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Bushnell-Haas Agar | Hydrocarbon Degrading Bacteria Medium | Ausamics AS-1161
$115.00 AUD
Documentation
BUSHNELL-HAAS AGAR
Catalogue No. AS-1161
www.ausamics.com.au
AS-1161 Carbon-Free Base Petroleum Microbiology Bioremediation Research Bushnell & Haas (1941)
Bushnell-Haas Agar
Carbon-free mineral salts agar for the isolation, enrichment, and study of hydrocarbon-degrading microorganisms
Bushnell-Haas Agar is a carbon-free mineral salts medium derived from the classic Bushnell and Haas (1941) approach for studying microbial hydrocarbon utilisation. The basal formulation supplies nitrogen, phosphorus, magnesium, calcium, and iron while intentionally omitting an added organic carbon source — making the experimental hydrocarbon the key added carbon and energy source.
AuSaMicS manufactures Bushnell-Haas Agar in Melbourne, Australia, to the original reference formulation, giving reliable selectivity for isolation, enrichment, and comparative screening of hydrocarbon-degrading isolates.
Performance Snapshot
Final pH7.0 ± 0.2
Carbon sourceNone (user-added)
Gel strengthFirm, 2.0% agar-equivalent
Precipitate after autoclavingYellow-orange — normal
Shelf life (powder)24 months
Formulation accuracy note: Bushnell-Haas Agar correctly contains anhydrous calcium chloride (CaCl₂) at 0.02 g/L. Some secondary sources list the dihydrate (CaCl₂·2H₂O) at the same mass, which would under-deliver calcium relative to the original Bushnell and Haas (1941) formula. AuSaMicS documentation reflects the verified anhydrous formulation, cross-checked against current HiMedia and peer-reviewed technical sources.
Testing Workflow
1
SampleSoil, water, fuel, or oil-impacted sample
2
Inoculate PlateStreak or spread onto poured agar
3
Add HydrocarbonOverlay liquid, or invert plate for volatiles
4
Incubate25–30°C, up to 1–2 weeks
5
ObserveColony growth = hydrocarbon-utilising isolate
Scientific Background & Principle
Origin
Bushnell-Haas Agar is the solid form of the classic mineral salts medium described by Bushnell and Haas (1941) and later recommended for the microbiological examination of fuels by the SIM Committee on Microbiological Deterioration of Fuels. It remains the standard reference medium for hydrocarbon-degradation isolation and screening work more than eight decades later.
Mechanism
The medium supplies nitrogen (ammonium nitrate), phosphorus and buffering capacity (mono- and dipotassium phosphate), and trace elements (magnesium, calcium, iron — with sulfate incidentally supplied via magnesium sulfate) — but contains no added carbon source. Agar (20 g/L) solidifies the medium for plate-based isolation and colony morphology observation. Because carbon is the one nutrient withheld, only organisms capable of metabolising an externally supplied hydrocarbon as their sole carbon and energy source can form colonies — growth itself is the diagnostic signal.
Why a Carbon-Free Basal Medium
By minimising alternative carbon inputs, the medium lets researchers test directly whether an inoculum can grow when a selected hydrocarbon is the principal added carbon source — useful for enrichment, comparative screening across substrates or isolates, and selection of candidates for downstream analytical or molecular workflows.
Key Applications
Isolation & Screening
Isolation of hydrocarbon-degrading microorganisms from soil, water, fuel-associated, and oil-impacted samples.
Bioremediation Research
Petroleum biodegradation studies and screening of candidate degrader strains.
Environmental Microbiology
Study of hydrocarbon-adapted microbial communities in contaminated sites.
Enrichment Prior to Confirmation
Enrichment step ahead of confirmatory analytical or molecular identification workflows.
