The aim of this study was to investigate the reproductive ecology including fecundity, reproductive season, timing of larval release, evaporative water loss during reproductive migration, and the salinity tolerance of first zoeal stage of Cardisoma carnifex in Houwan, Hengchun Peninsula. The average carapace width of ovigerous females was 70.5 ¡Ó 6.57 mm, with a size range of 56.0-93.7 mm. Fecundity intimates increased with size and the number of hatched larvae from individual female varied from 90,000 to 490,000. The reproductive season of C. carnifex began at the onset of the rainy season and ends approximately at the end of that season. The reproductive season was from June to Oct. in 2009 and May to Sept. in 2010. The peaks of larval release occurred in July in 2009, 2010. Some females had the capacity in spawning twice during a reproductive season. Ovigerous females releasing larvae synchronously. The relationship between larval release timing and environmental cycle is listed in decreasing order of importance: diel rhythm, semilunar rhythm, tidal amplitude rhythm, and a low correlation was found with the tidal rhythm. Ovigerous females of C. carnifex started to release larvae after the new and full moon, lasting for 6-8 days with peaks occurring 4-6 days after the new and full moon. The peaks of larval release occurred within 2-3 hours after the sunset time (i.e., between 1930H and 2130H). This species exhibits different larval release timing and behavior compare to other species of Gecarcinidae; ovigerous females of gecarcinid crabs follow a lunar rhythm but C. carnifex follow a semilunar rhythm. More than half of the ovigerous females stayed in the water for up to 10 min (average 5.5 ¡Ó 2.87 min) and could move to deep water (i.e., 120 cm) to release their larvae. A few females even released their larvae in the freshwater environment. For C. carnifex, a mean of 15¢H of its original weight was lost by the time they died and the osmolality of the hemolymph is 856 mOsm kg-1. Compare with the hemolymph osmolality of ovigerous females (694 ¡Ó 6.3 mOsm kg-1), the results indicated that the ovigerous females at Houwan may not experience severe water loss during larval release migration. The first zoeal stage of C. carnifex could not survive in the freshwater, all larvae died within 2 hours. However, at salinities large than 5‰, most larvae could survive for more than 3 days. The results show first zoeal stage of C. carnifex had adapted to the low salinity in the estuary.

4 Summary Seasonal metabolic adjustments and partitioning of evaporative water loss in Wahlberg’s epauletted fruit bat, Epomophorus wahlbergi Student: Ingrid A. Minnaar Supervisor: Prof. A. E. McKechnie Co-supervisors: Prof. N. C. Bennett, Prof. Christian T. Chimimba Department: Zoology and Entomology, University of Pretoria Degree: MSc: Zoology The capacity to thermoregulate over a wide range of TaS is critical for maintaining homeostasis in endotherms. Several aspects of the thermoregulatory properties of bats remain poorly studied when compared to other mammals and birds. I examined two specific aspects of thermoregulation in bats: the seasonal variation of maximum metabolic heat production and the partitioning of total evaporative water loss (TEWL) into respiratory and cutaneous components. I measured basal metabolic rate (BMR) and summit metabolism (Msum) in captive and wild Wahlberg’s epauletted fruit bats, Epomophorus wahlbergi, during summer and winter. I measured metabolic rate using flow-through respirometry, and elicited Msum by exposing bats to low temperatures in a helox (21% O2, 79% He) atmosphere. BMR decreased by 22-25% during winter in both captive and wild bats, with the BMR of captive bats 9-13% lower than the wild individuals across seasons. Msum was approximately seasonally stable in both captive and wild bats, but Msum in captive individuals was 13-18% higher than their wild conspecifics during both seasons. The ratio between Msum and BMR (i.e., metabolic expansibility) was greater in winter than during summer for both captive and wild bats. One likely explanation for the greater resting thermogenic capacity of the bats in captive individuals concerns their reduced activity levels; compared to wild, free-ranging bats, heat produced as a by-product of activity probably contributed far less to thermoregulation, apparently leading to an increase in resting heat production capacity in captive individuals. 