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• 1. [Article] The Biology of Eastern Kingbirds at Malheur National Wildlife Refuge: Survival, Reproduction, and Testosterone Secretion

  This dissertation presents the results of a study that I undertook to better understand the breeding biology of Eastern Kingbirds (hereafter, kingbirds) at Malheur National Wildlife Refuge in southeastern ...

  Citation × Citation Citation Abstract Copy Title: The Biology of Eastern Kingbirds at Malheur National Wildlife Refuge: Survival, Reproduction, and Testosterone Secretion Author: Redmond, Lucas J. Year: 2015 This dissertation presents the results of a study that I undertook to better understand the breeding biology of Eastern Kingbirds (hereafter, kingbirds) at Malheur National Wildlife Refuge in southeastern Oregon from 2003 to 2009. Kingbirds are long-distance migratory songbirds that breed across much of North America. This species is socially monogamous but, via frequent extra-pair copulations, is genetically polygamous. Kingbirds exhibit relatively high breeding site fidelity, often returning to the same tree to nest in subsequent years. Both members of a pair provide parental care, but there are often specific duties performed by both male and female kingbirds. For example, males typically perform much of the vigilant nest defense that this species is well known for and contribute, to an extent, to the feeding of nestlings. Females, on the other hand, are entirely responsible for incubating and brooding. Beginning in 2002, most adults within the population of kingbirds at Malheur had been banded with a unique combination of three colored plastic leg bands and an aluminum USFWS band. Also, as many nests were located in each year as possible, and any young that survived to fledging age were banded with a unique combination of leg bands. Considerable effort was then spent each year to locate as many banded individuals as possible, which allowed me to document adult and juvenile survival. Annual survival rates of adult male and female kingbirds in the population at Malheur did not differ and were relatively high at approximately 0.65. Juvenile survival rate was approximately 0.29, indicating that slightly less than one-third of nestlings survive the interval between fledging and their first potential breeding season. Resighting probability was high for both sexes, although higher for adult males (0.94) than for adult female (0.84). The latter finding is consistent with the higher site fidelity of males than of females. Resighting probabilities for juveniles were much lower at 0.68 and 0.40 for males and females, respectively. Again, this was expected because natal site fidelity is typically much lower than breeding site fidelity. Compared to most other reports, resighting probability and return rates of juvenile kingbirds was high, presumably because the riparian habitats where kingbirds breed at Malheur function as an ecological island surrounded by, for kingbirds, unsuitable high desert habitat. Thus, unless they disperse very long distances, the only option for juvenile kingbirds is to begin nesting on the refuge. The collection of blood samples from birds has become an increasingly common practice in ornithology. The data that can be gained from these samples allow a number of interesting questions to be asked such as understanding the genetic mating system of a species, patterns of hormone secretion, and discerning migratory pathways via stable isotope analysis. The volume of blood collected is usually small and was assumed to cause no long-term negative effects on sampled individuals. However, few studies have rigorously examined the effect of blood sampling on survival. I used a multistate mark-recapture analysis to assess the effect blood sampling on annual survival of kingbirds by combining the annual survival data described above with whether or not individuals were subjected to blood sample collection. The results of this analysis indicated that blood sampling had no effect on annual survival rates of kingbirds. Whether or not this is the case for other species remains to be seen. However, my results support the assumption that when done correctly, blood sampling has little to no long-term negative effects on birds. Comparative analyses of many bird species show that testosterone secretion exhibits fairly predictable patterns among breeding birds of different mating systems. Monogamous species reach a peak during mate attraction and the period of female fertility which is then followed by a sharp decline when young are in the nest. By contrast, males of polygynous species tend to maintain higher levels of testosterone throughout the breeding season to, presumably, maximize opportunities for acquiring additional mates. Kingbirds are socially monogamous but cryptically polygamous. However, because of high rates of extra-pair paternity, variation in reproductive success among males is much higher than what is expected for a monogamous species, and, instead, is similar to what has been reported for polygynous species. Therefore from 2005 to 2009 I measured testosterone concentrations from the blood samples collected from male kingbirds to determine the breeding season profile of testosterone secretion in kingbirds and to understand the factors that influence testosterone variation among individuals. Contrary to expected for a monogamous species, the testosterone profile of kingbirds did not exhibit the brief peak in testosterone followed by a precipitous decline. Testosterone peaked early in the season, but declined very slowly as the nest cycle progressed. I attributed this gradual decline in testosterone to the cryptically polygynous nature of the kingbird extra-pair mating system. I also found substantial variation in testosterone concentration among male kingbirds and was able to identify several factors contributing to this variation. As expected, testosterone declined as the breeding season progressed (independent of stage in nest cycle) and nest density increased, while increases in testosterone were correlated with the number of fertile females within the population. This suggests that male kingbirds were capable of modulating testosterone concentrations to both cope with an increase in conspecific density, but also to ready themselves for times when extra-pair copulations were more likely. Title: The Biology of Eastern Kingbirds at Malheur National Wildlife Refuge: Survival, Reproduction, and Testosterone Secretion Author: Redmond, Lucas J. Year: 2015 This dissertation presents the results of a study that I undertook to better understand the breeding biology of Eastern Kingbirds (hereafter, kingbirds) at Malheur National Wildlife Refuge in southeastern Oregon from 2003 to 2009. Kingbirds are long-distance migratory songbirds that breed across much of North America. This species is socially monogamous but, via frequent extra-pair copulations, is genetically polygamous. Kingbirds exhibit relatively high breeding site fidelity, often returning to the same tree to nest in subsequent years. Both members of a pair provide parental care, but there are often specific duties performed by both male and female kingbirds. For example, males typically perform much of the vigilant nest defense that this species is well known for and contribute, to an extent, to the feeding of nestlings. Females, on the other hand, are entirely responsible for incubating and brooding. Beginning in 2002, most adults within the population of kingbirds at Malheur had been banded with a unique combination of three colored plastic leg bands and an aluminum USFWS band. Also, as many nests were located in each year as possible, and any young that survived to fledging age were banded with a unique combination of leg bands. Considerable effort was then spent each year to locate as many banded individuals as possible, which allowed me to document adult and juvenile survival. Annual survival rates of adult male and female kingbirds in the population at Malheur did not differ and were relatively high at approximately 0.65. Juvenile survival rate was approximately 0.29, indicating that slightly less than one-third of nestlings survive the interval between fledging and their first potential breeding season. Resighting probability was high for both sexes, although higher for adult males (0.94) than for adult female (0.84). The latter finding is consistent with the higher site fidelity of males than of females. Resighting probabilities for juveniles were much lower at 0.68 and 0.40 for males and females, respectively. Again, this was expected because natal site fidelity is typically much lower than breeding site fidelity. Compared to most other reports, resighting probability and return rates of juvenile kingbirds was high, presumably because the riparian habitats where kingbirds breed at Malheur function as an ecological island surrounded by, for kingbirds, unsuitable high desert habitat. Thus, unless they disperse very long distances, the only option for juvenile kingbirds is to begin nesting on the refuge. The collection of blood samples from birds has become an increasingly common practice in ornithology. The data that can be gained from these samples allow a number of interesting questions to be asked such as understanding the genetic mating system of a species, patterns of hormone secretion, and discerning migratory pathways via stable isotope analysis. The volume of blood collected is usually small and was assumed to cause no long-term negative effects on sampled individuals. However, few studies have rigorously examined the effect of blood sampling on survival. I used a multistate mark-recapture analysis to assess the effect blood sampling on annual survival of kingbirds by combining the annual survival data described above with whether or not individuals were subjected to blood sample collection. The results of this analysis indicated that blood sampling had no effect on annual survival rates of kingbirds. Whether or not this is the case for other species remains to be seen. However, my results support the assumption that when done correctly, blood sampling has little to no long-term negative effects on birds. Comparative analyses of many bird species show that testosterone secretion exhibits fairly predictable patterns among breeding birds of different mating systems. Monogamous species reach a peak during mate attraction and the period of female fertility which is then followed by a sharp decline when young are in the nest. By contrast, males of polygynous species tend to maintain higher levels of testosterone throughout the breeding season to, presumably, maximize opportunities for acquiring additional mates. Kingbirds are socially monogamous but cryptically polygamous. However, because of high rates of extra-pair paternity, variation in reproductive success among males is much higher than what is expected for a monogamous species, and, instead, is similar to what has been reported for polygynous species. Therefore from 2005 to 2009 I measured testosterone concentrations from the blood samples collected from male kingbirds to determine the breeding season profile of testosterone secretion in kingbirds and to understand the factors that influence testosterone variation among individuals. Contrary to expected for a monogamous species, the testosterone profile of kingbirds did not exhibit the brief peak in testosterone followed by a precipitous decline. Testosterone peaked early in the season, but declined very slowly as the nest cycle progressed. I attributed this gradual decline in testosterone to the cryptically polygynous nature of the kingbird extra-pair mating system. I also found substantial variation in testosterone concentration among male kingbirds and was able to identify several factors contributing to this variation. As expected, testosterone declined as the breeding season progressed (independent of stage in nest cycle) and nest density increased, while increases in testosterone were correlated with the number of fertile females within the population. This suggests that male kingbirds were capable of modulating testosterone concentrations to both cope with an increase in conspecific density, but also to ready themselves for times when extra-pair copulations were more likely. Close

