Method for detecting viable count in pre-coated feed microecological preparation product

A technology of micro-ecological preparations and detection methods, applied in the preparation of test samples, measurement devices, color/spectral characteristics measurement, etc., can solve the problems of inapplicability, no indication of adenosine, and no consideration of bacterial cell damage, etc. , to achieve the effect of small influence of human factors, short detection cycle and good reproducibility

Active Publication Date: 2020-08-21
TIANJIN BIOFEED TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the traditional MTT method cannot uniformly disperse the tightly bound bacteria in the microcapsules without destroying its activity, and cannot accurately detect the number of live bacteria wrapped in the microcapsules. CN101620188A discloses a method using tetrazolium blue method to quickly The method for measuring the total number of viable bacteria in luminescent bacteria uses the MTT method to detect the number of viable bacteria at different concentrations to make a standard curve, and then uses the MTT method to detect the number of viable bacteria in the fermentation broth. However, this method is suitable for bacterial liquid, and the measurement parameters are not perfect. Bacterial ac

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  • Method for detecting viable count in pre-coated feed microecological preparation product
  • Method for detecting viable count in pre-coated feed microecological preparation product
  • Method for detecting viable count in pre-coated feed microecological preparation product

Examples

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Embodiment 1

[0044] In this embodiment, taking lactic acid bacteria as an example, a method for detecting the number of viable bacteria in a pre-coated microecological preparation product for feeding according to an embodiment of the present invention is described in detail. The principles and methods of other types of probiotics are the same as those of the examples here.

[0045] A method for detecting the number of live bacteria in coated lactic acid bacteria, comprising the following steps (taking the sample of coated lactic acid bacteria before the amount of bacteria is 1×1010 as an example):

[0046] Step 1: Draw the standard curve

[0047] (1) Cultivate free lactic acid bacteria for 21 hours, calculate the number of live bacteria by gradient dilution, and take an appropriate amount of bacterial liquid to centrifuge, add an equal volume of sterile water and resuspend in sterile water to obtain a bacterial suspension, and set aside;

[0048] (2) Take the bacterial solution from step ...

Embodiment 2

[0067] In this embodiment, taking lactic acid bacteria as an example, a method for detecting the number of viable bacteria in a pre-coated microecological preparation product for feeding according to an embodiment of the present invention is described in detail. The principles and methods of other types of probiotics are the same as those of the examples here.

[0068] A method for detecting the number of live bacteria in pre-coated lactic acid bacteria, comprising the following steps:

[0069] Step 1: Draw the standard curve

[0070] (1) Cultivate free lactic acid bacteria for 23 hours, calculate the number of live bacteria by gradient dilution, and take an appropriate amount of bacterial liquid to centrifuge, add an equal volume of sterile water and resuspend to obtain a bacterial suspension, and set aside;

[0071] (2) Take the bacterial solution from step (1), and after diluting different gradients, take 100 μL of the bacterial solution and add it to a 96-well microtiter ...

Embodiment 3

[0090] In this embodiment, taking Saccharomyces cerevisiae as an example, a method for detecting the number of viable bacteria in a pre-coated probiotic product for feeding according to an embodiment of the present invention is described in detail. The principles and methods of other types of probiotics are the same as those of the examples here.

[0091] A method for detecting the number of live bacteria in Saccharomyces cerevisiae, comprising the following steps:

[0092] Step 1: Draw the standard curve

[0093] (1) Cultivate free Saccharomyces cerevisiae for 38 hours, calculate the number of live bacteria by gradient dilution, and take an appropriate amount of bacterial liquid to centrifuge, add an equal volume of sterile water to resuspend to obtain a bacterial suspension, and set aside;

[0094] (2) Take the bacterial solution from step (1), and after diluting different gradients, take 100 μL of the bacterial solution and add it to a 96-well microtiter plate, and add 50 ...

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Abstract

The invention relates to a method for detecting viable count in a pre-coated feed microecological preparation product. The method comprises the following steps: (1), culturing free bacteria, carryingout gradient dilution to count the viable count, centrifuging a proper amount of bacterial liquid, and carrying out resuspension with sterile water to obtain a bacterial suspension for later use; (2),taking the bacterial liquid obtained in the (1), carrying out gradient dilution, reacting with an MTT solution to obtain a precipitate, dissolving the precipitate with DMSO, and carrying out colorimetry by using a microplate reader to measure a light absorption value; (3), preparing a standard curve by using the viable count of the free bacteria obtained in the (1) and the light absorption valuemeasured in the (2); (4), crushing and sieving a pre-coated sample, adding the crushed and sieved pre-coated sample into a conical flask added with glass beads, adding a capsule breaking solution, andbreaking the capsule to obtain a bacterial suspension; (5), carrying out ultrasonic treatment on the bacterial suspension to obtain a discrete bacterial suspension, centrifuging the discrete bacterial suspension, adding DMSO for resuspending, performing diluting to different gradients, and measuring a light absorption value in the same (2); and (6), calculating the viable count in a pre-coated microecological preparation according to the standard curve obtained in the (3).

Description

technical field [0001] The invention belongs to the technical field of detecting the number of viable microorganisms, and relates to a method for detecting the number of viable bacteria in pre-coated microecological preparation products for feeding. Background technique [0002] Microencapsulated culture of probiotics before fermentation means that free bacteria are encapsulated in capsules using microencapsulation technology before fermentation, and then fermented and cultured. Compared with free cultured probiotics, the strains have faster growth Advantages, effectively improve the tolerance of strains to high copper and gastrointestinal fluids, play a very good role in protecting microbial strains, ensure the play of their physiological functions, and solve the problem of insufficient potency of probiotics as feed additives Stability issue. [0003] A mature probiotic counting method should have the characteristics of good parallelism and good reproducibility of the coun...

Claims

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Application Information

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IPC IPC(8): G01N15/14G01N21/31G01N1/28G01N1/38
CPCG01N15/14G01N21/31G01N1/286G01N1/38G01N2015/1486
Inventor 李爽刘琼李阳汪玮王海燕张广民蔡辉益
Owner TIANJIN BIOFEED TECH CO LTD
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