5 At the other end of the thermal scale, knowledge of heat tolerance and the evaporative cooling mechanisms employed by bats in hot weather remains rudimentary. At high air temperatures (Ta), endotherms avoid overheating by dissipating heat via evaporative water loss. TEWL may be partitioned into cutaneous evaporative water loss (CEWL) and respiratory evaporative water loss (REWL). I quantified CEWL and REWL in E. wahlbergi at Tas of 10-40 °C using a latex mask. When Ta exceeded normothermic Tb, bats drastically increased their TEWL, metabolic rate and Tb. The relative contribution of CEWL to TEWL was the greatest at moderate Tas where it represented up to 80% of TEWL. REWL was the major route of evaporative cooling at the highest Ta: at Ta = 40 ºC, REWL represented 45% of TEWL. To avoid hyperthermia, E. wahlbergi greatly increased metabolic rate at high TaS to avoid hyperthermia, further compounding the need to cool down. REWL is thought to be less efficient as than CEWL in offloading heat at high TaS as panting increases metabolic heat, whereas CEWL occurs passively. There is a need for further studies to be conducted on the thermoregulatory capabilities of bats in varying environmental conditions, both intra- and interspecifically. / Dissertation (MSc)--University of Pretoria, 2013. / gm2014 / Zoology and Entomology / unrestricted

Understanding how organisms respond to climatic variability and novel conditions is becoming an increasingly important task for ecologists. For ectotherms in the northern hemisphere, the response to cold is of special interest, considering that poleward range expansion events and increasing variability of temperatures during winter are already being observed as consequences of a warming planet. Though direction of change in physiological variables in response to cold is well studied in ectotherms, the extent to which traits can change and the rate at which they can change is not. We compared the extent and rate of change in cold tolerance (CTmin) between two long-term captive populations of the Italian wall lizard (Podarcis siculus) during a lab cold-acclimation treatment. Heat tolerance (CTmax), thermal preference (Tpref), temperature dependent rates of oxygen consumption (SMRO2), and temperature dependent rates of water loss (EWL) were also compared between Italian wall lizards previously introduced to Long Island, NY and San Pedro, CA before and after the lab cold acclimation treatment. Because our study coincided with a cold snap during the spring 2018 season for the San Pedro, CA population, we also studied the effects of cold acclimatization on wild lizards from the CA population. After initial lab acclimation of the lizards to laboratory conditions, SMRO2 at 15°C and EWL at 10°C were higher in NY lizards compared to CA lizards. Lizards from the two populations did not differ in any other variables measured before the cold acclimation treatment. We found that lizards from the NY population experienced an 80% decrease in CTmin following a switch from 20°C:18°C to 17.5°C:16°C (12h light:12h dark) acclimation treatment. Lizards from the CA population did not decrease CTmin in response to the same cold acclimation treatment. Overall, NY lizards decreased CTmin, CTmax, and Tpref following cold acclimation, whereas CA lizards decreased CTmax only. Wild CA lizards decreased CTmax following the cold spring 2018 season in a manner similar to that of lab acclimated NY and CA lizards, suggesting that these lizards do not maintain a high CTmax when the environment is unlikely to expose them to high temperatures. Thermal sensitivity (Q10) of SMRO2 and EWL was lower in NY lizards, suggesting physiological adaptation to fluctuation in diurnal temperatures. The ratio of CO2 produced to O2 consumed (respiratory exchange ratio, RER) measured at 15°C increased in NY lizards following cold acclimation suggesting an increased use of carbohydrates and/or an increased production of lipids in the colder conditions. These responses in combination with the higher observed plasticity in NY lizards are in accordance with the climatic variability hypothesis, which predicts that organisms from more variable climates will be better adapted to physiologically respond to variable conditions. The higher capacity for physiological plasticity may explain the relatively high success of P. siculus in NY and other northern U.S. states. By describing the rate of change of CTmin during cold acclimation we hope to better understand how these lizards minimize the risk of low temperature exposure during winter. We ultimately hope to incorporate the rate at which cold tolerance can change into predictions of species distributions and hypothesis tests investigating the relationship between climatic variability and the rate at which animals can exhibit plasticity.

