Selective Bifidobacterium breve Culture Medium Using Sorbitol
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Solution Overview
Problem
Current methods for determining the viable cell count of Bifidobacterium breve in products containing multiple bifidobacterial species are challenging due to similar colony morphologies and the unavailability of sterile defibrinated blood, making it difficult to distinguish and quantify the breve species.
Innovation Solution
A culture medium containing sorbitol or mannitol as a sole sugar source, along with specific concentrations of peptone, meat extract, and yeast extract, is developed to selectively promote large colony formation by Bifidobacterium breve, allowing for easy identification and quantification.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a culture medium containing sterile defibrinated blood is used to determine viable cell count of bifidobacteria, then the measurement precision is improved, but the ease of manufacture deteriorates due to unavailability of sterile defibrinated blood in many countries
Solution Approach 1:
The invention replaces expensive and difficult-to-obtain sterile defibrinated blood with a simple, inexpensive alternative culture medium formulation that achieves the same measurement objective without requiring rare materials
Solution Approach 2:
The invention changes the chemical composition parameters of the culture medium by using specific concentrations of peptone (6.0-14.0 g/1000mL), meat extract (6.0-14.0 g/1000mL), and yeast extract (1.8-4.2 g/1000mL) to achieve selective growth of B. breve without blood components
2Measurement precision
If colony morphology is used to identify Bifidobacterium breve species, then the measurement precision is improved, but the device complexity increases due to requiring specialized knowledge for interpretation
Solution Approach 1:
The invention uses colony size (diameter ≥0.7mm) as a visual indicator that is easily distinguishable and does not require specialized knowledge for interpretation, replacing complex morphological analysis
Solution Approach 2:
The invention segments the identification criteria into a simple binary classification: colonies with diameter ≥0.7mm are counted as B. breve, while smaller colonies are not, eliminating the need for complex morphological evaluation
3Measurement precision
If a selective culture medium for B. breve is developed using sugar alcohols, then the measurement precision is improved, but the productivity decreases due to limited sugar source options
Solution Approach 1:
The invention changes the carbon source parameter from conventional sugars to sugar alcohols (sorbitol or mannitol) at controlled concentrations (≤4% by mass) to achieve selective growth of B. breve while maintaining formulation flexibility
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This method enables accurate determination of the viable cell count of Bifidobacterium breve even in the presence of other bifidobacterial species, without requiring sterile defibrinated blood, and is simple to prepare, providing a reliable and efficient means for product quality assessment.
Implementation Method 1
most bacterial cells of breve species may assimilate sorbitol and mannitol during the viable cell count through a culture method
Data Source
AI summary
There are provided a method of identifying a microorganism belonging to Bifidobacterium breve alone and measuring the viable cell count thereof, amongst the bacterial cells to be tested which include a microorganism belonging to the genus Bifidobacterium by using a culture medium that contains at least one sugar alcohol selected from sorbitol and mannitol as a sole sugar source; and also a culture medium which is useful as a selective medium to be used in the above measurement method and which is also easy to prepare.