• 2. [Article] Design and Synthesis of Novel Chloroquine-based Antimalarials

  Malaria is an infectious, often fatal disease that afflicts nearly 200 million people every year. The disease, characterized by recurring and extreme flu-like symptoms, is caused by the protozoan parasite ...

  Citation × Citation Citation Abstract Copy Title: Design and Synthesis of Novel Chloroquine-based Antimalarials Author: Murphy, Kevin Vincent Year: 2015 Malaria is an infectious, often fatal disease that afflicts nearly 200 million people every year. The disease, characterized by recurring and extreme flu-like symptoms, is caused by the protozoan parasite Plasmodium falciparum. Victims usually contract the disease through a mosquito vector. Chloroquine is a chemotherapeutic that was introduced in the 1940s. For many years the drug was the foremost treatment of malaria, being effective and producing few side effects. Unfortunately, tolerance to chloroquine developed when the parasite evolved a resistance mechanism. Newer drugs have been developed and implemented, but these medicines also show a decreasing effect with continued administration. It is imperative that a new pipeline of drugs be developed in order to combat the disease and anticipated resistance. Reversed chloroquines are a new class of multiple-ligand compounds that are active against chloroquine-sensitive and chloroquine-resistance malaria species. This thesis describes research targeted at the modification of lead reversed chloroquine molecules to discover new and effective moieties, as well as to improve pharmacokinetic-related properties. An especial emphasis of this project is the addition of a sulfonamide functional group to a reversed chloroquine. Preliminary evidence indicates that this is a promising direction for this line of research. Brief discussions of some reversed chloroquine characterization studies are included in appendices. Title: Design and Synthesis of Novel Chloroquine-based Antimalarials Author: Murphy, Kevin Vincent Year: 2015 Malaria is an infectious, often fatal disease that afflicts nearly 200 million people every year. The disease, characterized by recurring and extreme flu-like symptoms, is caused by the protozoan parasite Plasmodium falciparum. Victims usually contract the disease through a mosquito vector. Chloroquine is a chemotherapeutic that was introduced in the 1940s. For many years the drug was the foremost treatment of malaria, being effective and producing few side effects. Unfortunately, tolerance to chloroquine developed when the parasite evolved a resistance mechanism. Newer drugs have been developed and implemented, but these medicines also show a decreasing effect with continued administration. It is imperative that a new pipeline of drugs be developed in order to combat the disease and anticipated resistance. Reversed chloroquines are a new class of multiple-ligand compounds that are active against chloroquine-sensitive and chloroquine-resistance malaria species. This thesis describes research targeted at the modification of lead reversed chloroquine molecules to discover new and effective moieties, as well as to improve pharmacokinetic-related properties. An especial emphasis of this project is the addition of a sulfonamide functional group to a reversed chloroquine. Preliminary evidence indicates that this is a promising direction for this line of research. Brief discussions of some reversed chloroquine characterization studies are included in appendices. Close

• 3. [Article] Localized Ecological and Educational Effects of Environmental Service-Learning in Portland, Oregon

  Environmental service-learning is an intentional educational experience(s) wherein learners engage in meaningful activities designed to serve the environment. Environmental service-learning activities ...

