Fifty-nine female and six male little brown myotis (Myotis lucifugus) were radio-tagged during the summers of 2012, 2013, 2014, and 2015 in Yellowstone National Park. The grand models for daily maximum skin temperature (F98,154 = 1.55, P = 0.007), daily minimum skin temperature (F98,154 = 1.33, P = 0.05), and daily variation in skin temperature (F98,154 = 1.56, P = 0.006) were significant across roost type and reproductive condition class for adult females. Roosts were classified into Types A (warmest roosts), B (roosts with largest daily temperature variance), and C (stable and cool roosts) depending on differences in mean maximum, minimum, and variance in temperatures per day (P < 0.001). A total of 347 torpor bouts were recorded from 38 females across the 2012 to 2015 summer seasons. Bats across different reproductive classes and roost types used torpor at different hours of the day. My research suggests that adult female little brown myotis at high elevations in the Park extensively use and rely on building structures for roosting sites during the reproductive season, whereas males used primarily natural roosts.

The importance of social behaviour is evident in human society, but there are both costs and benefits associated with cooperation and sociality throughout the animal kingdom. At what point do the benefits outweigh the costs, and when do selective pressures favour sociality and colonization over solitude and independence? To investigate these questions, we have focused on an anomalous species of spider, Anelosimus studiosus, also known now as the northern social spider. Throughout its broad range, A. studiosus is solitary and aggressive, but recently, colonies of cooperative and social individuals have been observed at northern latitudes. This leads to two research questions: 1) what characteristics differentiate the two variants behaviourally, and, 2) how are they different physiologically? Colonies and individuals were collected from multiple populations throughout the Tennessee River watershed area and maintained in a laboratory environment for quantitative and qualitative assessment of behavioural traits as well as specific neurochemical analysis by high performance liquid chromatography with electrochemical detection. After classifying individuals as social or aggressive, I looked at the influence of factors such as age, reproductive state, nutritional state, and time of day on behaviour and neurophysiology. I found correlations between social behaviours and serotonin, aggressive behaviours and octopamine (invertebrate counterpart of norepinephrine), and several other compounds associated with an increase or decrease in aggression. These studies combine techniques from multiple disciplines to contribute to the greater understanding of the proximate control of social and aggressive behaviours as well as factors influencing the evolution of sociality.

Ectothermic animals are susceptible to temperature changes such as cold shock with seasons. To survive through a cold shock, ectotherms have developed unique strategies. My interest is focusing on the physiological function of during cold shock and prolonged cold exposure in the fruit fly (Drosophila melanogaster) and crayfish (Procambarus clarkii). I used Drosophila melanogaster as a model system to investigate cardiac function in response to modulators (serotonin, acetylcholine, octopamine, dopamine and a cocktail of modulators) in acute cold shock and chronic cold shock conditions as possible mechanism to regulate heart rate in the cold. To examine if the dampened heart rate in the cold could still be enhanced by modulators or calcium loading, modulators and light-sensitive channelrhodopsin proteins were utilized to stimulate the heart. This light induced cardiac activation increased heart rate in all conditions, and potentially can be used for cardiac therapy in mammals. Also, the acute and chronic cold conditioned heart showed responsiveness to the above mention modulators. In examining how synaptic transmission is influenced by acute and chronic cold, the crayfish neuromuscular junction was used as a model. This is a good model as there are high and low output synapses to be investigated. The low output neuromuscular junction was enhanced in response to acute cold. The high output nmj increased in synaptic response to acute cold. In addressing chronic cold conditions, the nmj were physiologically assayed in their response to acute warm changes as well as influence of serotonin and octopamine. In chronic cold condition, the synaptic output was varied in enhanced and dampened responses to an acute warm environment. These junctions were enhanced in their synaptic output by serotonin and octopamine (100nM). In assessing, by HPLC assay, octopamine concentration increased in chronic cold crayfish. This suggests compensation in synaptic transmission in cold acclimation possibility via endocrine responses.

