Accounting for missing ticks: Use (or lack thereof) of hierarchical models in tick ecology studies.

Ixodid (hard) ticks play important ecosystem roles and have significant impacts on animal and human health via tick-borne diseases and physiological stress from parasitism. Tick occurrence, abundance, activity, and key life-history traits are highly influenced by host availability, weather, microclimate, and landscape features. As such, changes in the environment can have profound impacts on ticks, their hosts, and the spread of diseases. Researchers recognize that spatial and temporal factors influence activity and abundance and attempt to account for both by conducting replicate sampling bouts spread over the tick questing period. However, common field methods notoriously underestimate abundance, and it is unclear how (or if) tick studies model the confounding effects of factors influencing activity and abundance. This step is critical as unaccounted variance in detection can lead to biased estimates of occurrence and abundance. We performed a descriptive review to evaluate the extent to which studies account for the detection process while modeling tick data. We also categorized the types of analyses that are commonly used to model tick data. We used hierarchical models (HMs) that account for imperfect detection to analyze simulated and empirical tick data, demonstrating that inference is muddled when detection probability is not accounted for in the modeling process. Our review indicates that only 5 of 412 (1 %) papers explicitly accounted for imperfect detection while modeling ticks. By comparing HMs with the most common approaches used for modeling tick data (e.g., ANOVA), we show that population estimates are biased low for simulated and empirical data when using non-HMs, and that confounding occurs due to not explicitly modeling factors that influenced both detection and abundance. Our review and analysis of simulated and empirical data shows that it is important to account for our ability to detect ticks using field methods with imperfect detection. Not doing so leads to biased estimates of occurrence and abundance which could complicate our understanding of parasite-host relationships and the spread of tick-borne diseases. We highlight the resources available for learning HM approaches and applying them to analyzing tick data.

Understanding drivers of mercury in lake trout (Salvelinus namaycush), a top-predator fish in southwest Alaska's parklands.

Mercury (Hg) is a widespread element and persistent pollutant, harmful to fish, wildlife, and humans in its organic, methylated form. The risk of Hg contamination is driven by factors that regulate Hg loading, methylation, bioaccumulation, and biomagnification. In remote locations, with infrequent access and limited data, understanding the relative importance of these factors can pose a challenge. Here, we assessed Hg concentrations in an apex predator fish species, lake trout (Salvelinus namaycush), collected from 14 lakes spanning two National Parks in southwest Alaska, U.S.A. We then examined factors associated with the variation in fish Hg concentrations using a Bayesian hierarchical model. We found that total Hg concentrations in water were consistently low among lakes (0.11-0.50 ng L-1). Conversely, total Hg concentrations in lake trout spanned a thirty-fold range (101-3046 ng g-1 dry weight), with median values at 7 lakes exceeding Alaska's human consumption threshold. Model results showed that fish age and, to a lesser extent, body condition best explained variation in Hg concentration among fish within a lake, with Hg elevated in older, thinner lake trout. Other factors, including plankton methyl Hg content, fish species richness, volcano proximity, and glacier loss, best explained variation in lake trout Hg concentration among lakes. Collectively, these results provide evidence that multiple, hierarchically nested factors control fish Hg levels in these lakes.

Gene Expression Profiles in Two Razor Clam Populations: Discerning Drivers of Population Status.

With rapidly changing marine ecosystems, shifts in abundance and distribution are being documented for a variety of intertidal species. We examined two adjacent populations of Pacific razor clams (Siliqua patula) in lower Cook Inlet, Alaska. One population (east) supported a sport and personal use fishery, but this has been closed since 2015 due to declines in abundance, and the second population (west) continues to support commercial and sport fisheries. We used gene expression to investigate potential causes of the east side decline, comparing razor clam physiological responses between east and west Cook Inlet. The target gene profile used was developed for razor clam populations in Alaska based on physiological responses to environmental stressors. In this study, we identified no differences of gene expression between east and west populations, leading to two potential conclusions: (1) differences in factors capable of influencing physiology exist between the east and west and are sufficient to influence razor clam populations but are not detected by the genes in our panel, or (2) physiological processes do not account for the differences in abundance, and other factors such as predation or changes in habitat may be impacting the east Cook Inlet population.

Using a structured decision analysis to evaluate bald eagle vital signs monitoring in Southwest Alaska National Parks.

