In vitro approach in aquatic organism cells to evaluate the cytotoxicity of microbial bioinput with herbicidal potential

• Grant number: 25/19228-9
• Support Opportunities: Scholarships in Brazil - Master
• Start date: October 01, 2025
• End date: May 31, 2027
• Field of knowledge: Biological Sciences - Biology
• Principal Investigator: Cláudia Maris Ferreira Mostério
• Grantee: Guilherme Felicioni Ferreira
• Host Institution: Instituto de Pesca. Agência Paulista de Tecnologia dos Agronegócios (APTA). São Paulo , SP, Brazil
• Associated research grant: 24/20944-8 - Evaluating the toxicity of microbial bio-inputs and their effects on aquatic organisms: a path towards clean agriculture, AP.R

Abstract

Growing concern about the impact of commercial pesticides on health and the environment has driven the production of agricultural bio-inputs, particularly microbial bioherbicides. These bioherbicides, developed from microorganisms such as fungi and bacteria, act by degrading the cell walls of weeds and are essential for more sustainable agriculture. Rich in enzymes with herbicidal potential, these bio-inputs can be used alone or in synergy with the microorganisms themselves. In line with the concept of circular economy, their production uses waste as substrates, reducing costs and minimizing environmental impacts. However, the ecological safety of these products still needs to be proven, because although they are considered environmentally friendly, uncertainties remain regarding possible toxic effects on non-target organisms, especially vertebrates. These in vitro tests on cell cultures comply with the 3Rs principles (reduction, replacement, and refinement) and the guidelines of Toxicology for the 21st Century, reducing the use of animals in experiments involving the testing of new products. To this end, fibroblast cell lines from fish and amphibians will be used to perform Trypan Blue Exclusion and Neutral Red Uptake tests (for membrane integrity), MTT Reduction (to assess cell viability and growth), Sulforhodamine B and Crystal Violet Assays (to assess cell viability and quantify nucleic acids), and XTT Assay (to assess cell respiration). These tests will help predict possible ecotoxicological risks and define safe levels of bioinput use. The results are expected to contribute to the development of safe and sustainable bio-inputs, promoting agricultural innovations in line with circular economy guidelines and offering natural and viable solutions for the food production sector.