Abstract

Irrigation water is a primary source of fresh produce contamination by bacteria during the preharvest, particularly in hydroponic systems where the control of pests and pathogens is a major challenge. In this work, we demonstrate the development of a Listeria biosensor using platinum interdigitated microelectrodes (Pt-IME). The sensor is incorporated into a particle/sediment trap for the real-time analysis of irrigation water in a hydroponic lettuce system. We demonstrate the application of this system using a smartphone-based potentiostat for rapid on-site analysis of water quality. A detailed characterization of the electrochemical behavior was conducted in the presence/absence of DNA and Listeria spp., which was followed by calibration in various solutions with and without flow. In flow conditions (100 mL samples), the aptasensor had a sensitivity of 3.37 ± 0.21 kΩ log-CFU−1 mL, and the LOD was 48 ± 12 CFU mL−1 with a linear range of 102 to 104 CFU mL−1. In stagnant solution with no flow, the aptasensor performance was significantly improved in buffer, vegetable broth, and hydroponic media. Sensor hysteresis ranged from 2 to 16% after rinsing in a strong basic solution (direct reuse) and was insignificant after removing the aptamer via washing in Piranha solution (reuse after adsorption with fresh aptamer). This is the first demonstration of an aptasensor used to monitor microbial water quality for hydroponic lettuce in real time using a smartphone-based acquisition system for volumes that conform with the regulatory standards. The aptasensor demonstrated a recovery of 90% and may be reused a limited number of times with minor washing steps.

Highlights

  • The Centers for Disease Control and Prevention (CDC) estimates that up to 48 million illnesses, 128,000 hospitalizations, and 3000 deaths in the United States are caused by foodborne pathogens each year [1]

  • The produce safety rule (PSR) is focused on generic Escherichia coli as an indicator organism, but current efforts in many research labs are underway to consider the direct measurement of pathogenic microorganism

  • We demonstrate the development of a Listeria biosensor using platinum interdigitated microelectrodes (Pt-IME) biofunctionalized with Listeria-specific aptamer (47-mer) and incorporate the sensor into a particle/sediment trap for real-time analysis of irrigation water in hydroponic media

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Summary

Introduction

The Centers for Disease Control and Prevention (CDC) estimates that up to 48 million illnesses, 128,000 hospitalizations, and 3000 deaths in the United States are caused by foodborne pathogens each year [1]. We demonstrate the development of a Listeria biosensor using platinum interdigitated microelectrodes (Pt-IME) biofunctionalized with Listeria-specific aptamer (47-mer) and incorporate the sensor into a particle/sediment trap for real-time analysis of irrigation water in hydroponic media. The resulting biosensor is capable of sensing Listeria spp. in buffer solution and real food (vegetable broth) in stagnant media, as well as in a high flow-through system of irrigation water in hydroponic systems at relevant concentrations to regulatory standards for assessing agricultural water quality. This biosensor has a high level of recovery and can be reused a number of times with minor washing steps

Experimental
Bacteria Strains and Culture
Electrochemical Characterization
Pt-IME Fabrication Procedure
In Silico Model of Pt-IME Capacitance
Biofunctionalization of Pt-IMEs with Aptamers
Protocol for Bacteria Detection
Hysteresis Testing
Analysis of Hydroponic Water
2.10. Statistical Analysis and Portfolio Analysis
Results and Discussion
In Silico Pt-IME Model
Electrochemical
Pt-IME
Adsorption
Hysteresis and Reusability
Reusability
Analysis of Hydroponic Water in Particle Trap Filter
Calibration of Pt-IME biosensor in hydroponic media forinnocua
Comparison of Listeria Sensors in the Literature
Conclusions
Full Text
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