Effects of nitrate and ammonium on assimilation of nitric oxide by Heterosigma akashiwo.

The harmful alga Heterosigma akashiwo possesses a hybrid nitrate reductase (NR) enzyme, NR2-2/2HbN, which has the potential to convert NO to nitrate for assimilation into biomass. In previous research, NR transcription in H. akashiwo was induced by nitrate while NR activity was inhibited by ammonium. Here, the capacity of H. akashiwo to use NO in the presence of nitrate and/or ammonium was investigated to understand the regulation of NO assimilation. Continuous cultures of H. akashiwo were acclimated to growth on nitrate, ammonium, or a mixture of both. Aliquots from these cultures were spiked with 15N-labeled NO. The expression of genes involved in nitrogen assimilation was evaluated, as well as nitrate reductase activity and assimilation of 15N-labeled nitrogen into algal biomass. Results showed that NO induced expression and activity of NR, and upregulated expression of GOGAT regardless of the presence of other inorganic nitrogen sources, while GS expression decreased over time. Furthermore, 15NO uptake and assimilation was significantly higher in cultures acclimated for growth on ammonium compared to cultures acclimated for growth on nitrate alone. Assimilation of NO may provide H. akashiwo with a competitive advantage in N-poor environments or areas with elevated NO.

Chattonella subsalsa (Raphidophyceae) growth and hemolytic activity in response to agriculturally-derived estuarine contaminants.

The potential for toxic contaminants and nutrient pollution to alter natural cycles of estuarine phytoplankton blooms is well known, yet few studies have examined how these combined stressors affect harmful algal species. Here, a robust testing protocol was developed to enable an ecotoxicological assessment of responses to commonly co-occurring estuarine contaminants by harmful algal bloom species. The population growth and toxicity (as cell density and hemolytic activity, respectively) of a cultured strain of the toxigenic raphidiophycean, Chattonella subsalsa, were assessed in two experiments (duration 10 days and 28 days) across a gradient of atrazine concentrations and N:P ratios simulating nutrient-rich versus nutrient-depleted regimes. The response of this large-celled, slowly growing alga to atrazine × nutrients depended on growth phase; atrazine was most inhibitory during early exponential population growth (day 10), whereas nutrient regime was a more important influence during later phases of growth (day 28). Without atrazine, toxicity toward fish was highest in low-P cultures. At atrazine levels >25 µg L-1, hemolytic activity was highest in low-N cultures, and increased with increasing atrazine concentration in all nutrient-limited cultures. Hemolytic activity varied inversely with atrazine concentration in N,P-replete conditions. Overall, atrazine inhibitory effects on population growth of this C. subsalsa strain depended on the growth phase and the nutrient regime; hemolytic activity was higher and further enhanced by atrazine in low N-P regimes; and atrazine inhibited hemolytic activity in nutrient-replete conditions. The data suggest that, depending on the growth phase and nutrient regime, atrazine can help promote toxic C. subsalsa blooms.

Imbalanced nutrient regimes increase Prymnesium parvum resilience to herbicide exposure.

The toxigenic haptophyte Prymnesium parvum is a mixotrophic phytoplankter with an extensive historic record of forming nearly monospecific, high-biomass, ecosystem-disrupting blooms, and it has been responsible for major fish kills in brackish waters and aquaculture facilities in many regions of the world. Little is known about how this species responds to commonly occurring environmental contaminants, or how nutrient (nitrogen, phosphorus) pollution may interact with environmentally relevant pesticide exposures to affect this harmful algal species. Here, standard algal toxicity bioassays from pesticide hazard assessments were used along with modified erythrocyte lysis assays to evaluate how atrazine exposures, imbalanced nutrient supplies, and salinity interact to influence the growth and toxicity in P. parvum isolates from three different regions. In nutrient-replete media, P. parvum 96 h IC50s ranged from 73.0 to 88.3 µg atrazine L-1 at salinity 10 and from 118 to >200 µg atrazine µg L-1 at salinity 20, and the response depended on the strain and the test duration. Relative hemolytic activity, used as an indication of toxicity, was a function of herbicide exposure, nutrient availability, salinity, geographic origin, and interactions among these factors. Highest levels of hemolytic activity were measured from a South Carolina strain in low-nitrogen media with high atrazine concentrations. Herbicide concentration was related to relative hemolytic activity, although a consistent relationship between growth phase and toxicity was not observed. Overall, these findings suggest that increasing chemical contamination is helping to promote ecosystem-disruptive, strongly mixotrophic algal blooms.

Assessment of atrazine toxicity to the estuarine phytoplankter, Dunaliella tertiolecta (Chlorophyta), under varying nutrient conditions.

Anthropogenic inputs of chemical environmental contaminants are frequently associated with developing harmful algal blooms, but little is known about how estuarine phytoplankton assemblages respond to multiple, co-occurring chemical stressors in chronically disturbed habitats. The goals of this research were to establish a robust protocol for testing the effects of atrazine on estuarine phytoplankton, and then to use that protocol to compare the effects of atrazine exposure with and without nutrient enrichment on a cosmopolitan estuarine/marine alga, Dunaliella tertiolecta (Chlorophyta). Atrazine sensitivity in nutrient-replete media (96-h growth inhibition [Formula: see text]) was 159.16 µg l-1, but sensitivity was influenced by exposure duration, and inhibitory effects of herbicide on algal growth decreased under imbalanced nutrient regimes and low nitrogen and phosphorus supplies. These findings advance knowledge about how nutrient regimes and herbicides interact to control estuarine phytoplankton population dynamics.