Lizabeth Bowen - Academia.edu (original) (raw)

Papers by Lizabeth Bowen

Frontiers for Young Minds

Oceans are changing and these changes are affecting the animals that live there. Animals respond ... more Oceans are changing and these changes are affecting the animals that live there. Animals respond differently to changes in water temperature, food availability, and contaminants. Those responses can be seen in their genes. A technique called transcriptomics allows scientists to see the response of an animal’s genes to its environment. We used transcriptomics to compare two populations of Pacific razor clams in Alaska (United States): one that has lots of clams and one that used to have lots but does not anymore. We were surprised when we did not find any differences in their gene responses! So, we had to think about what else might be influencing the number of clams in these two populations. As we “dug” for answers, we found out that there are differences between the populations that do not influence their genes but may impact their numbers, such as being eaten by predators.

Gene transcription analysis for diagnosing or monitoring wildlife health requires the ability to ... more Gene transcription analysis for diagnosing or monitoring wildlife health requires the ability to distinguish pathophysiological change from natural variation. Herein, we describe methodology for the development of quantitative real-time polymerase chain reaction (qPCR) assays to measure differential transcript levels of multiple immune function genes in the sea otter (Enhydra lutris); sea otter-specific qPCR primer sequences for the genes of interest are defined. We establish a ‘reference’ range of transcripts for each gene in a group of clinically healthy captive and free-ranging sea otters. The 10 genes of interest represent multiple physiological systems that play a role in immuno-modulation, inflammation, cell protection, tumour suppression, cellular stress response, xenobiotic metabolizing enzymes, antioxidant enzymes and cell–cell adhesion. The cycle threshold (CT) measures for most genes were normally distributed; the complement cytolysis inhibitor was the exception. The rela...

Ecological Monographs, 2021

Conservation Physiology

Increasing risk of pathogen spillover coupled with overall declines in wildlife population abunda... more Increasing risk of pathogen spillover coupled with overall declines in wildlife population abundance in the Anthropocene make infectious disease a relevant concern for species conservation worldwide. While emerging molecular tools could improve our diagnostic capabilities and give insight into mechanisms underlying wildlife disease risk, they have rarely been applied in practice. Here, employing a previously reported gene transcription panel of common immune markers to track physiological changes, we present a detailed analysis over the course of both acute and chronic infection in one wildlife species where disease plays a critical role in conservation, bighorn sheep (Ovis canadensis). Differential gene transcription patterns distinguished between infection statuses over the course of acute infection and differential correlation (DC) analyses identified clear changes in gene co-transcription patterns over the early stages of infection, with transcription of four genes—TGFb, AHR, IL...

Open-File Report, 2017

The re-colonization of the Santa Barbara channel by sea otters brings these ESA-listed marine mam... more The re-colonization of the Santa Barbara channel by sea otters brings these ESA-listed marine mammals closer to active oil and gas production facilities, shipping lanes and naturally occurring oil and gas seeps. However, the degree to which sea otters may actually be affected by human-caused oil spills or exposure to natural oil seeps is currently unknown. Between 2012 and 2014, the U.S. Geological Survey and collaborating agencies conducted a telemetry-based study of sea otters in Santa Barbara channel, in order to provide critical information for resource managers (specifically the Bureau of Ocean Energy Management, henceforth BOEM, and the U.S. Fish and Wildlife Service, henceforth USFWS) about the spatial ecology, population status, and potential population threats to sea otters in Santa Barbara Channel, with particular reference to exposure to manmade structures and sources of oil and natural gas. Analysis of spatial monitoring data using a Bayesian-based synoptic model allowed for description of sea otter home ranges, identification of hot-spots of use, and insights into habitat selection behavior by male and female sea otters. Important findings included the deeper modal depth preferred by males versus females, strong preferences by both sexes for areas with persistent kelp canopy, and greater use of soft-sediment areas by males. The synoptic model also provided the ability to predict population-level density distribution for each sex in new habitats: by calculating the value of these probability density distributions at the known locations of natural seeps, we were able to identify those seeps with higher potential for sea otter encounters. The relative probability of occurrence at locations near to some seeps was sufficiently high (about 1% likelihood of occurrence for some of our study animals) that one would anticipate occasional encounters. Data on male and female survival, reproductive success, activity budgets, and body condition all indicated that sea otters in Santa Barbara Channel are not resource limited, and thus we would expect to see continued strong population growth in this area. However, the principal cause of death for study animals was lethal bites by white sharks, suggesting that shark bite mortality represents the single biggest threat to continued population growth in the Santa Barbara Channel.

