Hemlock Woolly Adelgid and Elongate Hemlock Scale Induce Changes in Foliar and Twig Volatiles of Eastern Hemlock

Eastern hemlock (Tsuga canadensis) is in rapid decline because of infestation by the invasive hemlock woolly adelgid (Adelges tsugae; \u27HWA\u27) and, to a lesser extent, the invasive elongate hemlock scale (Fiorinia externa; \u27EHS\u27). For many conifers, induced oleoresin-based defenses play a central role in their response to herbivorous insects; however, it is unknown whether eastern hemlock mobilizes these inducible defenses. We conducted a study to determine if feeding by HWA or EHS induced changes in the volatile resin compounds of eastern hemlock. Young trees were experimentally infested for 3 years with HWA, EHS, or neither insect. Twig and needle resin volatiles were identified and quantified by gas chromatography/mass spectrometry. We observed a suite of changes in eastern hemlock\u27s volatile profile markedly different from the largely terpenoid-based defense response of similar conifers. Overall, both insects produced a similar effect: most twig volatiles decreased slightly, while most needle volatiles increased slightly. Only HWA feeding led to elevated levels of methyl salicylate, a signal for systemic acquired resistance in many plants, and benzyl alcohol, a strong antimicrobial and aphid deterrent. Green leaf volatiles, often induced in wounded plants, were increased by both insects, but more strongly by EHS. The array of phytochemical changes we observed may reflect manipulation of the tree\u27s biochemistry by HWA, or simply the absence of functional defenses against piercing-sucking insects due to the lack of evolutionary contact with these species. Our findings verify that HWA and EHS both induce changes in eastern hemlock\u27s resin chemistry, and represent the first important step toward understanding the effects of inducible chemical defenses on hemlock susceptibility to these exotic pests

A suite of methods for representing activity space in a healthcare accessibility study

BACKGROUND: "Activity space" has been used to examine how people's habitual movements interact with their environment, and can be used to examine accessibility to healthcare opportunities. Traditionally, the standard deviational ellipse (SDE), a Euclidean measure, has been used to represent activity space. We describe the construction and application of the SDE at one and two standard deviations, and three additional network-based measures of activity space using common tools in GIS: the road network buffer (RNB), the 30-minute standard travel time polygon (STT), and the relative travel time polygon (RTT). We compare the theoretical and methodological assumptions of each measure, and evaluate the measures by examining access to primary care services, using data from western North Carolina. RESULTS: Individual accessibility is defined as the availability of healthcare opportunities within that individual's activity space. Access is influenced by the shape and area of an individual's activity space, the spatial distribution of opportunities, and by the spatial structures that constrain and direct movement through space; the shape and area of the activity space is partly a product of how it is conceptualized and measured. Network-derived measures improve upon the SDE by incorporating the spatial structures (roads) that channel movement. The area of the STT is primarily influenced by the location of a respondent's residence within the road network hierarchy, with residents living near primary roads having the largest activity spaces. The RNB was most descriptive of actual opportunities and can be used to examine bypassing. The area of the RTT had the strongest correlation with a healthcare destination being located inside the activity space. CONCLUSION: The availability of geospatial technologies and data create multiple options for representing and operationalizing the construct of activity space. Each approach has its strengths and limitations, and presents a different view of accessibility. While the choice of method ultimately lies in the research question, interpretation of results must consider the interrelated issues of method, representation, and application. Triangulation aids this interpretation and provides a more complete and nuanced understanding of accessibility

TERPENE CHEMISTRY OF EASTERN HEMLOCKS RESISTANT TO HEMLOCK WOOLLY ADELGID

Recent studies have identified a small number of individual eastern hemlock trees that demonstrate relative resistance to the introduced sap-feeding insect, the hemlock woolly adelgid. Using gas chromatography, we compared concentrations of twenty-two terpenoids in susceptible and relatively resistant trees, both in the forest and in propagated cuttings in a common-garden setting. Terpenoid concentrations were higher in twig tissue of resistant versus susceptible trees, across six sampling dates and at both sites. These changes may be particularly important because the hemlock woolly adelgid feeds on twig tissue. Because the common-garden cuttings were free of herbivores, the higher terpenoid concentrations are interpreted as a constitutive defense. Increased levels of monoterpenes and sesquiterpenes imply an overall increase in the input of carbon precursors to both terpenoid synthesis pathways

