Thesis (M.A.)--Humboldt State University, 2009. / Includes bibliographical references (leaves 64-71). Also available via Humboldt Digital Scholar.

Thesis--University of Washington. / Vita. Bibliography: leaves [205]-227.

Graduation date: 1995

Thesis (Ph. D.)--Oregon State University, 1995. / Typescript (photocopy). Includes bibliographical references (leaves 211-237). Also available online.

As our knowledge of bivalves has increased, so has information about their parasites. Most studies of bivalve parasites have been devoted to groups infesting commercially important mollusks, notably trematodes and copepods. Another group of bivalve symbionts which has been known since the early nineteenth century is the brachyuran genus Pinnotheres Latreille, 1802 (commonly known as pea crabs). The generic name was given because of the belief that the crabs were predators upon the scallops in which they were most frequently found (Pearce, 1966). However, Stebbing (1893) suggested that instead of acting as predators, the crabs were in fact serving as guards for their hosts.

The intertidal zone is a dynamic environment that fluctuates with the 12.4-h tidal and 24-h light/dark cycle to predictably alter food availability, temperature, air exposure, wave action, oxygen partial pressure, and osmotic conditions. Intertidal sessile bivalves exhibit behavioral or physiological changes to minimize the persistent challenges of fluctuating environmental conditions, such as adjusting gaping behavior and heart rate. At the cellular level, transcriptomic studies on mussels’ baseline circadian and circatidal rhythms have determined that the circadian rhythm is the dominant transcriptional rhythm. However, as proteins reflect the basic molecular phenotype of an organism and their abundance may differ greatly from that of mRNA, these methods could fail to detect important cyclical changes in the proteome that cope with the regular stress of tidal rhythms. For this study, we acclimated intertidal Mytilus californianus to circadian (12:12 h light/dark cycle) and circatidal (6:6 h tidal cycle) conditions in a tidal simulator and sampled gill tissue from mussels every 2 h for 48 h for proteomic analysis. Approximately 86% of the proteins that were detected exhibited rhythmicity over the time course. The circadian cycle primarily determined the cyclic abundance of energy metabolism proteins, pivoting around the transition to the nighttime high tide. The tidal cycle contributed to alterations in cytoskeletal components, ER protein processing and vesicular trafficking, extracellular matrix and immune proteins, and oxidative stress and chaperoning proteins. We also found evidence that post-translational modifications may be important for driving these rhythms, as acetylation and phosphorylation motifs were enriched in the rhythmic proteins and we identified rhythms in elements of methylation, mitochondrial peptide processing, and acylation. These dynamic changes in proteins across numerous functional categories indicate that the combination of circadian and tidal cycles drive complex cellular changes to coordinate processes in a changing environment. This variation clearly shows that differential expression studies and biomonitoring efforts cannot assume a static baseline of cellular conditions in intertidal mussels.

This study deals with the reproductive ecology and larval development of Mytilus californianus. Mytilus californianus is the common mussel found along the more exposed west coast of North America from the Aleutian Islands south Isla Socorro, Mexico (Soot-Ryen, 1955) and was chosen because it is an important species in rocky intertidal communities (being the most abundant if not the most conspicuous) and forms the basis for diverse associations of numerous other species. The reproductive cycle, larval development to settlement, comparisons of reproduction in high and low populations and field observations on larval settlement are described.