Conservation Biology, 1997

Conservation Physiology

Manipulative experiments provide stronger evidence for identifying cause-and-effect relationships... more Manipulative experiments provide stronger evidence for identifying cause-and-effect relationships than correlative studies, but protocols for implementing temperature manipulations are lacking for large species in remote settings. We developed an experimental protocol for holding adult Chinook salmon (Oncorhynchus tshawytscha) and exposing them to elevated temperature treatments. The goal of the experimental protocol was to validate heat stress biomarkers by increasing river water temperature from ambient (~14°C) to a treatment temperature of 18°C or 21°C and then maintain the treatment temperature over 4 hours within a range of ±1.0°C. Our protocol resulted in a mean rate of temperature rise of 3.71°C h-1 (SD = 1.31) to treatment temperatures and mean holding temperatures of 18.0°C (SD = 0.2) and 21.0°C (SD = 0.2) in the low- and high-heat treatments, respectively. Our work demonstrated that manipulative experiments with large, mobile study species can be successfully developed in ...

Conservation Physiology

Chinook salmon (Oncorhynchus tshawytscha) declines are widespread and may be attributed, at least... more Chinook salmon (Oncorhynchus tshawytscha) declines are widespread and may be attributed, at least in part, to warming river temperatures. Water temperatures in the Yukon River and tributaries often exceed 18°C, a threshold commonly associated with heat stress and elevated mortality in Pacific salmon. Untangling the complex web of direct and indirect physiological effects of heat stress on salmon is difficult in a natural setting with innumerable system challenges but is necessary to increase our understanding of both lethal and sublethal impacts of heat stress on populations. The goal of this study was to characterize the cellular stress response in multiple Chinook salmon tissues after acute elevated temperature challenges. We conducted a controlled 4-hour temperature exposure (control, 18°C and 21°C) experiment on the bank of the Yukon River followed by gene expression (GE) profiling using a 3′-Tag-RNA-Seq protocol. The full transcriptome was analysed for 22 Chinook salmon in musc...

PeerJ

An emerging approach to ecosystem monitoring involves the use of physiological biomarker analyses... more An emerging approach to ecosystem monitoring involves the use of physiological biomarker analyses in combination with gene transcription assays. For the first time, we employed these tools to evaluate the Pacific razor clam (Siliqua patula), which is important both economically and ecologically, as a bioindicator species in the northeast Pacific. Our objectives were to (1) develop biomarker and gene transcription assays with which to monitor the health of the Pacific razor clam, (2) acquire baseline biomarker and gene transcription reference ranges for razor clams, (3) assess the relationship between physiological and gene transcription assays and (4) determine if site-level differences were present. Pacific razor clams were collected in July 2015 and 2016 at three sites within each of two national parks in southcentral Alaska. In addition to determining reference ranges, we found differences in biomarker assay and gene transcription results between parks and sites which indicate va...

PeerJ

Coastal regions worldwide face increasing management concerns due to natural and anthropogenic fo... more Coastal regions worldwide face increasing management concerns due to natural and anthropogenic forces that have the potential to significantly degrade nearshore marine resources. The goal of our study was to develop and test a monitoring strategy for nearshore marine ecosystems in remote areas that are not readily accessible for sampling. Mussel species have been used extensively to assess ecosystem vulnerability to multiple, interacting stressors. We sampled bay mussels (Mytilus trossulus) in 2015 and 2016 from six intertidal sites in Lake Clark and Katmai National Parks and Preserves, in south-central Alaska. Reference ranges for physiological assays and gene transcription were determined for use in future assessment efforts. Both techniques identified differences among sites, suggesting influences of both large-scale and local environmental factors and underscoring the value of this combined approach to ecosystem health monitoring.

Ecology and Evolution

The immune response is central in the defenses against infectious disease, but accurately measuri... more The immune response is central in the defenses against infectious disease, but accurately measuring immune function and targeted responses in nonmodel wildlife can be challenging (Bowden,

Open-File Report

Detecting populations of rare or cryptic species is essential for their conservation. For species... more Detecting populations of rare or cryptic species is essential for their conservation. For species like giant gartersnakes (Thamnophis gigas), conventional survey methods can be expensive and inefficient. These sampling difficulties might be overcome by modern techniques that detect deoxyribonucleic acid (DNA) shed by organisms into the environment (eDNA). We evaluated the efficacy of detecting giant gartersnake eDNA in water samples from the laboratory and at locations with known giant gartersnake populations in the Sacramento Valley of California, and failed to detect giant gartersnake DNA in most laboratory and all field samples. Aspects of giant gartersnake biology-such as highly keratinized skin and spending extensive time in the terrestrial environment, as well as hot, sunny, and turbid conditions in wetlands and canals of the Sacramento Valley-likely contributed to low detection probabilities. Although detection of eDNA shows promise under many conditions, further development is needed before sampling for eDNA is a viable option for detecting giant gartersnake populations.

