A novel antibody-based biomarker for chronic algal toxin exposure and sub-acute neurotoxicity.

Kathi A Lefebvre,John D Hansen,Elizabeth R Frame,Frances Gulland,Theo K Bammler,Frederico M Farin,Preston S Kendrick,Emma M Hiolski,Donald R Smith,Richard P Beyer,David J Marcinek,Brett Neilan

doi:10.1371/journal.pone.0036213

Abstract

The neurotoxic amino acid, domoic acid (DA), is naturally produced by marine phytoplankton and presents a significant threat to the health of marine mammals, seabirds and humans via transfer of the toxin through the foodweb. In humans, acute exposure causes a neurotoxic illness known as amnesic shellfish poisoning characterized by seizures, memory loss, coma and death. Regular monitoring for high DA levels in edible shellfish tissues has been effective in protecting human consumers from acute DA exposure. However, chronic low-level DA exposure remains a concern, particularly in coastal and tribal communities that subsistence harvest shellfish known to contain low levels of the toxin. Domoic acid exposure via consumption of planktivorous fish also has a profound health impact on California sea lions (Zalophus californianus) affecting hundreds of animals yearly. Due to increasing algal toxin exposure threats globally, there is a critical need for reliable diagnostic tests for assessing chronic DA exposure in humans and wildlife. Here we report the discovery of a novel DA-specific antibody response that is a signature of chronic low-level exposure identified initially in a zebrafish exposure model and confirmed in naturally exposed wild sea lions. Additionally, we found that chronic exposure in zebrafish caused increased neurologic sensitivity to DA, revealing that repetitive exposure to DA well below the threshold for acute behavioral toxicity has underlying neurotoxic consequences. The discovery that chronic exposure to low levels of a small, water-soluble single amino acid triggers a detectable antibody response is surprising and has profound implications for the development of diagnostic tests for exposure to other pervasive environmental toxins.

Highlights

Domoic acid (DA) is a small water-soluble marine algal toxin that is naturally produced by diatoms of the genus Pseudo-nitzschia during harmful algal blooms (HABs) [1]
The fact that DA is highly water soluble and rapidly excreted, led us to interpret that the increase in DA signal after 18 weeks and into recovery was likely due to the presence of a DAspecific antibody and not an indication of increased serum DA levels (Figure 1)
A DA specific antibody (IgM) present in zebrafish serum would bind to DA conjugated to the ELISA plate and compete with the BiosenseH labeled detection antibody, resulting in a decrease in absorbance just as DA in a sample competes with the labeled antibody in solution, causing decreased absorbance

Summary

Introduction

Domoic acid (DA) is a small water-soluble marine algal toxin that is naturally produced by diatoms of the genus Pseudo-nitzschia during harmful algal blooms (HABs) [1]. In addition to human health concerns, DA exposure via consumption of planktivorous fish has a significant impact on California sea lions along the US West Coast resulting in the stranding and/or deaths of hundreds of animals yearly [6,7]. These naturally exposed sea lions represent a valuable ‘‘sentinel’’ species for human health risks as related to DA exposure. The chronic syndrome has been observed in stranded sea lions even in the absence of DA producing algal blooms in the environment, suggesting that sub-lethal DA exposure leads to lasting neurologic effects in mammalian species [8]

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Results

Discussion

Conclusion