Liofilchem® MacConkey Agar with Bile Salts & Crystal Violet
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Specifications:
Application | Microbiology | ||
Storage Temperature | Room Temperature | ||
Product Type | Culture Medium | Forms | Powder |
Product Brand | Liofilchem | ||
Product Grade | Microbiology grade | ||
Liofilchem® MacConkey Agar is a selective and differential medium used for the detection and differentiation of Gram-negative enteric bacteria, particularly from samples of clinical, food, environmental, and pharmaceutical origins. It is widely utilized for identifying lactose-fermenting and non-lactose-fermenting organisms such as Escherichia coli, Klebsiella spp., Salmonella spp., and Shigella spp..
Intended Purpose
This medium is designed to:
- Selectively inhibit the growth of Gram-positive bacteria.
- Differentiate Gram-negative enteric bacteria based on their ability to ferment lactose.
- Support diagnostic workflows by providing presumptive identification of pathogens such as E. coli in feces, urine, foodstuffs, wastewater, and non-sterile pharmaceutical products.
It is a valuable diagnostic aid, although additional confirmatory tests are required for definitive results.
Key Features
- Selective and Differential:
- Crystal Violet and Bile Salts inhibit Gram-positive organisms, ensuring selective growth of Gram-negative bacteria.
- Lactose Fermentation is indicated by a color change due to acid production, differentiating lactose fermenters (pink colonies) from non-fermenters (colorless colonies).
- Wide Range of Applications:
- Effective for clinical samples (e.g., feces, urine).
- Supports food and environmental safety testing.
- Used in pharmaceutical microbiology for detecting E. coli contamination in non-sterile products.
- Standardized Formula:
- Complies with harmonized USP/EP/JP methods for microbial testing.
- Suitable for detecting E. coli and other Enterobacteriaceae from a variety of matrices.
- High Reliability:
- Consistent and reproducible results validated by quality control strains, including E. coli (ATCC® 8739), Salmonella Typhimurium (ATCC® 14028), and Shigella flexneri (ATCC® 12022).
Composition (Typical Formula)
Component | g/L | Function |
---|---|---|
Pancreatic Digest of Gelatin | 17.0 | Source of amino acids, nitrogen, and vitamins. |
Peptone from Meat and Casein | 3.0 | Nutrient supply for bacterial growth. |
Lactose | 10.0 | Fermentable carbohydrate for differentiation. |
Sodium Chloride | 5.0 | Maintains osmotic balance. |
Bile Salts | 1.5 | Inhibits Gram-positive bacteria. |
Agar | 15.0 | Solidifying agent. |
Neutral Red | 0.03 | pH indicator for lactose fermentation. |
Crystal Violet | 0.001 | Selective agent for Gram-negative bacteria. |
Final pH | 7.1 ± 0.2 | Optimal pH for bacterial growth. |
Preparation
For Dehydrated Medium:
- Suspend 51.5 g of powder in 1 L of distilled water.
- Heat to dissolve completely, boiling for 1 minute with frequent agitation.
- Autoclave at 121°C for 15 minutes.
- Cool to 45–50°C and pour into sterile Petri dishes.
For Pre-Bottled Medium:
- Melt the medium in a water bath at 100°C.
- Cool to 45–50°C, mix well, and aseptically distribute into Petri dishes.
Applications
- Clinical Microbiology:
- Identification of enteric pathogens from fecal, urine, or other biological samples.
- Presumptive identification of lactose-fermenting bacteria like E. coli.
- Food Microbiology:
- Detection of Salmonella and other pathogens in foodstuffs.
- Environmental Testing:
- Isolation of E. coli and other Gram-negative bacteria from wastewater.
- Pharmaceutical Microbiology:
- Detection of E. coli in non-sterile pharmaceutical products using harmonized methods.
Interpreting Results
Organism | Colony Appearance |
---|---|
E. coli | Pink colonies with or without bile precipitate. |
Klebsiella spp. | Pink, mucoid colonies. |
Salmonella spp., Shigella spp. | Colorless or clear colonies. |
Proteus spp. | Colorless colonies, may show swarming in non-selective areas. |
Gram-positive bacteria | Inhibited or extremely limited growth. |
Quality Control
Organism | Growth | Colony Appearance |
---|---|---|
E. coli (ATCC® 8739) | Good | Pink colonies with bile precipitation. |
Klebsiella pneumoniae (ATCC® 13883) | Good | Pink, mucoid colonies. |
Salmonella Typhimurium (ATCC® 14028) | Good | Colorless colonies. |
Enterococcus faecalis (ATCC® 29212) | Inhibited | Very small, opaque colonies (if any). |
Performance Characteristics
- Validated against standard QC strains.
- Reliable in differentiating lactose fermenters from non-fermenters.
Limitations
- May require additional confirmatory tests for definitive identification.
- Growth can vary based on specific organism requirements or environmental factors.