Monitoring programs can benefit from an adaptive monitoring approach, where key decisions about why, where, what, and how to monitor are revisited periodically in order to ensure programmatic relevancy.The National Park Service (NPS) monitors status and trends of vital signs to evaluate compliance with the NPS mission. Although abundant, The Southwest Alaska Network (SWAN) monitors bald eagles because of their inherent importance to park visitors and role as an important ecological indicator. Our goal is to identify an optimal monitoring program that may be standardized among participating parks.We gathered an expert panel of scientists and managers, and implemented a Delphi Process to gather information about the bald eagle monitoring program. Panelists generated a list of means objectives for the monitoring program: minimizing cost, minimizing effort, maximizing the ability to detect change in bald eagle populations, and maximizing the amount of accurate information collected about bald eagles.We used a swing-weighting technique to assign importance to each objective. Collecting accurate information about bald eagles was considered the most important means objective.Combining panelist-generated information with objective importance, we analyzed the scenarios and defined the optimal decision using linear value modeling. Through our analysis, we found that a "Comprehensive" monitoring scenario, comprised of all feasible monitoring metrics, is the optimal monitoring scenario. Even with greatly increased cost, the Comprehensive monitoring scenario remains the best solution.We suggest further exploration of the cost and effort required for the Comprehensive scenario, to determine whether it is in the parks' best interest to begin monitoring additional metrics.

Monitoring nearshore ecosystem health using Pacific razor clams (Siliqua patula) as an indicator species.

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 variation in both large-scale and local environmental conditions. Our intent is to employ these methods to evaluate Pacific razor clams as a bioindicator of nearshore ecosystem health. Links between the results of the biomarker and gene transcription assays were observed that support the applicability of both assays in ecosystem monitoring. However, we recognize the need for controlled studies to examine the range of responses in physiology and gene transcripts to different stressors.

Direct and indirect effects of temperature and prey abundance on bald eagle reproductive dynamics.

Understanding the mechanisms by which populations are regulated is critical for predicting the effects of large-scale perturbations. While discrete mortality events provide clear evidence of direct impacts, indirect pathways are more difficult to assess but may play important roles in population and ecosystem dynamics. Here, we use multi-state occupancy models to analyze a long-term dataset on nesting bald eagles in south-central Alaska with the goal of identifying both direct and indirect mechanisms influencing reproductive output in this apex predator. We found that the probabilities of both nest occupancy and success were higher in the portion of the study area where water turbidity was low, supporting the hypothesis that access to aquatic prey is a critical factor limiting the reproductive output of eagles in this system. As expected, nest success was also positively related to salmon abundance; however, the negative effect of spring warmth suggested that access to salmon resources is indirectly diminished in warm springs as a consequence of increased glacial melt. Together, these findings reveal complex interrelationships between a critical prey resource and large-scale weather and climate processes which likely alter the accessibility of resources rather than directly affecting resource abundance. While important for understanding bald eagle reproductive dynamics in this system specifically, our results have broader implications that suggest complex interrelationships among system components.

Physiological and gene transcription assays to assess responses of mussels to environmental changes.

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.

Nest use dynamics of an undisturbed population of bald eagles.

Management or conservation targets based on demographic rates should be evaluated within the context of expected population dynamics of the species of interest. Wild populations can experience stable, cyclical, or complex dynamics, therefore undisturbed populations can provide background needed to evaluate programmatic success. Many raptor species have recovered from large declines caused by environmental contaminants, making them strong candidates for ongoing efforts to understand population dynamics and ecosystem processes in response to human-caused stressors. Dynamic multistate occupancy models are a useful tool for analyzing species dynamics because they leverage the autocorrelation inherent in long-term monitoring datasets to obtain useful information about the dynamic properties of population or reproductive states. We analyzed a 23-year bald eagle monitoring dataset in a dynamic multistate occupancy modeling framework to assess long-term nest occupancy and reproduction in Lake Clark National Park and Preserve, Alaska. We also used a hierarchical generalized linear model to understand changes in nest productivity in relation to environmental factors. Nests were most likely to remain in the same nesting state between years. Most notably, successful nests were likely to remain in use (either occupied or successful) and had a very low probability of transitioning to an unoccupied state in the following year. There was no apparent trend in the proportion of nests used by eagles through time, and the probability that nests transitioned into or out of the successful state was not influenced by temperature or salmon availability. Productivity was constant over the course of the study, although warm April minimum temperatures were associated with increased chick production. Overall our results demonstrate the expected nesting dynamics of a healthy bald eagle population that is largely free of human disturbance and can be used as a baseline for the expected dynamics for recovering bald eagle populations in the contiguous 48 states.

