Short-term Detection of Imidacloprid in Streams after Applications in Forests.

Imidacloprid, a neonicotinoid insecticide, is a major component of hemlock woolly adelgid (HWA) [ (Annand)] management programs that are critical to protecting forest health in the eastern United States. However, the impact of imidacloprid soil applications in forests on some aquatic macroinvertebrate species by leaching into aquatic systems is uncertain. The time for residues from imidacloprid soil applications to migrate from treated hemlocks to nearby streams and the concentrations at which imidacloprid may occur after initial migration is unknown. The presence and concentration of imidacloprid in three streams adjacent to recently treated hemlock (soil drench >10 m from stream channels) were assessed in the Big South Fork National River and Recreation Area. Two standard water grabs were collected monthly for 1 yr from a location downstream from imidacloprid-treated areas. Samples were analyzed using liquid chromatography tandem mass spectrometry (limit of detection = 0.025 µg L). Imidacloprid was detected in all treatment streams during a single rain event that occurred 184 to 196 d after treatments, and concentrations ranged from 0.053 to 0.833 µg L. Imidacloprid was not detected on any other sampling date from treatment streams. All observed positive detections exceeded the USEPA freshwater invertebrate chronic endpoint (0.01 µg L). One stream sample exceeded the USEPA freshwater invertebrate acute endpoint (0.39 µg L). However, previous macroinvertebrate community assessments in streams with similar concentrations did not indicate negative effects to aquatic fauna. These findings help characterize the risk of imidacloprid treatments to stream macroinvertebrates within 1 yr of soil applications.

Spatial and temporal distribution of residues of imidacloprid and its insecticidal 5-hydroxy and olefin and metabolites in eastern hemlock (Pinales: Pinaceae) in the southern Appalachians.

Widespread mortality of eastern hemlock, Tsuga canadensis (L.) Carrière, resulting from infestation by hemlock woolly adelgid, Adelges tsugae Annand (Hemiptera: Adelgidae), has occurred throughout the native range of eastern hemlock within the eastern United States. Imidacloprid, a systemic insecticide, is one of the primary chemical compounds used to control hemlock woolly adelgid in both urban and, in a limited manner, in natural forest environments. The metabolism of imidacloprid in eastern hemlock produces 12 metabolites; two of these, imidacloprid 5-hydroxy and imidacloprid olefin, are considered toxicologically important metabolites. However, little is known about the persistence of these metabolites in eastern hemlock in the southern Appalachians. Concentrations ofimidacloprid, olefin, and 5-hydroxy were quantified by using HPLC/MS/MS techniques. Over the 3-yr study, concentrations of imidacloprid and consequent 5-hydroxy and olefin were highest in trees treated with a soil injection in the spring. Imidacloprid and 5-hydroxy concentrations in sap were highest at 12 mo posttreatment and in tissue at 15 mo posttreatment. Imidacloprid was detected through 36 mo posttreatment and 5-hydroxy was detected through 15 mo posttreatment. Olefin concentrations in both sap and tissue were highest at 36 mo posttreatment and were detected in high concentrations through 36 mo posttreatment. Concentrations of imidacloprid were highest in the bottom stratum of the canopy and lowest in the top stratum. Concentrations of olefin and 5-hydroxy were highest in the top stratum and lowest in the bottom stratum.

Spatial and temporal distribution of imidacloprid in eastern hemlock in the southern Appalachians.

The hemlock woolly adelgid, Adelges tsugae Annand (Hemiptera: Adelgidae), is an exotic insect species dramatically reducing populations of eastern hemlock, Tsuga canadensis (L.) Carrieré, throughout the eastern United States. Systemic imidacloprid is one of the primary chemicals used to successfully control infestations of the hemlock woolly adelgid. The concentration levels for this systemic insecticide present in the sap of eastern hemlocks were evaluated from three strata within the canopy over a two year time span. Enzyme-linked immunosorbent assays were conducted every three months posttreatment to assess imidacloprid concentration within the sap. The effect of application timing (fall versus spring) and application method (soil drench, soil injection, and tree injection) on the translocation of imidacloprid throughout the canopy, and the quantity of imidacloprid translocated in the sap of eastern hemlock branches and the terminal twig and needle tissue was determined. Concentration levels progressively declined from the bottom strata to the top strata of the canopy. This trend was consistent in all chemically treated trees. Tree injections provided the lowest concentration and the most nonuniform distribution of imidacloprid throughout the canopy. The highest insecticide concentrations within the tree across all strata over the two year period were consistently associated with the soil drench method followed by the soil injection method. Imidacloprid concentrations peaked between month 9 and 12 posttreatment, and then declined; however, at two years posttreatment, soil drench and soil injected trees contained concentrations reported as being effective for control of the hemlock woolly adelgid.

