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PPLO Broth | Mycoplasma Broth Base l AS-1333
$115.00 AUD
Documentation
PPLO BROTH BASE
Catalogue No. AS-1333
www.ausamics.com.au
AS-1333 Base Medium Without Crystal Violet Mycoplasma spp.
PPLO Broth (Mycoplasma Broth Base)
Basal culture medium for the isolation and cultivation of Mycoplasma species (pleuropneumonia-like organisms)
PPLO Broth Base is a nutrient-rich basal medium formulated for the cultivation of Mycoplasma species, prepared according to the formula described by Morton and Lecce (1953). Crystal violet, present in earlier PPLO formulations, is deliberately omitted here because it inhibits the growth of some Mycoplasma species.
As a base medium, it requires aseptic supplementation with sterile serum (commonly horse serum) and often fresh yeast extract to support the growth of these fastidious, sterol-requiring organisms. AuSaMicS manufactures this base in Melbourne, Australia, to the verified reference formulation.
Performance Snapshot
Final pH7.8 ± 0.2
Total weight21.0 g/L
Appearance (powder)Pale amber, free-flowing
Appearance (prepared)Yellow-coloured, clear
Shelf life (powder)24 months
Formulation accuracy note: This base medium contains no phenol red and is not pink or red in appearance — it is a pale amber powder producing a yellow-coloured, clear solution once prepared. Phenol red belongs to the fully-supplemented "PPLO Selective Broth" product tier (where it gives a visible red-to-yellow colour shift as a growth indicator), not to this base. Total dehydrated weight is verified at 21.0 g/L (Beef Heart Infusion 6.0g + Peptone 10.0g + Sodium Chloride 5.0g), cross-checked against HiMedia (M1586) and Neogen (NCM0113A) technical specifications, which agree exactly.
Testing Workflow
1
SpecimenSwab, tissue, or clinical sample
2
Inoculate BrothAdd specimen to prepared, supplemented base
3
Incubate35°C, 5-10% CO2, up to 30 days
4
MonitorWatch for turbidity / colour change if indicator supplement used
5
SubcultureTo PPLO Agar for colony confirmation
Principle of the Medium
Origin
PPLO Agar and Broth media trace to Morton, Smith and Leberman (1951), who formulated the original base for isolating and cultivating Mycoplasma species. Hayflick later modified the approach with the addition of yeast extract and unheated horse serum. Morton and Lecce (1953) subsequently removed crystal violet from the formulation after finding it inhibitory to some Mycoplasma — the resulting "without crystal violet" variant is the standard base used today.
Functional Components
Beef Heart Infusion and Peptone supply the amino acids, peptides, and nutrients required for Mycoplasma growth. Sodium Chloride maintains osmotic equilibrium, which is particularly important for Mycoplasma since they lack a rigid cell wall and are therefore more osmotically sensitive than walled bacteria. The base alone does not support growth to a complete endpoint — Mycoplasma require cholesterol and other membrane precursors they cannot synthesise themselves, which is why serum supplementation is essential.
Key Applications
Clinical Diagnostics
Isolation and identification of Mycoplasma pneumoniae and other clinically relevant species.
Cell Culture QC
Contamination screening for Mycoplasma in mammalian cell culture and biopharmaceutical production.
Veterinary Diagnostics
Detection of Mycoplasma bovis, M. hyopneumoniae, and other livestock/companion animal pathogens.
Research
Cultivation of Mycoplasma and L-form bacteria for research applications.
Formulation & Specifications
| Component | g/L |
|---|---|
| Beef Heart Infusion (from 50g fresh tissue) | 6.0 |
| Peptone | 10.0 |
| Sodium Chloride | 5.0 |
| Total | 21.0 g/L |
Technical Specifications
| Catalogue No. | AS-1333 |
| Final pH | 7.8 ± 0.2 @ 25°C |
| Contains crystal violet | No |
| Contains phenol red | No |
| Storage (powder) | 10–30°C, dry, dark |
| Storage (prepared) | 2–8°C, protected from light |
Laboratory Preparation
1
Suspend 21.0 g of dehydrated medium in 700 mL purified water.
2
Heat to boiling to dissolve the medium completely.
3
Sterilise by autoclaving at 121°C for 15 minutes.
4
Cool to 45–50°C and aseptically add 300 mL sterile horse serum or an equivalent enrichment supplement.