Warming drives a front of white spruce establishment near western treeline, Alaska.

Regional warming has led to increased productivity near the boreal forest margin in Alaska. To date, the effects of warming on seedling recruitment have received little attention, in spite of forecasted forest expansion. Here, we used stand structure and environmental data from 95 white spruce (Picea glauca) plots sampled across a longitudinal gradient in southwest Alaska to explore factors influencing spruce establishment and recruitment near western treeline. We used total counts of live seedlings, saplings, and trees, representing five life stages, to evaluate whether geospatial, climate, and measured plot covariates predicted abundance, using current abundance distributions as a surrogate for climate conditions in the past. We used generalized linear models to test the null hypothesis that conditions favorable for recruitment were similar along the environmental gradient represented by longitude, by exploring relationships between per-plot counts of each life stage and the covariates hypothesized to affect abundance. We also examined the relationship between growing degree days (GDD) and seedling establishment over a period of three decades using tree-ring chronologies obtained from cores taken at a subset of our sites (n = 30). Our results indicated that seedling, sapling, and tree abundance were positively correlated with temperature across the study area. The response to longitude was mixed, with earlier life stages (seedlings, saplings) most abundant at the western end of the gradient, and later life stages (trees) most abundant to the east. The differential relationship between longitude and life-stage abundance suggests a moving front of white spruce establishment through time, driven by changes in environmental conditions near the species' western range limit. Likewise, we found a positive relationship between periods of seedling establishment and GDD, suggesting that longer summers and/or greater heat accumulation might enhance establishment, consistent with the positive relationship we found between life-stage abundance and temperature.

Improving inference for aerial surveys of bears: The importance of assumptions and the cost of unnecessary complexity.

Obtaining useful estimates of wildlife abundance or density requires thoughtful attention to potential sources of bias and precision, and it is widely understood that addressing incomplete detection is critical to appropriate inference. When the underlying assumptions of sampling approaches are violated, both increased bias and reduced precision of the population estimator may result. Bear (Ursus spp.) populations can be difficult to sample and are often monitored using mark-recapture distance sampling (MRDS) methods, although obtaining adequate sample sizes can be cost prohibitive. With the goal of improving inference, we examined the underlying methodological assumptions and estimator efficiency of three datasets collected under an MRDS protocol designed specifically for bears. We analyzed these data using MRDS, conventional distance sampling (CDS), and open-distance sampling approaches to evaluate the apparent bias-precision tradeoff relative to the assumptions inherent under each approach. We also evaluated the incorporation of informative priors on detection parameters within a Bayesian context. We found that the CDS estimator had low apparent bias and was more efficient than the more complex MRDS estimator. When combined with informative priors on the detection process, precision was increased by >50% compared to the MRDS approach with little apparent bias. In addition, open-distance sampling models revealed a serious violation of the assumption that all bears were available to be sampled. Inference is directly related to the underlying assumptions of the survey design and the analytical tools employed. We show that for aerial surveys of bears, avoidance of unnecessary model complexity, use of prior information, and the application of open population models can be used to greatly improve estimator performance and simplify field protocols. Although we focused on distance sampling-based aerial surveys for bears, the general concepts we addressed apply to a variety of wildlife survey contexts.

Intravenously injected human apolipoprotein A-I rapidly enters the central nervous system via the choroid plexus.

BACKGROUND: Brain lipoprotein metabolism is dependent on lipoprotein particles that resemble plasma high-density lipoproteins but that contain apolipoprotein (apo) E rather than apoA-I as their primary protein component. Astrocytes and microglia secrete apoE but not apoA-I; however, apoA-I is detectable in both cerebrospinal fluid and brain tissue lysates. The route by which plasma apoA-I enters the central nervous system is unknown. METHODS AND RESULTS: Steady-state levels of murine apoA-I in cerebrospinal fluid and interstitial fluid are 0.664 and 0.120 µg/mL, respectively, whereas brain tissue apoA-I is ≈10% to 15% of its levels in liver. Recombinant, fluorescently tagged human apoA-I injected intravenously into mice localizes to the choroid plexus within 30 minutes and accumulates in a saturable, dose-dependent manner in the brain. Recombinant, fluorescently tagged human apoA-I accumulates in the brain for 2 hours, after which it is eliminated with a half-life of 10.3 hours. In vitro, human apoA-I is specifically bound, internalized, and transported across confluent monolayers of primary human choroid plexus epithelial cells and brain microvascular endothelial cells. CONCLUSIONS: Following intravenous injection, recombinant human apoA-I rapidly localizes predominantly to the choroid plexus. Because apoA-I mRNA is undetectable in murine brain, our results suggest that plasma apoA-I, which is secreted from the liver and intestine, gains access to the central nervous system primarily by crossing the blood-cerebrospinal fluid barrier via specific cellular mediated transport, although transport across the blood-brain barrier may also contribute to a lesser extent.

