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.