Here’s a professional, course-ready infographic-style content layout for Contamination Diagnostics and Troubleshooting in Aquilaria tissue culture, designed for COPI labs, SOP manuals, and training courses. You can later convert this directly into a visual poster or infographic.
Contamination Diagnostics & Troubleshooting in Agarwood Tissue Culture
1. Types of Contaminants
| Contaminant | Visual Signs | Growth Pattern | Likely Source |
|---|---|---|---|
| Bacteria | Cloudy or turbid media, slimy film, tiny colonies | Rapid, spreads throughout media | Explant surface, operator handling, water |
| Fungi | White, grey, black mycelium; spores | Slower than bacteria, aerial growth | Airborne spores, poor asepsis |
| Yeasts | Milky or turbid patches | Moderate growth | Explant tissue or water |
| Endophytic microbes | Delayed contamination (7–14 days), localized | Slow, often hidden in tissue | Internal explant contamination, resistant to surface sterilization |
2. Diagnostic Methods
- Visual Inspection
- Daily check for turbidity, discoloration, mycelium growth, bubbles.
- Microscopic Examination
- Differentiate bacteria from fungal spores.
- Media Response
- pH changes or color shifts in indicator media.
- Sample Testing (Optional)
- Swab tissue, plate on selective media for identification.
3. Troubleshooting Table
| Problem | Possible Cause | Corrective Action |
|---|---|---|
| Cloudy media, rapid spread | Bacterial contamination | Isolate culture, discard contaminated media, review sterilization |
| Mycelium on explant | Fungal contamination | Remove affected tissue, transfer clean portions, improve sterilization |
| Slow contamination | Endophytes | Pre-treat mother plant, optimize sterilization agents, consider HgCl₂ under safety protocols |
| Browning / necrosis | Over-sterilization or oxidative stress | Reduce sterilant exposure, rinse thoroughly, use antioxidants |
| Recurrent batch contamination | Airborne spores, operator error | Check LAF airflow, clean surfaces, retrain personnel |
4. Corrective Workflow (COPI SOP)
- Identify type – visual/microscopic confirmation
- Isolate affected culture – prevent spread
- Determine source – explant, media, environment, operator
- Decide action – discard or salvage tissue
- Document – date, batch, contaminant type, corrective steps
- Revise SOP / Retrain personnel – prevent recurrence
5. Preventive Measures
- Strict adherence to aseptic technique
- Regular LAF cabinet maintenance & HEPA filter replacement
- Use fresh sterilants & sterile consumables
- Monitor and quarantine new explants
- Maintain a contamination log for continuous improvement
6. Key Principle for COPI Course
Early identification and systematic troubleshooting of contamination ensures high survival, preserves genetic fidelity, and supports reproducible, high-quality Aquilaria propagation.
If you want, I can convert this into a print-ready infographic / lab poster with flow diagrams, icons for contaminant types, and stepwise troubleshooting arrows to make it visually intuitive for technicians and trainees.
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