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Commentary: Tracing the fate of metabolic substrates during changes in whole-body energy expenditure in mice
For small mammals, such as mice, cannulation procedures can be quite challenging, limiting research associated with tracing isotopically labelled substrates at the whole-animal level. When cannulation in mice is possible, assessment of substrate use is further limited to when mice are either under anesthesia or are at rest, as there are no studies directly quantifying substrate use during exercise in mice. The use of isotopic tracer techniques has greatly advanced our knowledge in understanding how metabolic substrates (carbohydrates, amino acids, and fatty acids) contribute to whole-body metabolism. However, research regarding tissue-specific fuel use contributions to whole-body energy expenditure in mice at varying metabolic intensities (i.e., exercise) is lacking, despite the popularity of using mice in a variety of metabolic models. In this commentary, we briefly discuss the methodologies, advantages, and disadvantages of using radiolabelled, positron emission, and stable isotopes with a specific focus on fatty acids. We highlight recent mouse studies that have used creative experimental designs employing the use of isotopic tracer techniques and we briefly discuss how these methodologies can be further pursued to deepen our understanding of substrate use during exercise. Lastly, we show findings of a recent study we performed using a radiolabelled fatty acid tracer (14C-bromopalmitic acid) to determine fatty acid uptake in 16 muscles, two brown and two white adipose tissue depots during submaximal exercise in deer mice
Regulation of muscle pyruvate dehydrogenase activity and fuel use during exercise in high-altitude deer mice
Adult, lab-reared, highland deer mice acclimate to hypoxia by increasing reliance on carbohydrates to fuel exercise. Yet neither the underlying mechanisms for this shift in fuel use nor the impact of lifetime hypoxia exposure experienced in high alpine conditions, are fully understood. Thus, we assessed the use of fuel during exercise in wild highland deer mice running in their native environment. We examined a key step in muscle carbohydrate oxidation - the regulation of pyruvate dehydrogenase (PDH) - during exercise at altitude in wild highlanders and in first generation (G1) lab-born and -raised highlanders acclimated to normoxia or hypoxia. PDH activity was also determined in the gastrocnemius ofG1 highlanders using an in situ muscle preparation.We found that wild highlanders had a high reliance on carbohydrates while running in their native environment, consistent with data from hypoxia-acclimated G1 highlanders. PDH activity in the gastrocnemius was similar post exercise between G1 and wild highlanders. However, when the gastrocnemius was stimulated at a light work rate in situ, PDH activity was higher in hypoxia-acclimated G1 highlanders and was associated with lower intramuscular lactate levels. These findings were supported by lower PDH kinase 2 protein production in hypoxia-acclimated G1 mice. Our findings indicate that adult phenotypic plasticity in response to low oxygen is sufficient to increase carbohydrate reliance during exercise in highland deer mice. Additionally, variation in PDH regulation with hypoxia acclimation contributes to shifts in whole-animal patterns of fuel use and is likely to improve exercise performance via elevated energy yield permole ofO2
Chronic cold exposure induces mitochondrial plasticity in deer mice native to high altitudes
Key points: Small mammals native to high altitude must sustain high rates of thermogenesis to cope with cold. Skeletal muscle is a key site of shivering and non-shivering thermogenesis, but the importance of mitochondrial plasticity in cold hypoxic environments remains unresolved. We examined high-altitude deer mice, which have evolved a high capacity for aerobic thermogenesis, to determine the mechanisms of mitochondrial plasticity during chronic exposure to cold and hypoxia, alone and in combination. Cold exposure in normoxia or hypoxia increased mitochondrial leak respiration and decreased phosphorylation efficiency and OXPHOS coupling efficiency, which may serve to augment non-shivering thermogenesis. Cold also increased muscle oxidative capacity, but reduced the capacity for mitochondrial respiration via complex II relative to complexes I and II combined. High-altitude mice had a more oxidative muscle phenotype than low-altitude mice. Therefore, both plasticity and evolved changes in muscle mitochondria contribute to thermogenesis at high altitude. Abstract: Small mammals native to high altitude must sustain high rates of thermogenesis to cope with cold and hypoxic environments. Skeletal muscle is a key site of shivering and non-shivering thermogenesis, but the importance of mitochondrial plasticity in small mammals at high altitude remains unresolved. High-altitude deer mice (Peromyscus maniculatus) and low-altitude white-footed mice (P. leucopus) were born and raised in captivity, and chronically exposed as adults to warm (25°C) normoxia, warm hypoxia (12 kPa O2), cold (5°C) normoxia, or cold hypoxia. We then measured oxidative enzyme activities, oxidative fibre density and capillarity in the gastrocnemius, and used a comprehensive substrate titration protocol to examine the function of muscle mitochondria by high-resolution respirometry. Exposure to cold in both normoxia or hypoxia increased the activities of citrate synthase and cytochrome oxidase. In lowlanders, this was associated with increases in capillary density and the proportional abundance of oxidative muscle fibres, but in highlanders, these traits were unchanged at high levels across environments. Environment had some distinct effects on mitochondrial OXPHOS capacity between species, but the capacity of complex II relative to the combined capacity of complexes I and II was consistently reduced in both cold environments. Both cold environments also increased leak respiration and decreased phosphorylation efficiency and OXPHOS coupling efficiency in both species, which may serve to augment non-shivering thermogenesis. These cold-induced changes in mitochondrial function were overlaid upon the generally more oxidative phenotype of highlanders. Therefore, both plasticity and evolved changes in muscle mitochondria contribute to thermogenesis at high altitudes