EcoHealth, Jan 22, 2017

Increasingly, population- and ecosystem-level health assessments are performed using sophisticate... more Increasingly, population- and ecosystem-level health assessments are performed using sophisticated molecular tools. Advances in molecular technology enable the identification of synergistic effects of multiple stressors on the individual physiology of different species. Brown bears (Ursus arctos) are an apex predator; thus, they are ideal candidates for detecting potentially ecosystem-level systemic perturbations using molecular-based tools. We used gene transcription to analyze 130 brown bear samples from three National Parks and Preserves in Alaska. Although the populations we studied are apparently stable in abundance and exist within protected and intact environments, differences in transcript profiles were noted. The most prevalent differences were among locations. The transcript patterns among groups reflect the influence of environmental factors, such as nutritional status, disease, and xenobiotic exposure. However, these profiles also likely represent baselines for each uniq...

Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases, 2018

We examined the associations between California sea lion MHC class II DRB (Zaca-DRB) configuratio... more We examined the associations between California sea lion MHC class II DRB (Zaca-DRB) configuration and diversity, and leptospirosis. As Zaca-DRB gene sequences are involved with antigen presentation of bacteria and other extracellular pathogens, we predicted that they would play a role in determining responses to these pathogenic spirochaetes. Specifically, we investigated whether Zaca-DRB diversity (number of genes) and configuration (presence of specific genes) explained differences in disease severity, and whether higher levels of Zaca-DRB diversity predicted the number of specific Leptospira interrogans serovars that a sea lion's serum would react against. We found that serum from diseased sea lions with more Zaca-DRB loci reacted against a wider array of serovars. Specific Zaca-DRB loci were linked to reactions with particular serovars. Interestingly, sea lions with clinical manifestation of leptospirosis that had higher numbers of Zaca-DRB loci were less likely to recover ...

Conservation Physiology

The analysis of blood constituents is a widely used tool to aid in monitoring of animal health an... more The analysis of blood constituents is a widely used tool to aid in monitoring of animal health and disease. However, classic blood diagnostics (i.e. hematologic and plasma biochemical values) often do not provide sufficient information to determine the state of an animal's health. Field studies on wild tortoises and other reptiles have had limited success in drawing significant inferences between blood diagnostics and physiological and immunological condition. However, recent research using gene transcription profiling in the threatened Mojave desert tortoise (Gopherus agassizii) has proved useful in identifying immune or physiologic responses and overall health. To improve our understanding of health and immune function in tortoises, we evaluated both standard blood diagnostic (body condition, hematologic, plasma biochemistry values, trace elements, plasma proteins, vitamin A levels) and gene transcription profiles in 21 adult tortoises (11 clinically abnormal; 10 clinically normal) from Clark County, NV, USA. Necropsy and histology evaluations from clinically abnormal tortoises revealed multiple physiological complications, with moderate to severe rhinitis or pneumonia being the primary cause of morbidity in all but one of the examined animals. Clinically abnormal tortoises had increased transcription for four genes (SOD, MyD88, CL and Lep), increased lymphocyte production, biochemical enzymes and organics, trace elements of copper, and decreased numbers of leukocytes. We found significant positive correlations between increased transcription for SOD and increased trace elements for copper, as well as genes MyD88 and Lep with increased inflammation and microbial insults. Improved methods for health assessments are an important element of monitoring tortoise population recovery and can support the development of more robust diagnostic measures for ill animals, or individuals directly impacted by disturbance.