Reduced reproduction has been shown to increase lifespan in many animals, yet the mechanisms behind this trade-off are mostly unknown. A previous study has shown that in the lubber grasshopper, Romalea microptera, ovariectomized (OVX) individuals have a 30% increase in lifespan relative to controls (Sham). In a separate study, an increase in fat body mass and a halting of ovarian growth were seen upon reduction of vitellogenin transcript via RNAi (VgRNAi). These data suggest that VgRNAi increases lifespan through the trade-off between reproduction and longevity and animals with combined ovariectomy and VgRNAi, might show additive physiological responses. In this study, we used two injection control groups for the VgRNAi treatment, namely buffer injection or injection with RNAi against a 90kDa hexamerin storage protein (Hex90RNAi). We have combined these manipulations to test lifespans upon: OVX & VgRNAi, OVX & Hex90RNAi, OVX & Buffer, Sham & VgRNAi, Sham & Hex90RNAi, and Sham & Buffer. Ovariectomy and VgRNAi exhibited similar reductions in feeding (~40%) and extensions in lifespan (13-21%) but showed differences in vitellogenin protein levels. This study also observed the effects of reduced reproduction on hexamerin storage proteins. We observed that upon ovariectomy and VgRNAi, hexamerins were increased, emphasizing the importance of protein in insect life extension. When methods to reduce reproduction were combined (OVX VgRNAi), no additive physiological responses were observed, suggesting ovariectomy and VgRNAi each extend lifespan by overlapping or convergent pathways.

As the earth’s climate changes due to anthropogenic emissions, it has increasingly become an imperative within the ecological community to understand existing species adaptations to climate change. Much focus has been paid to how a species might react to climate change, but the role of locally adapted traits and responsible environmental mechanisms have received less attention. Quantifying how sublethal (e.g. growth rates) and lethal (e.g. thermal tolerance) trait performance vary between populations can thus improve our understanding of how populations, and the entire species, will react to climate change. Here, I quantified the spatial patterns of performance of several traits in populations of the predatory marine snail Urosalpinx cinerea from across two thermal gradients on the Pacific and Atlantic coasts of North America. In chapter 2, I quantified local adaptation and plasticity of thermal tolerance, warming tolerance, and developmental traits of Urosalpinx. I found that while low latitude populations have evolved higher thermal tolerance than their low latitude counterparts, they also demonstrate negative plasticity in response to higher acclimation temperatures. This is likely a result of low latitude population adaptation to cooler developmental conditions. Further, low latitude populations live in environments much closer to their thermal maxima than high latitude counterparts, resulting in higher climate sensitivity in low latitudes. In chapter 3, I quantified growth and consumption rates of Urosalpinx via a common garden experiment. I found evidence for a novel pattern of trait adaptation, wherein high latitude populations tended to have higher trait performance at higher thermal optima than low latitude counterparts. This can be attributed to the maximizing of growth rate during short growing seasons at high latitudes. Together, these results demonstrate that local adaptation in endemic across two traits in Urosalpinx. I demonstrate that these traits tend to be adapted to aspects of the environment directly related to aspects of Urosalpinx phenology, and not to environmental means as is commonly assumed. These insights suggest that models of organismal performance under climate change must consider not only the potential for local adaptation in populations, but also the aspects of the environment to which these populations are evolved.

