Detection of cyanotoxins (microcystins/nodularins) in livers from estuarine and coastal bottlenose dolphins (Tursiops truncatus) from Northeast Florida

Microcystins/Nodularins (MCs/NODs) are potent hepatotoxic cyanotoxins produced by harmful algal blooms (HABs) that occur frequently in the upper basin of the St. Johns River (SJR), Jacksonville, FL, USA. Areas downstream of bloom locations provide critical habitat for an estuarine population of bottlenose dolphins (Tursiops truncatus). Since 2010, approximately 30 of these dolphins have stranded and died within this impaired watershed; the cause of death was inconclusive for a majority of these individuals. For the current study, environmental exposure to MCs/NODs was investigated as a potential cause of dolphin mortality. Stranded dolphins from 2013 to 2017 were categorized into estuarine (n = 17) and coastal (n = 10) populations. Because estuarine dolphins inhabit areas with frequent or recurring cyanoblooms, they were considered as a comparatively high-risk group for cyanotoxin exposure in relation to coastal animals. All available liver samples from estuarine dolphins were tested regardless of stranding date, and samples from coastal individuals that stranded outside of the known cyanotoxin bloom season were assessed as controls. The MMPB (2-methyl-3-methoxy-4-phenylbutiric acid) technique was used to determine total (bound and free) concentrations of MCs/NODS in liver tissues. Free MCs/NODs extractions were conducted and analyzed using ELISA and LC-MS/MS on MMPB-positive samples to compare test results. MMPB testing resulted in low-level total MCs/NODs detection in some specimens. The Adda ELISA produced high test values that were not supported by concurrent LC-MS/MS analyses, indicative of false positives. Our results indicate that both estuarine and coastal dolphins are exposed to MCs/NODs, with potential toxic and immune health impacts.