Environmental Toxicology and Chemistry, 2016

The authors quantified hepatic hydrocarbon-inducible cytochrome P4501A (CYP1A) expression, as eth... more The authors quantified hepatic hydrocarbon-inducible cytochrome P4501A (CYP1A) expression, as ethoxyresorufin-O-deethylase (EROD) activity, in wintering harlequin ducks (Histrionicus histrionicus) captured in Prince William Sound, Alaska (USA), during 2011, 2013, and 2014 (22-25 yr following the 1989 Exxon Valdez oil spill). Average EROD activity was compared between birds from areas oiled by the spill and those from nearby unoiled areas. The present study replicated studies conducted from 1998 to 2009 demonstrating that harlequin ducks using areas oiled in 1989 had elevated EROD activity, indicative of oil exposure, up to 2 decades post spill. In the present study, it was found that average EROD activity during March 2011 was significantly higher in wintering harlequin ducks captured in oiled areas relative to unoiled areas, which the authors interpret to indicate that harlequin ducks continued to be exposed to residual Exxon Valdez oil up to 22 yr after the original spill. However, the 2011 results also indicated reductions in exposure relative to previous years. Average EROD activity in birds from oiled areas was approximately 2 times that in birds from unoiled areas in 2011, compared with observations from 2005 to 2009, in which EROD activity was 3 to 5 times higher in oiled areas. It was also found that average EROD activity during March 2013 and March 2014 was not elevated in wintering harlequin ducks from oiled areas. The authors interpret these findings to indicate that exposure of harlequin ducks to residual Exxon Valdez oil abated within 24 yr after the original spill. The present study finalizes a timeline of exposure, extending over 2 decades, for a bird species thought to be particularly vulnerable to oil contamination in marine environments. Environ Toxicol Chem 2016;9999:1-7. Published 2016 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.

Journal of Marine Science and Engineering, 2016

Using a panel of genes stimulated by oil exposure in a laboratory study, we evaluated gene transc... more Using a panel of genes stimulated by oil exposure in a laboratory study, we evaluated gene transcription in blood leukocytes sampled from sea otters captured from 2006-2012 in western Prince William Sound (WPWS), Alaska, 17-23 years after the 1989 Exxon Valdez oil spill (EVOS). We compared WPWS sea otters to reference populations (not affected by the EVOS) from the Alaska Peninsula (2009), Katmai National Park and Preserve (2009), Clam Lagoon at Adak Island (2012), Kodiak Island (2005) and captive sea otters in aquaria. Statistically, sea otter gene transcript profiles separated into three distinct clusters: Cluster 1, Kodiak and WPWS 2006-2008 (higher relative transcription); Cluster 2, Clam Lagoon and WPWS 2010-2012 (lower relative transcription); and Cluster 3, Alaska Peninsula, Katmai and captive sea otters (intermediate relative transcription). The lower transcription of the aryl hydrocarbon receptor (AHR), an established biomarker for hydrocarbon exposure, in WPWS 2010-2012 compared to earlier samples from WPWS is consistent with declining hydrocarbon exposure, but the pattern of overall low levels of transcription seen in WPWS 2010-2012 could be related to other factors, such as food limitation, pathogens or injury, and may indicate an inability to mount effective responses to stressors. Decreased transcriptional response across the entire gene panel precludes the evaluation of whether or not individual sea otters show signs of exposure to lingering oil. However, related studies on sea otter demographics indicate that by 2012, the sea otter population in WPWS had recovered, which indicates diminishing oil exposure.

Veterinary Immunology and Immunopathology, 2016

Early identification of illness and/or presence of environmental and/or social stressors in free-... more Early identification of illness and/or presence of environmental and/or social stressors in free-ranging and domestic cetaceans is a priority for marine mammal health care professionals. Incorporation of leukocyte gene transcript analysis into the diagnostic tool kit has the potential to augment classical diagnostics based upon ease of sample storage and shipment, inducible nature and well-defined roles of transcription and associated downstream actions. Development of biomarkers that could serve to identify "insults" and potentially differentiate disease etiology would be of great diagnostic value. To this end, a modest number of peripheral blood leukocyte gene transcripts were selected for application to a domestic killer whale population with a focus on broad representation of inducible immunologically relevant genes. Normalized leukocyte transcript values, longitudinally acquired from 232 blood samples derived from 26 clinically healthy whales, were not visibly influenced temporally nor by sex or the specific Park in which they resided. Stability in leukocyte transcript number during periods of health enhances their potential use in diagnostics through identification of outliers. Transcript levels of two cytokine genes, IL-4 and IL-17, were highly variable within the group as compared to the other transcripts. IL-4 transcripts were typically absent. Analysis of transcript levels on the other genes of interest, on an individual animal basis, identified more outliers than were visible when analyzed in the context of the entire population. The majority of outliers (9 samples) were low, though elevated transcripts were identified for IL-17 from 2 animals and one each for Cox-2 and IL-10. The low number of outliers was not unexpected as sample selection was intentionally directed towards animals that were clinically healthy at the time of collection. Outliers may reflect animals experiencing subclinical disease that is transient and self-limiting. The immunologic knowledge derived from longitudinal immunologic studies in killer whales, as was the target of the present study, has the potential to improve diagnostics and health related decision making for this and other domestic and free-ranging cetacean species.

