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BCYE Agar (Buffered Charcoal Yeast Extract Agar) ISO 11731 Reference Medium for Isolation of Legionella Species
  • BCYE Agar (Buffered Charcoal Yeast Extract Agar) ISO 11731 Reference Medium for Isolation of Legionella Species
BCYE Agar (Buffered Charcoal Yeast Extract Agar)

BCYE Agar (Buffered Charcoal Yeast Extract Agar)

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BCYE Agar (Buffered Charcoal Yeast Extract Agar) | ISO 11731 Reference Medium | AS-1133
AuSaMicS Life Science • Legionella Reference Media

BCYE Agar (Buffered Charcoal Yeast Extract Agar)

ISO 11731 Reference Medium for Isolation of Legionella Species

Reference-grade charcoal-buffered yeast extract agar for primary isolation and cultivation of Legionella spp., especially Legionella pneumophila, from environmental and clinical reference samples. Suitable for both non-selective BCYE and selective BCYE workflows.

AS-1133 🧫 Legionella Reference Medium ✓ ISO 11731 Aligned ⚗️ BCYE / GVPC / MWY / BMPA Ready
Use: Legionella isolation Preparation: 36.0–38.0 g/L base Incubation: 35–36°C / 5–10% CO₂ / up to 14 days

Legionella Recovery Specialist

Activated Charcoal — neutralizes toxic by-products
ACES Buffer — precise pH control
L-Cysteine + Iron — essential growth requirements
α-Ketoglutarate — supports colony maturation
Gold-Standard Base Medium
Clinical + Water Microbiology

Overview

BCYE Agar (Buffered Charcoal Yeast Extract Agar) is the international reference solid culture medium for the primary isolation and cultivation of Legionella species, particularly Legionella pneumophila, from clinical and environmental samples.

Originally developed for fastidious Legionella recovery and later standardized in ISO 11731 workflows, BCYE Agar contains activated charcoal, ACES buffer, α-ketoglutarate, iron, and L-cysteine, all of which are essential for the successful culture of Legionella organisms.

AuSaMicS BCYE Agar is positioned as a reference-grade base medium suitable for both non-selective BCYE and selective BCYE variants, including workflows based on GVPC, MWY, and BMPA-style supplements.

Key Features & Benefits

Activated Charcoal Protection
Neutralizes toxic metabolites and protects sensitive Legionella cells.
Precise Buffering
ACES maintains critical pH stability for dependable culture performance.
Essential Growth Factors
Iron and L-cysteine satisfy the specific metabolic needs of Legionella.
Reference-Medium Status
Recognized as the gold-standard base medium in Legionella culture workflows.

Typical Applications

Environmental & Water Microbiology
Isolation of Legionella spp. from water systems, cooling towers, potable water networks, and plumbing-associated biofilm samples.

Clinical & Reference Laboratories
Recovery of Legionella from respiratory specimens, bronchoalveolar samples, and lung tissue in reference or validated laboratory workflows.

Regulatory & Surveillance Testing
Reference culture medium in Legionella monitoring and surveillance programs aligned with ISO 11731 and related laboratory standards.

Principle of the Medium

BCYE Agar provides a nutrient-rich and protective environment specifically tailored to the metabolic requirements of Legionella species:

  • Activated charcoal absorbs toxic peroxides and inhibitory compounds
  • Yeast extract supplies vitamins, peptides, and growth-supporting nutrients
  • ACES buffer maintains a tightly controlled pH, a critical factor for performance
  • Iron and L-cysteine are essential metabolic requirements for Legionella
  • α-Ketoglutarate stimulates growth and supports colony maturation

The finished medium appears black and opaque, which helps characteristic Legionella colony morphology stand out during prolonged incubation.

