Short-term responses of identified soil beneficial-bacteria to the insecticide fipronil: toxicological impacts.

Pesticides including insecticides are often applied to prevent distortion posed by plant insect pests. However, the application of these chemicals detrimentally affected the non-target organisms including soil biota. Fipronil (FIP), a broad-spectrum insecticide, is extensively used to control pests across the globe. The frequent usage calls for attention regarding risk assessment of undesirable effects on non-target microorganisms. Here, laboratory-based experiments were conducted to assess the effect of FIP on plant-beneficial bacteria (PBB); Rhizobium leguminosarum (Acc. No. PQ578652), Azotobacter salinestris (Acc. No. PQ578649) and Serratia marcescens (Acc. No. PQ578651). PBB synthesized growth regulating substances were negatively affected by increasing fipronil concentrations. For instance, at 100 µg FIPmL, a decrease in indole-3-acetic acid (IAA) synthesis by bacterial strains followed the order: A. salinestris (95.6%) S. marcescens (91.6%) > R. leguminosarum (87%). Also, exposure of bacteria cells to FIP hindered the growth and morphology of PBB observed as distortion, cracking, and aberrant structure under scanning electron microscopy (SEM). Moreover, FIP-treated and propidium iodide (PI)-stained bacterial cells displayed an insecticide dose-dependent increase in cellular permeability as observed under a confocal laser microscope (CLSM). Colony counts (log CFU mL) and growth of A. salinestris was completely inhibited at 150 µg FIPmL. The surface adhering ability (biofilm formation) of PBB was also disrupted/inhibited in a FIP dose-related manner. The respiration loss due to FIP was coupled with a reduction in population size. Fipronil at 150 µgmL decreased cellular respiration in A. salinestris (72%) S. marcescens (53%) and R. leguminosarum (85%). Additionally, biomarker enzymes; lactate dehydrogenase (LDH), lipid peroxidation (LPO), and oxidative stress (catalase; CAT) induced by FIP represented significant (p ≤ 0.05) toxicity towards PBB strains. Conclusively, fipronil suggests a toxic effect that emphasizes their careful monitoring in soils before application and their optimum addition in the soil-plant system. It is high time to prepare both target-specific and slow-released agrochemical formulation for crop protection with concurrent safeguarding of soils.