Frontiers for Young Minds

Oceans are changing and these changes are affecting the animals that live there. Animals respond ... more Oceans are changing and these changes are affecting the animals that live there. Animals respond differently to changes in water temperature, food availability, and contaminants. Those responses can be seen in their genes. A technique called transcriptomics allows scientists to see the response of an animal’s genes to its environment. We used transcriptomics to compare two populations of Pacific razor clams in Alaska (United States): one that has lots of clams and one that used to have lots but does not anymore. We were surprised when we did not find any differences in their gene responses! So, we had to think about what else might be influencing the number of clams in these two populations. As we “dug” for answers, we found out that there are differences between the populations that do not influence their genes but may impact their numbers, such as being eaten by predators.

Gene transcription analysis for diagnosing or monitoring wildlife health requires the ability to ... more Gene transcription analysis for diagnosing or monitoring wildlife health requires the ability to distinguish pathophysiological change from natural variation. Herein, we describe methodology for the development of quantitative real-time polymerase chain reaction (qPCR) assays to measure differential transcript levels of multiple immune function genes in the sea otter (Enhydra lutris); sea otter-specific qPCR primer sequences for the genes of interest are defined. We establish a ‘reference’ range of transcripts for each gene in a group of clinically healthy captive and free-ranging sea otters. The 10 genes of interest represent multiple physiological systems that play a role in immuno-modulation, inflammation, cell protection, tumour suppression, cellular stress response, xenobiotic metabolizing enzymes, antioxidant enzymes and cell–cell adhesion. The cycle threshold (CT) measures for most genes were normally distributed; the complement cytolysis inhibitor was the exception. The rela...

Ecological Monographs, 2021

Conservation Physiology

Increasing risk of pathogen spillover coupled with overall declines in wildlife population abunda... more Increasing risk of pathogen spillover coupled with overall declines in wildlife population abundance in the Anthropocene make infectious disease a relevant concern for species conservation worldwide. While emerging molecular tools could improve our diagnostic capabilities and give insight into mechanisms underlying wildlife disease risk, they have rarely been applied in practice. Here, employing a previously reported gene transcription panel of common immune markers to track physiological changes, we present a detailed analysis over the course of both acute and chronic infection in one wildlife species where disease plays a critical role in conservation, bighorn sheep (Ovis canadensis). Differential gene transcription patterns distinguished between infection statuses over the course of acute infection and differential correlation (DC) analyses identified clear changes in gene co-transcription patterns over the early stages of infection, with transcription of four genes—TGFb, AHR, IL...

Open-File Report, 2017

The re-colonization of the Santa Barbara channel by sea otters brings these ESA-listed marine mam... more The re-colonization of the Santa Barbara channel by sea otters brings these ESA-listed marine mammals closer to active oil and gas production facilities, shipping lanes and naturally occurring oil and gas seeps. However, the degree to which sea otters may actually be affected by human-caused oil spills or exposure to natural oil seeps is currently unknown. Between 2012 and 2014, the U.S. Geological Survey and collaborating agencies conducted a telemetry-based study of sea otters in Santa Barbara channel, in order to provide critical information for resource managers (specifically the Bureau of Ocean Energy Management, henceforth BOEM, and the U.S. Fish and Wildlife Service, henceforth USFWS) about the spatial ecology, population status, and potential population threats to sea otters in Santa Barbara Channel, with particular reference to exposure to manmade structures and sources of oil and natural gas. Analysis of spatial monitoring data using a Bayesian-based synoptic model allowed for description of sea otter home ranges, identification of hot-spots of use, and insights into habitat selection behavior by male and female sea otters. Important findings included the deeper modal depth preferred by males versus females, strong preferences by both sexes for areas with persistent kelp canopy, and greater use of soft-sediment areas by males. The synoptic model also provided the ability to predict population-level density distribution for each sex in new habitats: by calculating the value of these probability density distributions at the known locations of natural seeps, we were able to identify those seeps with higher potential for sea otter encounters. The relative probability of occurrence at locations near to some seeps was sufficiently high (about 1% likelihood of occurrence for some of our study animals) that one would anticipate occasional encounters. Data on male and female survival, reproductive success, activity budgets, and body condition all indicated that sea otters in Santa Barbara Channel are not resource limited, and thus we would expect to see continued strong population growth in this area. However, the principal cause of death for study animals was lethal bites by white sharks, suggesting that shark bite mortality represents the single biggest threat to continued population growth in the Santa Barbara Channel.