Conifer Responses to a Stylet-Feeding Invasive Herbivore and Induction with Methyl Jasmonate: Impact on the Expression of Induced Defenses and a Native Folivore

Trees attacked by multiple herbivores need to defend themselves against dynamic biotic challenges; appropriate responses to one stressor can elicit hormonal responses that are antagonistic to another. Hemlock (Tsuga canadensis) infestation by hemlock woolly adelgid (HWA; Adelges tsugae) results in the accumulation of the defensive hormone salicylic acid. We explored the potential for HWA infestation to interfere with anti‐folivore‐induced defence signalling and its implications for a native folivore (hemlock looper; Lambdina fiscellaria). Hemlocks were infested with HWA and/or sprayed with methyl jasmonate; foliar defences were analyzed and foliage quality for looper larvae was assessed. Both treatments activated foliar defensive traits, including a HWA‐mediated increase in peroxidase activity and an accumulation of cell wall‐bound phenolics and lignin, as well as a methyl jasmonate‐mediated increase in lipoxygenase activity. The two treatments had an additive effect on other defensive traits and both treatments negatively affected looper performance. These results suggest that salicylic acid and jasmonic acid are not strictly antagonistic in conifers and that both have a role in anti‐folivore defence signalling. The present study illustrates the need for a better understanding of hormone signalling, cross‐talk and induced responses in conifers

A new way to estimate disease prevalence from random partial-mouth samples

Standard partial-mouth estimators of chronic periodontitis that define an individual’s disease status solely in terms of selected sites underestimate prevalence. This study proposes an improved prevalence estimator based on randomly sampled sites and evaluates its accuracy in a well characterized population cohort

Failure Under Stress: The Effect of the Exotic Herbivore \u3cem\u3eAdelges tsugae\u3c/em\u3e on Biomechanics of \u3cem\u3eTsuga canadensis\u3c/em\u3e

Background and Aims Exotic herbivores that lack a coevolutionary history with their host plants can benefit from poorly adapted host defences, potentially leading to rapid population growth of the herbivore and severe damage to its plant hosts. The hemlock woolly adelgid (Adelges tsugae) is an exotic hemipteran that feeds on the long-lived conifer eastern hemlock (Tsuga canadensis), causing rapid mortality of infested trees. While the mechanism of this mortality is unknown, evidence indicates that A. tsugae feeding causes a hypersensitive response and alters wood anatomy. This study investigated the effect of A. tsugae feeding on biomechanical properties at different spatial scales: needles, twigs and branches. Methods Uninfested and A. tsugae-infested samples were collected from a common garden experiment as well as from naturally infested urban and rural field sites. Tension and flexure mechanical tests were used to quantify biomechanical properties of the different tissues. In tissues that showed a significant effect of herbivory, the potential contributions of lignin and tissue density on the results were quantified. Key Results Adelges tsugae infestation decreased the abscission strength, but not flexibility, of needles. A. tsugae feeding also decreased mechanical strength and flexibility in currently attacked twigs, but this effect disappeared in older, previously attacked branches. Lignin and twig tissue density contributed to differences in mechanical strength but were not affected by insect treatment. Conclusions Decreased strength and flexibility in twigs, along with decreased needle strength, suggest that infested trees experience resource stress. Altered growth patterns and cell wall chemistry probably contribute to these mechanical effects. Consistent site effects emphasize the role of environmental variation in mechanical traits. The mechanical changes measured here may increase susceptibility to abiotic physical stressors in hemlocks colonized by A. tsugae. Thus, the interaction between herbivore and physical stresses is probably accelerating the decline of eastern hemlock, as HWA continues to expand its range

Seasonal variation in effects of herbivory on foliar nitrogen of a threatened conifer

