Development of a rapid field test is needed capable of determining if field supplies of water are safe to drink by the warfighter during a military operation. The present study sought to assess the effectiveness of handheld assays (HHAs) in detecting ricin and Staphylococcal Enterotoxin B (SEB) in water. Performance of HHAs was evaluated in formulated tap water with and without chlorine, reverse osmosis water (RO) with chlorine, and RO with bromine. Each matrix was prepared, spiked with ricin or SEB at multiple concentrations, and then loaded onto HHAs. HHAs were allowed to develop and then read visually. Limits of detection (LOD) were determined for all HHAs in each water type. Both ricin and SEB were detected by HHAs in formulated tap water at or below the suggested health effect levels of 455？ng/mL and 4.55？ng/mL, respectively. However, in brominated or chlorinated waters, LODs for SEB increased to approximately 2,500？ng/mL. LODs for ricin increased in chlorinated water, but still remained below the suggested health effect level. In brominated water, the LOD for ricin increased to approximately 2,500？ng/mL. In conclusion, the HHAs tested were less effective at detecting ricin and SEB in disinfected water, as currently configured. 1. Introduction The protection of water supplies and water distribution infrastructure such as reservoirs is of great concern for those defending against bioterrorism. The US military is concerned because there exist multiple biological warfare agents (BWAs) that could be intentionally introduced into the water supply which could have serious implications for military operations as well as civilians located within a theater of operations. To protect potable water supplies, the Department of Defense (DoD) seeks to develop a rapid field test that would determine if field supplies of water are safe to drink by the warfighter during a military operation. The US armed services have fielded biological defense systems that are designed to protect against a large scale attack via BWAs dispersed through the air. These defensive systems utilize high-efficiency aerosol collectors that impinge the sampled air into a liquid buffer which is tested for BWAs using antibody-based handheld assays (HHAs). Civilian first responders use similar versions of these antibody-based HHAs for testing suspicious powder samples suspected to contain BWAs. Depending on which manufacturer of HHA is being tested, the technology can be low cost, requires no electrical power, and requires minimal to no sample preparation to test a sample. It was an HHA panel
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