Influence of Imidacloprid and Horticultural Oil on Spider Abundance on Eastern Hemlock in the Southern Appalachians.

Hemlock woolly adelgid, Adelges tsugae Annand (Hemiptera: Adelgidae), is an exotic pest of eastern hemlock, Tsuga canadensis (L.) Carrière (Pinales: Pinaceae), in the eastern United States. Two commonly used insecticides to manage adelgid are imidacloprid, a systemic neonicotinoid insecticide, and horticultural oil, a refined petroleum oil foliar spray. We have investigated the influence of imidacloprid and horticultural oil on spider abundance at different canopy strata in eastern hemlock. In total, 2,084 spiders representing 11 families were collected from the canopies of eastern hemlock. In beat-sheet and direct observation samples, the families Theridiidae, Araneidae, Salticidae, and Anyphaenidae were the most abundant. Significantly higher numbers of spiders were recorded on untreated control trees compared with trees treated with imidacloprid using soil drench and soil injection applications. Spider abundance in trees injected with imidacloprid and horticultural oil applications did not significantly differ from control trees. Spider abundance was significantly greater in the top and middle strata of the canopy than in the bottom stratum, where imidacloprid concentrations were the highest. Regression analysis showed that spider abundance was inversely associated with imidacloprid concentration. This research demonstrates that imidacloprid, when applied with selected methods, has the potential to result in reductions of spider densities at different strata. However, slight reductions in spider abundance may be an acceptable short-term ecological impact compared with the loss of an untreated hemlock and all the associated ecological benefits that it provides. Future studies should include investigations of long-term impact of imidacloprid on spiders associated with eastern hemlock.

First Report of Geosmithia morbida in North Carolina: The Pathogen Involved in Thousand Cankers Disease of Black Walnut.

In the past decade, black walnut (Juglans nigra) trees throughout western North America have suffered from widespread branch dieback and canopy loss, causing substantial tree mortality (2,3). The fungus, Geosmithia morbida, vectored by the walnut twig beetle (WTB), Pityophthorus juglandis, has been associated with this devastating disease known as Thousand Cankers Disease (TCD) (2,3). In August of 2012, branch samples from TCD symptomatic black walnut trees (5 to 10 cm in diameter and 15 to 30 cm long) were collected on the North Carolina side of the Great Smoky Mountain National Park (GRSM) in Cataloochee Cove (35°37.023' N, 83°07.351' W) and near the Big Creek Campground (35°45.290' N, 83°06.473' W), in Haywood County. Five symptomatic trees near the Big Creek Campground and three from Cataloochee Cove displayed typical TCD signs including progressive crown thinning, branch flagging, and branch dieback; however, insect holes were not observed. Samples were double bagged in Ziploc plastic bags, sealed in a 19-liter plastic bucket, and transported to the University of Tennessee. Outer bark was removed from the samples and small, elliptical, necrotic cankers were observed. Wood chips (3 to 4 mm2) from cankers were excised and placed on 1/10 strength potato dextrose agar amended with 30 mg/liter streptomycin sulfate and 30 mg/liter chlortetracycline HCL and incubated on a 12-h dark/light cycle at 22°C for 5 to 7 days. Fungal isolates were tentatively identified as G. morbida by using culture morphology, and characteristics of conidiophores and conidia (2). The isolated fungus from the Cataloochee Cove location was grown in 1/10 strength potato dextrose broth at room temperature for 2 weeks. Isolates from Big Creek Campground were contaminated and were not analyzed further. Fungal colonies were tan to light yellow. Conidia were tan, subcylindrical, and catenulate. Conidiophores were multibranched, verticillate, and verrucose. To verify the morphological data, DNA was extracted from fungal mycelia using DNeasy Plant Mini Kit (Qiagen, Valencia, CA) according to the manufacturer's published protocol. Isolates from Cataloochee Cove were characterized using ITS1 and ITS4 universal primers (4). The putative G. morbida isolate (GenBank Accession No. KC461929) had ITS sequences that were 100% identical to the G. morbida type isolate CBS124663 (FN434082.1) (2). Additionally, fungal DNA from Cataloochee Cove was amplified using G. morbida-specific microsatellite loci (GS04, GS27, and GS36) (1). PCR products were analyzed with the QIAxcel Capillary Electrophoresis System (Qiagen) and were similar to those previously published (2). To date, all confirmed cases of TCD in the native range of black walnut have been in urban areas, along rural roadsides and/or fence rows. The report in North Carolina is the first finding of G. morbida, the causal agent of TCD, in a forest setting. References: (1) D. Hadziabdic et al. Conserv. Genet. Resources 4:287, 2012. (2) M. Kolarik et al. Mycologia 103:325, 2011. (3) N. Tisserat et al. Plant Health Progr. doi:10.1094/PHP-2011-0630-01-BR, 2011. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, CA, 1990.