Community response of insects associated with eastern hemlock to imidacloprid and horticultural oil treatments.

The hemlock woolly adelgid, Adelges tsugae Annand, is an invasive species reducing the populations of eastern hemlock, Tsuga canadensis L. Carrière, throughout the eastern United States. Systemic imidacloprid and horticultural oil are the primary chemicals used to control infestations of this invasive pest; however, the impact of these two chemicals on nontarget canopy insects is unknown. This study was initiated in November 2005 to assess the effects of (1) imidacloprid soil drench, (2) imidacloprid soil injection, (3) imidacloprid tree injections, and (4) horticultural oil applications on multiple levels of organization (composition, overall specimen abundance and species richness, guild specimen abundance and species richness, and individual species) within the phytophagous and transient canopy insect community. Community composition differed significantly among treatments based on analysis of similarity. Mean species richness and specimen abundance were significantly reduced by one or more treatments. Soil drench applications significantly reduced species richness for the detritivore and phytophaga guilds. Furthermore, specimen abundance for species in the detritivore, fungivore, phytophaga, scavenger, and transient phytophaga guilds was significantly lower in the soil drench treatment. This trend was consistent in all insect guilds examined, with the exception of the hematophaga guild that was not significantly lower than for species on the control trees. Of the 293 species documented to be associated with eastern hemlocks, 33 species were found to be directly effected by one or more of the chemical treatments.

Assessment of Alternative Candidate Subcortical Insect Vectors From Walnut Crowns in Habitats Quarantined for Thousand Cankers Disease.

Thousand cankers disease (TCD) results from the combined activity of the fungal pathogen, Geosmithia morbida Kolarík, Freeland, Utley, and Tisserat and its principle vector, Pityophthorus juglandis (Blackman) (Coleoptera: Curculionidae: Scolytinae) in Juglans L. spp. and Pterocarya Kunth spp. host plants. TCD has been reported from the eastern and western United States. To evaluate potential for other beetle species to vector the fungus in east Tennessee, specimens were collected using ethanol-baited traps that were suspended beneath crowns of TCD-symptomatic trees. Associations of G. morbida with insect species collected in traps were assessed in an unsuccessful, preliminary culture-based fungal assay, and then with a molecular-based detection method. For culture-based assays, rinsate from washed, individual insects was plated on nutrient media and growing colonies were subcultured to obtain axenic G. morbida cultures for identification. For the molecular-based method, G. morbida presence was detected by amplifying the previously developed, species-specific microsatellite locus GS004. Capillary electrophoresis was used to detect the amplified amplicons and representative reactions were validated using Sanger sequencing. Eleven beetle species were found to carry G. morbida, including Cnestus mutilatus (Blandford), Dryoxylon onoharaensum (Murayama), Hylocurus rudis (LeConte), Monarthrum fasciatum (Say), Monarthrum mali (Fitch), Xyleborinus saxesenii (Ratzeburg), Xylosandrus crassiusculus (Motschulsky), Xylosandrus germanus (Blandford) (all Coleoptera: Curculionidae: Scolytinae), Stenomimus pallidus (Boheman) (Coleoptera: Curculionidae: Cossoninae), Oxoplatypus quadridentatus (Olivier) (Coleoptera: Curculionidae: Platypodinae), and Xylops basilaris (Say) (Coleoptera: Bostrichidae). These findings raise concerns that alternative subcortical insect species that already occur within quarantined habitats can sustain incidence of introduced G. morbida and contribute to spread within the native range of black walnut, Juglans nigra L., in the eastern United States.

A novel molecular toolkit for rapid detection of the pathogen and primary vector of thousand cankers disease.