Caffeine, a biologically active drug with many known molecular targets, is recognized as a contaminant of aquatic systems including marine systems. Although the concentrations of caffeine reported from aquatic systems are low (ng/L to μ/L), harmful ecological effects not detected by traditional toxicity tests could occur as a result of caffeine contamination. The goals of this thesis project were to: 1) quantify concentrations of caffeine occurring in the coastal ocean off of the Oregon Coast and to determine if higher levels were correlated with proximity to caffeine pollution sources; and 2) investigate the sub-lethal cellular toxicity of environmentally relevant concentrations of caffeine on Mytilus californianus, an intertidal mussel that is one of the dominant species of the Oregon Coast. Caffeine was analyzed at 14 locations in the coastal ocean off of the Oregon Coast. Sampling locations were stratified between populated areas identified as having sources of caffeine pollution and sites located in sparsely populated areas not in proximity to major caffeine pollution sources. Caffeine concentrations were also measured in the major water body discharging near sampling locations. Caffeine was detected in water from the coastal ocean off of the Oregon Coast at concentrations ranging from below the reporting limit to 44.7 ng/L. The occurrence and concentrations of caffeine in the coastal ocean did not correspond well with pollution threats from population density and point and non-point sources, but did correspond with a storm event occurrence. Caffeine concentrations in rivers and estuaries draining to the coast ranged from below the reporting limit to 152.2 ng/L. Hsp70, a molecular biomarker of cellular stress, was used to investigate the sub-lethal toxicity of environmentally relevant concentrations of caffeine to the mussel M. californianus. Hsp70 concentrations in the gill and mantle tissue of mussels exposed to 0.05, 0.2, and 0.5 μ/L of caffeine for 10, 20, and 30 days were compared to basal levels in control mussels. Hsp70 in the gill tissue of M. californianus had an initial attenuation of the stress protein followed by a significant albeit moderate up-regulation relative to controls in all but the 0.5 μ/L treatment. Hsp70 in the mantle tissue of mussels exposed to caffeine did not differ from control mussels. This Study confirms the presence of caffeine in Oregon's coastal ocean and provides laboratory evidence that environmentally relevant concentrations of caffeine can exert an effect on M. californianus gill tissue at the molecular-level.

Contaminants such as pharmaceuticals are of increasing concern due to their ubiquitous use and persistence in surface waters worldwide. Limited attention has been paid to the effects of pharmaceuticals on marine life, despite widespread detection of these contaminants in the marine environment. Of the existing studies, the majority assess the negative effects of pharmaceuticals over an exposure period of 30 days or less and focus on cellular and subcellular biomarkers. Longer studies are required to determine if chronic contaminant exposure poses risks to marine life at environmentally relevant concentrations. Also scarce in the literature is examination of whole organism effects to identify potential community-level consequences. Two long-term studies with the antidepressant pharmaceutical, fluoxetine (the active constituent in Prozac®) were conducted to determine whether nominal concentrations detected in estuarine and coastal environments affect organism health and interactions. First, we measured whole organism metrics in the California mussel, Mytilus californianus over a period of 107 days. Specifically, we measured algal clearance rates, growth, and condition indices for both reproductive and overall health. We found that fluoxetine negatively affects all measured characteristics, however many effects are mediated by length of exposure. Perhaps the most notable result was that mussels spiked with fluoxetine cleared less algae after 30 days of exposure. Reduced growth and condition indices likely are a consequence of improper nutrition among fluoxetine-treated mussels. Any level of fluoxetine significantly affected the gonadosomatic index after 47 days. The results from this study on mussels fill an important data gap, highlighting organism-level effects of chronic exposure periods; such data more explicitly identify the impacts of pharmaceuticals and other contaminants on marine communities and ecosystems. Fluoxetine has also been documented to affect the behavior of fish and invertebrates, including freshwater and marine bivalves, crustaceans, and fish. Given that other crustaceans exhibited increased activity levels under fluoxetine exposure, we hypothesized that this would subject them to greater predation risk. In our second exposure study, we assessed whether a similar range of fluoxetine concentrations used in the mussel study altered the risk behavior of the Oregon mud crab, Hemigrapsus oregonensis, in response to a common predator, the red rock crab, Cancer productus. We conducted this study for 60 days, conducting day and night behavioral trials (with and without predators) four times a week. We found that crabs exposed to any amount of fluoxetine (3 or 30 ng/L) had increased activity levels relative to controls; however behaviors of 3 ng/L-spiked crabs were not always significantly different from controls. Among control crabs, day and night trials yielded similar results, where a clear response to the addition of the predator was observed. Crabs dosed with fluoxetine exhibited more foraging and active behaviors in the presence of the predator. Additionally, crabs spiked with fluoxetine at 30 ng/L had the greatest risk of mortality either by predation by red rock crabs or due to more aggressive behaviors among conspecifics. The results of this study shed light on a particularly unexplored area of contaminants research: how do psychoactive pharmaceuticals affect animal behavior when exposed to the low concentrations persisting in the aquatic environment for a prolonged period of time?