Physiology

Stressors are often an inescapable part of an organism's life. While the effects of many stressors have been well studied individually, potential interactions between stressors exist that may result in greater than additive negative effects. Stressors may be linked by conflicting demands on energy budgets, interfering with important physiological pathways, or necessitating incompatible adaptive responses. Using Ranavirus (FV3) and larval dragonfly predators (Anax spp.) in a 2x2 factorial experiment on green frog (Lithobates clamitans) tadpoles, I investigate the interactions in behaviour, morphology, and metabolism when both stressors were applied in concert. I demonstrate that activity and feeding are reduced additively by both stressors, and tadpoles increase distance between conspecifics in FV3-exposed tanks, but only in the absence of predators. I also note decreases in mass, and a non-significant marginal increase in metabolic rate of tadpoles exposed to FV3. Interestingly, I provide evidence that FV3 can compromise morphometric responses through antagonistic interactions with perceived predation risk exposure, which may result in significantly elevated mortality even when either stressor is present in sub-lethal quantities. Thus, I conclude that sub-lethal exposure to stressors can nonetheless have substantial impacts on organisms and a more integrative approach to examining the impacts of stressors on individual physiology and fitness is necessary.

Author Keywords: Behaviour, Interaction, Morphology, Predation Risk, Ranavirus, Tadpoles

Salamanders are capable of tissue regeneration throughout all life-stages, which requires the dedifferentiation of mature cells to regrow lost tissues. Dedifferentiation is promoted by degradation of the extracellular matrix by matrix metalloproteases, as well as lysosomal degradation of intracellular and cell-surface proteins that mark cells as part of a mature lineage. Salamanders are also capable of developing cannibalistic phenotypes, plastic traits that are elicited by environmental stressors that result in elevated circulating glucocorticoid (e.g., corticosterone) levels that underlie many fundamental adaptive changes in morphology. Interestingly, the direct effect of corticosterone on regeneration and the cannibalistic phenotype have yet to be examined. In the present thesis, axolotls (Ambystoma mexicanum) were exposed to exogenous corticosterone and 50% of the distal tail tissue was removed. The effects of high corticosterone levels on matrix metalloprotease (MMP-2, MMP-9) and lysosomal acid phosphatase (LAP) activity were assessed; these are two classes of enzymes which are markers of extracellular matrix and intracellular remodeling during regeneration, respectively. We found that elevated corticosterone levels inhibited tissue regeneration, by prolonging the dedifferentiation phase as indicated by increased LAP and reduced MMP-2 and MMP-9 activity. Elevated corticosterone levels also promoted the cannibalistic morphology and this effect was strongest among smaller individuals.

Author Keywords: amphibian, cannibalistic morphology, corticosterone, dedifferentiation, regeneration, stress

Biological differences between island and mainland conspecifics have been well studied, but few studies have addressed differences in stress physiology. Stressors, such as predation and competition for resources, cause the release of glucocorticoids (GCs). Characteristics of island wildlife, called "island syndrome", are attributed to low levels of predators and competitors. I tested the hypothesis that island syndrome includes differences in GC levels between island and mainland rodents using two approaches; first, using white-footed mice (Peromyscus leucopus) from a near-shore archipelago (Thousand Islands, Ontario) and the nearby mainland; second, using study-skins of deer mice (Peromyscus maniculatus) from two archipelagos offshore of Vancouver Island, British Columbia. White-footed mice in the near-shore archipelago did not show characteristics of island syndrome, or changes in GC levels (feces and hair); however deer mice from both archipelagos in British Columbia were heavier and had lower hair GCs for their size than Vancouver Island mice.

