Bacteria may play a role in regulating harmful algal blooms, but little is known about the biochemical and physiological changes associated with cell death induced by algicidal bacteria. Previous work characterized an algicidal exudate (IRI-160AA) produced by Shewanella sp. IRI-160 that is effective against dinoflagellates, while having little to no effect on other phytoplankton species in laboratory culture experiments. The objective of this study was to evaluate biochemical changes associated with cell death and impacts on the cell cycle in three dinoflagellate species (Prorocentrum minimum, Karlodinium veneficum and Gyrodinium instriatum) after exposure to IRI-160AA. In this study, IRI-160AA induced cell cycle arrest in all dinoflagellates examined. Several indicators for programmed cell death (PCD) that are often observed in phytoplankton in response to a variety of stressors were also evaluated. Cell death was accompanied by significant increases in DNA degradation, intra- and extracellular ROS concentrations and DEVDase (caspase-3 like) protease activity, which have been associated with PCD in other phytoplankton species. Overall, results of this investigation provide strong evidence that treatment with the bacterial algicide, IRI-160AA results in cell cycle arrest and induces biochemical changes consistent with stress-related cell death responses observed in other phytoplankton.

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The algicide, IRI-160AA, induces mortality in dinoflagellates but not other species of algae, suggesting that a shared characteristic or feature renders this class of phytoplankton vulnerable to the algicide. In contrast to other eukaryotic species, the genome of dinoflagellates is stabilized by high concentrations of divalent cations and transition metals and contains large amounts of DNA with unusual base modifications. These distinctions set dinoflagellates apart from other phytoplankton and suggest that the nucleus may be a dinoflagellate-specific target for IRI-160AA. In this study, morphological and ultrastructural changes in three dinoflagellate species, Prorocentrum minimum, Karlodinium veneficum and Gyrodinium instriatum, were evaluated after short-term exposure to IRI-160AA using super resolution structured illumination microscopy (SR-SIM) and transmission electron microscopy (TEM). Exposure to the algicide resulted in cytoplasmic membrane blebbing, differing chloroplast morphologies, nuclear expansion, and chromosome expulsion and/or destabilization. TEM analysis showed that chromosomes of algicide-treated K. veneficum appeared electron dense with fibrous protrusions. In algicide-treated P. minimum and G. instriatum, chromosome decompaction occurred, while for P. minimum, nuclear expulsion was also observed for several cells. Results of this investigation demonstrate that exposure to the algicide destabilizes dinoflagellate chromosomes, although it was not clear if the nucleus was the primary target of the algicide or if the observed effects on chromosomal structure were due to downstream impacts. In all cases, changes in cellular morphology and ultrastructure were observed within two hours, suggesting that the algicide may be an effective and rapid approach to mitigate dinoflagellate blooms.

Macronutrients and trace metals are incorporated into phytoplankton during growth and regenerated back into the water column when phytoplankton decay, a process that contributes to the distributions of dissolved trace metals and macronutrients in depth profiles. To study this, we incubated mixed Gulf of Mexico phytoplankton assemblages and monocultures of the diatom Pseudo-nitzschia dolorosa and the dinoflagellate Karenia brevis in the dark. Over 6 months, macronutrients (phosphate, silicic acid, nitrate + nitrite, nitrite, ammonium), chlorophyll-a, particulate organic carbon and nitrogen, and prokaryotes were monitored alongside dissolved manganese (Mn), iron (Fe), cobalt (Co), nickel (Ni), copper (Cu), zinc (Zn), cadmium (Cd), and lead (Pb). Results were compared to depth profiles to evaluate the role of regeneration in trace metal cycling. In contrast to water-column distributions, silicic acid and phosphate were closely coupled in experiments containing diatoms, indicating a shared regeneration pathway. Nitrification and nitrifying prokaryotes were only observed near the end of a subset of the experiments. Of the trace metals, Cd was most tightly coupled with phosphate. Regeneration of Mn was followed by rapid drawdown, consistent with Mn-oxide formation. Iron (Fe), Cu, and Pb typically remained low until Mn was depleted, suggesting either scavenging to Mn-oxides or otherwise delayed regeneration of these elements. Cobalt (Co) and Ni were largely conservative, but behaved like nutrients in the experiment using more offshore water low in Cd and Zn. Although experimental conditions were limited in their representation of the water column, these incubations provide novel insight into macronutrient and trace metal regeneration in the oceans. At the base of the marine food web, phytoplankton require the macronutrients carbon (C), nitrogen (N), and phosphorous (P) to build cellular organic matter, and, for diatoms, silicon (Si) to build frustules (Redfield 1934; Brzezinski 1985). Phytoplankton also assimilate a suite of trace metals into their cells, including manganese (Mn), iron (Fe), cobalt (Co), nickel (Ni), copper (Cu), zinc (Zn), cadmium (Cd), and lead (Pb) (

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