Immunological Tradeoffs and the Impacts of Urbanization on the Reproductive Ecology and Physiology of the Side-Blotched Lizard (\u3cem\u3eUta stansburiana\u3c/em\u3e)

Investing resources into reproduction can limit energy available to other competing demands, such as fighting off an infection; yet, both processes are necessary for organisms to survive and pass on their genes to the next generation. These strategies often follow patterns associated with lifespan, such that shorter-lived animals are more likely to invest more resources into reproduction over survival, and vice versa in long-lived animals. However, environmental change caused by urbanization can disrupt these relationships, and the within- and transgenerational costs of urbanization on females and offspring are unknown. I address these uncertainties in three research chapters to better understand the effects of urbanization on reproductive investment in female Side-blotched Lizards (Uta stansburiana), a small and abundant species of reptile found throughout the western United States. In my second chapter, I examine general variation in female immunity and stress and how it relates to egg number, egg mass, and egg yolk immunity and stress. I found oxidative stress and immunity vary in females depending on how many eggs they produced, and that larger eggs had lower yolk stress levels. I built upon this by examining how metabolism differs across the reproductive cycle and tested whether simulating an infection in females affected immunity, stress, and metabolism. Metabolism was higher at the onset of reproduction and decreased until ovulation, and females differentially responded to infection depending on their stage in the reproductive cycle, which may suggest limited resources underly these findings. In my final chapter, I investigated the impacts of urbanization on female and egg yolk physiology and tested whether simulating infection altered female investment into egg yolk. I found immunity and stress in females and eggs were only apparent in rural females with ectoparasites, but not in urban females and eggs. Fertilization rates were lower in urban populations, which also influenced egg yolk physiology. Differential physiological investment can drastically alter offspring traits; therefore, it is imperative to develop a better understanding of the transgenerational costs of inhabiting an urban environment

What Does the Snake Eat? Breadth, Overlap, and Non-Native Prey in the Diet of Three Sympatric Natricine Snakes

We investigated diet breadth and diet overlap in three sympatric snakes of similar body size: Dekay’s Brownsnakes (Storeria dekayi), Red-bellied Snakes (S. occipitomaculata), and sub-adult Common Gartersnakes (Thamnophis sirtalis), by examining recently consumed prey (n = 388) collected from wild-caught snakes (n = 263) in northern Illinois. Storeria occipitomaculata were dietary specialists, feeding nearly exclusively on slugs. Storeria dekayi fed predominately on slugs but also consumed snails and earthworms. Sub-adult T. sirtalis fed predominately on earthworms but also consumed frogs and small mammals. Diet overlap was extensive between Storeria species but relatively low between Storeria and Thamnophis. It is noteworthy that the two most abundant prey types, slugs and earthworms, are non-native. These non-native prey occur in high numbers, which may ameliorate competitive interactions and influence grassland snake abundance and persistence regionally

Structure of anhydrotetracycline-bound Tet(X6) reveals the mechanism for inhibition of type 1 tetracycline destructases

Inactivation of tetracycline antibiotics by tetracycline destructases (TDases) remains a clinical and agricultural threat. TDases can be classified as type 1 Tet(X)-like TDases and type 2 soil-derived TDases. Type 1 TDases are widely identified in clinical pathogens. A combination therapy of tetracycline and a TDase inhibitor is much needed to rescue the clinical efficacy of tetracyclines. Anhydrotetracycline is a pan-TDase inhibitor that inhibits both type 1 and type 2 TDases. Here, we present structural, biochemical, and phenotypic evidence that anhydrotetracycline binds in a substrate-like orientation and competitively inhibits the type 1 TDase Tet(X6) to rescue tetracycline antibiotic activity as a sacrificial substrate. Anhydrotetracycline interacting residues of Tet(X6) are conserved within type 1 TDases, indicating a conserved binding mode and mechanism of inhibition. This mode of binding and inhibition is distinct from anhydrotetracycline\u27s inhibition of type 2 TDases. This study forms the framework for development of next-generation therapies to counteract enzymatic tetracycline resistance

Invasive frogs show persistent physiological differences to elevation and acclimate to colder temperatures