5
Mix well and dispense into sterile tubes or flasks as required.
Supplementation Guide
- Horse Serum: aseptically add after autoclaving and cooling — reference procedure uses 300 mL per 700 mL base (30% v/v of final volume); consult your specific protocol, as ratios in the 10–30% v/v range are reported across different sources.
- Yeast Extract: a fresh 25% w/v yeast extract solution is commonly added alongside serum to supply B-complex vitamins and enhance growth.
- Selective agents (optional): penicillin, thallium acetate, or amphotericin B may be added to suppress bacterial or fungal contaminants where a selective formulation is required.
- This base medium as supplied does not include serum, yeast extract, or antibiotics — all must be added by the user according to their validated protocol.
Limitations & Technical Considerations
- Base medium only — will not support Mycoplasma growth without serum and yeast extract supplementation added by the user.
- No visual growth indicator — since the base contains no phenol red, colour-change monitoring requires either adding an indicator yourself or subculturing to agar for colony confirmation.
- Slow growth — some Mycoplasma species require up to 30 days incubation; do not discard cultures as negative before this window has elapsed.
- Ureaplasma urealyticum may be inhibited at this medium's pH; a lower-pH modification is typically used for Ureaplasma recovery.
Cross-Reference / Equivalent Products
| Brand | Equivalent Product | Catalogue No. |
|---|---|---|
| BD Difco | PPLO Broth without Crystal Violet | 255420 |
| BD BBL | Mycoplasma Broth Base (PPLO Broth Base) | 211458 |
| Oxoid (Thermo Scientific) | Mycoplasma Broth Base (similar formula, not exact) | CM0403 |
| Merck / Sigma-Aldrich | Mycoplasma Broth | M0535 |
| HiMedia | PPLO Modified Broth Base w/o CV | M1586 |
| Neogen | PPLO Broth Base without Crystal Violet | NCM0113A |
| AuSaMicS | PPLO Broth Base | AS-1333 |
Ordering Information
| Catalogue No. | Pack Size | Approx. Yield | Format |
|---|---|---|---|
| AS-1333-500 | 500 g | ≈ 23.8 L prepared broth (base only, before serum) | Dehydrated powder, bottle |
| AS-1333-2K5 | 2.5 kg | ≈ 119 L prepared broth (base only, before serum) | Dehydrated powder, bottle |
Yield calculated at 21.0 g/L reconstitution rate for the base only; final supplemented volume will be greater once serum is added (e.g., 700 mL base + 300 mL serum = 1000 mL complete medium, per the reference procedure).
Frequently Asked Questions
What is PPLO Broth Base used for?
It is a basal medium for the isolation and cultivation of Mycoplasma species (pleuropneumonia-like organisms, PPLO), used in clinical, veterinary, and cell culture contamination testing.
Why does PPLO Broth Base not contain crystal violet?
Crystal violet, present in earlier PPLO formulations, was found to inhibit the growth of some Mycoplasma species. The Morton and Lecce (1953) formula removes it for this reason.
Does PPLO Broth Base contain phenol red?
No. The base medium does not contain phenol red. Phenol red is used in the fully-supplemented PPLO Selective Broth product, where it gives a visible red-to-yellow colour shift as a growth indicator.
What supplements does PPLO Broth Base need before use?
Aseptic addition of a sterile serum supplement (commonly horse serum) and often fresh yeast extract after autoclaving and cooling, since Mycoplasma require cholesterol and other growth factors they cannot synthesise themselves.
What colour should prepared PPLO Broth Base be?
A pale amber to yellow-coloured, clear solution. A pink or red colour is not expected from this base medium.
Common Search Queries This Product Answers
Researchers and procurement teams typically find this page searching for:
- "PPLO broth base composition" / "Mycoplasma broth base formula"
- "PPLO broth without crystal violet"
- "Mycoplasma culture medium Australia"
- "does PPLO broth contain phenol red"
- "buy PPLO broth base Australia Melbourne"
- "mycoplasma cell culture contamination testing medium"
Technical Data Sheet
| Product Name | PPLO Broth (Mycoplasma Broth Base) |
| Synonyms | PPLO Broth Base without Crystal Violet |
| Catalogue Number | AS-1333 |
| Format | Dehydrated powder |
| Manufacturer | AuSaMicS Pty Ltd, Melbourne, Australia |
Intended Use
Basal medium for the isolation and cultivation of Mycoplasma species (pleuropneumonia-like organisms), requiring aseptic supplementation with serum and yeast extract to become a complete growth medium.
