturers have also strengthened monitoring measures for key production links and control points. Both require the detection of the total number of bacteria in the product. "Food Hygiene Microbiological Inspection" (GB / T4789-2008) stipulates the inspection method of the total number of bacterial colonies in many types of foods. Only active lactic acid bacteria products do not have a unified inspection method for the total number of colonies. [0003] Lactic acid bacteria is a general term for a class of bacteria that can produce large amounts of lactic acid from fermentable sugars. Its common features are Gram stain positive, catalase negative, facultative anaerobic, co...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/06
Inventor 马德良蔡芷荷吴清平何天文
Owner GUANGDONG HUANKAI MICROBIAL SCI & TECH
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