  Citation × Citation Citation Abstract Copy Title: Localized Ecological and Educational Effects of Environmental Service-Learning in Portland, Oregon Author: Braun, Steven Matthew Year: 2015 Environmental service-learning is an intentional educational experience(s) wherein learners engage in meaningful activities designed to serve the environment. Environmental service-learning activities vary according to their learning and service goals and include ecomanagement, persuasion, legal action, economic action and political action. The purpose of this mixed methods research was to explore the ecological and educational impacts of grades 6-12 environmental education, with special attention to environmental service-learning throughout Portland, Oregon. Ecological impacts considered restoration and conservation outcomes of several environmental service-learning programs including plant communities, soils, litter removal and trail maintenance. Educational outcomes considered aspects of environmental literacy including locus of control, environmental sensitivity, indicated environmentally responsible behaviors, investigating environmental issues and knowledge of physical systems. The relative influence of some significant life experiences on youths' response to environmental education, including environmental service-learning, was also considered. Telephone surveys were used to gather data from 22 Portland metropolitan area environmental education programs. Data included 2014 annual biophysical impacts (e.g., area of invasive species removed, pounds of litter removed) and information on programming (e.g., length of program, % time outside). Eleven programs administered a 33-question environmental literacy assessment to participants of their programs (n=393). The assessment included the New Environmental Paradigm, the Inclusion of Nature in Self, questions from Environmental Identity Scale and self-constructed questions. One 8th grade program was identified for a detailed case study. In this 8th grade programs, slight variations in educational activities occurred among three treatment groups which varied the amount of time youth spent engaged in ecomanagement. Youth from the three treatment groups and a control group were administered the environmental literacy assessment at the beginning and end of the program. Qualitative data for the youth in the treatment groups were gathered to further consider how environmental literacy was impacted by participation in the program. Stronger associational correlations to environmental literacy occurred for the percentage of time an environmental education program spent outdoors rather than the percentage of time an environmental education program engaged in environmental service-learning (e.g., "With other people, I can work to make a positive impact on the environment." rho: .276 vs. "I have the skills necessary to make a positive impact on the environment" rho: .176). Random forests indicated that environmental education program features and some significant life experiences could predict collapsed environmental literacy variables (locus of control, environmental sensitivity and environmentally responsible behaviors). 22.4% of the variance in a collapsed environmental sensitivity variable was explained by nine predictor variables; those variables with the strongest influence were youth response to "Before this program, how frequently did you spend time in the outdoors," age and the presence of a positive adult role model who cares for the environment. Youth participating in environmental education programs showed higher environmental literacy than control groups (e.g., "I feel an important part of my life would be missing if I couldn't get out and enjoy nature from time to time" U: 3642.500, p: 0.025). Youth with significant formative life experiences (e.g., those indicating previous environmental education or a positive adult role model that cares for the environment) responded better (higher environmental literacy) to environmental education than those youth without ("I pay special attention to things outdoors" chi 10.633, p: 0.031). This research provides insight on the efficacy of environmental service-learning. Environmental service-learning positively affected environmental literacy, but outdoor environmental education was more effective in terms of environmental literacy. Results corroborate the body of literature regarding significant life experiences. Further, results suggest that significant life experiences are a critical development milestone necessary for youth to respond to environmental education on a developmental trajectory to empowered environmentally literate citizens. Title: Localized Ecological and Educational Effects of Environmental Service-Learning in Portland, Oregon Author: Braun, Steven Matthew Year: 2015 Environmental service-learning is an intentional educational experience(s) wherein learners engage in meaningful activities designed to serve the environment. Environmental service-learning activities vary according to their learning and service goals and include ecomanagement, persuasion, legal action, economic action and political action. The purpose of this mixed methods research was to explore the ecological and educational impacts of grades 6-12 environmental education, with special attention to environmental service-learning throughout Portland, Oregon. Ecological impacts considered restoration and conservation outcomes of several environmental service-learning programs including plant communities, soils, litter removal and trail maintenance. Educational outcomes considered aspects of environmental literacy including locus of control, environmental sensitivity, indicated environmentally responsible behaviors, investigating environmental issues and knowledge of physical systems. The relative influence of some significant life experiences on youths' response to environmental education, including environmental service-learning, was also considered. Telephone surveys were used to gather data from 22 Portland metropolitan area environmental education programs. Data included 2014 annual biophysical impacts (e.g., area of invasive species removed, pounds of litter removed) and information on programming (e.g., length of program, % time outside). Eleven programs administered a 33-question environmental literacy assessment to participants of their programs (n=393). The assessment included the New Environmental Paradigm, the Inclusion of Nature in Self, questions from Environmental Identity Scale and self-constructed questions. One 8th grade program was identified for a detailed case study. In this 8th grade programs, slight variations in educational activities occurred among three treatment groups which varied the amount of time youth spent engaged in ecomanagement. Youth from the three treatment groups and a control group were administered the environmental literacy assessment at the beginning and end of the program. Qualitative data for the youth in the treatment groups were gathered to further consider how environmental literacy was impacted by participation in the program. Stronger associational correlations to environmental literacy occurred for the percentage of time an environmental education program spent outdoors rather than the percentage of time an environmental education program engaged in environmental service-learning (e.g., "With other people, I can work to make a positive impact on the environment." rho: .276 vs. "I have the skills necessary to make a positive impact on the environment" rho: .176). Random forests indicated that environmental education program features and some significant life experiences could predict collapsed environmental literacy variables (locus of control, environmental sensitivity and environmentally responsible behaviors). 22.4% of the variance in a collapsed environmental sensitivity variable was explained by nine predictor variables; those variables with the strongest influence were youth response to "Before this program, how frequently did you spend time in the outdoors," age and the presence of a positive adult role model who cares for the environment. Youth participating in environmental education programs showed higher environmental literacy than control groups (e.g., "I feel an important part of my life would be missing if I couldn't get out and enjoy nature from time to time" U: 3642.500, p: 0.025). Youth with significant formative life experiences (e.g., those indicating previous environmental education or a positive adult role model that cares for the environment) responded better (higher environmental literacy) to environmental education than those youth without ("I pay special attention to things outdoors" chi 10.633, p: 0.031). This research provides insight on the efficacy of environmental service-learning. Environmental service-learning positively affected environmental literacy, but outdoor environmental education was more effective in terms of environmental literacy. Results corroborate the body of literature regarding significant life experiences. Further, results suggest that significant life experiences are a critical development milestone necessary for youth to respond to environmental education on a developmental trajectory to empowered environmentally literate citizens. Close

