TPGY Broth Base w/o Trypsin | Tryptone Peptone Glucose Yeast Extract | AS‑1366
$37.00 AUD
AuSaMicS Life Science • Anaerobic Enrichment Culture Media
Tryptone Peptone Glucose Yeast Extract (TPGY) Broth Base
w/o Trypsin — Clostridium botulinum & Anaerobic Spore-Former Enrichment
The gold-standard enrichment broth for recovery and cultivation of Clostridium botulinum and anaerobic spore-forming bacteria from food, environmental, and clinical specimens. Formulated without trypsin — trypsin is added separately as required for neurotoxin activation in botulism testing workflows per FDA BAM Chapter 17.
Cat #: AS-1366 FDA BAM Ch. 17 Compliant ✓ Australian Stock Without Trypsin
🚀 Same-Day Dispatch: Australian Stock 🧪 Botulism Testing: FDA BAM / AOAC 977.26 🔬 Incubation: 35–37°C / 5–7 days anaerobic
🏋 Botulism Enrichment Specialist
🛒 No Trypsin Included — Add separately as required
🧬 Highly Nutritious Formula — Supports fastidious anaerobes
📋 Complete Package — COA + SDS included
⚡ Fast Supply — Ships within 24h
📞 Expert Support — Protocol optimization
Static & Agitated Anaerobic Culture
Validated for both static enrichment and gentle agitation anaerobic protocols
🔬 Technical Overview & Biochemistry
TPGY Broth Base is a rich, non-selective enrichment medium formulated for isolation and cultivation of Clostridium botulinum and other obligate anaerobic spore-forming bacteria from food, environmental, and clinical specimens. Supplied without trypsin — add trypsin separately for neurotoxin activation per FDA BAM Chapter 17.
Nutritional Components:
Tryptone — Enzymatic casein digest; rich peptides and amino acids for fastidious anaerobes
Peptone — Meat-derived protein digest; supplementary nitrogen and micronutrients
Glucose (Dextrose) — Fermentable carbohydrate; drives anaerobiosis via CO₂ production
Yeast Extract — B-vitamins, nucleotide precursors, trace minerals for obligate anaerobes
Sodium Thioglycollate — Reduces Eh; creates the reduced microenvironment required for spore germination
Tryptone — Enzymatic casein digest; rich peptides and amino acids for fastidious anaerobes
Peptone — Meat-derived protein digest; supplementary nitrogen and micronutrients
Glucose (Dextrose) — Fermentable carbohydrate; drives anaerobiosis via CO₂ production
Yeast Extract — B-vitamins, nucleotide precursors, trace minerals for obligate anaerobes
Sodium Thioglycollate — Reduces Eh; creates the reduced microenvironment required for spore germination
Why Without Trypsin? Trypsin activates the progenitor toxin complex into its fully toxic form prior to mouse bioassay or ELISA confirmation. Supplying the base without trypsin gives laboratories full flexibility to add it at the precise workflow stage.
⚙️ Mode of Action
TPGY Broth provides the combined nutritional, reductive, and osmotic conditions for C. botulinum spore germination and vegetative growth under anaerobic incubation. Glucose fermentation drives down dissolved O₂ and redox potential, complementing sodium thioglycollate to create a strictly anaerobic microenvironment. Yeast extract supplies growth cofactors that overcome biosynthetic limitations of obligate anaerobes, significantly enhancing recovery from heavily processed food matrices where spore populations may be low-density or sub-lethally stressed.
📊 Anaerobic Enrichment Media Comparison
🧬 Applications
🧪 Food Safety — Botulism Testing
Primary enrichment broth for C. botulinum detection in low-acid canned foods, meat products, fish, honey, and fermented foods per FDA BAM Chapter 17 and AOAC 977.26. Trypsin added post-enrichment for neurotoxin activation prior to mouse bioassay or ELISA confirmation.
🛒 Wound & Infant Botulism
Anaerobic enrichment of wound swabs, tissue samples, and stool for C. botulinum recovery in wound botulism and infant botulism case investigation. Used in reference public health and clinical microbiology laboratories.