Chapter 1: Animals can respond to extreme climate by behaviorally avoiding it, or by physiologically coping with it. We understand behavioral thermoregulation and physiological thermal tolerances, but water balance has largely been neglected. Climate change includes both global warming and changes in precipitation regimes, so improving our understanding of organismal water balance is increasingly urgent. We assessed the hydric physiology of endangered Blunt-nosed Leopard Lizards (Gambelia sila) by measuring cutaneous evaporative water loss (CEWL), plasma osmolality, body mass, and body condition throughout their active season. On average, G. sila had low CEWL that is likely desert-adaptive, and high plasma osmolality that is indicative of dehydration. Given that our study was in a drought year, it is reasonable to believe that every lizard measured was dehydrated to a degree. We hypothesized that throughout the G. sila active season, as their habitat got hotter and drier, G. sila would become increasingly dehydrated and watertight. Instead, CEWL and plasma osmolality showed minimal change for females and nonlinear change for males, which we hypothesize is connected to sex-specific reproductive behaviors and changes in food availability. We also measured thermoregulation and microhabitat use, expecting that more hydrated lizards would have higher body temperature, better thermoregulatory accuracy, and spend more time aboveground. However, we found no effect of CEWL, plasma osmolality, body mass, or body condition on these thermal and behavioral metrics. We posit either that G. sila tolerate dehydration to maintain activity during their brief active season, or that because every lizard was dehydrated due to the drought, they all experienced equally constrained thermoregulation and microhabitat use. Finally, G. sila spend considerable time underground in burrows, and we believe burrows serve as essential hydric, not only thermal, refugia. Our findings suggest that these lizards might benefit from artificial humid refugia and supplemental hydration, especially during drought. Chapter 2: Testing acclimation plasticity informs our understanding of functional biodiversity and applies to conservation management amidst our rapidly changing climate. While there is a wealth of research on the plasticity of thermal and hydric physiology in response to temperature acclimation, there is a comparative gap for research on acclimation to different hydric regimes, as well as the interaction between water and temperature. We sought to fill this gap by acclimating Western Fence Lizards (Sceloporus occidentalis) to experimental climate conditions (crossed design of Hot or Cool, Dry or Humid) for eight days, and measuring cutaneous evaporative water loss (CEWL), plasma osmolality, hematocrit, and body condition before and after acclimation under common conditions. CEWL changed plastically in response to the different climates, with lizards acclimated to Hot Humid conditions experiencing the greatest increase in CEWL. Change in CEWL among individuals was negatively related to treatment vapor pressure deficit. Plasma osmolality, hematocrit, and body condition all showed greater changes in response to temperature than to humidity or vapor pressure deficit. CEWL and plasma osmolality were positively related across treatment groups before acclimation and within treatment groups after acclimation, but the two variables showed different responses to acclimation, suggesting that they are interrelated but governed by different mechanisms. This study is among just a small number of studies that assess more than one metric of hydric physiology and that test the interactive effects of temperature and humidity. Such measurements will be essential for predictive models of activity and survival for animals under climate change.