Global biodiversity is declining as a direct result of anthropogenic climate change. Ectothermic species have become focal organisms for studying the ecological effects of altered climates due to the clear relationship between environmental temperatures and ectotherms’ basic physiological functions. Historically, examinations of these effects have focused heavily on heliothermic lizards, and most others have tended to focus on single populations or sympatric species within a single community. Addressing the longterm energetic implications of environmental temperature variation will provide valuable insight into the cascading physiological effects that certain populations or species may experience as a result of altered climates. In this study, we used thermal and behavioral data collected between 2010 and 2017 from four distinct populations of Pacific rattlesnakes (Crotalus oreganus) on the Central Coast of California. Two of these populations occupy thermally mild, coastal habitats while the other two occupy more thermally dynamic, inland habitats. Using operative temperature models, surgically implanted temperature loggers, and radiotelemetry, we collected data on the thermal microhabitats available within each of these study sites as well as field active body temperatures for 85 individual snakes. With the addition of a lab-derived preferred body temperature range, we determined the thermal quality of each site and the thermoregulatory accuracy of snakes from each population. Field behavioral observations, gathered from snakes at all four sites simultaneously during the year 2017, revealed how snakes utilize the thermal landscape and adjust thermoregulatory behavior to mitigate the effects of different climates. Although overall thermal quality was best at coastal sites, thermal quality of the microhabitats within each site varied greatly. Consistent with findings in other squamate reptiles, inland snakes thermoregulated more accurately, despite being in more thermally constrained environments. Despite the fact that coastal snakes had lower mean field active body temperatures, the preferred body temperatures of snakes were the same across all four sites. However, field active body temperatures were consistently lower than the preferred range, suggesting there are additional variables that influence thermoregulatory behavior. Using established equations estimating the resting metabolic rates of snakes based on body mass and temperature, we calculated resting metabolic rate and annual baseline maintenance energy expenditure for each population. Coastal snakes, which had lower field active body temperatures, had overall lower metabolic rates than inland snakes, but upon correcting for mass, snakes at neither coastal nor inland sites differed in metabolic rates. Therefore, the majority of the differences observed in metabolic rates are driven by body size and not field-active body temperature. Inland snakes need, on average, approximately 1.6x more food annually than coastal snakes. Due to overall low resting metabolic rates, this translates to snakes at all sites needing less than one ground squirrel (their most common food item on the Central Coast) per year to fuel basic physiological functions. Finally, we used conservative predictive climate change models allowing either 1°C or 2°C increases to predict changes in the thermal quality of each site and ensuing changes in snake metabolic rates and maintenance energy expenditure. Due to the relatively high preferred body temperature of C. oreganus, thermal quality of the environment will actually increase under these climate models; due to an increase in ambient temperature, the proportion of hourly temperatures that fall within the preferred body temperature range will increase. If snake body temperature were to increase as the climate warms, a theoretical increase in body temperature of 1 and 2°C would have a low impact on the overall energetic needs of snakes, still allowing them to meet baseline maintenance energetic needs with only one large meal a year. Furthermore, we expect small increases in ambient temperature to have little impact on rattlesnakes because they are fairly precise thermoregulators, maintaining fairly constant body temperatures regardless of their thermal surroundings. Overall, our results show that studying the thermal ecology of multiple populations of a single species can reveal fine-scale information about the relationship between the thermal landscape and both ectotherm behavior and physiological processes. Additionally, our findings show that some species of large-bodied reptiles may be robust to modest thermal perturbations under conservative climate change predictions.

Amidst the amphibian extinction crisis, in situ and ex situ amphibian species conservation initiatives utilize assisted reproductive technologies for optimal genetic management of captive and wild populations. Development of effective, simple, low-cost methods for obtaining gametes for artificial fertilization, sperm biobanking, or natural breeding alleviates obstacles for institutions or programs to apply reproductive technologies. Objectives herein include investigating the efficacy of alternative hormones for inducing spermiation, an alternative hormone administration route for inducing ovulation, exploring the physiological effects of hormone therapy, and utilizing sperm from deceased animals for biobanking.

Glucose is transported in crustacean hepatopancreas and intestine by Na+-dependent co-transport, while Na+-dependent D-fructose influx has only been described for the hepatopancreas. It is still unclear if the two sugars are independently transported by two distinct cotransporter carrier systems. In this study lobster (Homarus americanus) hepatopancreas brush border membrane vesicles (BBMV) were used to characterize, in detail, the cation-dependency of both D-[3H] glucose and D-[3H] fructose influxes, while in vitro perfused intestines were employed to determine the nature of cation-dependent sugar transport in this organ. Over the sodium concentration range of 0-100 mM, both 3H-D-glucose and 3H-D-fructose influxes (0.1 mM; 1 min uptakes) by hepatopancreatic BBMV were hyperbolic functions of [Na+], exhibiting Km values of 2.30 ± 0.59 and 2.58 ± 0.95 mM, respectively. D-[3H] glucose and fructose influxes by hepatopancreatic BBMV over a potassium concentration range of 15-100 mM were hyperbolic functions of [K+], exhibiting Km values of 9.85 ± 0.41 and 12.6 ± 0.80 mM respectively. Both sugars displayed significant (p < 0.01) Na+/K+-dependent and Na+-independent uptake processes. Transepithelial 25 μM D-[3H] glucose and D-[3H] fructose fluxes across lobster intestine over a luminal sodium and potassium concentration range of 0 – 50 mM and 5-100 mM, respectively, were hyperbolic functions of luminal [Na+] and [K+]. As with hepatopancreatic sugar transport, transepithelial intestinal sugar transport exhibited both significant (p < 0.01) Na+/K+-dependent and Na+-independent processes. Results suggest that both D-glucose and D-fructose are transported by a single carrier process in each organ with sodium being the preferred cation for both sugars in the hepatopancreas, and potassium being the preferred cation for both sugars in the intestine.