🌍 Environmental & Soil Monitoring
Recovery of C. botulinum spores from soil, sediment, compost, and water samples. Used in ecological surveys, agricultural land risk assessments, and avian botulism investigations.
🔬 Anaerobic Research
General enrichment and maintenance of obligate anaerobes and Clostridium spp. in research settings. Supports anaerobe physiology, sporulation studies, toxin gene expression, and industrial anaerobic fermentation.
Additional Laboratory Applications:
- Infant Botulism: Stool enrichment for C. botulinum spore recovery
- NATA Accredited Labs: Botulism reference testing panels
- Outbreak Investigation: Foodborne botulism source tracing
- Pharmaceutical QC: Supplementary anaerobe sterility testing
- Aquaculture: Sediment and fish tissue C. botulinum monitoring
- Industrial Fermentation: Anaerobic process organism cultivation
📋 Recommended Procedure (FDA BAM Chapter 17 / AOAC 977.26)
🧪 Method A — Food Sample Enrichment
- Homogenise 10 g food sample in 90 mL TPGY Broth (1:10 dilution)
- Transfer aliquots into TPGY Broth tubes
- Heat-shock duplicate aliquots at 80°C / 15 min (spore selection)
- Incubate heated and unheated at 35°C under strict anaerobic conditions for 5–7 days
- For toxin testing: add trypsin (0.1% final) to turbid cultures; 37°C / 30 min before ELISA/bioassay
🔬 Method B — Environmental Enrichment
- Suspend 1–5 g soil/sediment in 20 mL TPGY Broth
- Mix; allow large particles to settle (10 min)
- Transfer supernatant to fresh TPGY Broth tubes
- Heat-shock 80°C / 15 min for spore selection
- Incubate 35–37°C anaerobic / 7–14 days; subculture to isolation agar
🪲 Trypsin Activation Protocol (for Toxin Testing)
Trypsin Source: Use trypsin from porcine pancreas (activity: 1:250); add to 0.1–0.5% (w/v) final concentration in enrichment culture
Activation Conditions: 37°C for 30–60 min; gentle mixing. Do NOT overheat — excessive heat inactivates the toxin
Neutralisation: After activation, add soybean trypsin inhibitor (SBTI) if required before downstream testing to stop trypsin activity
Safety Note: Handle trypsin-activated cultures as Biosafety Level 2 (BSL-2) material; follow institutional botulinum toxin handling procedures
⚠️ Critical: Strict anaerobic conditions are required throughout incubation. Prepare and dispense TPGY Broth freshly before use; pre-reduce medium in anaerobic chamber if available. For spore-selective testing, heat-shock at 80°C / 15 min is mandatory to destroy vegetative cells.
💡 Protocol Optimization Guidelines
Preparation:
Dissolve 33.0 g/L in purified water. Heat gently to dissolve. Autoclave 121°C / 15 min. Cool to 48–50°C before dispensing to tubes.
Dissolve 33.0 g/L in purified water. Heat gently to dissolve. Autoclave 121°C / 15 min. Cool to 48–50°C before dispensing to tubes.
pH (25°C):
7.2 ± 0.2 after sterilisation
7.2 ± 0.2 after sterilisation
Pre-reduction:
For optimal spore recovery, pre-reduce broth in anaerobic jar or chamber for 24 h before inoculation, especially with dry or aged samples.
For optimal spore recovery, pre-reduce broth in anaerobic jar or chamber for 24 h before inoculation, especially with dry or aged samples.
Storage (prepared broth):
2–8°C, use within 2 weeks. Dehydrated powder: below 25°C, dry and dark. Discard if medium shows turbidity or discolouration before use.
2–8°C, use within 2 weeks. Dehydrated powder: below 25°C, dry and dark. Discard if medium shows turbidity or discolouration before use.
