Seasonal Assessment of Supercooling Points for Two Introduced and One Native Laricobius spp. (Coleoptera: Derodontidae), Predators of Adelgidae.

The hemlock woolly adelgid, Adelges tsugae Annand, is an invasive insect that threatens hemlock species in eastern North America. Several species from the genus Laricobius are predators of A. tsugae in its native areas of Asia and the western United States. Two Laricobius species have been released as biological control agents: Laricobius nigrinus Fender, and Laricobius osakensis Montgomery and Shiyake. Laricobius rubidus LeConte is an adelgid predator native to the Eastern United States, where it can feed and complete development on A. tsugae opportunistically. Laboratory assays were conducted to assess the cold hardiness of these three Laricobius species, including two distinct populations of L. osakensis, by measuring the supercooling points of each species from November 2016 through March 2017. This information may be useful for choosing the best-suited biological control agent for a particular region to control A. tsugae. There was a significant difference between the overall mean supercooling point of L. rubidus compared to the other Laricobius spp. There were also significant differences of supercooling points between L. rubidus and both strains of L. osakensis in January, and significant differences between L. rubidus and all other strains in February. L. rubidus appear better adapted to cold extremes in the eastern U.S. than imported Laricobius spp.

Cold Tolerance of Halyomorpha halys (Hemiptera: Pentatomidae) Across Geographic and Temporal Scales.

The brown marmorated stink bug, Halyomorpha halys (Stål), is native to eastern Asia and is presently invading North America. Little is known about the exposure to and effects of winter temperatures in newly invaded regions on H. halys The overwintering habitats that this species utilizes vary greatly in their thermal buffering capacity. They naturally overwinter in aggregations beneath loose bark on trees and in cliff outcroppings, but will also commonly aggregate in buildings. Effects of cold temperatures such as mortality and freezing have yet to be quantified in the invading population. We report that H. halys is chill intolerant (i.e., dies before reaching its freezing point), and that the degree of cold tolerance of populations in North America differs by season, sex, and acclimation location. The mean winter supercooling point (± SEM) of individuals acclimated in Minnesota was -17.06 °C ± 0.13 and in Virginia was -13.90 °C ± 0.09. By using laboratory assays of lower lethal temperatures and ambient air temperature records, we accurately forecasted mortality for field experiments in Minnesota and Virginia. Temperature refugia provided by human-built structures are likely crucial for overwintering survival during atypically cold winters and possibly contribute to the northern geographic range expansion of this economically damaging insect in the temperate climates of North America.

Fitness and physiology of Adelges tsugae (Hemiptera: Adelgidae) in relation to the health of the eastern hemlock.

The hemlock woolly adelgid, Adelges tsugae Annand is an invasive insect that frequently causes hemlock (Tsuga spp.) mortality in the eastern United States. Studies have shown that once healthy hemlocks become infested by the adelgid, nutrients are depleted from the tree, leading to both tree decline and a reduction of the adelgid population. Since A. tsugae is dependent on hemlock for nutrients, feeding on trees in poor health may affect the ability of the insect to obtain necessary nutrients and may consequently affect their physiological and population health. Trees were categorized as lightly or moderately impacted by A. tsugae based on quantitative and qualitative tree health measurements. Population health of A. tsugae on each tree was determined by measuring insect density and peak mean fecundity; A. tsugae physiological health was determined by measuring insect biomass, total carbon, carbohydrate, total nitrogen, and amino nitrogen levels. Adelges tsugae from moderately impacted trees exhibited significantly greater fecundity than from lightly impacted trees. However, A. tsugae from lightly impacted hemlocks contained significantly greater levels of carbohydrates, total nitrogen, and amino nitrogen. While the results of the physiological analysis generally support our hypothesis that A. tsugae on lightly impacted trees are healthier than those on moderately impacted trees, this was not reflected in the population health measurements. Adelges tsugae egg health in response to tree health should be verified. This study provides the first examination of A. tsugae physiological health in relation to standard A. tsugae population health measures on hemlocks of different health levels.

Physiology of environmental adaptations and resource acquisition in cockroaches.

Cockroaches are a group of insects that evolved early in geological time. Because of their antiquity, they for the most part display generalized behavior and physiology and accordingly have frequently been used as model insects to examine physiological and biochemical mechanisms involved with water balance, nutrition, reproduction, genetics, and insecticide resistance. As a result, a considerable amount of information on these topics is available. However, there is much more to be learned by employing new protocols, microchemical analytical techniques, and molecular biology tools to explore many unanswered questions.

Characterization of physical and chemical defenses in the hemlock woolly adelgid.

The invasive hemlock woolly adelgid (Adelges tsugae Annand, Hemiptera: Sternorrhyncha: Adelgidae) causes significant mortality to eastern and Carolina hemlock (Tsuga canadensis Carrière and T. caroliniana Engelmann, respectively) throughout the eastern United States. Adelges tsugae produces vast quantities of a wax covering that surrounds most of the instars as well as the adult and eggs. Using direct probe EI mass spectrometry, this wax covering was characterized as a diketoester wax, 17-oxohexatriacontanyl 11-oxotriacontanoate, and accounted for ≈ 42 % of A. tsugae total biomass. The presence of the anthraquinone, chrysophanol, and its precursor anthrone, chrysarobin, in A. tsugae has only been briefly described. Further study confirmed these compounds in all A. tsugae life stages. Additionally, several predatory beetles in use or under consideration as biological control agents for this species appear to ingest and excrete these compounds when feeding on A. tsugae. The production of both a physical and a chemical defense may represent a significant energy investment by A. tsugae.