The coqui frog (Eleutherodactylus coqui) was introduced to the island of Hawai’i in the 1980s and has spread across much of the island. Concern remains that this frog will continue to expand its range and invade higher elevation habitats where much of the island’s endemic species are found. We determined whether coqui thermal tolerance and physiology change along Hawai’i’s elevational gradients. We measured physiological responses using a short-term experiment to determine baseline tolerance and physiology by elevation, and a long-term experiment to determine the coqui’s ability to acclimate to different temperatures. We collected frogs from low, medium, and high elevations. After both the short and long-term experiments, we measured critical thermal minimum (CTmin), blood glucose, oxidative stress, and corticosterone levels. CTmin was lower in high elevation frogs than low elevation frogs after the short acclimation experiment, signifying that they acclimate to local conditions. After the extended acclimation, CTmin was lower in frogs acclimated to cold temperatures compared to warm-acclimated frogs and no longer varied by elevation. Blood glucose levels were positively correlated with elevation even after the extended acclimation, suggesting glucose may also be related to lower temperatures. Oxidative stress was higher in females than males, and corticosterone was not significantly related to any predictor variables. The extended acclimation experiment showed that coquis can adjust their thermal tolerance to different temperatures over a 3-week period, suggesting the expansion of coqui into higher elevation habitats may still be possible, and they may not be as restricted by cold temperatures as previously thought

Noroviruses Co-opt the Function of Host Proteins VAPA and VAPB for Replication via a Phenylalanine-Phenylalanine-Acidic-Tract-Motif Mimic in Nonstructural Viral Protein NS1/2.

The Norovirus genus contains important human pathogens, but the role of host pathways in norovirus replication is largely unknown. Murine noroviruses provide the opportunity to study norovirus replication in cell culture and in small animals. The human norovirus nonstructural protein NS1/2 interacts with the host protein VAMP-associated protein A (VAPA), but the significance of the NS1/2-VAPA interaction is unexplored. Here we report decreased murine norovirus replication in VAPA- and VAPB-deficient cells. We characterized the role of VAPA in detail. VAPA was required for the efficiency of a step(s) in the viral replication cycle after entry of viral RNA into the cytoplasm but before the synthesis of viral minus-sense RNA. The interaction of VAPA with viral NS1/2 proteins is conserved between murine and human noroviruses. Murine norovirus NS1/2 directly bound the major sperm protein (MSP) domain of VAPA through its NS1 domain. Mutations within NS1 that disrupted interaction with VAPA inhibited viral replication. Structural analysis revealed that the viral NS1 domain contains a mimic of the phenylalanine-phenylalanine-acidic-tract (FFAT) motif that enables host proteins to bind to the VAPA MSP domain. The NS1/2-FFAT mimic region interacted with the VAPA-MSP domain in a manner similar to that seen with bona fide host FFAT motifs. Amino acids in the FFAT mimic region of the NS1 domain that are important for viral replication are highly conserved across murine norovirus strains. Thus, VAPA interaction with a norovirus protein that functionally mimics host FFAT motifs is important for murine norovirus replication.IMPORTANCE Human noroviruses are a leading cause of gastroenteritis worldwide, but host factors involved in norovirus replication are incompletely understood. Murine noroviruses have been studied to define mechanisms of norovirus replication. Here we defined the importance of the interaction between the hitherto poorly studied NS1/2 norovirus protein and the VAPA host protein. The NS1/2-VAPA interaction is conserved between murine and human noroviruses and was important for early steps in murine norovirus replication. Using structure-function analysis, we found that NS1/2 contains a short sequence that molecularly mimics the FFAT motif that is found in multiple host proteins that bind VAPA. This represents to our knowledge the first example of functionally important mimicry of a host FFAT motif by a microbial protein

Molecular mechanisms of cell death: recommendations of the Nomenclature Committee on Cell Death 2018.