Conservation Biology, 1997

Conservation Physiology

Manipulative experiments provide stronger evidence for identifying cause-and-effect relationships... more Manipulative experiments provide stronger evidence for identifying cause-and-effect relationships than correlative studies, but protocols for implementing temperature manipulations are lacking for large species in remote settings. We developed an experimental protocol for holding adult Chinook salmon (Oncorhynchus tshawytscha) and exposing them to elevated temperature treatments. The goal of the experimental protocol was to validate heat stress biomarkers by increasing river water temperature from ambient (~14°C) to a treatment temperature of 18°C or 21°C and then maintain the treatment temperature over 4 hours within a range of ±1.0°C. Our protocol resulted in a mean rate of temperature rise of 3.71°C h-1 (SD = 1.31) to treatment temperatures and mean holding temperatures of 18.0°C (SD = 0.2) and 21.0°C (SD = 0.2) in the low- and high-heat treatments, respectively. Our work demonstrated that manipulative experiments with large, mobile study species can be successfully developed in ...

Conservation Physiology

Chinook salmon (Oncorhynchus tshawytscha) declines are widespread and may be attributed, at least... more Chinook salmon (Oncorhynchus tshawytscha) declines are widespread and may be attributed, at least in part, to warming river temperatures. Water temperatures in the Yukon River and tributaries often exceed 18°C, a threshold commonly associated with heat stress and elevated mortality in Pacific salmon. Untangling the complex web of direct and indirect physiological effects of heat stress on salmon is difficult in a natural setting with innumerable system challenges but is necessary to increase our understanding of both lethal and sublethal impacts of heat stress on populations. The goal of this study was to characterize the cellular stress response in multiple Chinook salmon tissues after acute elevated temperature challenges. We conducted a controlled 4-hour temperature exposure (control, 18°C and 21°C) experiment on the bank of the Yukon River followed by gene expression (GE) profiling using a 3′-Tag-RNA-Seq protocol. The full transcriptome was analysed for 22 Chinook salmon in musc...

PeerJ

An emerging approach to ecosystem monitoring involves the use of physiological biomarker analyses... more An emerging approach to ecosystem monitoring involves the use of physiological biomarker analyses in combination with gene transcription assays. For the first time, we employed these tools to evaluate the Pacific razor clam (Siliqua patula), which is important both economically and ecologically, as a bioindicator species in the northeast Pacific. Our objectives were to (1) develop biomarker and gene transcription assays with which to monitor the health of the Pacific razor clam, (2) acquire baseline biomarker and gene transcription reference ranges for razor clams, (3) assess the relationship between physiological and gene transcription assays and (4) determine if site-level differences were present. Pacific razor clams were collected in July 2015 and 2016 at three sites within each of two national parks in southcentral Alaska. In addition to determining reference ranges, we found differences in biomarker assay and gene transcription results between parks and sites which indicate va...

PeerJ

Coastal regions worldwide face increasing management concerns due to natural and anthropogenic fo... more Coastal regions worldwide face increasing management concerns due to natural and anthropogenic forces that have the potential to significantly degrade nearshore marine resources. The goal of our study was to develop and test a monitoring strategy for nearshore marine ecosystems in remote areas that are not readily accessible for sampling. Mussel species have been used extensively to assess ecosystem vulnerability to multiple, interacting stressors. We sampled bay mussels (Mytilus trossulus) in 2015 and 2016 from six intertidal sites in Lake Clark and Katmai National Parks and Preserves, in south-central Alaska. Reference ranges for physiological assays and gene transcription were determined for use in future assessment efforts. Both techniques identified differences among sites, suggesting influences of both large-scale and local environmental factors and underscoring the value of this combined approach to ecosystem health monitoring.

Ecology and Evolution

The immune response is central in the defenses against infectious disease, but accurately measuri... more The immune response is central in the defenses against infectious disease, but accurately measuring immune function and targeted responses in nonmodel wildlife can be challenging (Bowden,

Open-File Report

Detecting populations of rare or cryptic species is essential for their conservation. For species... more Detecting populations of rare or cryptic species is essential for their conservation. For species like giant gartersnakes (Thamnophis gigas), conventional survey methods can be expensive and inefficient. These sampling difficulties might be overcome by modern techniques that detect deoxyribonucleic acid (DNA) shed by organisms into the environment (eDNA). We evaluated the efficacy of detecting giant gartersnake eDNA in water samples from the laboratory and at locations with known giant gartersnake populations in the Sacramento Valley of California, and failed to detect giant gartersnake DNA in most laboratory and all field samples. Aspects of giant gartersnake biology-such as highly keratinized skin and spending extensive time in the terrestrial environment, as well as hot, sunny, and turbid conditions in wetlands and canals of the Sacramento Valley-likely contributed to low detection probabilities. Although detection of eDNA shows promise under many conditions, further development is needed before sampling for eDNA is a viable option for detecting giant gartersnake populations.