Thousand Cankers Disease (TCD) of Juglans and Pterocarya (Juglandaceae) involves a fungal pathogen, Geosmithia morbida, and a primary insect vector, Pityophthorus juglandis. TCD was described originally from dying Juglans nigra trees in the western United States (USA), but it was reported subsequently from the eastern USA and northern Italy. The disease is often difficult to diagnose due to the absence of symptoms or signs on the bark surface of the host. Furthermore, disease symptoms can be confused with those caused by other biotic and abiotic agents. Thus, there is a critical need for a method for rapid detection of the pathogen and vector of TCD. Using species-specific microsatellite DNA markers, we developed a molecular protocol for the detection of G. morbida and P. juglandis. To demonstrate the utility of the method for delineating TCD quarantine zones, we tested whether geographical occurrence of symptoms and signs of TCD was correlated with molecular evidence for the presence of the cryptic TCD organisms. A total of 1600 drill cores were taken from branch sections collected from three regions (n = 40 trees for each location): California-J. hindsii (heavy disease incidence); Tennessee-J. nigra (mild disease incidence); and outside the known TCD zone (Missouri-J. nigra, no record of the disease). California samples had the highest incidence of the TCD organisms (85%, 34/40). Tennessee had intermediate incidence (42.5%, 17/40), whereas neither organism was detected in samples from Missouri. The low cost molecular protocol developed here has a high degree of sensitivity and specificity, and it significantly reduces sample-processing time, making the protocol a powerful tool for rapid detection of TCD.

Insect guild structure associated with eastern hemlock in the southern Appalachians.

An assessment of the insect guild structure associated with immature and mature eastern hemlock, Tsuga canadensis (L.) Carrière, at high and low elevations was made before the invasion by the hemlock woolly adelgid, Adelges tsugae (Annand), and elongate hemlock scale, Fiorinia externa Ferris. Guild dynamics were determined among 243 insect species associated with eastern hemlock in the southern Appalachians. Detritivore, hematophage, herbivore, fungivore, parasitoid, predator, scavenger, and transient guild assignments were made. The herbivore and transient guilds were subdivided into chewers, sapsuckers, flower feeders, pollen feeders, pollen/nectar/sap feeders, and seed feeders. A significant association was found between guilds on immature and mature hemlocks at low elevations, but no significant association was found at high elevations. There was also a strong association of phytophagous insects on immature hemlocks and scavengers on mature hemlocks. The observed transient, scavenger, and predator guilds had a significantly lower co-occurrence among species indicating segregation of the species within these guilds. The herbivore guild had a significantly higher co-occurrence among species than the simulated index indicating aggregation of species within this guild across sites. Detritivore, hematophage, and parasitoid guilds did not differ significantly from the simulated community. These results document a diverse and dynamic insect community on eastern hemlock before invasion by hemlock woolly adelgid and elongate hemlock scale.

Emergence, Seasonality, and Hybridization of Laricobius nigrinus (Coleoptera: Derodontidae), an Introduced Predator of Hemlock Woolly Adelgid (Hemiptera: Adelgidae), in the Tennessee Appalachians.

From 2010 through 2013, adult emergence and seasonality of Laricobius nigrinus Fender, an introduced predatory species native to western North America, as well as hybridization with the native species Laricobius rubidus (LeConte), were evaluated using emergence traps and beat-sheet sampling in areas of previous release against hemlock woolly adelgid, Adelges tsugae Annand. The shortest emergence period of adult L. nigrinus was 7 wk beginning 22 October 2010, and the longest emergence was 15 wk beginning 17 October 2012. Native L. rubidus also were collected from emergence traps placed on the ground surface and beat-sheet samples all 3 yr, with emergence of L. rubidus initiating later than L. nigrinus each season. Seasonality of both Laricobius species was similar across a 44-mo study period. Adult L. nigrinus were present from October through April, and larvae of Laricobius spp. were collected from February to May. The average number of L. nigrinus from emergence traps was significantly greater than the average number of beetles collected from beat-sheet samples in 2010, while the converse was observed during 2012. Hybridization between L. nigrinus and L. rubidus was documented from 10.75% of specimens collected during 2010 and 2011, indicating periodic interbreeding between the introduced and native species. These findings suggest emergence trapping may be a useful method to assess establishment, population densities, and seasonality of Laricobius species in areas of release to enhance their use in management of A. tsuage.