📋 Technical Specifications
| Catalogue Number | AS-1366 |
| Common Name | TPGY Broth / TPGY Medium |
| Format | Dehydrated powder |
| Trypsin | Not included — add separately |
| Reconstitution | ~33.0 g/L in purified water |
| pH (25°C) | 7.2 ± 0.2 |
| Incubation Temp | 35–37°C |
| Incubation Time | 5–7 days (food); 7–14 days (environmental) |
| Atmosphere | Strictly anaerobic |
| Growth Indicator | Turbidity / gas production |
| Sterilisation | Autoclave 121°C / 15 min |
| HS Code | 3821.00.00 |
✓ Quality Control Performance
| Test Organism | ATCC | Expected Result | CFU Spec |
|---|---|---|---|
| Clostridium sporogenes | 19404 | Good growth, turbidity | ≤100 CFU |
| Clostridium perfringens | 13124 | Good growth, turbidity | ≤100 CFU |
| Bacteroides fragilis | 25285 | Good growth | ≤100 CFU |
| Escherichia coli | 25922 | Inhibited under anaerobic conditions | ≤100 CFU |
🧪 Typical Formulation (per litre)
| Tryptone | 17.0 g |
| Peptone | 5.0 g |
| Yeast Extract | 5.0 g |
| Glucose (Dextrose) | 4.0 g |
| Sodium Thioglycollate | 1.0 g |
* Formulation may vary slightly between lot numbers. Refer to the current COA for exact composition. Trypsin is NOT included.
📜 Standards & Regulatory Compliance
- ✓ FDA BAM Chapter 17 — Clostridium botulinum enrichment in foods
- ✓ AOAC 977.26 — Clostridium botulinum and its toxins in foods
- ✓ USDA FSIS MLG 10 — Detection of C. botulinum in meat and poultry products
- ✓ AFNOR NF V08-062 — Anaerobic enrichment of food samples
- ✓ NATA Accreditation — Suitable for NATA-accredited food and reference laboratories
📝 Alternative Names
TPGY Broth Base (w/o Trypsin)
Tryptone Peptone Glucose Yeast Extract Broth
TPGY Medium
Clostridium botulinum Enrichment Broth
Anaerobic Spore-Former Enrichment Broth
🔄 Cross-Reference / Equivalent Products
Cross-references are provided for convenience. Catalogue numbers are trademarks of their respective owners. AuSaMicS products are manufactured and quality-tested independently.
📚 Literature & References
- Dezfulian M, Hatheway CL, Volpe JP, Dowell VR Jr. (1984). Combination selective medium for isolation of Clostridium botulinum from human faeces. J Clin Microbiol. 20(2):74–75.
- Hauschild AHW, Dodds KL. (1993). Clostridium botulinum: Ecology and Control in Foods. Marcel Dekker, New York.
- FDA Bacteriological Analytical Manual (BAM) Chapter 17: Clostridium botulinum. U.S. Food and Drug Administration. Current edition.
- AOAC International (1995). AOAC Official Method 977.26: Clostridium botulinum and its toxins in foods. AOAC Int., Gaithersburg, MD.
- USDA FSIS Microbiology Laboratory Guidebook (MLG) 10.00: Detection of Clostridium botulinum in meat and poultry products. Current edition.
- Dowell VR Jr, McCroskey LM, Hatheway CL, Lombard GL, Hughes JM, Merson MH. (1977). Coproexamination for botulinal toxin and Clostridium botulinum. JAMA. 238(18):1829–1832.
🧬 Complete Anaerobic & Botulism Testing System
Clostridium & Anaerobe Media Range
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Need Clostridium botulinum Testing Method Support?
Our food and anaerobic microbiology specialists can assist with FDA BAM Chapter 17 workflows, trypsin activation protocols, mouse bioassay and ELISA toxin detection, anaerobic incubation system selection, heat-shock spore enrichment, and NATA documentation for botulism reference testing.
For laboratory, research, and industrial use only. Not for food, feed, household, cosmetic, therapeutic, or personal use. Results should be interpreted by qualified laboratory personnel in accordance with applicable standard methods. Strictly anaerobic incubation conditions are required for valid results. Trypsin is not included and must be sourced separately; follow all applicable biosafety procedures when handling potentially toxigenic cultures.
AuSaMicS Pty Ltd | ABN: 56 676 640 467 | 31 Longview CT, Thomastown VIC 3074, Australia
www.ausamics.com.au | support@ausamics.com.au | +61 412 520 598
Same-day dispatch • Australian stock • Full documentation included