EcoHealth, Jan 22, 2017

Increasingly, population- and ecosystem-level health assessments are performed using sophisticate... more Increasingly, population- and ecosystem-level health assessments are performed using sophisticated molecular tools. Advances in molecular technology enable the identification of synergistic effects of multiple stressors on the individual physiology of different species. Brown bears (Ursus arctos) are an apex predator; thus, they are ideal candidates for detecting potentially ecosystem-level systemic perturbations using molecular-based tools. We used gene transcription to analyze 130 brown bear samples from three National Parks and Preserves in Alaska. Although the populations we studied are apparently stable in abundance and exist within protected and intact environments, differences in transcript profiles were noted. The most prevalent differences were among locations. The transcript patterns among groups reflect the influence of environmental factors, such as nutritional status, disease, and xenobiotic exposure. However, these profiles also likely represent baselines for each uniq...

Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases, 2018

We examined the associations between California sea lion MHC class II DRB (Zaca-DRB) configuratio... more We examined the associations between California sea lion MHC class II DRB (Zaca-DRB) configuration and diversity, and leptospirosis. As Zaca-DRB gene sequences are involved with antigen presentation of bacteria and other extracellular pathogens, we predicted that they would play a role in determining responses to these pathogenic spirochaetes. Specifically, we investigated whether Zaca-DRB diversity (number of genes) and configuration (presence of specific genes) explained differences in disease severity, and whether higher levels of Zaca-DRB diversity predicted the number of specific Leptospira interrogans serovars that a sea lion's serum would react against. We found that serum from diseased sea lions with more Zaca-DRB loci reacted against a wider array of serovars. Specific Zaca-DRB loci were linked to reactions with particular serovars. Interestingly, sea lions with clinical manifestation of leptospirosis that had higher numbers of Zaca-DRB loci were less likely to recover ...

Conservation Physiology

The analysis of blood constituents is a widely used tool to aid in monitoring of animal health an... more The analysis of blood constituents is a widely used tool to aid in monitoring of animal health and disease. However, classic blood diagnostics (i.e. hematologic and plasma biochemical values) often do not provide sufficient information to determine the state of an animal's health. Field studies on wild tortoises and other reptiles have had limited success in drawing significant inferences between blood diagnostics and physiological and immunological condition. However, recent research using gene transcription profiling in the threatened Mojave desert tortoise (Gopherus agassizii) has proved useful in identifying immune or physiologic responses and overall health. To improve our understanding of health and immune function in tortoises, we evaluated both standard blood diagnostic (body condition, hematologic, plasma biochemistry values, trace elements, plasma proteins, vitamin A levels) and gene transcription profiles in 21 adult tortoises (11 clinically abnormal; 10 clinically normal) from Clark County, NV, USA. Necropsy and histology evaluations from clinically abnormal tortoises revealed multiple physiological complications, with moderate to severe rhinitis or pneumonia being the primary cause of morbidity in all but one of the examined animals. Clinically abnormal tortoises had increased transcription for four genes (SOD, MyD88, CL and Lep), increased lymphocyte production, biochemical enzymes and organics, trace elements of copper, and decreased numbers of leukocytes. We found significant positive correlations between increased transcription for SOD and increased trace elements for copper, as well as genes MyD88 and Lep with increased inflammation and microbial insults. Improved methods for health assessments are an important element of monitoring tortoise population recovery and can support the development of more robust diagnostic measures for ill animals, or individuals directly impacted by disturbance.