Mode of Action
Beef Heart Infusion and Peptone supply amino acids and peptides required for Mycoplasma growth. Sodium Chloride maintains osmotic equilibrium, particularly important given Mycoplasma's lack of a rigid cell wall. The base alone is nutritionally incomplete for Mycoplasma — these organisms require exogenous cholesterol and other membrane precursors, supplied by serum supplementation, since their minimal genome cannot synthesise these components.
Formulation (per litre)
| Ingredient | Quantity | Function |
|---|---|---|
| Beef Heart Infusion (from 50g fresh tissue) | 6.0 g | Nutrient source for Mycoplasma growth |
| Peptone | 10.0 g | Amino acid / nitrogen source |
| Sodium Chloride | 5.0 g | Osmotic stabiliser |
Total: 21.0 g/L. Final pH: 7.8 ± 0.2 at 25°C. Contains neither crystal violet nor phenol red.
Formulation verified against HiMedia (M1586) and Neogen (NCM0113A) technical specifications, which independently agree on total weight (21.0 g/L) and composition. Both confirm the prepared medium is yellow-coloured and clear — not pink or red.
Preparation
Suspend 21.0 g in 700 mL purified water. Heat to boiling to dissolve completely. Autoclave at 121°C for 15 minutes. Cool to 45–50°C and aseptically add 300 mL sterile horse serum (or equivalent enrichment supplement). Mix well and dispense into sterile tubes or flasks.
Incubation Conditions
| Parameter | Condition |
|---|---|
| Temperature | 35°C (22–35°C for saprophytic strains) |
| Time | Up to 30 days; do not discard as negative before this period |
| Atmosphere | 5–10% CO2 (most strains); some anaerobic saprophytic strains |
Quality Control — Cultural Response
| Organism | ATCC No. | Expected Result (supplemented medium) |
|---|---|---|
| Mycoplasma bovis | 25523 | Good to luxuriant growth |
| Mycoplasma pneumoniae | 15531 | Good to luxuriant growth |
| Escherichia coli | 25922 | Inhibited (differential control) |
| Staphylococcus aureus | 25923 | Inhibited (differential control) |
Storage & Shelf Life
| Dehydrated medium | 10–30°C, dry, tightly sealed — 24 months |
| Prepared medium | 2–8°C, protected from light |
Literature & References
- Morton, H.E., Lecce, J.G. (1953). Journal of Bacteriology, 66:646.
- Morton, H.E., Smith, P.F., Leberman, P.R. (1951). American Journal of Syphilis, Gonorrhea and Venereal Diseases, 35:361.
- Hayflick, L., Chanock, R.M. (1965). Mycoplasma Species of Man. Bacteriological Reviews, 29(2). American Society for Microbiology.
For laboratory use only. Not intended for use in the diagnosis of disease in humans without appropriate 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 | PPLO Broth Base, AS-1333 |
| Recommended Use | Laboratory culture medium (dehydrated) |
| Manufacturer | AuSaMicS Pty Ltd, Melbourne, Victoria, Australia |
| GHS Classification | Not classified as hazardous per Safe Work Australia criteria |
| Signal Word | None allocated |
Hazard Summary
Not classified as a hazardous chemical or dangerous good under Safe Work Australia / GHS criteria in its dehydrated form. May cause mechanical irritation to eyes and respiratory tract as a fine powder. Contains components of animal origin (Beef Heart Infusion) — handle with standard laboratory precautions.
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 | PPLO Broth (Mycoplasma Broth Base) |
| Catalogue Number | AS-1333 |
| 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) | Pale amber, free-flowing | Conforms |
| Final pH (25°C) | 7.8 ± 0.2 | Conforms |
| Colour/clarity (prepared) | Yellow-coloured, clear solution | Conforms |
| M. bovis ATCC 25523 recovery (supplemented) | Good to luxuriant growth | Conforms |
| E. coli ATCC 25922 (differential control) | Inhibited | 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 use only — not intended for use in the diagnosis of disease in humans without appropriate validation. Users should verify suitability for their specific application prior to use.