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Bushnell-Haas Broth | Hydrocarbon Degrading Bacteria Medium | Ausamics AS-1162
$115.00 AUD
Documentation
BUSHNELL-HAAS BROTH
Catalogue No. AS-1162
www.ausamics.com.au
AS-1162 Carbon-Free Base Petroleum Microbiology Bioremediation Bushnell & Haas (1941)
Bushnell-Haas Broth
Bushnell-Haas Medium — for the isolation, enumeration, and study of hydrocarbon-degrading microorganisms
Bushnell-Haas Broth is a carbon-free mineral salts medium used for the isolation, enumeration, and study of hydrocarbon-degrading microorganisms. Originally developed by Bushnell and Haas (1941), it provides essential inorganic nutrients while deliberately lacking a carbon source, requiring supplementation with a hydrocarbon such as mineral oil, kerosene, paraffin wax, diesel, or gasoline to support microbial growth.
AuSaMicS manufactures Bushnell-Haas Broth in Melbourne, Australia, to the original reference formulation, giving reliable selectivity and consistent lot-to-lot performance for petroleum microbiology and bioremediation workflows.
Performance Snapshot
Final pH7.0 ± 0.2
Carbon sourceNone (user-added)
ReadoutTurbidity in liquid culture
Precipitate after autoclavingYellow-orange — normal
Shelf life (powder)24 months
Testing Workflow
1
SampleSoil, water, fuel, or oil-spill sample
2
InoculateAdd sample to sterile broth
3
Add HydrocarbonAseptically add sterile hydrocarbon source
4
Incubate25–30°C, up to 1–2 weeks
5
ObserveTurbidity = hydrocarbon-degrading growth
Scientific Background & Principle
Origin
Bushnell-Haas Broth was described by Bushnell and Haas (1941) in their classic study of hydrocarbon utilisation by soil microorganisms, and was 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 studies 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) required for microbial growth — but contains no carbon source. This is the defining design feature: because carbon is the one nutrient withheld, only organisms capable of metabolising an externally supplied hydrocarbon as their sole carbon and energy source can grow. Growth itself, observed as turbidity in the liquid medium, becomes the diagnostic signal for hydrocarbon-degrading capability — no additional indicator or stain is needed.
The Yellow-Orange Precipitate
A white precipitate typically forms before sterilisation, turning yellow to orange after autoclaving. This is a normal and expected characteristic of the ferric chloride and phosphate components reacting under heat, and does not indicate contamination or reduced performance.
Key Applications
Bioremediation Research
Degradation studies of total petroleum hydrocarbons (TPH) and polycyclic aromatic hydrocarbons (PAHs).
Petroleum Microbiology
Detection and enumeration of hydrocarbon-utilising microorganisms in soil, water, fuel, and oil-spill samples.
Fuel Quality Control
Microbiological quality control of aviation fuels, lubricants, and other petroleum-based products.
Environmental Monitoring
Monitoring fuel contamination and microbial degradation in impacted sites.
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 |
| Total | 3.27 g/L |
Technical Specifications
| Catalogue No. | AS-1162 |
| HS Code | 3821.00 |
| Format | Dehydrated powder |
| Final pH | 7.0 ± 0.2 @ 25°C |
| Storage (powder) | 10–30°C, dry, dark |
| Storage (prepared) | 2–8°C, use ≤ 2 weeks |
Laboratory Preparation
1
Suspend 3.27 g of dehydrated medium in 1 litre of distilled or purified water.
2
Heat gently with frequent agitation until completely dissolved.
3
Dispense into suitable tubes or flasks.
4
Sterilise by autoclaving at 121°C for 15 minutes.
5
Cool, then aseptically add a sterile hydrocarbon source and inoculate the sample.
Working With Hydrocarbon Supplements
- Liquid hydrocarbons (mineral oil, kerosene, diesel) are typically layered onto the surface of inoculated broth or agar.
- Volatile hydrocarbons (e.g., gasoline) are often added to the lid of an inverted plate rather than directly into the medium, to test vapour-phase utilisation.
- Always add hydrocarbon supplements aseptically and work in a well-ventilated area or fume hood.
- Dispose of hydrocarbon-contaminated materials according to laboratory biosafety and chemical waste procedures.
Limitations & Technical Considerations
- Requires a hydrocarbon supplement — the base broth alone will not support growth; failure to add a carbon source is the most common cause of a false-negative result.
- Not a quantitative TPH/PAH assay by itself — turbidity indicates degrading activity is present, but quantifying the rate or extent of degradation requires additional analytical methods (e.g., GC-FID, gravimetric analysis).
- Extended incubation may be needed — hydrocarbon-degrading organisms can grow slowly; 1–2 weeks of incubation at 25–30°C is common, longer than typical nutrient-rich media.
- Yellow-orange precipitate is normal — do not discard a batch or assume contamination based on this colour change alone.