Optimizations for time and effort in long-term monitoring: a case study using a multidecadal terrestrial salamander monitoring program
Long-term monitoring programs can identify environmental trends or reveal limitations to protocols, as long as their results are analysed appropriately. While monitoring programs are not necessarily hypothesis-driven, their data are important for conservation and can guide improvements to monitoring programs. Here, we present a case study using dynamic occupancy models to guide the optimization of time and effort in a long-term terrestrial salamander monitoring program. To ensure a detailed analysis, we analysed the available long-term data to first identify estimates of occupancy and detection parameters for the salamanders. Using these estimates, we created simulations to identify the optimal number of years for monitoring and the optimal allocation of spatial and temporal survey replicates. Our data support previous claims that monitoring programs should be allowed to run for at least a decade. We also found that in order to obtain accurate estimates of species occupancy, programs should consider appropriate partitioning of monitoring effort across spatial and temporal scales. We show how analyses of long-term monitoring datasets are valuable not only for trend detection but also for the development of templates to guide the design and optimization of similar programs
Identification of most spectrally distinguishable phenological stage of invasive Phramites australis in Lake Erie wetlands (Canada) for accurate mapping using multispectral satellite imagery
Phragmites australis (Cav.) Trin. ex Steudel subspecies australis is one of the worst plant invaders in wetlands of North America. Remote sensing is the most cost-effective method to track its spread given its widespread distribution and rapid colonization rate. We hypothesize that the morphological and/or physiological features associated with different phenological states of Phragmites can influence their reflectance signal and thus affect mapping accuracies. We tested this hypothesis by comparing classification accuracies of cloud-free images acquired by Landsat 7, Landsat 8, and Sentinel 2 at roughly monthly intervals over a calendar year for two wetlands in southern Ontario. We used the Support Vector Machines classification and employed field observations and image acquired from unmanned aerial vehicle (8 cm) to perform accuracy assessments. The highest Phragmites producer’s, user’s, and overall accuracy (96.00, 91.11, and 88.56% respectively) were provided by images acquired in late summer and fall period. During this period, green, Near Infrared, and Short-Wave Infrared bands generated more unique reflectance signals for Phragmites. Both Normalized Difference Vegetation Index and Normalized Difference Water Index showed significant difference between Phragmites and the most confused classes (cattail; Typha latifolia L., and meadow marsh) during the late summer and fall period. Since meadow marsh separated out best from Phragmites and cattail in the February image, we used it to mask the meadow marsh in the July image to reduce confusion. The unique reflectance signal of Phragmites in late summer and fall is likely due to prolonged greenness of Phragmites when compared to other wetland vegetation, large, distinct inflorescence, and the water content of Phragmites during this period
Effects of European common reed on Blanding's turtle spatial ecology
European common reed (Phragmites australis; common reed) is an aggressive invader of North American wetlands that forms homogenous patches and replaces native flora. Dense patches of common reed generally provide poor habitat for many species, although specific effects on at-risk turtles are largely unknown. We created 3 predictive scenarios to relate the amount of common reed in the landscape to amount of effective habitat for Blanding's turtles (Emydoidea blandingii) and investigated the spatial ecology of 46 adult Blanding's turtles using Euclidean distance analysis within 2 wetland complexes in southern Ontario, Canada. At the home-range scale, we identified a positive association between turtle home ranges and common reed. At the individual scale, turtle radio-locations were significantly farther from common reed patches than from random points, consistent with the hypothesis that they avoided common reed patches locally. When we analyzed habitat selection by sex, results were similar to population-level results except for nesting females. During nesting migrations, females did not avoid common reed patches at the individual scale but instead interacted with common reed, potentially placing themselves at risk of being stranded within dense patches. Our results are consistent with our dynamic home range plus saturation hypothesis that invasion of common reed reduces the amount of effective habitat for at-risk turtles in wetlands because Blanding's turtles significantly avoided common reed patches at individual scales. Management of common reed is an important step to restore habitat for Blanding's turtles and future research is needed to determine best restoration practices. © 2018 The Wildlife Society
