FusaPrime™ Formulation Guide

Product Type: Biological Fungal Inoculant
Application: Agarwood Resin Induction (Aquilaria spp.)

1. Formulation Objective

To develop a stable, high-viability liquid inoculant containing a resin-inducing Fusarium oxysporum strain that:

• Maintains ≥1×10⁶ CFU/mL viability
• Ensures uniform injection performance
• Provides 6-month refrigerated shelf life
• Supports consistent resin induction across field conditions

2. Core Formulation Architecture

• Active Biological Phase
  • Microorganism: Fusarium oxysporum (resin-inducing strain)
  • Target Concentration:
    • Standard: 1×10⁶ CFU/mL
    • Premium: 1×10⁷–10⁸ CFU/mL

Must be:

• Non-lethal to host tree
• High colonization efficiency
• Stable under storage conditions

• Carrier System (Liquid Base)
  • Sterile Water (USP-grade): 95–99%
  • Function:
    • Suspension medium
    • Injection flow optimization
    • Maintains low viscosity (1–5 cP)

• Nutrient Micro-Support System

(Optional but recommended for viability stability)

Component	Function	Range
Peptone	Nitrogen source	0.1–0.3%
KH₂PO₄ / K₂HPO₄	Buffer system	0.05–0.2%
Glucose (minimal)	Energy source	0.05–0.1%

Keep nutrients low to avoid:

• Overgrowth during storage
• Premature metabolic activity

• Stabilization System

Component	Function	Range
Glycerol	Osmotic stabilizer	0.1–0.3%
Trehalose (optional)	Cell membrane protection	0.05–0.2%

Critical for:

• Cold-chain stability
• Viability retention during transport

• pH Control System
  • Target pH: 5.5 – 6.5
  • Adjust using:
    • Citric acid (downward)
    • NaOH (upward)

Optimal for:

• Fungal stability
• Contamination resistance

3. Formulation Process Flow

• Step 1 – Base Solution Preparation
  • Prepare sterile water
  • Dissolve:
    • Nutrient buffer components
    • Stabilizers
  • Adjust pH to 5.8–6.2

• Step 2 – Sterilization
  • Autoclave at 121°C for 15–20 minutes
  • Cool to 25–30°C before inoculation

• Step 3 – Inoculation
  • Introduce seed culture (5–10% v/v)
  • Perform under laminar flow / aseptic conditions

• Step 4 – Controlled Growth Phase
  • Incubate:
    • 25–28°C
    • 48–72 hours
  • Agitation:
    • Lab: 100–150 rpm
    • Industrial: controlled impeller

Target: Achieve desired CFU without overgrowth

• Step 5 – Stabilization Phase
  • Stop active growth at target CFU
  • Add final stabilizer adjustment (if needed)
  • Gently mix to avoid shear stress

• Step 6 – Filtration / Clarification (Optional)
  • Remove large mycelial clumps (if needed)
  • Maintain injectable consistency

• Step 7 – Final QC Adjustment
  • Verify:
    • CFU concentration
    • pH
    • Sterility

• Step 8 – Aseptic Filling
  • Fill into sterile containers
  • Seal immediately
  • Store at 4–10°C

4. Critical Control Points (CCPs)

Stage	Parameter	Risk	Control
Inoculation	Sterility	Contamination	Laminar flow
Growth	CFU density	Overgrowth	Timed incubation
Formulation	pH	Viability loss	Continuous monitoring
Packaging	Exposure	Contamination	Aseptic filling
Storage	Temperature	CFU decline	Cold chain

5. Stability Profile

Condition	Expected Stability
4–10°C	6 months
25°C	2–4 weeks
>30°C	Rapid viability loss

6. Formulation Optimization Options

• High-Performance Variant
  • CFU: ≥10⁷ CFU/mL
  • Slightly higher stabilizer content
  • Used for high-value plantations

• Extended Shelf-Life Variant
  • Add trehalose + glycerol blend
  • Optional mild preservative (if compliant)

• Rapid Induction Variant (R&D)
  • Micro-nutrient enrichment
  • Designed for faster colonization (experimental)

7. Compatibility Notes

• Fully compatible with:
  • BarIno™ FusaBlaze™ (enzymatic booster)
  • BarIno™ FusaTrinity™ (dual induction system)
• Should NOT be mixed directly with strong chemicals before injection

8. Documentation Requirements (GMP)

Each batch must include:

• Raw material traceability
• CFU certificate
• pH record
• Sterility report
• Batch number & expiry

9. Technical Positioning Insight

This formulation is designed to:

• Act as a biological ignition phase
• Enable predictable resin initiation
• Serve as the foundation for premium sequential induction systems

10. Formulation Summary

Parameter	Specification
CFU	10⁶–10⁸ CFU/mL
pH	5.5–6.5
Appearance	Clear–slightly turbid
Shelf Life	6 months (cold)
Application	Injectable

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