• 4. [Article] The Boiling Springs Lake Metavirome: Charting the Viral Sequence-Space of an Extreme Environment Microbial Ecosystem

  Viruses are the most abundant organisms on Earth, yet their collective evolutionary history, biodiversity and functional capacity is not well understood. Viral metagenomics offers a potential means of ...

  Citation × Citation Citation Abstract Copy Title: The Boiling Springs Lake Metavirome: Charting the Viral Sequence-Space of an Extreme Environment Microbial Ecosystem Author: Diemer, Geoffrey Scott Year: 2014 Viruses are the most abundant organisms on Earth, yet their collective evolutionary history, biodiversity and functional capacity is not well understood. Viral metagenomics offers a potential means of establishing a more comprehensive view of virus diversity and evolution, as vast amounts of new sequence data becomes available for comparative analysis.Metagenomic DNA from virus-sized particles (smaller than 0.2 microns in diameter) was isolated from approximately 20 liters of sediment obtained from Boiling Springs Lake (BSL) and sequenced. BSL is a large, acidic hot-spring (with a pH of 2.2, and temperatures ranging from 50°C to 96°C) located in Lassen Volcanic National Park, USA. BSL supports a purely microbial ecosystem comprised largely of Archaea and Bacteria, however, the lower temperature regions permit the growth of acid- and thermo-tolerant Eukarya. This distinctive feature of the BSL microbial ecosystem ensures that virus types infecting all domains of life will be present. The metagenomic sequence data was used to characterize the types of viruses present within the microbial ecosystem, to ascertain the extent of genetic diversity and novelty comprising the BSL virus assemblage, and to explore the genomic and structural modalities of virus evolution.Metagenomic surveys of natural virus assemblages, including the survey of BSL, have revealed that the diversity within the virosphere far exceeds what has currently been determined through the detailed study of viruses that are relevant to human health and agriculture. The number of as-yet-uncharacterized virus protein families present in the BSL assemblage was estimated by clustering analysis. Genomic context analysis of the predicted viral protein sequences in the BSL dataset indicates that most of the putative uncharacterized proteins are endemic or unique to BSL, and are largely harbored by known virus types. A comparative metagenomic analysis approach identified a set of conserved, yet uncharacterized BSL protein sequences that are commonly found in other similar and dissimilar environments.New sequence data from metagenomic surveys of natural virus assemblages was also used to better characterize and define known virus protein families, as some of the viruses found in the BSL environment represent distant relatives of well-characterized isolates. By comparing viral genes and protein sequences from these highly divergent species, it is possible to better understand the dynamics of adaptation and evolution in the virosphere. Additionally, as structures of virus proteins continue to be experimentally determined by X-ray crystallography and cryo-electron microscopy, a merger of structural and metagenomic sequence data allows the opportunity to observe the structural dynamics underlying virus protein evolution.Capsid (structural) proteins from two distinct Microviridae strains; a globally ubiquitous and highly sequence-diverse virus family, were identified in, and isolated from the BSL metagenomic DNA sample. These BSL capsid protein sequences, along with several other homologous sequences derived from metagenomic surveys and laboratory isolates, were mapped to the solved structure of a closely related capsid protein from the Spiroplasma phage-4 microvirus. Patterns of amino acid sequence conservation, unveiled by structure-based homology modeling analysis, revealed that the protein sequences within this family exhibit a remarkable level of plasticity, while remaining structurally and functionally congruent.Lateral gene transfer is thought to have had a significant impact on the genomic evolution and adaptation of virus families. Genomic context analysis was also utilized to identify interviral gene transfer within the BSL virus assemblage. An ostensibly rare interviral gene transfer event, having transpired between single-stranded RNA and DNA virus types, was detected in the BSL metagenome. Similar genomes were subsequently detected in other ecosystems around the globe. The discovery of this new virus genome dramatically underscores the scope and importance of genetic mobility and genomic mosaicism as major forces driving the evolution of viruses.The analyses conducted herein demonstrate the many ways in which viral metagenomic sequence data may be utilized to not only evaluate the composition of a natural virus assemblage, but to discover new viral genes, and to better understand the dynamics of both genomic and structural evolution within the virosphere. Title: The Boiling Springs Lake Metavirome: Charting the Viral Sequence-Space of an Extreme Environment Microbial Ecosystem Author: Diemer, Geoffrey Scott Year: 2014 Viruses are the most abundant organisms on Earth, yet their collective evolutionary history, biodiversity and functional capacity is not well understood. Viral metagenomics offers a potential means of establishing a more comprehensive view of virus diversity and evolution, as vast amounts of new sequence data becomes available for comparative analysis.Metagenomic DNA from virus-sized particles (smaller than 0.2 microns in diameter) was isolated from approximately 20 liters of sediment obtained from Boiling Springs Lake (BSL) and sequenced. BSL is a large, acidic hot-spring (with a pH of 2.2, and temperatures ranging from 50°C to 96°C) located in Lassen Volcanic National Park, USA. BSL supports a purely microbial ecosystem comprised largely of Archaea and Bacteria, however, the lower temperature regions permit the growth of acid- and thermo-tolerant Eukarya. This distinctive feature of the BSL microbial ecosystem ensures that virus types infecting all domains of life will be present. The metagenomic sequence data was used to characterize the types of viruses present within the microbial ecosystem, to ascertain the extent of genetic diversity and novelty comprising the BSL virus assemblage, and to explore the genomic and structural modalities of virus evolution.Metagenomic surveys of natural virus assemblages, including the survey of BSL, have revealed that the diversity within the virosphere far exceeds what has currently been determined through the detailed study of viruses that are relevant to human health and agriculture. The number of as-yet-uncharacterized virus protein families present in the BSL assemblage was estimated by clustering analysis. Genomic context analysis of the predicted viral protein sequences in the BSL dataset indicates that most of the putative uncharacterized proteins are endemic or unique to BSL, and are largely harbored by known virus types. A comparative metagenomic analysis approach identified a set of conserved, yet uncharacterized BSL protein sequences that are commonly found in other similar and dissimilar environments.New sequence data from metagenomic surveys of natural virus assemblages was also used to better characterize and define known virus protein families, as some of the viruses found in the BSL environment represent distant relatives of well-characterized isolates. By comparing viral genes and protein sequences from these highly divergent species, it is possible to better understand the dynamics of adaptation and evolution in the virosphere. Additionally, as structures of virus proteins continue to be experimentally determined by X-ray crystallography and cryo-electron microscopy, a merger of structural and metagenomic sequence data allows the opportunity to observe the structural dynamics underlying virus protein evolution.Capsid (structural) proteins from two distinct Microviridae strains; a globally ubiquitous and highly sequence-diverse virus family, were identified in, and isolated from the BSL metagenomic DNA sample. These BSL capsid protein sequences, along with several other homologous sequences derived from metagenomic surveys and laboratory isolates, were mapped to the solved structure of a closely related capsid protein from the Spiroplasma phage-4 microvirus. Patterns of amino acid sequence conservation, unveiled by structure-based homology modeling analysis, revealed that the protein sequences within this family exhibit a remarkable level of plasticity, while remaining structurally and functionally congruent.Lateral gene transfer is thought to have had a significant impact on the genomic evolution and adaptation of virus families. Genomic context analysis was also utilized to identify interviral gene transfer within the BSL virus assemblage. An ostensibly rare interviral gene transfer event, having transpired between single-stranded RNA and DNA virus types, was detected in the BSL metagenome. Similar genomes were subsequently detected in other ecosystems around the globe. The discovery of this new virus genome dramatically underscores the scope and importance of genetic mobility and genomic mosaicism as major forces driving the evolution of viruses.The analyses conducted herein demonstrate the many ways in which viral metagenomic sequence data may be utilized to not only evaluate the composition of a natural virus assemblage, but to discover new viral genes, and to better understand the dynamics of both genomic and structural evolution within the virosphere. Close

