Tetrodotoxin (TTX) is a potent neurotoxin that acts by blocking the pore region of voltage-gated sodium channels in nerve and muscle tissue. This causes paralysis, and often death due to asphyxiation. Interestingly, TTX is found in an array of organisms ranging from bacterial species to vertebrates. Further, TTX is found in both aquatic and terrestrial environments. This range of taxa and environments has led to three common lines of study for ecological research on this toxin: production, predation, and identification of novel TTX bearing taxa. I began my research by also refining a Competitive Inhibition Enzymatic Immunoassay technique for fast, easy, and inexpensive quantification of TTX. I then focused on the three previously mentioned areas of research. Female newts (Taricha granulosa) are known to endow their eggs with TTX in order to protect them from predation. I looked at whether females allocated TTX to their eggs evenly over three years in captivity and compared those levels to TTX levels in eggs directly after capture. I found that eggs had lower levels of TTX following initial capture, but those levels did not change over the next three years. This provides evidence that TTX is endogenously produced in this species. Because of the high levels of TTX in newts, there are few known predators. I observed river otters feeding on newts in a high elevation lake in Oregon. I found that these newts have very low levels of TTX, and that in general high elevation populations in Oregon have low levels of TTX relative to low elevation populations. Finally, I documented TTX in two species of terrestrial flatworm (Bipalium adventitium and Bipalium kewense). Tetrodotoxin has never before been identified in a terrestrial invertebrate species. Further, I found evidence that suggests that TTX is used for both defense and prey capture in these worms. These studies add to our understanding of the evolution of TTX and how it influences interactions between organisms and their biotic and abiotic environments.

I investigated the ecological functions of halogenated secondary metabolites from the red alga Asparagopsis armata, their localisation in specialised cells and also their cost of production. A. armata produces large amounts of halogenated metabolites ( &lt 20 ??g / mg dry weight) that are sequestered in gland cells, as was demonstrated with light, epifluorescence and transmission electron microscopy. Cellular structures were identified that likely assist the release of metabolites from the gland cells to the algal surface. The halogenated metabolites of A. armata have multiple ecological roles, functioning as both inhibitors of bacterial fouling and as herbivore deterrents. Their activity against bacteria and herbivores was measured by a novel test in which the metabolites were manipulated in A. armata by omitting bromide ions from the culture media. This technique prevented the production of halogenated metabolites, but did not impact on other aspects of algal biology. Algae lacking halogenated metabolites (bromide [-] algae) had higher densities of epiphytic bacteria than those that continued to produce metabolites (bromide [+] algae). Bioassays with pure compounds against individual bacterial isolates further supported an inhibitory role for the halogenated metabolites against epiphytic bacteria, and also indicated an affect on bacterial community structure as well as abundance. Bromide (+) A. armata produced halogenated metabolites that also deterred feeding by two herbivores (an amphipod and an abalone), but not a third (an opisthobranch mollusc). A novel outcome from these feeding assays was the demonstration of a relationship between herbivore size and consumption of the chemically defended A. armata by the abalone Haliotis rubra. In addition to the fitness benefits gained from chemical defence, there were also costs for allocating resources to secondary metabolites. These costs were only detected under limiting light resources, consistent with predictions of the plant defence models. The integration of chemical analyses and cellular measures of chemical defence proved essential in elucidating resource allocation to chemical defence in the filamentous stage of A. armata. This thesis highlights that the simple relationships between growth and defence in filamentous algae can provide an excellent model for studies of the ecology and evolution of chemical defences in marine algae.

Marine consumers and their prey display plasticity that affects the outcomes of their dynamic interactions as well as community structure and ecosystem function. Aquatic chemical signals induced plasticity in consumers and prey from a broad range of taxonomy (phytoplankton to fishes), sizes (microscopic to macroscopic), and habitats (pelagic to benthic), and this complex plasticity strongly affected consumer-prey interactions. Two fishes,

Thesis (M.S.)--Oregon State University, 2010. / Printout. Includes bibliographical references (leaves 45-54). Also available on the World Wide Web.

Thesis (M.S. in Bio.)--School of Biology, Georgia Institute of Technology, 2004. Directed by Julia Kubanek. / Includes bibliographical references (leaves 42-48).

Thesis (M. Phil.)--Hong Kong University of Science and Technology, 2003. / Includes bibliographical references. Also available in electronic version. Access restricted to campus users.