Formulation & Specifications
| Component | g/L |
|---|---|
| Magnesium Sulfate (MgSO₄·7H₂O) | 0.20 |
| Calcium Chloride, anhydrous (CaCl₂) | 0.02 |
| Monopotassium Phosphate (KH₂PO₄) | 1.00 |
| Dipotassium Phosphate (K₂HPO₄) | 1.00 |
| Ammonium Nitrate (NH₄NO₃) | 1.00 |
| Ferric Chloride (FeCl₃·6H₂O) | 0.05 |
| Agar | 20.00 |
| Total | 23.27 g/L |
Technical Specifications
| Catalogue No. | AS-1161 |
| HS Code | 3821.00 |
| Format | Dehydrated powder |
| Final pH | 7.0 ± 0.2 @ 25°C |
| Storage (powder) | 10–30°C, dry, dark |
| Storage (plates) | 2–8°C, use ≤ 2 weeks |
Laboratory Preparation
1
Suspend 23.27 g of dehydrated medium in 1 litre of distilled or purified water.
2
Heat with gentle agitation and bring to boiling until completely dissolved.
3
Dispense into suitable laboratory containers.
4
Sterilise by autoclaving at 121°C for 15 minutes.
5
Cool, pour plates, and use according to your validated laboratory method.
Working With Hydrocarbon Supplements
- The hydrocarbon is not supplied as part of the dehydrated medium and must be sourced separately.
- Liquid hydrocarbons (mineral oil, kerosene, diesel, paraffin) are typically layered onto the surface of inoculated agar.
- Volatile hydrocarbons (e.g., gasoline) are often added to the lid of an inverted plate to test vapour-phase utilisation.
- Hydrocarbons vary substantially in solubility, volatility, and toxicity — delivery geometry and evaporation control can materially affect exposure, especially for volatile compounds.
- Always add hydrocarbon supplements aseptically and follow the substrate's own SDS and risk controls.
Research Guidance for Stronger Experimental Design
Controls That Improve Interpretation
- No-carbon inoculated control — checks for carry-over nutrients and endogenous reserves.
- Abiotic substrate control — helps identify non-biological loss or physical changes to the hydrocarbon.
- Positive growth control — validates inoculum viability in a separate, suitable medium.
- Replicates — improve confidence when comparing substrates or isolates.
Variables Worth Reporting
- Hydrocarbon identity, purity, and concentration
- Liquid, surface-overlay, or vapour-phase delivery method
- Inoculum source and preparation
- Temperature, incubation time, and atmosphere
- Growth endpoint and any confirmatory analytical method used
For publication-quality work: treat growth on Bushnell-Haas Agar as one line of evidence. Where the research question concerns actual biodegradation, pair microbiological observations with an analytical measurement of substrate depletion and/or metabolites (e.g., GC-FID, gravimetric analysis).
Limitations & Technical Considerations
- Requires a hydrocarbon supplement — the base agar alone will not support growth; this is the most common cause of a false-negative result.
- Growth does not prove complete degradation — colony formation indicates hydrocarbon-utilisation capability, not mineralisation or complete chemical breakdown.
- Extended incubation may be needed — hydrocarbon-degrading organisms can grow slowly; 1–2 weeks at 25–30°C is common.
- Yellow-orange precipitate after autoclaving is normal — do not discard a batch or assume contamination based on this colour change alone.
Cross-Reference / Equivalent & Related Products
| Brand | Equivalent Product |
|---|---|
| BD Difco | Bushnell-Haas Agar |
| HiMedia | Bushnell Haas Agar (M349) |
| Sigma-Aldrich | Bushnell Haas Agar (B4926) |
| AuSaMicS | Bushnell-Haas Agar (AS-1161) · companion liquid form: Bushnell-Haas Broth (AS-1162) |
Ordering Information
| Catalogue No. | Pack Size | Approx. Yield | Format |
|---|---|---|---|
| AS-1161-500 | 500 g | ≈ 21.5 L prepared medium | Dehydrated powder, bottle |
| AS-1161-2K5 | 2.5 kg | ≈ 107 L prepared medium | Dehydrated powder, bottle |
Yield calculated at 23.27 g/L reconstitution rate. Hydrocarbon supplement is not included and must be sourced and added separately by the user.
Frequently Asked Questions
What is Bushnell-Haas Agar used for?
It isolates, enriches, and studies hydrocarbon-degrading microorganisms from soil, water, fuel, and oil-impacted samples, and is used in bioremediation research, petroleum microbiology, and environmental monitoring.