Invasive herbivores can dramatically impact the nitrogen (N) economy of native hosts. In deciduous species, most N is stored in stem tissues, while in evergreen conifer species N is stored in needles, making them potentially more vulnerable to herbivory. In eastern forests of the USA, the long-lived, foundational conifer eastern hemlock (Tsuga canadensis) is under the threat of extirpation by the invasive hemlock woolly adelgid (HWA: Adelges tsugae). We assessed the impact of HWA infestation on the patterns of seasonal foliar N availability in hemlock planted in a deciduous forest understory. Over the course of a year, we sampled needles and twigs and measured N, carbon (C), C:N ratio, and total protein concentrations. Tissue sampling events were timed to coincide with key life-history transitions for HWA to determine the association between HWA development and feeding with these foliar nutrients. In uninfested trees, needle and twig N concentrations fluctuated across seasons, indicating the potential importance of N storage and remobilization for the N economy of eastern hemlock. Although N levels in HWA-infested trees also cycled annually, the degree to which N concentrations fluctuated seasonally in tissues was significantly affected by HWA feeding. These fluctuations exceeded N levels observed in control trees and coincided with HWA feeding. HWA feeding generally increased N concentrations but did not affect protein levels, suggesting that changes in N do not occur via adelgid-induced protein breakdown. Herbivore-induced mobilization of N to feeding sites and its rapid depletion may be a significant contributor to eastern hemlock mortality in US forests

Facilitation between invasive herbivores: hemlock woolly adelgid increases gypsy moth preference for and performance on eastern hemlock

Interactions between invertebrate herbivores with different feeding modes are common on long‐lived woody plants. In cases where one herbivore facilitates the success of another, the consequences for their shared host plant may be severe. Eastern hemlock (Tsuga canadensis), a canopy‐dominant conifer native to the eastern U.S., is currently threatened with extirpation by the invasive stylet‐feeding hemlock woolly adelgid (Adelges tsugae). The effect of adelgid on invasive hemlock‐feeding folivores remains unknown. This study evaluated the impact of feeding by hemlock woolly adelgid on gypsy moth (Lymantria dispar) larval preference for, and performance on, eastern hemlock. To assess preference, 245 field‐grown hemlocks were surveyed for gypsy moth herbivory damage and laboratory paired‐choice bioassays were conducted. To assess performance, gypsy moth larvae were reared to pupation on adelgid‐infested or uninfested hemlock foliage, and pupal weight, proportional weight gain, and larval period were analysed. Adelgid‐infested hemlocks experienced more gypsy moth herbivory than did uninfested control trees, and laboratory tests confirmed that gypsy moth larvae preferentially feed on adelgid‐infested hemlock foliage. Gypsy moth larvae reared to pupation on adelgid‐infested foliage gained more weight than larvae reared on uninfested control foliage. These results suggest that the synergistic effect of adelgid and gypsy moth poses an additional threat to eastern hemlock that may increase extirpation risk and ecological impact throughout most of its range

A Potential Biofilm Metabolite Signature for Caries Activity - A Pilot Clinical Study

BACKGROUND: This study's aim was to compare the dental biofilm metabolite-profile of caries-active (N=11) or caries-free (N=4) children by gas chromatography-mass spectrometry (GC/MS) analyses. METHODS: Samples collected after overnight fasting, with or without a previous glucose rinse, were combined for each child based on the caries status of the site, re-suspended in ethanol and analyzed by GC/MS. RESULTS: Biofilm from caries-active sites exhibited a different chromatographic profile compared to caries-free sites. Qualitative and quantitative analysis suggested a special cluster of branched alcohols and esters present at substantially higher intensity in biofilms of caries-active sites. CONCLUSIONS: This pilot study indicates that there are metabolites present in the biofilm which have the potential to provide a characteristic metabolomics signature for caries activity

Review and Recommendations for Zero-Inflated Count Regression Modeling of Dental Caries Indices in Epidemiological Studies

Over the past five to ten years, zero-inflated count regression models have been increasingly applied to the analysis of dental caries indices (e.g., DMFT, dfms, etc). The main reason for that is linked to the broad decline in children’s caries experience, such that dmf and DMF indices more frequently generate low or even zero counts. This article specifically reviews the application of zero-inflated Poisson and zero-inflated negative binomial regression models to dental caries, with emphasis on the description of the models and the interpretation of fitted model results given the study goals. The review finds that interpretations provided in the published caries research are often imprecise or inadvertently misleading, particularly with respect to failing to discriminate between inference for the class of susceptible persons defined by such models and inference for the sampled population in terms of overall exposure effects. Recommendations are provided to enhance the use as well as the interpretation and reporting of results of count regression models when applied to epidemiological studies of dental caries