• 5. [Article] Agricultural Management Decisions Impact Isoprene Emission and Physiology of Arundo donax, an Emerging Bioenergy Crop

  Arundo donax (Giant Reed) is quickly being developed as a rapidly-growing, robust, and highly productive bioenergy crop, with large scale cultivation of this species planned for the Columbia River basin ...

  Citation × Citation Citation Abstract Copy Title: Agricultural Management Decisions Impact Isoprene Emission and Physiology of Arundo donax, an Emerging Bioenergy Crop Author: Maxfield, Jason Charles Year: 2014 Arundo donax (Giant Reed) is quickly being developed as a rapidly-growing, robust, and highly productive bioenergy crop, with large scale cultivation of this species planned for the Columbia River basin of the Pacific Northwest (USA). Despite its potential as a next generation biomass crop, relatively few studies have examined the physiological performance of A. donax under agricultural conditions. Unlike traditional crops, A. donax is known to be a high-emitter of the volatile compound isoprene, which may significantly impact regional air quality, but it has not been widely cultivated in North America and little is known about how this species will perform in the Pacific Northwest. Over two field seasons, we measured isoprene fluxes from A. donax plants in both greenhouse conditions and in an agricultural field setting under a variety of conditions and fertilizer treatments. We also measured several other attributes of A. donax productivity and leaf physiology including chlorophyll content, photosynthesis rate, stomatal conductance, specific leaf mass, water use efficiency and gas exchange. We found that A. donax physiologically performs well under cultivation in the Columbia River basin, but that it also emits isoprene at significantly higher rates than previous reports indicate. We also found that both isoprene emission and leaf physiology were highly affected by agricultural management decisions, including nitrogen and irrigation management. Our findings indicate that crop management strategies can be developed that simultaneously seek to minimize isoprene emission while maximizing biomass production in this newly emerging bioenergy crop. Title: Agricultural Management Decisions Impact Isoprene Emission and Physiology of Arundo donax, an Emerging Bioenergy Crop Author: Maxfield, Jason Charles Year: 2014 Arundo donax (Giant Reed) is quickly being developed as a rapidly-growing, robust, and highly productive bioenergy crop, with large scale cultivation of this species planned for the Columbia River basin of the Pacific Northwest (USA). Despite its potential as a next generation biomass crop, relatively few studies have examined the physiological performance of A. donax under agricultural conditions. Unlike traditional crops, A. donax is known to be a high-emitter of the volatile compound isoprene, which may significantly impact regional air quality, but it has not been widely cultivated in North America and little is known about how this species will perform in the Pacific Northwest. Over two field seasons, we measured isoprene fluxes from A. donax plants in both greenhouse conditions and in an agricultural field setting under a variety of conditions and fertilizer treatments. We also measured several other attributes of A. donax productivity and leaf physiology including chlorophyll content, photosynthesis rate, stomatal conductance, specific leaf mass, water use efficiency and gas exchange. We found that A. donax physiologically performs well under cultivation in the Columbia River basin, but that it also emits isoprene at significantly higher rates than previous reports indicate. We also found that both isoprene emission and leaf physiology were highly affected by agricultural management decisions, including nitrogen and irrigation management. Our findings indicate that crop management strategies can be developed that simultaneously seek to minimize isoprene emission while maximizing biomass production in this newly emerging bioenergy crop. Close

• 6. [Article] Mimicking the Bioactivation of the Antitubercular Agent Ethionamide using Peracetic Acid

  Tuberculosis still affects a large number of the world’s population and drug resistance is becoming an increasing problem. Second line of defense antitubercular agents such as ethionamide (ETD) are thus ...

