The use of flow cytometric applications to measure the effects of PAHs on growth, membrane integrity, and relative lipid content of the benthic diatom, Nitzschia brevirostris.

This laboratory study measured the direct effects of three polycyclic aromatic hydrocarbon (PAH) compounds (naphthalene, pyrene, and benzo(a)pyrene) upon cell growth, membrane integrity, and BODIPY-stained lipid fluorescence intensity of the benthic diatom Nitzschia brevirostris using flow cytometry as an analysis tool. Previous field and laboratory studies have reported reductions in algal populations following PAH exposure, but specific, functional responses of the microalgae to these pollutants could not be revealed by cell numbers alone. Using flow-cytometric measurements, we confirmed that maximal cell densities in PAH-exposed diatom cultures were significantly lower compared to controls; however, we also discovered increases in lipids and cells with compromised membranes in PAH-exposed cultures. These results highlight new tools for measuring the direct effects of organic pollutants upon the physiology of taxa comprising microphytobenthic communities important in estuarine food webs.

Immunomodulation in eastern oysters, Crassostrea virginica, exposed to a PAH-contaminated, microphytobenthic diatom.

The trophic transfer of sediment-associated pollutants is a growing concern in shellfish harvesting areas. Previous studies have examined the role of phytoplankton in the transport of organic contaminants to bivalve species, but little information on microphytobenthic communities and their role as contaminant vectors exists. Polycyclic aromatic hydrocarbons (PAHs) are organic compounds formed during natural and industrial processes; they are termed "persistent organic pollutants" because they are only slowly degraded by natural processes. This study examined the transfer of PAH compounds (naphthalene, pyrene, and benzo(a)pyrene) by a microphytobenthic diatom to the eastern oyster, a commercially important shellfish species, to determine if dietary accumulation is a route of contaminant exposure capable of inducing physiological responses. PAH compounds were adsorbed to a diatom culture (Nitzschia brevirostris) in a range of concentrations (5, 125, 625, and 1000 µg L(-1)), and eastern oysters were exposed experimentally to the contaminated diatom cultures to assess possible effects upon oyster hemocytes and selected immune-defense functions. A preliminary experiment was designed to identify individual effects of several PAH compounds (naphthalene, pyrene, and benzo(a)pyrene) on hemocyte viability and phagocytic activity. Results from this experiment revealed that the most-toxic compound, benzo(a)pyrene, at the highest concentration, stimulated an increase in agranular hemocyte counts. A follow-up study examined the effects of benzo(a)pyrene on hemocyte viability, adhesion, phagocytosis, and reactive oxygen species (ROS). These studies showed the ability of this benthic diatom to transport PAHs to the eastern oyster and to cause immunomodulation. Hemocyte responses to dietary PAH exposure included an increase in circulating hemocytes and increased production of reactive oxygen species by these cells.