Author Keywords: Glucocorticoids, Island rule, Island syndrome, Peromyscus, Stress physiology

Early developmental stages of cold-adapted ectotherms such as brook trout

(Salvelinus fontinalis) are at risk of mortality with increasing water temperatures because

of their sensitivity to changes in their environment. I studied the mass and routine

metabolic rate (RMR) of wild-origin brook trout eggs, alevin and young fry reared at

normal (5°C) and elevated (9°C) temperatures for the duration of the study or at

mismatched temperatures. This setup determined if preconditioning acclimation for one

temperature benefits or hinders the organism later in life. Three levels of biological

organization (ancestry, population, family) were studied using Akaike's Information

Criterion (AIC) to identify models that best accounted for variation in the data. Family,

mass and temperature were most important in predicting body mass and mass-adjusted

RMR, although population and ancestral-level differences were also detected at some life

stages. Strong variation in body mass and mass-adjusted RMR among families may

indicate adaptive potential within brook trout populations to respond to increases in water

temperature with climate change.

Author Keywords: Acclimation, AIC, Brook trout (Salvelinus fontinalis), Environmental matching, Routine metabolic rate, Temperature

The predator vs. prey dynamic is an omnipresent factor in ecological systems that may drive changes in life history patterns in prey animals through behavioural, morphological, and physiological changes. Predation risk can have profound effects on the life history events of an animal, and is influenced by the neuroendocrine stress response. Activation of the hypothalamic-pituitary-adrenal/interrenal axis, and the induction of stress hormones (e.g., corticosterone (CORT)) have been shown to mediate the onset of inducible anti-predator defensive traits including increased tail-depth, and reduced activity. The predator-prey relationship between dragonfly nymphs and tadpoles can be a powerful model system for understanding mechanisms that facilitate changes in the stress response in accordance with altered severity of risk. It has been well demonstrated early in tadpole ontogeny that increased corticosterone (CORT) levels, observed within three weeks of predator exposure, are correlated with increased tail depth morphology. However, the reactivity of the stress response in relation to the growth modulation in developing prey has yet to be fully explored. Accordingly, this thesis assessed the stress and growth response processes in tadpoles that were continuously exposed to perceived predation risk later in ontogeny. Continuous exposure of prey to predation risk for three weeks significantly increased CORT levels, and tail depth. However, tadpoles exposed to six weeks of predation risk acclimated to the presence of the predator, which was observed as a significant reduction of stressor-induced CORT levels. In addition, although increased tail depth has been attributed to predator defense, predator-naïve tadpoles began to display similar tail depth morphology as treated tadpoles at the six week time point. Thus, this thesis suggests that the stress response in lower vertebrate systems (e.g., tadpoles) may operate in a similarly complex manner to that observed in higher vertebrates (e.g., rats), for which severity of risk associated with the stressor aids in defining activity of the stress response. Moreover, the lack of morphological difference between treatments among tadpoles exposed later in ontogeny suggests that the mechanisms for inducing defenses are normal morphological traits in the development of the animal. This thesis paves the way for future research to elucidate the relationship between the neuroendocrine stress response and hormonal pathways involved in growth modulation in the presence of environmental pressures.

Author Keywords: Acclimation, Corticosterone, Growth Modulation, Predation Risk, R. pipiens, Tadpole

Northern and southern flying squirrels (Glaucomys sabrinus and G. volans, respectively) are experiencing a climate change induced increase in range overlap, resulting in recent hybridization. We investigated the occurrence of heterospecific communal nesting, a potential facilitator of hybridization, and aimed to confirm the presence of torpor, a potential barrier to hybridization, in flying squirrels. In wild-caught captive squirrels, we conducted a paired nest choice experiment and found that heterospecific nesting did occur, but in a lower frequency than conspecific nesting. Ambient temperature did not affect the frequency of grouped nesting. We attempted to induce torpor in flying squirrels in a laboratory through cold exposure while measuring metabolic rate and body temperature. Strong evidence of torpor was not observed, and metabolic rate remained unchanged with season. We conclude that torpor is not a barrier to hybridization in flying squirrels, but resistance to heterospecific nesting may indicate the existence of one.

Author Keywords: heterospecific group, hybridization, northern flying squirrel, social thermoregulation, southern flying squirrel, torpor