  Citation × Citation Citation Abstract Copy Title: Mimicking the Bioactivation of the Antitubercular Agent Ethionamide using Peracetic Acid Author: Logan, Isabelle E. Year: 2016 Tuberculosis still affects a large number of the world’s population and drug resistance is becoming an increasing problem. Second line of defense antitubercular agents such as ethionamide (ETD) are thus becoming increasingly important. Since ethionamide is a prodrug, its metabolization through oxidation by a monooxygenase was successfully mimicked using peracetic acid. The reaction was found to be biphasic, with a fast initial oxidation to yield ethionamide sulfoxide (ETD-SO), which was relatively stable and was successfully isolated. We derived a bimolecular rate constant of 3.08 ± 0.72 × 102 M-1 s-1 for this initial phase of the reaction. In the slow second phase, further oxidation yielded 2-ethylisonicotinamide, where zero order kinetics were observed. Electrospray ionization mass spectroscopy (ESI-MS) was used to determine the identities of the products. No sulfinic nor sulfonic acids were detected, indicating that the sulfur-carbon bond was cleaved at the sulfenic acid stage, resulting in the release of an unstable sulfur monoxide species, which dimerized in solution to form dithionite. Title: Mimicking the Bioactivation of the Antitubercular Agent Ethionamide using Peracetic Acid Author: Logan, Isabelle E. Year: 2016 Tuberculosis still affects a large number of the world’s population and drug resistance is becoming an increasing problem. Second line of defense antitubercular agents such as ethionamide (ETD) are thus becoming increasingly important. Since ethionamide is a prodrug, its metabolization through oxidation by a monooxygenase was successfully mimicked using peracetic acid. The reaction was found to be biphasic, with a fast initial oxidation to yield ethionamide sulfoxide (ETD-SO), which was relatively stable and was successfully isolated. We derived a bimolecular rate constant of 3.08 ± 0.72 × 102 M-1 s-1 for this initial phase of the reaction. In the slow second phase, further oxidation yielded 2-ethylisonicotinamide, where zero order kinetics were observed. Electrospray ionization mass spectroscopy (ESI-MS) was used to determine the identities of the products. No sulfinic nor sulfonic acids were detected, indicating that the sulfur-carbon bond was cleaved at the sulfenic acid stage, resulting in the release of an unstable sulfur monoxide species, which dimerized in solution to form dithionite. Close

• 7. [Article] Interactions Among Fuel Management, Species Composition, Bark Beetles, and Climate Change and the Potential Effects on Forests of the Lake Tahoe Basin

  Climate-driven increases in wildfires, drought conditions, and insect outbreaks are critical threats to forest carbon stores. In particular, bark beetles are important disturbance agents although their ...

  Citation × Citation Citation Abstract Copy Title: Interactions Among Fuel Management, Species Composition, Bark Beetles, and Climate Change and the Potential Effects on Forests of the Lake Tahoe Basin Author: Scheller, Robert M., Kretchun, Alec M., Loudermilk, E. Louise, Hurteau, Matthew D., Weisberg, Peter J., Skinner, Carl Year: 2017 Climate-driven increases in wildfires, drought conditions, and insect outbreaks are critical threats to forest carbon stores. In particular, bark beetles are important disturbance agents although their long-term interactions with future climate change are poorly understood. Droughts and the associated moisture deficit contribute to the onset of bark beetle outbreaks although outbreak extent and severity is dependent upon the density of host trees, wildfire, and forest management. Our objective was to estimate the effects of climate change and bark beetle outbreaks on ecosystem carbon dynamics over the next century in a western US forest. Specifically, we hypothesized that (a) bark beetle outbreaks under climate change would reduce net ecosystem carbon balance (NECB) and increase uncertainty and (b) these effects could be ameliorated by fuels management. We also examined the specific tree species dynamics—competition and release—that determined NECB response to bark beetle outbreaks. Our study area was the Lake Tahoe Basin (LTB), CA and NV, USA, an area of diverse forest types encompassing steep elevation and climatic gradients and representative of mixed-conifer forests throughout the western United States. We simulated climate change, bark beetles, wildfire, and fuels management using a landscape-scale stochastic model of disturbance and succession. We simulated the period 2010–2100 using downscaled climate projections. Recurring droughts generated conditions conducive to large-scale outbreaks; the resulting large and sustained outbreaks significantly increased the probability of LTB forests becoming C sources over decadal time scales, with slower-than-anticipated landscape-scale recovery. Tree species composition was substantially altered with a reduction in functional redundancy and productivity. Results indicate heightened uncertainty due to the synergistic influences of climate change and interacting disturbances. Our results further indicate that current fuel management practices will not be effective at reducing landscape-scale outbreak mortality. Our results provide critical insights into the interaction of drivers (bark beetles, wildfire, fuel management) that increase the risk of C loss and shifting community composition if bark beetle outbreaks become more frequent. Title: Interactions Among Fuel Management, Species Composition, Bark Beetles, and Climate Change and the Potential Effects on Forests of the Lake Tahoe Basin Author: Scheller, Robert M., Kretchun, Alec M., Loudermilk, E. Louise, Hurteau, Matthew D., Weisberg, Peter J., Skinner, Carl Year: 2017 Climate-driven increases in wildfires, drought conditions, and insect outbreaks are critical threats to forest carbon stores. In particular, bark beetles are important disturbance agents although their long-term interactions with future climate change are poorly understood. Droughts and the associated moisture deficit contribute to the onset of bark beetle outbreaks although outbreak extent and severity is dependent upon the density of host trees, wildfire, and forest management. Our objective was to estimate the effects of climate change and bark beetle outbreaks on ecosystem carbon dynamics over the next century in a western US forest. Specifically, we hypothesized that (a) bark beetle outbreaks under climate change would reduce net ecosystem carbon balance (NECB) and increase uncertainty and (b) these effects could be ameliorated by fuels management. We also examined the specific tree species dynamics—competition and release—that determined NECB response to bark beetle outbreaks. Our study area was the Lake Tahoe Basin (LTB), CA and NV, USA, an area of diverse forest types encompassing steep elevation and climatic gradients and representative of mixed-conifer forests throughout the western United States. We simulated climate change, bark beetles, wildfire, and fuels management using a landscape-scale stochastic model of disturbance and succession. We simulated the period 2010–2100 using downscaled climate projections. Recurring droughts generated conditions conducive to large-scale outbreaks; the resulting large and sustained outbreaks significantly increased the probability of LTB forests becoming C sources over decadal time scales, with slower-than-anticipated landscape-scale recovery. Tree species composition was substantially altered with a reduction in functional redundancy and productivity. Results indicate heightened uncertainty due to the synergistic influences of climate change and interacting disturbances. Our results further indicate that current fuel management practices will not be effective at reducing landscape-scale outbreak mortality. Our results provide critical insights into the interaction of drivers (bark beetles, wildfire, fuel management) that increase the risk of C loss and shifting community composition if bark beetle outbreaks become more frequent. Close

