Development of a Novel Bioassay and Portable Spectrometer to Assess Inorganic Arsenic Bioavailability in the Environment
Creators: name: Pothier, Martin
Publisher: name: Université d'Ottawa / University of Ottawa
Year: 2020
Subjects: arsenic
biosensor
bioavailability
DOM
photooxidation
water quality

description: Arsenic is a notorious poison due to its high toxicity, worldwide distribution, and lack of any taste and colour once dissolved. The abundance of arsenic in Earth’s crust makes that it can naturally find its way into food and drinking water. Rapid and reliable detection of arsenic, directly in the field, is critical to support evidence-based decision-making in choosing irrigation or drinking water sources. Current cost-effective colourimetric techniques are associated with poor accuracy, health risks, and unacceptable levels of false negatives. Arsenic-specific cellular sensors, or biosensors, may present an inexpensive, safe, and renewable alternative, yet they have long been criticized for unsatisfactory sensing performance, and inconsistency of the outcome. This, in addition to the lack of suitable instruments capable of measuring the signals produced by these biosensors, has led to very few solutions reaching market. The goal of my thesis research was to test hypotheses that improve our fundamental understanding of As species biogeochemistry in simple and complex environmental matrices to then develop a new arsenic monitoring interface, one that would be both simple and accessible to the general public. Using a combination of wild-type and mutant strains, I managed to detail both the internal regulation of arsenic, and the external drivers of arsenic bioavailability. I started by designing a defined exposure protocol that achieved, for the first time, equimolar uptake of over 94% of the added As(III) and As(V) into the cells. By developing this control early into my thesis, I then worked to reintroduce commonly found constituents of environmental waters that are thought to impact arsenic uptake. This direct testing approach uncovered fundamentals of environmental arsenic redox chemistry such as As(III) photooxidation in solution, environmental ligand exchanges, and biological transport pathways. Simplifying a complex exposure protocol for use by the general public required automation of the data analysis steps. This consists of several hundred lines of code, capable of analyzing, normalizing and stabilizing biosensor output to improve the consistency and robustness of this system. These algorithms were then integrated into a new arsenic monitoring interface, one that was built and designed specifically for dehydrated biosensors. This portable, low-cost spectrometer achieved a fluorescent detection range that rivals expensive and sophisticated laboratory equipment at a fraction of the price, and without the need for a computer to compile the measurements. In contrast to highly criticized colorimetric techniques, the biosensor exposure protocol exceeds in operational use, reliability and detection limit. At its core, my thesis research provides a new and complete arsenic testing solution, one capable of measuring both As(III) and As(V) at levels relevant to the World Health Organization and Canadian guidelines for arsenic content in water (10 µg/L). It also provides a new method capable of selectively discriminating between arsenic species, thereby providing an inexpensive and high-throughput arsenic speciation method. I hope this work will help kickstart development of a marketable solution that empowers individuals to test and to monitor the quality of their water sources.