Potential feeding deterrents found in hemlock woolly adelgid, Adelges tsugae.

The nonnative hemlock woolly adelgid (Adelges tsugae Annand, Hemiptera: Sternorrhyncha: Adelgidae) has been a significant mortality agent of eastern hemlock (Tsuga canadensis Carriere) throughout a large portion of its geographic range. During a study investigating adelgid vigor in relation to host health, it was noted that adelgid extracts ranged from a yellow to a deep red color. Analysis by GC-MS identified the presence of the anthraquinone, chrysophanol and its anthrone precursor, chrysarobin in the extract. These compounds are predator deterrents in several other insects, including chrysomelid beetles. It is hypothesized that these compounds serve a similar purpose in the hemlock woolly adelgid.

Evaluation of ELISA for imidacloprid detection in eastern hemlock (Tsuga canadensis) wood and needle tissues.

BACKGROUND: Imidacloprid is the primary insecticide used against the exotic invasive insect hemlock woolly adelgid, Adelges tsugae Annand, a pest of eastern hemlock [Tsuga canadensis (L.) Carrière] trees in the eastern United States. A competitive enzyme-linked immunosorbent assay (ELISA) was evaluated for quantification of imidacloprid in eastern hemlock wood and needle tissues. RESULTS: Matrix effects in the form of false positives and overestimated imidacloprid concentrations were observed in both wood and needle extracts. Tissues required a 100-1000-fold dilution with water in order to reduce matrix effects. Standard curves in 1% wood or needle extract were not significantly different from standard curves prepared in water. Matrix effects were more pronounced at concentrations in the lower working range of the kit, with recovery of 5 microg L(-1) imidacloprid more accurate than recovery of 0.2 microg L(-1). CONCLUSION: ELISA remains a valuable tool for semi-quantitative imidacloprid detection within the hemlock system because of its sensitivity, cost and ease of use. However, a 1000-fold dilution of hemlock tissue extract is recommended to ensure accurate imidacloprid determinations.

The fate of 14C-diazinon in compost, compost-amended soil, and uptake by earthworms.

A process for disposing of pesticide rinsates using sorption onto organic matter followed by composting is being evaluated. As a part of this evaluation process, we have studied the bioavailability of composted delta-2-14C-diazinon and its degradation products to earthworms (Eisenia foetida Savigny) in 30 and 60 d compost amended soil. After 60 d of composting there was considerable degradation of diazinon (95%) and a corresponding increase in the primary hydrolysis product, 2-isopropyl-4-methyl-6-hydroxypyrimidine (IMHP) as determined by high performance thin layer chromatography (HPTLC). Approximately 50% of the radioactivity became incorporated into the non-extractable fractions associated with composted organic matter with no measurable amounts of 14CO2 produced during the 60-day composting period. Following addition of the composted materials to soil, diazinon leading to 50% mortality after 14 d of exposure; continued to slowly degrade and become increasingly sorbed/entrapped within the soil-compost matrix. Soil amended with 30-d composted diazinon was toxic to earthworms whereas, no mortality was observed in those earthworms exposed to the 60-d composted diazinon. However, earthworms exposed to 30-d and 60-d composted diazinon were found to have similar levels of radioactivity in their tissues. The majority of the radioactivity in earthworms exposed 60-d composted diazinon was either unextractably bound within the earthworm tissue or was not acetone soluble. Most of the radioactivity that could be extracted with acetone was not separated by the two HPTLC methods we used. This study demonstrates that composting high concentrations of diazinon can greatly reduce toxicity and the amount of diazinon that is bioavailable to a representative soil macroinvertebrate (E. foetida).

The structural basis for water exchange between the female cockroach (Blattella germanica) and her ootheca.

Female German cockroaches usually carry their oothecae until they hatch. The success of embryogenesis may be dependent on the water-balance relationship between females and their developing oothecae. Oothecae detached from females early in embryogenesis often fail to develop, especially in low-humidity environments. Experiments reported here using tritiated water have confirmed the transport of a significant amount of water to the ootheca from the female during embryogenesis; 18% of the tritiated water injected into gravid females was recovered in their oothecae after 24 h. We describe a structural basis of water absorption by the oothecae. An area located on the proximal end of the ootheca (adjacent to the 'escutcheon-shaped vaginal imprint') contains small pores that penetrate the oothecal covering to access regions of the chorion lying beneath these pores. Experiments using microparabiotic chambers to examine transport of tritiated water and water-soluble materials across the escutcheon region of the oothecal covering, along with dark-field birefringent microscopy of the chorion, support the hypothesis that this chorionic network is capable of conducting water throughout the interior of the ootheca. Furthermore, the structural arrangement and intimate association of the female vestibulum with the oothecal pore field contained in the region of the escutcheon-shaped vaginal imprint appear to provide an efficient conduit to the chorion. The overall structural relationship might be a means for maintaining water balance between females and their oothecae during embryonic development. Evidence presented here supports observations that this species represents an important link in the transition from oviparity to ovoviviparity by internalization of cockroach oothecae.