• 8. [Article] Measuring and Managing Resistance and Resilience Under Climate Change in Northern Great Lake Forests (USA)

  Context: Climate change will have diverse and interacting effects on forests over the next century. One of the most pronounced effects may be a decline in resistance to chronic change and resilience to ...

  Citation × Citation Citation Abstract Copy Title: Measuring and Managing Resistance and Resilience Under Climate Change in Northern Great Lake Forests (USA) Author: Duveneck, Matthew Joshua, Scheller, Robert M. Year: 2016 Context: Climate change will have diverse and interacting effects on forests over the next century. One of the most pronounced effects may be a decline in resistance to chronic change and resilience to acute disturbances. The capacity for forests to persist and/or adapt to climate change remains largely unknown, in part because there is not broad agreement how to measure and apply resilience concepts. Objectives: We assessed the interactions of climate change, resistance, resilience, diversity, and alternative management of northern Great Lake forests. Methods: We simulated two landscapes (northern Minnesota and northern lower Michigan), three climate futures (current climate, a low emissions trajectory, and a high emissions trajectory), and four management regimes [business as usual, expanded forest reserves, modified silviculture, and climate suitable planting (CSP)]. We simulated each scenario with a forest landscape simulation model. We assessed resistance as the change in species composition over time. We assessed resilience and calculated an index of resilience that incorporated both recovery of pre-fire tree species composition and aboveground biomass within simulated burned areas. Results: Results indicate a positive relationship between diversity and resistance within low diversity areas. Simulations of the high emission climate future resulted in a decline in both resistance and resilience. Conclusions: Of the management regimes, the CSP regime resulted in some of the greatest resilience under climate change although our results suggest that differences in forest management are largely outweighed by the effects of climate change. Our results provide a framework for assessing resistance and resilience relevant and valuable to a broad array of ecological systems. Title: Measuring and Managing Resistance and Resilience Under Climate Change in Northern Great Lake Forests (USA) Author: Duveneck, Matthew Joshua, Scheller, Robert M. Year: 2016 Context: Climate change will have diverse and interacting effects on forests over the next century. One of the most pronounced effects may be a decline in resistance to chronic change and resilience to acute disturbances. The capacity for forests to persist and/or adapt to climate change remains largely unknown, in part because there is not broad agreement how to measure and apply resilience concepts. Objectives: We assessed the interactions of climate change, resistance, resilience, diversity, and alternative management of northern Great Lake forests. Methods: We simulated two landscapes (northern Minnesota and northern lower Michigan), three climate futures (current climate, a low emissions trajectory, and a high emissions trajectory), and four management regimes [business as usual, expanded forest reserves, modified silviculture, and climate suitable planting (CSP)]. We simulated each scenario with a forest landscape simulation model. We assessed resistance as the change in species composition over time. We assessed resilience and calculated an index of resilience that incorporated both recovery of pre-fire tree species composition and aboveground biomass within simulated burned areas. Results: Results indicate a positive relationship between diversity and resistance within low diversity areas. Simulations of the high emission climate future resulted in a decline in both resistance and resilience. Conclusions: Of the management regimes, the CSP regime resulted in some of the greatest resilience under climate change although our results suggest that differences in forest management are largely outweighed by the effects of climate change. Our results provide a framework for assessing resistance and resilience relevant and valuable to a broad array of ecological systems. Close

• 9. [Article] Prospectus for Future Research: Temperature Effects on Green Macroalgae

  Research on temperature change has received attention from scientists. Not only has temperature change attracted scientists who study the Earth’s weather and climate, but also those who study plants and ...

