Harmful Algal Bloom Toxicity in Lithobates catesbeiana Tadpoles

Harmful algal blooms (HAB) have become a major health concern worldwide, not just to humans that consume and recreate on contaminated waters, but also to the fauna that inhabit the environments surrounding affected areas. HABs contain heterotrophic bacteria, cyanobacterial lipopolysaccharide, and cyanobacterial toxins such as microcystins, that can cause severe toxicity in many aquatic species as well as bioaccumulation within various organs. Thus, the possibility of trophic transference of this toxin through the food chain has potentially important health implications for other organisms in the related food web. While some species have developed adaptions to attenuate the toxic effects of HAB toxins, there are still numerous species that remain vulnerable, including Lithobates catesbeiana (American bullfrog) tadpoles. In the current study we demonstrate that acute, short-term exposure of tadpoles to HAB toxins containing 1 µg/L (1 nmol/L) of total microcystins for only 7 days results in significant liver and intestinal toxicity within tadpoles. Exposed tadpoles had increased intestinal diameter, decreased intestinal fold heights, and a constant number of intestinal folds, indicating pathological intestinal distension, similar to what is seen in various disease processes, such as toxic megacolon. HAB-toxin-exposed tadpoles also demonstrated hepatocyte hypertrophy with increased hepatocyte binucleation consistent with carcinogenic and oxidative processes within the liver. Both livers and intestines of HAB-toxin-exposed tadpoles demonstrated significant increases in protein carbonylation consistent with oxidative stress and damage. These findings demonstrate that short-term exposure to HAB toxins, including microcystins, can have significant adverse effects in amphibian populations. This acute, short-term toxicity highlights the need to evaluate the influence HAB toxins may have on other vulnerable species within the food web and how those may ultimately also impact human health.

Olfactory epithelium ontogenesis and function in postembryonic North American Bullfrog (Rana (Lithobates) catesbeiana) tadpoles

During metamorphosis, the olfactory system remodelling in anuran tadpoles — to transition from detecting waterborne odorants to volatile odorants as frogs — is extensive. How the olfactory system transitions from the larval to frog form is poorly understood, particularly in species that become (semi-)terrestrial. We investigated the ontogeny and function of the olfactory epithelium of North American Bullfrog (Rana (Lithobates) catesbeiana Shaw, 1802) tadpoles at various stages of postembryonic development. Changes in sensory components observable at the epithelial surface were examined by scanning electron microscopy. Functionality of the developing epithelium was tested using a neurophysiological technique (electro-olfactography (EOG)), and behaviourally, using a choice maze to assess tadpole response to olfactory stimuli (algae extract, amino acids). The youngest (premetamorphic) tadpoles responded behaviourally to an amino acid mixture despite having underdeveloped olfactory structures (cilia, olfactory knobs) and no EOG response. The consistent appearance of olfactory structures in older (prometamorphic) tadpoles coincided with reliably obtaining EOG responses to olfactory stimuli. However, as tadpoles aged further, and despite indistinguishable differences in sensory components, behavioural- and EOG-based olfactory responses were drastically reduced, most strongly near metamorphic climax. This work demonstrates a more complex relationship between structure and function of the olfactory system during tadpole life history than originally thought.