I investigated how competitor species respond to chemical cues released from the red tide dinoflagellate Karenia brevis. K. brevis produces a mix of unstable, relatively polar, allelopathic organic molecules that are produced and released at low concentrations. The production of these compounds also varies greatly within and among strains of K. brevis. The majority of these compounds caused sublethal reductions in competitor growth. In laboratory experiments, these compounds inhibited the growth of competitors Asterionellopsis glacialis, Skeletonema grethae, Prorocentrum minimum, and Akashiwo sanguinea, although each species was susceptible to a different sub-set of K. brevis compounds. Cell physiological state and population densities were important in dictating the susceptibility of competitors to allelopathy: phytoplankton were most susceptible to K. brevis allelopathy when in earlier growth stages (rather than later stages) and in lower cell concentrations. However, these compounds have limited negative effects on natural, mixed populations of competitors from both near and offshore environments, and competitors from inshore and offshore environments appear to respond similarly to K. brevis allelopathy. In the sensitive competitor, Thalassiosira pseudonana, allelopathic compounds ultimately caused a reshuffling of cellular nitrogen pools, altered carbon storage and impaired osmotic regulation as determined using a nuclear magnetic resonance (NMR) based metabolomics approach. By characterizing the pool of primary metabolites present in the cell after exposure to K. brevis cues, we inferred which metabolic pathways may be affected by allelopathy. For instance, concentrations of betaine and the aromatic metabolite homarine were suppressed, indicating that K. brevis allelopathy may disrupt this competitor’s ability to osmoregulate. Exposure to K. brevis cues enhanced the concentrations of glutamate and the fatty acid caprylate/caprate in T. pseudonana, suggesting that protein degradation was enhanced and that energy metabolism was altered. This contrasts with the response to K. brevis allelopathy of the diatom Asterionellopsis glacialis, which was much more resistant to chemical cues produced by K. brevis, likely through as yet unidentified detoxification pathways. Overall, my dissertation research provides insight into how species-specific, antagonistic interactions among phytoplankton competitors can affect community structure through direct or indirect mechanisms, highlights the potential role of allelopathy in the maintenance of K. brevis blooms, and uses a novel tool set (i.e., metabolomics) to determine the molecular targets of K. brevis allelopathy. It further demonstrates that planktonic communities are complex and dynamic ecological systems and that interspecific interactions between phytoplankton can have unexpected, cascading impacts in marine systems.

I investigated the ecological functions of halogenated secondary metabolites from the red alga Asparagopsis armata, their localisation in specialised cells and also their cost of production. A. armata produces large amounts of halogenated metabolites ( &lt 20 ??g / mg dry weight) that are sequestered in gland cells, as was demonstrated with light, epifluorescence and transmission electron microscopy. Cellular structures were identified that likely assist the release of metabolites from the gland cells to the algal surface. The halogenated metabolites of A. armata have multiple ecological roles, functioning as both inhibitors of bacterial fouling and as herbivore deterrents. Their activity against bacteria and herbivores was measured by a novel test in which the metabolites were manipulated in A. armata by omitting bromide ions from the culture media. This technique prevented the production of halogenated metabolites, but did not impact on other aspects of algal biology. Algae lacking halogenated metabolites (bromide [-] algae) had higher densities of epiphytic bacteria than those that continued to produce metabolites (bromide [+] algae). Bioassays with pure compounds against individual bacterial isolates further supported an inhibitory role for the halogenated metabolites against epiphytic bacteria, and also indicated an affect on bacterial community structure as well as abundance. Bromide (+) A. armata produced halogenated metabolites that also deterred feeding by two herbivores (an amphipod and an abalone), but not a third (an opisthobranch mollusc). A novel outcome from these feeding assays was the demonstration of a relationship between herbivore size and consumption of the chemically defended A. armata by the abalone Haliotis rubra. In addition to the fitness benefits gained from chemical defence, there were also costs for allocating resources to secondary metabolites. These costs were only detected under limiting light resources, consistent with predictions of the plant defence models. The integration of chemical analyses and cellular measures of chemical defence proved essential in elucidating resource allocation to chemical defence in the filamentous stage of A. armata. This thesis highlights that the simple relationships between growth and defence in filamentous algae can provide an excellent model for studies of the ecology and evolution of chemical defences in marine algae.

Thesis (Ph. D.)--University of California, San Diego, 1998. / Vita. Includes bibliographical references (p. 223-227).

Thesis (Ph.D.)--Hong Kong University of Science and Technology, 2005. / Includes bibliographical references. Also available in electronic version.