Long-term habitat changes in a protected area: Implications for herpetofauna habitat management and restoration
Point Pelee National Park, located at the southern-most tip of Canada's mainland, historically supported a large number of herpetofauna species; however, despite nearly a century of protection, six snake and five amphibian species have disappeared, and remaining species-At-risk populations are thought to be in decline. We hypothesized that long-Term changes in availability and distribution of critical habitat types may have contributed to the disappearance of herpetofauna. To track habitat changes we used aerial image data spanning 85 years (1931±2015) and manually digitized and classified image data using a standardized framework. Change-detection analyses were used to evaluate the relative importance of proportionate loss and fragmentation of 17 habitat types. Marsh habitat diversity and aquatic connectivity has declined since 1931. The marsh matrix transitioned from a graminoid and forb shallow marsh interspersed with water to a cattail dominated marsh, altering critical breeding, foraging, and overwintering habitat. Reduced diversity of marsh habitats appears to be linked to the expansion of invasive Phragmites australis, which invaded prior to 2000. Loss of open habitats such as savanna and meadow has reduced availability of high quality thermoregulation habitat for reptiles. Restoration of the northwestern region and tip of Point Pelee National Park to a mixed landscape of shallow wetlands (cattail, graminoid, forb, open water) and eradication of dense Phragmites stands should improve habitat diversity. Our results suggest that long-Term landscape changes resulting from habitat succession and invasive species can negatively affect habitat suitability for herpetofauna and protection of land alone does not necessarily equate to protection of sensitive herpetofauna
UNMANNED AERIAL VEHICLES PRODUCE HIGH-RESOLUTION, SEASONALLY-RELEVANT IMAGERY FOR CLASSIFYING WETLAND VEGETATION
With recent advances in technology, personal aerial imagery acquired with unmanned aerial vehicles (UAVs) has transformed the way ecologists can map seasonal changes in wetland habitat. Here, we use a multi-rotor (consumer quad-copter, the DJI Phantom 2 Vision+) UAV to acquire a high-resolution (< 8 cm) composite photo of a coastal wetland in summer 2014. Using validation data collected in the field, we determine if a UAV image and SWOOP (Southwestern Ontario Orthoimagery Project) image (collected in spring 2010) differ in their classification of type of dominant vegetation type and percent cover of three plant classes: submerged aquatic vegetation, floating aquatic vegetation, and emergent vegetation. The UAV imagery was more accurate than available SWOOP imagery for mapping percent cover of submergent and floating vegetation categories, but both were able to accurately determine the dominant vegetation type and percent cover of emergent vegetation. Our results underscore the value and potential for affordable UAVs (complete quad-copter system < $3,000 CAD) to revolutionize the way ecologists obtain imagery and conduct field research. In Canada, new UAV regulations make this an easy and affordable way to obtain multiple high-resolution images of small (< 1.0 km2) wetlands, or portions of larger wetlands throughout a year
Wave exposure and hydrologic connectivity create diversity in habitat and zooplankton assemblages at nearshore Long Point Bay, Lake Erie
During an 11-day period in August 2008, we visited 102 sites along the nearshore (~60 km) of Long Point Bay. The purpose of our study was to evaluate the effects of wave exposure and hydrologic connectivity on zooplankton distributions. Long Point is located within the UNESCO Long Point Biosphere Reserve (26,250. ha) and encompasses the largest wetland complex in the Great Lakes system. We sampled for zooplankton, aquatic vegetation, temperature, specific conductance, pH, dissolved oxygen, dissolved organic carbon, water clarity, total nitrogen and depth. We evaluated the impacts of exposure using wind and fetch data to calculate a Relative Exposure Index (REI). Ordination techniques revealed a large variation in physical disturbance, water clarity, nutrient concentrations, water chemistry and aquatic vegetation that explained the distribution pattern of zooplankton at the 102 sites. Gradients of REI are strongly positively correlated with environmental variables, such as pH, dissolved oxygen and temperature and highly negatively correlated with conductivity and dissolved organic carbon. Visual inspection of the ordination site scores revealed the 102 sites clustering into six main groups based on spatial location and degree of surface-water connectivity to Long Point Bay. Sheltered sites (low REI) have much higher abundance of zooplankton whereas sites that have high REI scores are characterized by relatively low zooplankton abundance with a high prevalence of Polyarthra sp. This is the largest study on the distribution pattern of zooplankton in Long Point Bay, and it highlights the importance of wave exposure and hydrologic connectivity in structuring the zooplankton community. © 2012 Elsevier B.V