Environmental Toxicology and Chemistry, 2016

The authors quantified hepatic hydrocarbon-inducible cytochrome P4501A (CYP1A) expression, as eth... more The authors quantified hepatic hydrocarbon-inducible cytochrome P4501A (CYP1A) expression, as ethoxyresorufin-O-deethylase (EROD) activity, in wintering harlequin ducks (Histrionicus histrionicus) captured in Prince William Sound, Alaska (USA), during 2011, 2013, and 2014 (22-25 yr following the 1989 Exxon Valdez oil spill). Average EROD activity was compared between birds from areas oiled by the spill and those from nearby unoiled areas. The present study replicated studies conducted from 1998 to 2009 demonstrating that harlequin ducks using areas oiled in 1989 had elevated EROD activity, indicative of oil exposure, up to 2 decades post spill. In the present study, it was found that average EROD activity during March 2011 was significantly higher in wintering harlequin ducks captured in oiled areas relative to unoiled areas, which the authors interpret to indicate that harlequin ducks continued to be exposed to residual Exxon Valdez oil up to 22 yr after the original spill. However, the 2011 results also indicated reductions in exposure relative to previous years. Average EROD activity in birds from oiled areas was approximately 2 times that in birds from unoiled areas in 2011, compared with observations from 2005 to 2009, in which EROD activity was 3 to 5 times higher in oiled areas. It was also found that average EROD activity during March 2013 and March 2014 was not elevated in wintering harlequin ducks from oiled areas. The authors interpret these findings to indicate that exposure of harlequin ducks to residual Exxon Valdez oil abated within 24 yr after the original spill. The present study finalizes a timeline of exposure, extending over 2 decades, for a bird species thought to be particularly vulnerable to oil contamination in marine environments. Environ Toxicol Chem 2016;9999:1-7. Published 2016 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.

Journal of Marine Science and Engineering, 2016

Using a panel of genes stimulated by oil exposure in a laboratory study, we evaluated gene transc... more Using a panel of genes stimulated by oil exposure in a laboratory study, we evaluated gene transcription in blood leukocytes sampled from sea otters captured from 2006-2012 in western Prince William Sound (WPWS), Alaska, 17-23 years after the 1989 Exxon Valdez oil spill (EVOS). We compared WPWS sea otters to reference populations (not affected by the EVOS) from the Alaska Peninsula (2009), Katmai National Park and Preserve (2009), Clam Lagoon at Adak Island (2012), Kodiak Island (2005) and captive sea otters in aquaria. Statistically, sea otter gene transcript profiles separated into three distinct clusters: Cluster 1, Kodiak and WPWS 2006-2008 (higher relative transcription); Cluster 2, Clam Lagoon and WPWS 2010-2012 (lower relative transcription); and Cluster 3, Alaska Peninsula, Katmai and captive sea otters (intermediate relative transcription). The lower transcription of the aryl hydrocarbon receptor (AHR), an established biomarker for hydrocarbon exposure, in WPWS 2010-2012 compared to earlier samples from WPWS is consistent with declining hydrocarbon exposure, but the pattern of overall low levels of transcription seen in WPWS 2010-2012 could be related to other factors, such as food limitation, pathogens or injury, and may indicate an inability to mount effective responses to stressors. Decreased transcriptional response across the entire gene panel precludes the evaluation of whether or not individual sea otters show signs of exposure to lingering oil. However, related studies on sea otter demographics indicate that by 2012, the sea otter population in WPWS had recovered, which indicates diminishing oil exposure.

Veterinary Immunology and Immunopathology, 2016

Early identification of illness and/or presence of environmental and/or social stressors in free-... more Early identification of illness and/or presence of environmental and/or social stressors in free-ranging and domestic cetaceans is a priority for marine mammal health care professionals. Incorporation of leukocyte gene transcript analysis into the diagnostic tool kit has the potential to augment classical diagnostics based upon ease of sample storage and shipment, inducible nature and well-defined roles of transcription and associated downstream actions. Development of biomarkers that could serve to identify "insults" and potentially differentiate disease etiology would be of great diagnostic value. To this end, a modest number of peripheral blood leukocyte gene transcripts were selected for application to a domestic killer whale population with a focus on broad representation of inducible immunologically relevant genes. Normalized leukocyte transcript values, longitudinally acquired from 232 blood samples derived from 26 clinically healthy whales, were not visibly influenced temporally nor by sex or the specific Park in which they resided. Stability in leukocyte transcript number during periods of health enhances their potential use in diagnostics through identification of outliers. Transcript levels of two cytokine genes, IL-4 and IL-17, were highly variable within the group as compared to the other transcripts. IL-4 transcripts were typically absent. Analysis of transcript levels on the other genes of interest, on an individual animal basis, identified more outliers than were visible when analyzed in the context of the entire population. The majority of outliers (9 samples) were low, though elevated transcripts were identified for IL-17 from 2 animals and one each for Cox-2 and IL-10. The low number of outliers was not unexpected as sample selection was intentionally directed towards animals that were clinically healthy at the time of collection. Outliers may reflect animals experiencing subclinical disease that is transient and self-limiting. The immunologic knowledge derived from longitudinal immunologic studies in killer whales, as was the target of the present study, has the potential to improve diagnostics and health related decision making for this and other domestic and free-ranging cetacean species.