Description
Urea Broth for Enterobacteria Identification
Urea Broth serves as a vital tool in microbiology, specifically designed to aid in the identification of Enterobacteria. This specialized medium excels at differentiating Proteus from Salmonella and Shigella in clinical samples. By using Urea Broth, laboratories enhance their diagnostic capabilities and achieve reliable results.
Approximate Formula
The effectiveness of Urea Broth lies in its precise formulation. The ingredients per liter include:
- Pancreatic Digest of Casein: 1.0 g
- Potassium Dihydrogen Phosphate: 2.0 g
- Dextrose: 1.0 g
- Sodium Chloride: 5.0 g
- Urea: 20.0 g
- Phenol Red: 0.012 g
This combination promotes optimal growth conditions for bacteria. Notably, the final pH of the medium stands at 6.9±0.2 at 25°C, which ensures suitable conditions for microbial activity.
Composition and Functionality
Each ingredient plays a critical role in the performance of Urea Broth. For instance, pancreatic digest supplies essential nitrogen compounds, while potassium dihydrogen phosphate maintains stable pH levels. During incubation, when organisms metabolize urea, they produce ammonia. Consequently, this reaction leads to an alkaline environment, resulting in a visible color shift to red-pink. This change indicates urease production, which serves as a key marker for identifying Enterobacteria.
Preparation Instructions
To prepare Urea Broth, mix 29.01 g of the medium in one liter of distilled or deionized water. Ensure thorough mixing to dissolve the powder completely. After filtering and sterilizing, dispense the medium into sterile tubes. It is crucial to avoid autoclaving the medium to maintain its effectiveness.
Storage and Shelf Life
For optimal performance, store Urea Broth tightly capped in its original container and maintain temperatures between 5-30°C. The dehydrated medium boasts a shelf life of three years from the manufacturing date. Additionally, prepared medium should remain protected from direct light at 2-8°C.
Quality Control
Quality assurance remains essential in microbiological testing. Inoculate the prepared medium and incubate at 36±1°C for 18-24 hours. This step ensures that Broth meets performance criteria effectively.
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