Over the past decade, the Nomenclature Committee on Cell Death (NCCD) has formulated guidelines for the definition and interpretation of cell death from morphological, biochemical, and functional perspectives. Since the field continues to expand and novel mechanisms that orchestrate multiple cell death pathways are unveiled, we propose an updated classification of cell death subroutines focusing on mechanistic and essential (as opposed to correlative and dispensable) aspects of the process. As we provide molecularly oriented definitions of terms including intrinsic apoptosis, extrinsic apoptosis, mitochondrial permeability transition (MPT)-driven necrosis, necroptosis, ferroptosis, pyroptosis, parthanatos, entotic cell death, NETotic cell death, lysosome-dependent cell death, autophagy-dependent cell death, immunogenic cell death, cellular senescence, and mitotic catastrophe, we discuss the utility of neologisms that refer to highly specialized instances of these processes. The mission of the NCCD is to provide a widely accepted nomenclature on cell death in support of the continued development of the field

Recovery from Discrete Wound Severities in Side-Blotched Lizards (Uta stansburiana): Implications for Energy Budget, Locomotor Performance, and Oxidative Stress

Wounding events (predation attempts, competitive combat) result in injuries and/or infections that induce integrated immune responses for the recovery process. Despite the survival benefits of immunity in this context, the costs incurred may require investment to be diverted from traits contributing to immediate and/or future survival, such as locomotor performance and oxidative status. Yet, whether trait constraints manifest likely depends on wound severity and the implications for energy budget. For this study, food intake, body mass, sprint speed, and oxidative indices (reactive oxygen metabolites, antioxidant capacity) were monitored in male side-blotched lizards (Uta stansburiana) healing from cutaneous wounds of discrete sizes (control, small, large). Results indicate that larger wounds induced faster healing, reduced food consumption, and led to greater oxidative stress over time. Granted wounding did not differentially affect body mass or sprint speed overall, small wounded lizards with greater wound area healed had faster sprint speeds while large-wounded lizards with greater wound area healed had slower sprint speeds. During recovery from either wound severity, however, healing and sprint performance did not correspond with food consumption, body mass loss, nor oxidative status. These findings provide support that energy budget, locomotor performance, and oxidative status of a reptile are linked to wound recovery to an extent, albeit dependent on wound severity

Data_Sheet_1_Patch-level processes of vegetation underlying site-level restoration patterns in a megatidal salt marsh.pdf

Vegetation patterns during salt marsh restoration reflect underlying processes related to colonization, reproduction, and interactions of halotolerant plants. Examining both pattern and process during recovery is valuable for understanding and managing salt marsh restoration projects. We present a decade of vegetation dynamics during salt marsh restoration (2011–2020) at a study site in the Bay of Fundy with megatidal amplitudes, strong currents, cold winter temperatures, and ice. We mainly investigated reproduction (asexual and sexual) and associated spread rates of Spartina grasses, and their health-related states (stem density, canopy height, and percent flowering) which help inform the probability of processes occurring. We also estimated modes of colonization and began quantifying the effects of interspecific interactions and environmental conditions on plant state. Spartina pectinata was the only pastureland plant to survive dike-breaching and saltwater intrusion in 2010; however, it was stunted compared to reference plants. Spartina pectinata patches remained consistent initially, before decreasing in size, and disappearing by the fifth year (2015). This early dynamic may provide initial protection to a developing salt marsh before Spartina alterniflora becomes established. Spartina alterniflora first colonized the sites in year 2 (2012), likely via deposition of rhizomal material, and then spread asexually before seedlings (sexual reproduction) appeared in year 4 (2014). Vegetation cover subsequently increased greatly until near-complete in year 9 (2019). The early successional dynamics of S. pectinata and S. alterniflora occurred spatially independently of each other, and likely contributed to sediment retention, creating an improved environment for S. patens, the dominant high marsh species in our region. Spartina patens have been slowly spreading into restoration sites from high elevation areas since year 6 (2016). We expect that competition between S. alterniflora and S. patens will result in the typical distinct zonation between high and low marsh zones. A next study will use the quantified processes for spatial-explicit modeling to simulate patterns of vegetation recovery, and to evaluate different salt marsh restoration strategies for the Bay of Fundy and elsewhere. Thus, proper identification and quantification of pattern-building processes in salt marsh vegetation recovery, the focus of our present study, was an essential step.</p