Chromogenic media
Chromogenic Cronobacter Isolation Agar (CCI) ISO
For the isolation of presumptive Cronobacter spp. in food products and environmental samples.
- Cat. No.
- LC-1446
- Category
- Chromogenic media
- Format
- 500 g
- Storage
- 2 to 8 °C
- Minimum guaranteed shelf life
- 12 months
- Standards
- ISO 22964
Chromogenic Cronobacter Isolation Agar (CCI) is a selective medium for the detection of Cronobacter spp. in food products and ingredients intended for human consumption and the feeding of animals, and environmental samples in the area of food production and food handling.
Key features
Applications
Selective isolation of Cronobacter.
Industry
Food.
Standards
ISO 22964.
Preparation
32.4 g in one liter of distilled water; boil for one minute, sterilize in autoclave at 121 °C for 15 minutes.
Principles and uses
ISO 22964:2016 describes a horizontal method for the detection of Cronobacter spp. and recommends this medium for the isolation of Cronobacter spp.
Tryptone provides nitrogen, vitamins, minerals and amino acids essential for growth. Yeast extract is a source of vitamins, particularly the B-group, essential for bacterial growth. Sodium chloride supplies essential electrolytes for transport and osmotic balance. Sodium deoxycholate inhibits the accompanying Gram-positive flora. Sodium thiosulfate increases the selectivity and the recovery of Cronobacter and Enterobacter species. 5-Bromo-4-chloro-3-indolyl-alpha-D-glucopyranoside is the chromogenic substrate.
Cronobacter (formerly Enterobacter sakazakii) is currently considered an emerging pathogen responsible for severe meningitis and necrotic enterocolitis in un-weaned babies that can be the cause of a mortality rate between 40-80%.
The pathogenicity of Cronobacter for un-weaned babies makes it necessary to review the manufacturing process of the milk-based products specialized for babies, guaranteeing the absence of the bacteria in the final product.
Additional prevention measures at a hospital include the sanitary hygiene of the prepared food and reducing the time between the preparation and its administration, to impede the multiplication of microorganisms.
Technical specifications
| Solubility | Without residues |
|---|---|
| Appearance | Fine powder |
| Color of the dehydrated medium | Beige |
| Color of the prepared medium | Slightly amber |
| Final pH (25 °C) | 7.3 ± 0.2 |
| Amount per litre | 32.4 g |
| Storage | 2 to 8 °C |
Formula
| Ingredient | g/L |
|---|---|
| Bacteriological agar | 15 |
| Ferric ammonium citrate | 1 |
| Sodium chloride | 5 |
| Sodium deoxycholate | 0.25 |
| Sodium thiosulfate | 1 |
| Yeast extract | 3 |
| Tryptic digest of casein | 7 |
| 5-bromo-4-chloro-3-indolyl-alpha-D-glucopyranoside | 0.15 |
Preparation and use
Preparation
Suspend 32.4 grams of the medium in one liter of distilled water. Mix well and dissolve by heating with frequent agitation. Boil for one minute until complete dissolution. Sterilize in autoclave at 121 °C for 15 minutes. Cool to 50 °C, homogenize gently and dispense into Petri dishes in amounts of 15 mL.
According to ISO 22964
- Pre-enrich the test portion in a non-selective medium such as Buffered Peptone Water BPW (LC-1402).
- Incubate at a temperature of 34-38 °C for 18 ± 2 h.
- Inoculate the culture obtained in BPW in a selective enrichment medium: Selective Broth for Cronobacter (CSB) (LC-2143).
- Incubate at a temperature of 41.5 ± 1 °C for 24 ± 2 h.
- Sow and identify the colonies in the Chromogenic Cronobacter Isolation Agar (CCI).
- Incubate at a temperature of 41.5 ± 1 °C for 24 ± 2 h.
- For confirmation, typical colonies are selected from chromogenic agar, purified on a non-selective agar such as TSA (LC-1068) and characterized biochemically.
Quality control
Microbiological test
Incubation conditions: 41.5 ± 1 °C / 24 ± 2 h.
Inoculation conditions: productivity quantitative (100 ± 20 CFU, min. 50 CFU) / selectivity (10⁴-10⁶ CFU) / specificity (10³-10⁴ CFU).
| Microorganism | Specification | Characteristic reaction |
|---|---|---|
| Enterobacter cloacaeATCC 13047 | Growth (1-2) | The colonies do not have green or greenish-blue color |
| Staphylococcus aureusATCC 25923 | Total inhibition (0) | |
| Cronobacter sakazakiiATCC 29544 | Good growth (2) | Blue-green colonies of small to medium size (1-3 mm) |
| Cronobacter muytjensiiATCC 51329 | Good growth (2) | Blue-green colonies of small to medium size (1-3 mm) |
Bibliography
- ISO 22964:2016 Microbiology of the food chain - Horizontal method for the detection of Cronobacter spp.
- GUILLAUME-Gentil, O., Sonnard, V. Kandahai, M.C., Mauragg, J.D. and Jootsen, H. A simple and Rapid Cultural Method for Detection of Enterobacter sakazakii in environmental samples. Journal of Food Protection, 68 (1), 2005, pp. 64-69.
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