What is the difference between Bushnell-Haas Agar and Bushnell-Haas Broth?
They share the same mineral salts base formulation. The agar version is solidified with 20 g/L agar for plate-based isolation and colony observation, while the broth is a liquid form used for turbidimetric enrichment and screening.
Does Bushnell-Haas Agar contain calcium chloride dihydrate or anhydrous?
Anhydrous calcium chloride (CaCl₂) at 0.02 g/L — consistent with the original Bushnell and Haas (1941) formula and current commercial technical documentation.
How do you add the hydrocarbon source?
For liquid hydrocarbons, layer the substrate onto the surface of inoculated agar. For volatile hydrocarbons such as gasoline, invert the plate and add the hydrocarbon to the lid.
Does growth on this medium prove hydrocarbon biodegradation?
Growth indicates hydrocarbon-utilisation capability but does not by itself prove complete chemical degradation. Pair microbiological growth with an analytical measurement of substrate depletion or metabolites for publication-quality biodegradation claims.
Common Search Queries This Product Answers
Researchers and procurement teams typically find this page searching for:
- "Bushnell-Haas Agar composition" / "Bushnell Haas Medium formula"
- "hydrocarbon degrading bacteria agar isolation"
- "carbon-free mineral salts agar for bioremediation"
- "TPH PAH biodegradation screening medium"
- "petroleum microbiology culture medium Australia"
- "buy Bushnell-Haas Agar Australia Melbourne"
Technical Data Sheet
| Product Name | Bushnell-Haas Agar |
| Synonyms | Bushnell-Haas Medium |
| Catalogue Number | AS-1161 |
| Format | Dehydrated powder |
| Manufacturer | AuSaMicS Pty Ltd, Melbourne, Australia |
Intended Use
Carbon-free mineral salts agar for the isolation, enrichment, and study of hydrocarbon-degrading microorganisms in soil, water, fuel, and oil-impacted samples.
Mode of Action
The formulation supplies nitrogen, phosphorus, and trace elements (magnesium, calcium, iron) but deliberately contains no added carbon source. Agar solidifies the medium for plate-based isolation. Only organisms able to metabolise an externally supplied hydrocarbon as their sole carbon and energy source can form colonies.
Formulation (per litre)
| Ingredient | Quantity | Function |
|---|---|---|
| Magnesium Sulfate (MgSO4.7H2O) | 0.20 g | Trace nutrient, magnesium source |
| Calcium Chloride, anhydrous (CaCl2) | 0.02 g | Trace nutrient, calcium source |
| Monopotassium Phosphate (KH2PO4) | 1.00 g | Phosphorus source, buffering |
| Dipotassium Phosphate (K2HPO4) | 1.00 g | Phosphorus source, buffering |
| Ammonium Nitrate (NH4NO3) | 1.00 g | Nitrogen source |
| Ferric Chloride (FeCl3.6H2O) | 0.05 g | Trace nutrient, iron source |
| Agar | 20.00 g | Solidifying agent |
Final pH: 7.0 ± 0.2 at 25°C. Total: 23.27 g/L. Contains no carbon source — supplied by the user as a hydrocarbon supplement.
Calcium chloride is anhydrous (CaCl2), consistent with the original Bushnell and Haas (1941) formula and current HiMedia/peer-reviewed technical documentation — not the dihydrate form at the same mass.
Preparation
Suspend 23.27 g in 1 L distilled or purified water. Heat with gentle agitation and bring to boiling to dissolve completely. Dispense into suitable containers. Autoclave at 121°C for 15 minutes. Cool and pour plates.