Impairment of adenosine A3 receptor activity disrupts neutrophil migratory capacity and impacts innate immune function in vivo.

Adenosine possesses potent anti-inflammatory properties which are partly mediated by G(i) -coupled adenosine A3 receptors (A3Rs). A3R agonists have shown clinical benefit in a number of inflammatory conditions although some studies in A3R-deficient mice suggest a pro-inflammatory role. We hypothesised that, in addition to cell signalling effects, A3R compounds might inhibit neutrophil chemotaxis by disrupting the purinergic feedback loop controlling leukocyte migration. Human neutrophil activation triggered rapid upregulation of surface A3R expression which was disrupted by pre-treatment with either agonist (Cl-IB-MECA) or antagonist (MRS1220). Both compounds reduced migration velocity and neutrophil transmigration capacity without impacting the response to chemokines per se. Similar effects were observed in murine neutrophils, while cells from A3R-deficient mice displayed a constitutively impaired migratory phenotype indicating compound-induced desensitisation and genetic ablation had the same functional outcome. In a dextran sodium sulphate-induced colitis model, A3R-deficient mice exhibited reduced colon pathology and decreased tissue myeloperoxidase levels at day 8 - consistent with reduced neutrophil recruitment. However, A3R-deficient mice were unable to resolve the dextran sodium sulphate-induced inflammation and had elevated numbers of tissue-associated bacteria by day 21. Our data indicate that A3Rs play a role in neutrophil migration and disrupting this function has the potential to adversely affect innate immune responses.

Use of Trial 1 of the Test of Memory Malingering (TOMM) as a screening measure of effort: suggested discontinuation rules.

Trial 1 of the Test of Memory Malingering (TOMM) has been suggested as a screening tool, with several possible cut-off scores proposed. The purpose of the present study was to replicate the utility of previously suggested cut-off scores and to characterize neuropsychological profiles of persons who "pass" the TOMM but obtain Trial 1 scores < 45 and of persons with cognitive disorders. A total of 229 veterans were administered the TOMM as part of a neuropsychological evaluation. Trial 1 scores ≥ 41 and ≤ 25 showed good utility as discontinuation scores for adequate and poor effort, respectively, beyond which administration of additional trials were unnecessary. Findings suggest better Trial 1 performance is significantly related to better speeded mental flexibility and memory.

Additive effect of the combination of glatiramer acetate and minocycline in a model of MS.

There have been significant advances in the treatment of multiple sclerosis (MS) in recent years, but further improvement in therapy is required as not all patients have responded optimally. An approach to enhancing MS treatment is to combine drugs that impact on different aspects of the disease process. We have described that the tetracycline derivative, minocycline, attenuates the severity of experimental autoimmune encephalomyelitis (EAE), a model of MS. Here, we have evaluated the combination of minocycline and glatiramer acetate (GA), a current therapy in MS, on the course of EAE in mice. This combination resulted in a significant reduction of disease severity and disease burden with attenuation of the inflammation, axonal loss and demyelination.

The anchoring protein RACK1 links protein kinase Cepsilon to integrin beta chains. Requirements for adhesion and motility.

Integrin affinity is modulated by intracellular signaling cascades, in a process known as "inside-out" signaling, leading to changes in cell adhesion and motility. Protein kinase C (PKC) plays a critical role in integrin-mediated events; however, the mechanism that links PKC to integrins remains unclear. Here, we report that PKCepsilon positively regulates integrin-dependent adhesion, spreading, and motility of human glioma cells. PKCepsilon activation was associated with increased focal adhesion and lamellipodia formation as well as clustering of select integrins, and it is required for phorbol 12-myristate 13-acetate-induced adhesion and motility. We provide novel evidence that the scaffolding protein RACK1 mediates the interaction between integrin beta chain and activated PKCepsilon. Both depletion of RACK1 by antisense strategy and overexpression of a truncated form of RACK1 which lacks the integrin binding region resulted in decreased PKCepsilon-induced adhesion and migration, suggesting that RACK1 links PKCepsilon to integrin beta chains. Altogether, these results provide a novel mechanistic link between PKC activation and integrin-mediated adhesion and motility.