Organophosphorus (OP) compounds are neurotoxins that are among the most widely used pesticides in agriculture in the United States. In this application, a new integrated point-of-care smartphone/resistive nanosensor device is developed for onsite rapid and sensitive detection of exposure to OP pesticides from a drop of finger-stick blood among a sample of farmworkers. The nanosensor leverages the transport properties of a multiwalled carbon nanotube/polyaniline nanofiber (MWCNT/PAnNF) nanocomposite film on a gold interdigitated electrode and acetylcholinesterase/butyrylcholinesterase (AChE/BChE) hydrolysis of their respective substrates generating protons doping PAnNFs, thereby increasing the conductance of the film. As such, a conductance change can be used to quantify cholinesterase activity, enabling assessment of acute/chronic OP poisoning. Additionally, a mobile app was developed for the nanosensor to process, display, track, and share results. Under optimal conditions, the nanosensor demonstrated exceptional sensitivity with the detection limits of 0.11 U/mL for AChE and 0.093 U/mL for BChE, physiologically relevant dynamic ranges of 2.0-18.0 U/mL for AChE and 0.5-5.0 U/mL for BChE in whole blood, and high reproducibility with the relative standard variation of <4%. The nanosensor was further validated with widely used radiometric and Ellman's methods, utilizing both in vitro pesticide-spiked blood samples and blood samples from 22 farmworkers. The results between this nanosensor and those two methods demonstrated a strong agreement. This platform provides a new avenue for the simple, rapid, and sensitive biomonitoring of OP pesticide exposure.

Organophosphate (OP) pesticides remain a worldwide health concern due to their acute or chronic poisoning and widespread use in agriculture around the world. There is a need for robust and field-deployable tools for onsite detection of OP pesticides in food and water. Herein, we present an integrated smartphone/resistive biosensor for simple, rapid, reagentless, and sensitive monitoring of OP pesticides in food and environmental water. The biosensor leverages the hydrolytic activity of acetylcholinesterase (AChE) to its substrate, acetylcholine (ACh), and unique transport properties of polyaniline nanofibers (PAnNFs) of chitosan/AChE/PAnNF/carbon nanotube (CNT) nanocomposite film on a gold interdigitated electrode. The principle of the sensor relies on OP inhibiting AChE, thus, reducing the rate of ACh hydrolysis and consequently decreasing the rate of protons doping the PAnNFs. Such resulted decrease in conductance of PAnNF can be used to quantify OP pesticides in a sample. A mobile app for the biosensor was developed for analyzing measurement data and displaying and sharing testing results. Under optimal conditions, the biosensor demonstrated a wide linear range (1 ppt–100 ppb) with a low detection limit (0.304 ppt) and high reproducibility (RSD <5%) for Paraoxon-Methyl (PM), a model analyte. Furthermore, the biosensor was successfully applied for analyzing PM spiked food/water samples with an average recovery rate of 98.3% and provided comparable results with liquid chromatography-mass spectrometry. As such, the nanosensing platform provides a promising tool for onsite rapid and sensitive detection of OP pesticides in food and environmental water.

Farmworkers experience disproportionate risks of pesticide exposure, yet only two U.S. states, California and Washington, currently mandate routine cholinesterase biomonitoring programs for agricultural workers. Although regulatory and resource limitations influence implementation of worker protection programs, operational barriers also complicate repeated biomonitoring among highly mobile and underserved farmworker populations. The objective of this preliminary study was to evaluate the operational feasibility of recruiting, retaining, and repeatedly monitoring farmworkers using a novel, rapid, portable cholinesterase analysis device under routine agricultural field conditions. Guided by an evidence-based feasibility framework, a multidisciplinary team administered surveys and collected blood samples from 25 farmworkers in rural North Carolina at three time points during a single agricultural season. Participation and visit completion rates were high, although worker mobility, changing job assignments, environmental conditions, and temperature-sensitive samples created logistical challenges. Experienced bilingual promotoras de salud played a critical role in coordination, communication, scheduling, and retention. Findings provide preliminary evidence that repeated field-based cholinesterase biomonitoring protocols are feasible and acceptable when supported by community-based outreach organizations with Spanish speaking health workers, structured logistics, and culturally responsive engagement strategies. Evaluation of the analytical performance is beyond the scope of this report, however, these results provide practical insights for strengthening occupational health surveillance and future implementation studies designed to protect agricultural workers.