Incubation Conditions
| Parameter | Condition |
|---|---|
| Temperature | 25–30°C |
| Time | Up to 1–2 weeks (hydrocarbon-degrading organisms may grow slowly) |
| Atmosphere | Aerobic |
| Endpoint | Colony formation on hydrocarbon-supplemented plate |
Quality Control — Cultural Response
| Organism / Control | ATCC No. | Test Condition | Expected Result |
|---|---|---|---|
| Pseudomonas aeruginosa | 27853 | Plate + sterile mineral oil overlay, inoculated | Colony growth within 1 week at 25–30°C |
| Uninoculated control | — | Plate + mineral oil, no inoculum | No growth (sterility control) |
| Negative growth control | — | Inoculated plate, no hydrocarbon added | No significant growth (confirms carbon-free selectivity) |
Storage & Shelf Life
| Dehydrated medium | 10–30°C, tightly closed, protected from moisture/light — 24 months |
| Prepared plates | 2–8°C, inverted — use within 2 weeks |
Research Design Notes
Growth on this medium is best interpreted as one line of evidence for hydrocarbon-utilisation capability, not proof of complete degradation. For biodegradation claims, pair microbiological growth with an analytical measurement of substrate depletion (e.g., GC-FID) or metabolite formation. Recommended controls include a no-carbon inoculated control, an abiotic substrate control, and a positive growth control in a separate suitable medium.
Literature & References
- Bushnell, L.D., Haas, H.F. (1941). The utilization of certain hydrocarbons by microorganisms. Journal of Bacteriology, 41(5), 653–673. DOI: 10.1128/jb.41.5.653-673.1941.
- SIM Committee on Microbiological Deterioration of Fuels. Recommended procedures for the microbiological examination of fuels.
For laboratory research and industrial use only. Not intended for food, pharmaceutical, or veterinary applications. Not for diagnostic use without proper validation. AuSaMicS Pty Ltd accepts no liability for results obtained outside the stated preparation and storage conditions.
Safety Data Sheet — Summary
This is a summary view. The full 16-section GHS-compliant Safety Data Sheet (Australian format) is provided as a downloadable Word document alongside this page.
| Product Name | Bushnell-Haas Agar, AS-1161 |
| Recommended Use | Laboratory culture medium (dehydrated) |
| Manufacturer | AuSaMicS Pty Ltd, Melbourne, Victoria, Australia |
| GHS Classification | Not classified as hazardous per Safe Work Australia criteria (dehydrated powder) |
| Signal Word | None allocated |
Hazard Summary
The dehydrated medium is not classified as hazardous under Safe Work Australia / GHS criteria but may be irritating to eyes, skin, and the respiratory tract as a fine powder. Note: any hydrocarbon supplement added by the user carries its own hazards and must be handled per its own SDS — those hazards are outside the scope of this base medium's SDS.
First Aid
| Route | Response |
|---|---|
| Inhalation | Move to fresh air. Seek medical attention if irritation persists. |
| Skin Contact | Wash with soap and water. |
| Eye Contact | Rinse thoroughly with water for several minutes. Seek medical advice if irritation persists. |
| Ingestion | Rinse mouth. Seek medical advice if unwell. |
Full SDS (16 GHS sections, handling, disposal, transport and regulatory information) is provided as a separate Word document — see downloads.
Certificate of Analysis — Sample Format
Each batch is supplied with a lot-specific Certificate of Analysis. A representative COA is provided as a downloadable Word document; the batch-specific COA accompanies your shipment.
| Product Name | Bushnell-Haas Agar |
| Catalogue Number | AS-1161 |
| Batch/Lot Number | Assigned per production batch |
| Manufacture Date | Assigned per production batch |
| Retest / Expiry Date | 24 months from manufacture |
Quality Control Results (Representative)
| Test | Specification | Result |
|---|---|---|
| Appearance (powder) | Fine, homogeneous, free-flowing | Conforms |
| Final pH (25°C) | 7.0 ± 0.2 | Conforms |
| Gel firmness | Firm, comparable with 2.0% agar gel | Conforms |
| Precipitate after autoclaving | Yellow-orange (normal) | Conforms |
| P. aeruginosa ATCC 27853 + mineral oil | Colony growth within 1 week | Conforms |
| Sterility (uninoculated control) | No growth | Conforms |
This tab shows a representative format. The lot-specific Certificate of Analysis issued with your order carries the actual batch number, manufacture/expiry dates, and signed QC results.
The information provided is accurate to the best of AuSaMicS Pty Ltd's knowledge at the time of publication. Product specifications may be updated without notice. For laboratory research and industrial use only — not intended for food, pharmaceutical, or veterinary applications, and not for diagnostic use without proper validation. Users should verify suitability for their specific application prior to use.