Cross-Reference / Equivalent Products
| Brand | Equivalent Product |
|---|---|
| BD Difco | Bushnell-Haas Broth |
| HiMedia | Bushnell Haas Broth (M350) |
| Micro Master Laboratories | Bushnell Haas Broth (DM051) |
| AuSaMicS | Bushnell-Haas Broth (AS-1162) |
Ordering Information
| Catalogue No. | Pack Size | Approx. Yield | Format |
|---|---|---|---|
| AS-1162-500 | 500 g | ≈ 153 L prepared medium | Dehydrated powder, bottle |
| AS-1162-2K5 | 2.5 kg | ≈ 765 L prepared medium | Dehydrated powder, bottle |
Yield calculated at 3.27 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 Broth used for?
It isolates, enumerates, and studies hydrocarbon-degrading microorganisms in soil, water, fuel, and oil-spill samples, and is widely used in bioremediation research, petroleum microbiology, and fuel quality control.
Why does Bushnell-Haas Broth have no carbon source?
It is deliberately formulated without a carbon source so that only microorganisms capable of metabolising an added hydrocarbon as their sole carbon and energy source will grow — making growth itself the diagnostic signal.
Why does a precipitate form after autoclaving?
A white precipitate before sterilisation becoming yellow to orange after autoclaving is a normal characteristic of this medium and does not affect its performance.
How do you add the hydrocarbon source?
After sterilising and cooling the base broth, aseptically add a sterile hydrocarbon (e.g., mineral oil, kerosene, diesel, or paraffin). Volatile hydrocarbons such as gasoline are often added to the lid of an inverted plate instead.
What organisms grow in Bushnell-Haas Broth?
Hydrocarbon-utilising microorganisms such as Pseudomonas aeruginosa and other Pseudomonas species are classic examples, along with other environmental bacteria able to use hydrocarbons as their sole carbon and energy source.
Common Search Queries This Product Answers
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Technical Data Sheet
| Product Name | Bushnell-Haas Broth |
| Synonyms | Bushnell-Haas Medium |
| Catalogue Number | AS-1162 |
| Format | Dehydrated powder |
| Manufacturer | AuSaMicS Pty Ltd, Melbourne, Australia |
Intended Use
Carbon-free mineral salts medium for the isolation, enumeration, and study of hydrocarbon-degrading microorganisms in soil, water, fuel, and oil-spill samples, and for microbiological quality control of petroleum-based products.
Mode of Action
The formulation supplies nitrogen, phosphorus, and trace elements (magnesium, calcium, iron) but deliberately contains no carbon source. Only organisms able to metabolise an externally supplied hydrocarbon (e.g., mineral oil, kerosene, diesel, or paraffin) as their sole carbon and energy source can grow. Growth is observed as turbidity in the liquid medium, which serves as the diagnostic endpoint without need for additional indicators.
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 |
Final pH: 7.0 ± 0.2 at 25°C. Total: 3.27 g/L. Contains no carbon source — this must be supplied by the user as a hydrocarbon supplement.
Preparation
Suspend 3.27 g in 1 L distilled or purified water. Heat gently with frequent agitation until completely dissolved. Dispense into tubes or flasks. Autoclave at 121°C for 15 minutes. After cooling, aseptically add a sterile hydrocarbon source before or after inoculation.
A white precipitate before sterilisation becoming yellow to orange after autoclaving is a normal characteristic of this medium and does not affect performance.
Incubation Conditions
| Parameter | Condition |
|---|---|
| Temperature | 25–30°C |
| Time | Up to 1–2 weeks (hydrocarbon-degrading organisms may grow slowly) |
| Atmosphere | Aerobic |
| Endpoint | Turbidity in liquid culture indicates hydrocarbon-degrading growth |
Quality Control — Cultural Response
| Organism | ATCC No. | Test Condition | Expected Result |
|---|---|---|---|
| Pseudomonas aeruginosa | 27853 | Broth + sterile mineral oil, inoculated | Turbidity/growth within 1 week at 25–30°C |
| Uninoculated control | — | Broth + mineral oil, no inoculum | No turbidity (sterility control) |
| Negative growth control | — | Inoculated broth, no hydrocarbon added | No significant turbidity (confirms carbon-free selectivity) |
Storage & Shelf Life
| Dehydrated medium | 10–30°C, tightly closed, protected from moisture/light — 24 months |
| Prepared medium | 2–8°C — use within 2 weeks; discard if turbid, contaminated, or pH shifts significantly |
Literature & References
- Bushnell, L.D., Haas, H.F. (1941). The utilization of certain hydrocarbons by microorganisms. Journal of Bacteriology, 41(5), 653–673.
- SIM Committee on Microbiological Deterioration of Fuels. Recommended procedures for the microbiological examination of fuels.
- American Public Health Association. Standard Methods for the Examination of Water and Wastewater (latest edition).
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 Broth, AS-1162 |
| 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 (mineral oil, kerosene, diesel, petrol) 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 Broth |
| Catalogue Number | AS-1162 |
| 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 |
| Precipitate after autoclaving | Yellow-orange (normal) | Conforms |
| P. aeruginosa ATCC 27853 + mineral oil | Turbidity/growth within 1 week | Conforms |
| Sterility (uninoculated control) | No growth | Conforms |
| Selectivity (inoculated, no hydrocarbon) | No significant turbidity | 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.