Hypothesised causes of the observed world-wide decline of amphibian populations are varied and in some cases contentious. Insufficient information relating to the autecology of many amphibian species can cause erroneous speculations regarding critical habitat requirements and hence management programs designed to enhance population viability are often unsuccessful. Most amphibians display a bi-phasic life history that involves occupation of an aquatic breeding habitat and terrestrial habitats that are used for foraging, and shelter from predation and environmental stress. However, the focus of most amphibian research is centred on the breeding habitat, with limited research being conducted into the terrestrial habitat requirements of most amphibian species. Barakula State Forest is a large continuous area of open woodland situated in the semi-arid region of Queensland. The forest supports 21 species of endemic anurans, many of which use ephemeral waterbodies for breeding. This area is, therefore, an ideal location to test the relative importance of terrestrial habitat on the distribution of a suite of frogs that display different morphological and physiological characteristics. On the landscape scale, the attributes of the terrestrial environment at three survey areas within Barakula were similar. However, at the patch scale, ground truthing showed there were considerable variations in vegetation and ground cover attributes within and between each survey site. Measured properties of the soil also tended to vary within and between sites. Soil texture ranged from sandy to heavy clay, soil pH ranged from 3.9 to 6.4 and soil moisture varied considerably. Agar models, used for testing evaporative moisture loss at different microhabitats, retained significantly higher levels of moisture when positioned in the buried microhabitat during summer, but in winter, models that were placed under leaf litter retained higher levels of moisture. Variations in levels of moisture loss at the five different microhabitats were evident within and between the survey sites. Despite a prolonged drought, 1844 native frogs representing 17 species were pitfall trapped. Members from the family Myobatrachidae comprised 94% of these captures, and burrowing species accounted for 75% of total captures. Species were not randomly distributed within or between the survey sites. Vegetation attributes and soil properties played a significant role in influencing the catch rates and traplines that supported similar vegetation and soil attributes also tended to catch similar species. Capture rates of six of the seven burrowing species were significantly influenced by soil properties. When given a choice of four different microhabitats created in enclosures, individuals from five species showed varying responses to habitat choice during night time activity. During daylight all species tended to avoid bare areas and burrowing species tended to burrow under some form of cover. Pseudophryne bibronii metamorphs showed a significant avoidance to soils with high pH. The number of Limnodynastes ornatus metamorphs was significantly and positively correlated with moisture levels surrounding a breeding area. Limnodynastes ornatus metamorphs tended to avoid areas that did not support some form of cover. Embryos from the terrestrial egg laying P. bibronii translocated to sites with varying levels of soil pH, suffered increased mortality where the soil pH was &gt4.8. In the laboratory, embryonic survival was not significantly different between the four pH treatments. There was a significant influence of fungal infection on survival rates and ranked fungal infection was significantly different between the four pH treatments. The terrestrial environment at the three survey sites has provided sufficient protection from environmental elements to allow a large diversity of anurans to persist for long periods without access to permanent water. Management must consider the importance of the non-breeding habitat when defining buffer zones, restoration programs and conservation strategies to ensure that the complete set of ecological requirements for frog species are provided.

Anfíbios anuros, de uma forma geral, são caracterizados por baixa resistência cutânea à perda de água por evaporação, altas taxas de reidratação a partir da pele e alta tolerância à desidratação. Entretanto, diferenças interespecíficas destes parâmetros do balanço hídrico podem estar associadas a características ambientais das áreas de ocorrência. No Brasil, encontramos biomas com características climáticas contrastantes e uma rica anurofauna, consistindo em um modelo interessante para o estudo da evolução do balanço hídrico em anuros. Através de uma abordagem comparativa, este estudo testou as hipóteses de que, quando comparadas às espécies de anuros que habitam ambientes mais mésicos, espécies de ambientes mais xéricos deveriam apresentar: (1) maior resistência à perda de água por evaporação, (2) maior taxa de reidratação, (3) menor sensibilidade do desempenho locomotor à desidratação e (4) maior sensibilidade do desempenho locomotor à temperaturas mais baixas e menor sensibilidade deste a temperaturas mais altas. As taxas de reidratação e a resistência da pele à perda de água por evaporação apresentaram relação alométrica respectivamente positiva e negativa com a massa corpórea, indicando a evolução de mecanismos compensatórios das modificações das relações entre área de superfície e volume corpóreo com a variação de tamanho. Anuros de ambientes caracterizados por maior restrição em disponibilidade hídrica e temperaturas mais altas apresentaram maiores taxas de reidratação, menor sensibilidade do desempenho locomotor a temperaturas mais altas e maior sensibilidade deste a temperaturas mais