Standard Operating Procedures (SOP) Manual

Here is a Draft Standard Operating Procedures (SOP) Manual for an In-House Microbial Inoculant Laboratory, specifically designed for Agarwood inoculant research and production.

🌿 Crown MycoResin Biotech Inc.

In-House Microbial Inoculant Laboratory
Standard Operating Procedures (SOP) Manual
Version 1.0 | July 2025

Table of Contents

  1. Introduction
  2. Laboratory Workflow Overview
  3. Personnel Responsibilities
  4. General Laboratory Safety
  5. Aseptic Techniques
  6. Microbial Culture Preparation
  7. Inoculant Formulation and Mixing
  8. Quality Control and Testing
  9. Packaging and Labeling
  10. Documentation and Record-Keeping
  11. Waste Management
  12. Equipment Maintenance
  13. Emergency Protocols
  14. Annexes (Forms, Templates, Logs)

1. Introduction

This SOP manual defines procedures and safety guidelines for the production of microbial inoculants (biotic and chemical-biotic blends) used in Agarwood resin induction. The lab focuses on cultivating Fusarium oxysporum, enzyme-producing microbes, and preparing stabilized liquid formulations.

2. Laboratory Workflow Overview

  1. Strain Revival & Culture Maintenance
  2. Spore Suspension or Enzyme Extraction
  3. Formulation of Inoculants (e.g., BarIno™ Series)
  4. Quality Control & Microbial Enumeration
  5. Packaging (1-liter bottles)
  6. Storage & Distribution

3. Personnel Responsibilities

RoleResponsibilities
Lab ManagerSupervise operations, maintain records
MicrobiologistCulture handling, identification, safety
Lab TechnicianMedia prep, inoculation, formulation
QA OfficerMonitor QC parameters, documentation
Sanitation StaffClean workspace, dispose of waste

4. General Laboratory Safety

  • Always wear lab coat, gloves, and face mask.
  • Disinfect workspaces with 70% ethanol before and after work.
  • No food or drinks allowed.
  • Handle pathogenic cultures in biosafety cabinet.
  • Dispose of used media and glassware according to biosafety protocols.

5. Aseptic Techniques

  • Work near an open flame or in a laminar flow hood.
  • Sterilize tools using autoclave or alcohol flame.
  • Avoid unnecessary talking or movement while culturing.
  • Label all materials clearly with name, date, and organism.

6. Microbial Culture Preparation

6.1. Revival of Fusarium oxysporum Spores

  • Use PDA or Czapek-Dox Agar.
  • Incubate at 28°C for 5–7 days.
  • Check for sporulation using microscope.

6.2. Spore Suspension

  • Scrape mature spores with sterile saline + Tween 80 (0.01%).
  • Filter to remove hyphal debris.
  • Standardize spore count (10⁶–10⁷ spores/mL) using hemocytometer.

7. Inoculant Formulation and Mixing

Example: BarIno™ BioFusion™

  • Mix spore suspension + enzyme filtrate + carrier (e.g., AgarStart™ base).
  • Adjust pH (6.5–7.0) and osmotic concentration.
  • Add stabilizers (e.g., glycerol, molasses).
  • Store in sterile bottles under cool, dark conditions.

8. Quality Control and Testing

  • Microbial purity: streak test on selective media.
  • Spore viability: germination test.
  • Contamination check: 24–48h culture monitoring.
  • pH, EC, and viscosity consistency check.
  • Random sampling before release.

9. Packaging and Labeling

  • Use sterile 1-liter HDPE bottles.
  • Label includes:
    • Product name (e.g., BarIno™ FusaPrime™)
    • Strain and formulation details
    • Date of production and expiry
    • Storage instructions
  • Batch number traceability is mandatory.

10. Documentation and Record-Keeping

Maintain logs for:

  • Media preparation
  • Culture dates and batches
  • Formulation details
  • QC results
  • Packaging & dispatch records
  • Waste disposal

11. Waste Management

  • Autoclave biological waste before disposal.
  • Use biohazard bags for contaminated disposables.
  • Dispose of chemical waste per local DENR/EMB regulations.
  • Keep a log of all waste outputs.

12. Equipment Maintenance

  • Autoclave: Weekly validation using indicator strips.
  • pH meter: Calibrate weekly.
  • Laminar hood: UV sterilization daily.
  • Refrigerator: Temp log maintained daily.
  • Shaker incubators: Monthly calibration.

13. Emergency Protocols

  • Spill response: Cover with bleach-soaked tissue, wait 15 min, wipe and discard.
  • Exposure: Wash skin immediately, report to safety officer.
  • Power failure: Transfer cultures to backup incubator if >2 hrs.
  • Fire/chemical hazard: Follow facility evacuation procedures.

14. Annexes

  • Media Preparation Sheets
  • Daily Sanitation Checklist
  • Inoculant Batch Record Sheet
  • QC Test Forms
  • Label Templates
  • Waste Disposal Log
  • Equipment Calibration Record

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