Storage and Shelf Life
- Dehydrated Medium: 4 years at 10–30°C in a dry environment.
- Pre-Bottled Medium: 2 years.
- Prepared Plates: 6 months at 10–25°C away from light.
Safety and Precautions
- For in vitro diagnostic use only.
- Handle clinical specimens and reagents as potentially infectious.
- Dispose of all waste following local and national regulations.
- Use appropriate personal protective equipment (PPE).
MacConkey Agar is a widely used selective and differential medium designed to isolate and differentiate Gram-negative enteric bacteria based on their lactose fermentation capabilities. Various formulations exist, each tailored for specific applications. Below is a comparative analysis of Liofilchem® MacConkey Agar with Bile Salts & Crystal Violet, Oxoid MacConkey Agar No. 2, Oxoid MacConkey Agar No. 3, and other common brands.
1. Composition
Component | Liofilchem® MacConkey Agar | Oxoid MacConkey Agar No. 2 | Oxoid MacConkey Agar No. 3 | Other Common Brands (e.g., Sigma-Aldrich) |
---|---|---|---|---|
Pancreatic Digest of Gelatin | 17.0 g/L | 20.0 g/L | 20.0 g/L | 17.0 g/L |
Peptone from Meat | 1.5 g/L | — | — | — |
Peptone from Casein | 1.5 g/L | — | — | — |
Lactose | 10.0 g/L | 10.0 g/L | 10.0 g/L | 10.0 g/L |
Bile Salts | 1.5 g/L | 5.0 g/L | 1.5 g/L | 1.5 g/L |
Sodium Chloride | 5.0 g/L | 5.0 g/L | 5.0 g/L | 5.0 g/L |
Neutral Red | 0.03 g/L | 0.075 g/L | 0.03 g/L | 0.03 g/L |
Crystal Violet | 0.001 g/L | — | 0.001 g/L | 0.001 g/L |
Agar | 15.0 g/L | 12.0 g/L | 15.0 g/L | 13.5 g/L |
Final pH (at 25°C) | 7.1 ± 0.2 | 7.4 ± 0.2 | 7.1 ± 0.2 | 7.1 ± 0.2 |
2. Selectivity and Differentiation
Feature | Liofilchem® MacConkey Agar | Oxoid MacConkey Agar No. 2 | Oxoid MacConkey Agar No. 3 | Other Common Brands (e.g., Sigma-Aldrich) |
---|---|---|---|---|
Selective Agents | Bile Salts, Crystal Violet | Bile Salts | Bile Salts, Crystal Violet | Bile Salts, Crystal Violet |
Inhibits Gram-Positive Bacteria | Yes | Yes | Yes | Yes |
Differentiates Lactose Fermenters | Yes | Yes | Yes | Yes |
3. Applications
Application | Liofilchem® MacConkey Agar | Oxoid MacConkey Agar No. 2 | Oxoid MacConkey Agar No. 3 | Other Common Brands (e.g., Sigma-Aldrich) |
---|---|---|---|---|
Clinical Specimens | Yes | Yes | Yes | Yes |
Food Testing | Yes | Yes | Yes | Yes |
Water Testing | Yes | Yes | Yes | Yes |
Pharmaceutical Testing | Yes | Yes | Yes | Yes |
4. Key Differences
- Oxoid MacConkey Agar No. 2: Contains a higher concentration of bile salts (5.0 g/L) and lacks crystal violet, making it particularly useful for recognizing enterococci in the presence of coliforms and non-lactose fermenters.
- Oxoid MacConkey Agar No. 3: Incorporates both bile salts and crystal violet, providing enhanced selectivity against Gram-positive bacteria and improved differentiation between coliforms and non-lactose fermenting organisms.
- Liofilchem® MacConkey Agar: Features a balanced composition with both bile salts and crystal violet, suitable for a broad range of applications, including compliance with harmonized USP/EP/JP methods for detecting E. coli in non-sterile pharmaceutical products.
5. Considerations for Selection
- Target Organisms: Choose a formulation based on the specific Gram-negative bacteria of interest and the presence of Gram-positive flora in the sample.
- Sample Type: Consider the origin of the sample (clinical, food, water, pharmaceutical) to select the most appropriate medium.
- Regulatory Compliance: Ensure the medium aligns with relevant standards and guidelines for the intended application.
In summary, while all these MacConkey Agar formulations serve the fundamental purpose of isolating and differentiating Gram-negative enteric bacteria, variations in their composition, particularly concerning bile salts and crystal violet concentrations, influence their selectivity and suitability for specific applications.
Liofilchem® MacConkey Agar is an essential tool in microbiology labs for the isolation and differentiation of Gram-negative enteric bacteria. Its selective and differential properties make it suitable for applications ranging from clinical diagnostics to food and environmental safety testing. With its compliance to USP/EP/JP standards, it ensures reliability and reproducibility across a wide range of testing scenarios.
- Pack Size: 100g 500g 5kg