Laser Biospeckles Analysis for Rapid Evaluation of Organic Pollutants in Water

Rapid evaluation of water toxicity requires biological methods capable of detecting sub-lethal physiological changes without depending on chemical identification. Conventional microscopy-based bioassays are limited by low throughput and difficulties in observing small, transparent and fast-moving microorganisms. This study applies a laser-biospeckle, non-imaging microbioassay to assess the motility responses of Paramecium caudatum and Euglena gracilis exposed to two organic pollutants, trichloroacetic acid (TCAA) and acephate. Dynamic speckle patterns were recorded using a 638 nm laser diode (Thorlabs Inc., Tokyo, Japan) and a CCD camera (Gazo Co., Ltd., Tokyo, Japan) at 60 fps for 120 s. Correlation time, derived from temporal cross-correlation analysis, served as a quantitative indicator of motility. Exposure to TCAA (0.1–50 mg/L) produced strong concentration-dependent inhibition, with correlation time increasing up to 16-fold at 500× PL in P. caudatum (p < 0.01), whereas E. gracilis showed a delayed response, with significant inhibition only above 250× PL. In contrast, acephate exposure (0.036–3.6 mg/L) induced motility enhancement in both species, reflected by decreases in correlation time of up to 57% in P. caudatum and 40% in E. gracilis at 100× PL. Acute trends diminished after 24–48 h, indicating time-dependent physiological adaptation. These results demonstrate that biospeckled-derived correlation time sensitively captures both inhibitory and stimulatory behavioral responses, enabling real-time, high-throughput water toxicity screening without microscopic imaging. The method shows strong potential for integration into automated water-quality monitoring systems.