Methods of Quality Control and Techniques of
Biopesticides

Ajeet Kumar

► To cite this version:

Ajeet Kumar. Methods of Quality Control and Techniques of Biopesticides. Master. Bio-pesticides and Bio-fertilizers, DR. RAJENDRA PRASAD CENTRAL AGRICULTURAL UNIVERSITY PUSA, SAMASTIPUR-848125, BIHAR, INDIA, India. 2025, pp.1-4. [hal-05124497](https://hal.science/hal-05124497v1)

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Distributed under a Creative Commons Attribution 4.0 International License
Dr. Rajendra Prasad Central Agricultural University
Pusa (Samastipur) - 848125, Bihar

*A lecture notes*

*On*

*Methods of Quality Control and Techniques of Biopesticides*

<figure><img src="image_2.png" /></figure>

COURSE TITLE: Bio-pesticides and Bio-fertilizers

*(Elective course)*

COURSE NO.: ASS-307; CREDIT HOUR: 3 (2+1)

YEAR/SEMESTER: 3rd YEAR / 6th SEMESTER

Dr. AJEET KUMAR

Assistant Professor-cum-Scientist (Soil Science)

Department of Microbiology

Collage of Basic Sciences and Humanities (CBS&H)

Dr. Rajendra Prasad Central Agricultural University, Pusa (Samastipur)

June 2025
# Methods of Quality Control and Techniques of Biopesticides

Dr. AJEET KUMAR<fnref n="1" />

## 1. Introduction

Quality control (QC) in biopesticide production ensures **efficacy**, **safety**, and **shelf-life** of microbial agents before commercial use. Standardized QC techniques are essential to comply with **regulatory guidelines** and maintain **bioefficacy** across production batches.

## 2. Importance of Quality Control in Biopesticides

* Ensures **purity**, **viability**, and **correct strain identity**

* Prevents contamination with unwanted microorganisms

* Assures **consistency** in field performance

* Necessary for **registration** under Insecticide Act (1968), CIB & RC (India)

## 3. Quality Parameters for Common Biopesticides

<table><thead><tr><td>Biopesticide Type</td><td>Minimum Viable Count</td><td>Purity (Contaminants)</td><td>Moisture (%)</td><td>pH Range</td></tr></thead><tbody><tr><td>Bacillus thuringiensis</td><td>1 × 10<sup>9</sup> CFU/g (solid) or ml (liquid)</td><td>&lt; 0.1%</td><td>&lt; 10%</td><td>6.5-7.5</td></tr><tr><td>Trichoderma spp.</td><td>2 × 10<sup>6</sup> CFU/g</td><td>&lt; 0.1%</td><td>&lt; 20%</td><td>6.0-7.5</td></tr><tr><td>Beauveria bassiana</td><td>1 × 10<sup>7</sup> spores/g</td><td>&lt; 0.1%</td><td>&lt; 10%</td><td>6.0-7.5</td></tr><tr><td>NPV (HaNPV)</td><td>1 × 10<sup>9</sup> PIB/ml</td><td>No bacterial/fungal growth</td><td>Liquid</td><td>5.5-6.5</td></tr></tbody></table>

## 4. Quality Control Techniques

### A. Viability Testing

* **Bacteria**: Serial dilution and plate count method on Nutrient Agar

* **Fungi**: Spore germination on PDA plates (viable if ≥ 90% spores germinate)

* **Nematodes**: Visual movement under microscope in distilled water

* **Viruses**: Occlusion bodies counted using a hemocytometer

### B. Purity Testing

<footnote n="1">¹Assistant Professor-cum-Scientist (Soil Science-Soil Microbiology, Soil fertility, Soil Chemistry), Department of Microbiology, College of Basic Sciences & Humanities (CBS&H), RPCAU, Pusa-848125, Bihar. E-mail: ajeet.sri@rpcau.ac.in</footnote>
* Check for contamination using general growth media:

* **Nutrient Agar, Potato Dextrose Agar (PDA)**

* Observe colony morphology and identify contaminating species

* Should not exceed 0.1% contamination

## C. Moisture Content Analysis

* **Gravimetric Method:** Dry 1g sample at 105°C for 6 hrs

* Moisture content = (Initial weight – Dry weight) / Initial weight × 100

* Lower moisture ensures longer shelf-life

## D. pH Testing

* Use **digital pH meter** or indicator strips

* Adjusted with buffer solutions or stabilizers

## E. Pathogen Identification & Authentication

* **Microscopy:** Spore or bacterial morphology

* **Molecular Techniques:**

* PCR for strain-specific genes

* DNA barcoding or sequencing

* RAPD or AFLP fingerprinting

## F. Bioassay for Efficacy (Potency Test)

* Example: *Bt* tested against 3rd instar *Helicoverpa armigera* larvae

* Parameters:

* LC₅₀ (lethal concentration)

* LT₅₀ (lethal time)

* Compare with reference strain

## 5. Packaging and Labeling Standards

* Must be moisture-proof, UV-protective, and properly labeled with:

* Active ingredient

* Viable cell/spore count

* Expiry date

* Batch number

* Application method

## 6. Regulatory Guidelines in India

* **CIB & RC** – Central Insecticides Board and Registration Committee

* Must meet BIS (Bureau of Indian Standards) specifications
* QA/QC labs should be accredited under ISO/IEC 17025

**7. Diagram: Quality Control Workflow**

[Raw Strain] → [Media Optimization] → [Fermentation] → [Harvesting] → [Viability Test] → [Contamination Check] → [Packaging] → [Labeling] → [Release]

**8. Challenges in Quality Control**

* Variation in raw materials

* Lack of trained personnel in bioassay

* Inadequate storage conditions

* Field inconsistency despite lab QC

**9. Future Technologies in QC**

* Automated colony counters for viable counts

* Real-time PCR for rapid identity testing

* Digital imaging for nematode movement

* Portable bioassay kits for field evaluation