Juan González, researcher at the Molecular Biology Unit of Futureco Bioscience and lead scientist behind the development of the ISO 17025-certified method for viable spore quantification in NoFly, shares insights into the innovation. This breakthrough not only sets a new benchmark in biofungicide quality control, but also demonstrates how advanced technologies like automated particle counting can transform accuracy, reproducibility, and regulatory compliance in microbial crop protection products.
What was the most significant technical challenge you encountered while developing the automated particle counting technology, and what approach did you take to resolve it?
The greatest challenge was the selection and use of the fluorophore. With few practical references, we conducted extensive tests with a wide range of differential fluorescent dyes and conditions until we identified a reliable combination. We also observed that, in aqueous solution, NOFLY spores gradually lose their staining capacity due to a reduction in the spore’s metabolic rate, so solubilization and staining times became critical. We resolved this by optimizing dye chemistry, contact time, and sample handling, ensuring the method supports consistent biofungicide quality control.
Achieving ISO 17025 accreditation is a rigorous process. What specific aspects of the method’s validation were most critical in securing certification, and how does this set it apart from traditional quality control approaches?
Reproducibility and repeatability across analysts, days, and batches were decisive. Biological materials are heterogeneous, and batch-to-batch variability can interfere with measurement. We validated precision and accuracy using certified reference materials and multiple NOFLY manufacturing lots. The protocol includes predefined control points with strict verification checks, and we run technical replicates and duplicate samples for each measurement to ensure robustness. This approach reinforces robust biofungicide quality control, surpassing traditional plate count methods.
The method was benchmarked using certified reference materials. Why was this step important, and what does it demonstrate about the accuracy and reliability of your results?
An early comparison showed our automated method consistently reading slightly higher than colony-forming units (CFU) (expected when colonies fuse during growth). We therefore validated against a certified reference material (CRM) with a value traceable to international standards. As no CRM exists for Cordyceps spores or other entomopathogenic fungi, we used Candida albicans of similar size and shape. Repeated analyses by different analysts and conditions produced recovery values close to 100%, evidencing accuracy. We also tested NOFLY within its formulation matrix, confirming precision and reproducibility in real product conditions—critical factors in biofungicide quality control.
Does this technology have the potential for broader application across different types of microbial formulations? What are your plans for future applications?
Yes. The approach quantifies viable spherical yeast and fungal cells. While validation today covers Cordyceps fumosorosea (technical ingredient and NOFLY), the method is applicable to products based on spores of similar size and morphology, e.g., Trichoderma, Beauveria, Metarhizium, and even yeast-based fermentations in wine and brewing. This demonstrates that the technology can extend biofungicide quality control standards across multiple microbial products.
How do you anticipate this certification will influence quality control standards throughout the microbial formulation industry?
It raises the bar for certified, rapid, and objective viability counts. ISO 17025 accreditation validates batch-to-batch consistency, strengthens confidence with regulators and customers, and accelerates logistics: minutes-level QC supports faster batch release and smoother customs transit for live products. The speed also enables in-process adjustments and tighter stock control, improvements that traditional plate counts cannot match.