baixas, indicando padrões de adaptação fisiológica a características ambientais. A variação em sensibilidade do desempenho locomotor à desidratação apresentou padrões mais complexos de relação com variáveis ambientais, sendo esta relação dependente dos ambientes e grupos filogenéticos amostrados / Anuran amphibians, in general, are characterized by low skin resistance to evaporative water loss, high rates of hydration through the skin and high tolerance to dehydration. However, interspecific differences of these water balance traits can be related to environmental characteristics of the areas of occurrence. In Brazil, there are biomes with contrasting climatic conditions and rich anuran fauna, which constitute an interesting model for studies on water balance evolution in anurans. Through a comparative approach, this study tested the hypotheses that, when compared to anuran species that inhabit more mesic environments, species from xeric environments should show: (1) increased resistance to evaporative water loss, (2) higher rates of rehydration, (3) low sensitivity of locomotor performance to dehydration and (4) increased sensitivity of the locomotor performance at lower temperatures and lower sensitivity of it at higher temperatures. Rehydration rates and skin resistance to evaporative water loss showed, respectively, positive and negative allometric relations with body mass, indicating the evolution of compensatory mechanisms to changes in the relationship between surface area and volume with body size variation. Anurans from environments characterized by higher restriction in water availability and higher temperatures had higher rates of hydration, lower sensitivity of locomotor performance at higher temperatures and higher sensitivity of it at lower temperatures, indicating patterns of physiologic adaptation to environmental characteristics. The variation in sensitivity of the locomotor performance to dehydration showed more complex relation patterns with environmental variables. These relations were dependent on the environments and phylogenetic groups sampled

Anfíbios anuros, de uma forma geral, são caracterizados por baixa resistência cutânea à perda de água por evaporação, altas taxas de reidratação a partir da pele e alta tolerância à desidratação. Entretanto, diferenças interespecíficas destes parâmetros do balanço hídrico podem estar associadas a características ambientais das áreas de ocorrência. No Brasil, encontramos biomas com características climáticas contrastantes e uma rica anurofauna, consistindo em um modelo interessante para o estudo da evolução do balanço hídrico em anuros. Através de uma abordagem comparativa, este estudo testou as hipóteses de que, quando comparadas às espécies de anuros que habitam ambientes mais mésicos, espécies de ambientes mais xéricos deveriam apresentar: (1) maior resistência à perda de água por evaporação, (2) maior taxa de reidratação, (3) menor sensibilidade do desempenho locomotor à desidratação e (4) maior sensibilidade do desempenho locomotor à temperaturas mais baixas e menor sensibilidade deste a temperaturas mais altas. As taxas de reidratação e a resistência da pele à perda de água por evaporação apresentaram relação alométrica respectivamente positiva e negativa com a massa corpórea, indicando a evolução de mecanismos compensatórios das modificações das relações entre área de superfície e volume corpóreo com a variação de tamanho. Anuros de ambientes caracterizados por maior restrição em disponibilidade hídrica e temperaturas mais altas apresentaram maiores taxas de reidratação, menor sensibilidade do desempenho locomotor a temperaturas mais altas e maior sensibilidade deste a temperaturas mais baixas, indicando padrões de adaptação fisiológica a características ambientais. A variação em sensibilidade do desempenho locomotor à desidratação apresentou padrões mais complexos de relação com variáveis ambientais, sendo esta relação dependente dos ambientes e grupos filogenéticos amostrados / Anuran amphibians, in general, are characterized by low skin resistance to evaporative water loss, high rates of hydration through the skin and high tolerance to dehydration. However, interspecific differences of these water balance traits can be related to environmental characteristics of the areas of occurrence. In Brazil, there are biomes with contrasting climatic conditions and rich anuran fauna, which constitute an interesting model for studies on water balance evolution in anurans. Through a comparative approach, this study tested the hypotheses that, when compared to anuran species that inhabit more mesic environments, species from xeric environments should show: (1) increased resistance to evaporative water loss, (2) higher rates of rehydration, (3) low sensitivity of locomotor performance to dehydration and (4) increased sensitivity of the locomotor performance at lower temperatures and lower sensitivity of it at higher temperatures. Rehydration rates and skin resistance to evaporative water loss showed, respectively, positive and negative allometric relations with body mass, indicating the evolution of compensatory mechanisms to changes in the relationship between surface area and volume with body size variation. Anurans from environments characterized by higher restriction in water availability and higher temperatures had higher rates of hydration, lower sensitivity of locomotor performance at higher temperatures and higher sensitivity of it at lower temperatures, indicating patterns of physiologic adaptation to environmental characteristics. The variation in sensitivity of the locomotor performance to dehydration showed more complex relation patterns with environmental variables. These relations were dependent on the environments and phylogenetic groups sampled