Complete Formulation & Functional Profile

Base Medium (Typical – per Liter)

Ingredient Concentration Function Technical Role
Yeast extract 10.0 g Nutrient source Provides vitamins and amino acids
Activated charcoal 2.0 g Protective adsorbent Neutralizes toxic by-products
ACES buffer 10.0 g Buffering agent Maintains critical pH stability
α-Ketoglutarate (potassium salt) 1.0 g Growth stimulant Enhances colony development
Agar 13.0–15.0 g Solidifying matrix Supports prolonged incubation and colony observation

Supplements Added After Sterilization

Supplement Concentration Importance
L-Cysteine HCl·H₂O 0.4 g Essential for Legionella metabolism and growth
Iron(III) pyrophosphate (soluble) 0.25 g Essential trace factor supporting recovery
Critical Parameter: Final pH at 25 °C should be 6.9 ± 0.1 for optimal performance.

Legionella Culture Media Comparison

Medium Role Strength Limitation Best Use
BCYE Agar Reference base medium Gold-standard recovery of fastidious Legionella May require selective variant in contaminated samples Primary isolation and confirmation workflows
GVPC BCYE Selective Legionella isolation Improved selectivity for environmental matrices Higher selective pressure may reduce some recoveries Water and highly contaminated samples
MWY Agar Highly selective variant Strong differentiation/selectivity in some workflows More specialized supplement system Surveillance and difficult matrices
BMPA / Related selective BCYE variants Selective plating Adaptable for specific lab protocols Method-dependent supplement handling Reference and custom Legionella workflows

Selection Guide

Choose non-selective BCYE if:
• Sample contamination is low to moderate
• Maximum recovery is the main priority
• You need the classical reference base medium
Choose selective BCYE if:
• Samples are heavily contaminated
• Water or environmental matrices require stronger inhibition of background flora
Choose BCYE as base if:
• Your workflow needs flexibility across GVPC, MWY, or BMPA-type variants

Typical Colony Appearance

  • Grey-white to iridescent blue-green colonies
  • Characteristic “ground-glass” appearance
  • Cut-glass texture under stereomicroscopy

Technical Specifications

Cat. No. AS-1133
Medium Type Reference-grade BCYE agar base
Preparation 36.0–38.0 g in 950 mL purified water
Critical pH 6.9 ± 0.1 at 25 °C
Incubation 35–36 °C / 5–10% CO₂ / high humidity
Duration Up to 14 days
Appearance Black, opaque medium
Dehydrated Storage Below 30 °C, dry, moisture protected
Prepared Plates 2–8 °C, sealed, light protected, use within 4 weeks
HS Code 3821.00.00

Preparation Protocol

  1. 1 Suspend 36.0–38.0 g of BCYE base in 950 mL purified water.
  2. 2 Add ACES buffer and α-ketoglutarate; mix thoroughly.
  3. 3 Adjust pH to 6.9 at 50 °C using 1 N KOH.
  4. 4 Autoclave at 121 °C for 15 minutes.
  5. 5 Cool to 50 °C, then aseptically add filter-sterilized L-cysteine and iron solutions.
  6. 6 Mix well and pour plates.

Pack Sizes & Suggested Pricing

100 g AUD 68
250 g AUD 145
500 g AUD 245
1 kg AUD 420
5 kg AUD 1620

Quality & Compliance

  • Conforms to ISO 11731 reference-medium expectations
  • Suitable for regulated and accredited laboratories
  • Manufactured under controlled conditions for batch consistency
  • Certificate of Analysis (COA) available
  • Safety Data Sheet (SDS) available upon request

Storage & Stability

  • Dehydrated base: Store below 30 °C, dry, protected from moisture
  • Prepared plates: Store at 2–8 °C, sealed and protected from light
  • Recommended use: Within 4 weeks of preparation

Intended Use & Biosafety

For laboratory environmental, clinical reference, and research microbiology use only.

Legionella species are classified as Risk Group 2 organisms. Work must be conducted by trained personnel using appropriate biosafety practices.

Not for therapeutic or direct clinical diagnostic use without appropriate validation.

AuSaMicS Compliance & Supply Notes

Supplied for laboratory and research workflows. Batch-specific documentation may include Certificate of Analysis, Technical Data Sheet, and Safety Data Sheet depending on customer requirements.

For laboratory and research use only. Not for household, food, therapeutic, or direct clinical use without validation.

AuSaMicS Pty Ltd
31 Longview CT, Thomastown, VIC 3074, Australia
ABN 56676640467
www.ausamics.com.au
support@ausamics.com
+61 412 520 598

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