Factors affecting establishment and recovery of Sasajiscymnus tsugae (Coleoptera: Coccinellidae), an introduced predator of hemlock woolly adelgid (Hemiptera: Adelgidae) on eastern hemlock (Pinales: Pinaceae).

To reduce populations of hemlock woolly adelgid, Adelges tsugae Annand (Hemiptera: Adelgidae), >500,000 Sasajiscymnus tsugae (Sasaji and McClure) (Coleoptera: Coccinellidae) have been released in the Great Smoky Mountains National Park since 2002. To determine factors affecting establishment and recovery of these predatory beetles, 65 single release sites were sampled using beat sheets from 2008 to 2012. Several abiotic and biotic factors were evaluated for their association with establishment and recovery of S. tsugae. Information on predatory beetle releases (location, year of release, number released, and season of release), topographic features (elevation, slope, Beers transformed aspect, and topographic relative moisture index), and temperature data (minimum and maximum temperatures 1 d after release and average minimum and maximum temperatures 7 d after release) were obtained from Great Smoky Mountains National Park personnel. These factors were evaluated using stepwise logistic regression and Pearson correlation. S. tsugae was recovered from 13 sites 2 to 10 yr after release, and the greatest number was recovered from 2002 release sites. Regression indicated establishment and recovery was negatively associated with year of release and positively associated with the average maximum temperature 7 d after release and elevation (generally, recovery increased as temperatures increased). Several significant correlations were found between presence and number of S. tsugae and year of release, season of release, and temperature variables. These results indicate that releases of S. tsugae should be made in warmer (≍10-25°C) temperatures and monitored for at least 5 yr after releases to enhance establishment and recovery efforts.

Host utilization of field-caged native and introduced thistle species by Rhinocyllus conicus.

Rhinocyllus conicus Fröelich was introduced from Europe into North America as a biological control agent of the exotic weed Carduus nutans L. Concern exists over the feeding of this weevil on at least 25 species of native Cirsium thistles. Beginning in 2008, cage studies isolating adults of R. conicus on buds and flower heads of all eight thistle species (native and introduced) recorded from Tennessee were conducted to test if R. conicus could use these species for reproduction and what impacts larval feeding of R. conicus may have on seed production. Larvae of R. conicus completed development in heads of the native species C. carolinianum (Walter) Fernald and Schubert. and C. horridulum Michaux, and significant reductions in seed numbers of both species occurred during 2008. Rhinocyllus conicus oviposited on both C. carolinianum and C. horridulum at significantly greater levels than the introduced species C. arvense (L.) Scopoli and C. vulgare (Savi) Tenore. Infested heads of C. carolinianum contained numbers of R. conicus per centimeter of plant head width similar to Ca. nutans in 2008, and both native species contained numbers of R. conicus per centimeter of plant head width similar to C. arvense and C. vulgare in 2009. Body length was similar between R. conicus reared on native thistles and its target host Ca. nutans. This report is the first documentation of R. conicus feeding and reproducing on C. carolinianum and C. horridulum. Although R. conicus has been observed only on introduced thistles in naturally occurring populations in this region, the utilization of C. carolinianum and C. horridulum as host species in controlled conditions warrants continued monitoring of field populations and further investigation into factors that may influence nontarget feeding in the future.

Spatial prediction of habitat overlap of introduced and native thistles to identify potential areas of nontarget activity of biological control agents.

Nontarget feeding of Rhinocyllus conicus Fröelich and Trichosirocalus horridus (Panzer) on native North American thistles in the genus Cirsium has been documented. Some species of these native thistles have shown greater infestation levels of R. conicus in populations that are in close proximity to the target plant species, Carduus nutans L. In 2005 a study was initiated to identify areas of potential nontarget feeding by R. conicus and T. horridus on thistle species by predicting habitats of two known introduced hosts [C. nutans and Cirsium vulgare (Savi) Tenore] and two native species [Cirsium carolinianum (Walter) Fernald and Schubert and C. discolor (Muhlenberg ex Willdenow) Sprengel] using Mahalanobis distance (D(2)). Cumulative frequency graphs showed that the D(2) models for all four plant species effectively identified site conditions that contribute to the presence of the respective species. Poisson regression showed an association between D(2) values and plant counts at field-test sites for C. nutans and C. carolinianum. However, negative binomial regression detected no association between D(2) values and plant counts for C. discolor or C. vulgare. Chi-square analysis indicated associations between both weevil species and sites where C. vulgare and Carduus nutans were found, but not between the weevil and native thistle species. Habitats of C. nutans and Cirsium carolinianum overlapped in ≈12% of the study area. Data-based habitat models may provide a powerful tool for land managers and scientists to monitor native plant populations for nontarget feeding by introduced biological control agents.