Crustacean muscle fibers are some of the largest cells in the animal kingdom, with fiber diameters in the giant acorn barnacle (Balanus nubilus) exceeding 3 mm. Sessile animals with extreme muscle sizes and that live in the hypoxia-inducing intertidal zone – like B. nubilus – represent ideal models for probing the effects of oxygen limitation on muscle cells. We investigated changes in metabolism and structure of B. nubilus muscle in response to: normoxic immersion, anoxic immersion, or air emersion, for acute (6h) or chronic (6h exposures twice daily for 2wks) time periods. Following exposure, we immediately measured hemolymph pO2, pCO2, pH, Na+, Cl-, K+, and Ca+ then excised tergal depressor (TD) and scutal adductor (SA) muscles to determine citrate synthase (CS) activity, lactate dehydrogenase (LDH) activity, and D-lactate levels. We also prepared a subset of SA and TD muscles from the chronic barnacles for histological analysis of fiber diameter (Feret’s), cross-sectional area (CSA), mitochondrial distribution and relative density, as well as nuclear distribution and myonuclear domain size. There was a significant decrease in hemolymph pO2 and pCO2 following acute and chronic anoxic immersion, whereas air emersion pO2 and pCO2 was comparable to normoxic levels. Fiber CSA and diameter did not change significantly in either tissue, while myonuclear domain size in SA muscle was significantly lower in the anoxic and emersion groups than the normoxic control. Neither CS, nor LDH activity, showed any significant treatment effect in either tissue, whereas both muscles had significantly higher D-lactate levels after air emersion following acute (though not chronic) exposure. Thus far, our findings indicate that B. nubilus experience a general reduction in aerobic metabolism under anoxia, emersion is only mildly oxygen-limiting, and that muscle plasticity is occurring during chronic emersion and anoxia.

Teleost fishes incorporate renal ammonia excretion as part of a greater acid-base regulatory system. However, the transport mechanisms employed by the renal epithelium to excrete ammonia are relatively unknown. I hypothesized that, under metabolic acidosis, increased renal ammonia excretion would be the product of tubular secretion and involve a Na+/NH4+ exchange metabolon mediated through Rhesus (Rh) glycoproteins. To induce metabolic acidosis, goldfish (Carassius auratus) were exposed to a low pH environment (pH 4.0; 48-h). There was a clear signal of metabolic acidosis: a reduction in both plasma [HCO3-] and blood pH with no influence on plasma PCO2. Goldfish demonstrated an elevation in total plasma [ammonia] with a reduction in PNH3 under acidosis. Metabolic acidosis induced higher rates of urinary excretion of acidic equivalents in the form of both NH4+ and titratable acidity-HCO3- (TA-HCO3-) excretion. Urinary Na+ excretion was not affected by acidosis and urine [Na+] did not correlate with urinary [ammonia]. Alanine aminotransferase activity in the kidney was higher in acidotic goldfish. Glomerular filtration rate and urine flow rate were not affected by acidosis. Increased renal NH4+ excretion was due to increased secretion, and not increased filtration, of ammonia. There was a corresponding elevation in Rhcg1b mRNA expression but no change in renal Na+ reabsorption. My data support a secretion-based mechanism of teleost renal ammonia transport. This system is Na+ independent and is likely mediated by Rh glycoproteins and H+ ATPase, involving a parallel H+/NH3 secretion mechanism. To investigate effects of metabolic acidosis on elasmobranch fish, Pacific spiny dogfish (Squalus acanthias suckleyi) were infused with an acidic saline (125 mM HCl/375 mM NaCl; 3 ml/kg/h; 24-h). The results are preliminary, with no marked effects of HCl infusion on plasma acid-base or N-status, but increased branchial NHE2 and lower renal NHE3 protein expressions. These data are summarized in an Appendix. / Thesis / Master of Science (MSc)