  Citation × Citation Citation Abstract Copy Title: Prospectus for Future Research: Temperature Effects on Green Macroalgae Author: Gonzalez Pelayo, Ana Year: 2016 Research on temperature change has received attention from scientists. Not only has temperature change attracted scientists who study the Earth’s weather and climate, but also those who study plants and animals on land and in oceans. The research being conducted looks at how these temperature changes affect the anatomy and physiology of organisms, but other research also studies how terrestrial and aquatic ecosystems have changed and the types of responses that have occurred as a result. For many years, research in terrestrial ecosystems has dominated research in aquatic ecosystems. Limited research has been conducted in aquatic ecosystems involving green macroalgae, which are primary producers and an essential part of these ecosystems. Within the last fifteen years, research regarding the effects of temperature change on green macroalgae has shown that temperature affects photosynthesis and biochemistry. Overall, photosynthesis is almost completely inhibited after sufficient exposure due to damage in Photosystem II. There is also a substantial change in the production of chemical molecules known as reactive oxygen species that caused physical damage to the algal cells as well as a change in structure for proteins that is essential for function. I reviewed the literature within the past fifteen years, regarding the effects of temperature on a variety of green macroalgae species. The results indicated that after sufficient exposure, the photosynthetic rate of macroalgae is reduced significantly and that the change in chemistry lead to alterations in the physiology of green macroalgae. In this thesis, I propose two possible directions for algae research that involves genetic sequencing of a model species to identify a possible temperature tolerance gene, as well as the potential use of green macroalgae in photobioreactors as a source of renewable energy. Title: Prospectus for Future Research: Temperature Effects on Green Macroalgae Author: Gonzalez Pelayo, Ana Year: 2016 Research on temperature change has received attention from scientists. Not only has temperature change attracted scientists who study the Earth’s weather and climate, but also those who study plants and animals on land and in oceans. The research being conducted looks at how these temperature changes affect the anatomy and physiology of organisms, but other research also studies how terrestrial and aquatic ecosystems have changed and the types of responses that have occurred as a result. For many years, research in terrestrial ecosystems has dominated research in aquatic ecosystems. Limited research has been conducted in aquatic ecosystems involving green macroalgae, which are primary producers and an essential part of these ecosystems. Within the last fifteen years, research regarding the effects of temperature change on green macroalgae has shown that temperature affects photosynthesis and biochemistry. Overall, photosynthesis is almost completely inhibited after sufficient exposure due to damage in Photosystem II. There is also a substantial change in the production of chemical molecules known as reactive oxygen species that caused physical damage to the algal cells as well as a change in structure for proteins that is essential for function. I reviewed the literature within the past fifteen years, regarding the effects of temperature on a variety of green macroalgae species. The results indicated that after sufficient exposure, the photosynthetic rate of macroalgae is reduced significantly and that the change in chemistry lead to alterations in the physiology of green macroalgae. In this thesis, I propose two possible directions for algae research that involves genetic sequencing of a model species to identify a possible temperature tolerance gene, as well as the potential use of green macroalgae in photobioreactors as a source of renewable energy. Close

• 10. [Article] Intestinal Microbiota Diversity of Pre-Smolt Steelhead (Oncorhynchus mykiss) Across Six Oregon and Washington Hatcheries

  The Pacific Northwest is known for its once-abundant wild salmonid populations that have been in decline for more than 50 years due to habitat destruction and commercial overexploitation. To compensate, ...

  Citation × Citation Citation Abstract Copy Title: Intestinal Microbiota Diversity of Pre-Smolt Steelhead (Oncorhynchus mykiss) Across Six Oregon and Washington Hatcheries Author: Yildirimer, Christina Carrell Year: 2017 The Pacific Northwest is known for its once-abundant wild salmonid populations that have been in decline for more than 50 years due to habitat destruction and commercial overexploitation. To compensate, federal and state agencies annually release hundreds of thousands of hatchery-reared fish into the wild. However, accumulating data indicate that hatchery fish have lower fitness in natural environments, and that hatchery rearing negatively influences return rates of anadromous salmonids. Recently, mounting evidence revealed that the richness and diversity of intestinal microbial species influence host health. We examined the gut microbiota of pre-migratory hatchery-reared steelhead (Oncorhynchus mykiss) to assess microbial community diversity. The Cascade Mountains serve as an allopatric border between two distinct clades of steelhead that show significant differences in genomic and mitochondrial diversity. We identified differences in core microbiota of hatchery-reared fish that correlate with this divergent phylogeographic distribution. Steelhead sampled from hatcheries east of the Cascades had overall greater core gut microbiota diversity. These differences were found despite similarities in diet and rearing conditions. In addition to taxonomic variation across the geographic divide, we identified significant differences in metabolic pathways using PICRUSt gene prediction software. Our analysis revealed significant enrichment of genes associated with lipid metabolism in the gut microbiome of western fish. 8 of 19 individual lipid metabolism pathways were more prominent in western populations. Lipids are a vital nutritional component for teleost species involved in migration and subsequent return for spawning in natal environments. We hypothesize that the observed differences in lipid metabolism across this phylogenetic divide results from an increased ability of eastern Cascade (O. m. gairdneri) fish to utilize lipids taken in via the diet. This increased absorption and utilization would make lipids less available for the intestinal microbiota of the eastern fish, as evidenced by the lower abundance of lipid metabolism genes in the east. Our research utilizes information from the microbiome to understand the phenotypic implications occurring in segregated populations of hatchery-reared steelhead, further confirming elements of coevolution between an organism and its internal environment. Title: Intestinal Microbiota Diversity of Pre-Smolt Steelhead (Oncorhynchus mykiss) Across Six Oregon and Washington Hatcheries Author: Yildirimer, Christina Carrell Year: 2017 The Pacific Northwest is known for its once-abundant wild salmonid populations that have been in decline for more than 50 years due to habitat destruction and commercial overexploitation. To compensate, federal and state agencies annually release hundreds of thousands of hatchery-reared fish into the wild. However, accumulating data indicate that hatchery fish have lower fitness in natural environments, and that hatchery rearing negatively influences return rates of anadromous salmonids. Recently, mounting evidence revealed that the richness and diversity of intestinal microbial species influence host health. We examined the gut microbiota of pre-migratory hatchery-reared steelhead (Oncorhynchus mykiss) to assess microbial community diversity. The Cascade Mountains serve as an allopatric border between two distinct clades of steelhead that show significant differences in genomic and mitochondrial diversity. We identified differences in core microbiota of hatchery-reared fish that correlate with this divergent phylogeographic distribution. Steelhead sampled from hatcheries east of the Cascades had overall greater core gut microbiota diversity. These differences were found despite similarities in diet and rearing conditions. In addition to taxonomic variation across the geographic divide, we identified significant differences in metabolic pathways using PICRUSt gene prediction software. Our analysis revealed significant enrichment of genes associated with lipid metabolism in the gut microbiome of western fish. 8 of 19 individual lipid metabolism pathways were more prominent in western populations. Lipids are a vital nutritional component for teleost species involved in migration and subsequent return for spawning in natal environments. We hypothesize that the observed differences in lipid metabolism across this phylogenetic divide results from an increased ability of eastern Cascade (O. m. gairdneri) fish to utilize lipids taken in via the diet. This increased absorption and utilization would make lipids less available for the intestinal microbiota of the eastern fish, as evidenced by the lower abundance of lipid metabolism genes in the east. Our research utilizes information from the microbiome to understand the phenotypic implications occurring in segregated populations of hatchery-reared steelhead, further confirming elements of coevolution between an organism and its internal environment. Close