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A predator, Laricobius osakensis Montgomery and Shiyake (Coleoptera: Derodontidae), is being mass-produced and released for the biological control of the invasive hemlock woolly adelgid (HWA), Adelges tsugae Annand (Hemiptera: Adelgidae). To better understand and predict the seasonality of this predator in North America, the development and reproduction of L. osakensis were evaluated at constant temperatures ranging from 5 to 22°C. The predicted seasonal biology was compared with data from field collections. L. osakensis did not complete development from egg to adult at the two lowest temperatures tested, 5 and 8°C, but did so at the highest temperature of 22°C. The minimum development thresholds were estimated for eggs (4.2°C), first (1.8°C), second (5.5°C), third (4.6°C), and fourth instar (4.1°C), prepupa (3.6°C), and pupa (7.5°C). Oviposition rates were significantly greater at 5 and 10°C than at 20 and 25°C. Head capsule width significantly increased for each of the four larval instars with a mean of 0.19, 0.26, 0.35, and 0.44 mm, respectively. Laboratory and field data were used to develop a phenology forecasting model to predict the occurrence of all developmental stages of L. osakensis. This model will allow land managers to more accurately predict the optimal timing for L. osakensis larval sampling throughout its established range.

Laurel wilt is a destructive vascular disease of trees in the laurel family (Lauraceae) caused by a nonnative insect/pathogen complex. This study monitored the recent spread and impact of laurel wilt in sassafras (Sassafras albidum [Nutt.] Nees) from the Gulf-Atlantic Coastal Plain region of the southeastern United States (US) into the adjacent Piedmont/Sandhills and Mountain regions. Laurel wilt was detected at thirteen of forty-six sassafras sites including seven outside the Coastal Plain. Compared to nondiseased sites, sassafras mortality due to laurel wilt increased rapidly from 2018 to 2020 and occurred in all diameter classes monitored (≥ 5 cm diameter at breast height, dbh). Flight trapping for the laurel wilt vector, the redbay ambrosia beetle (RAB, Xyleborus glabratus Eichhoff), with α-copaene lures did not enhance early detection of latent laurel wilt infections. Seasonal flight activity of the RAB in the Piedmont and Mountains suggested two generations per year with little to no flight from December through March. Study Implications Sassafras is the most widely-distributed lauraceous tree species in the US and is valued for its ecological functions, durable wood, and numerous cultural uses. Our results strongly suggest the potential for substantial loss of the existing sassafras resource as laurel wilt continues spreading through the eastern US. Survey traps baited with current lures for the primary insect vector are unlikely to improve early detection of new laurel wilt infections. The georeferenced network of plots established for this project provides baseline data for future monitoring efforts and could be revisited to look for evidence of sassafras resistance to the laurel wilt pathogen.

Following the adventive arrival, subsequent spread, and ensuing impact of Adelges tsugae Annand (Hemiptera: Adelgidae), the hemlock woolly adelgid (HWA) in the eastern United States, a robust initiative was launched with the goal of decreasing ecosystem impacts from the loss of eastern hemlock (Pinales: Pinaceae). This initiative includes the use of biological control agents, including Laricobius spp. (Insecta: Coleoptera). Laboratory production of these agents is limited by subterranean mortality and early emergence. Therefore, the subterranean survivorship and timing of emergence of a mixture of Laricobius spp. was investigated. PVC traps internally lined with a sticky card and covered with a mesh screen were inserted into the soil to measure the percent emergence of adults based on the number of larvae placed within. The number of emerged adults in the field and laboratory-reared larval treatments was adjusted based on emergence numbers in the control and used as the response variable. Independent variables included in the final model were: treatment (field-collected vs. laboratory-reared), organic layer depth (cm), soil pH, and April-to-December mean soil moisture. No differences were found in survivorship between field-collected and laboratory-reared treatments. As pH and organic layer increased survivorship decreased, significantly. Although the majority of emergence occurred in the fall, emergence also occurred in spring and summer. The occurrence of spring and summer emergence and low survivorship (17.1 +/- 0.4%) in the field across all treatments suggests that these are characteristics of Laricobius spp. field biology in their introduced range and not artifacts of the laboratory rearing process. United States Department of Agriculture Forest ServiceUnited States Department of Agriculture (USDA)United States Forest Service [15-CA-11420004-0260] Published version We thank E. Lane, A. Hillen, R. Anderson, C. Jubb, and C. Preston for their technical assistance in the field and in the laboratory, and the University of Virginia's Mountain Lake Biological Station for allowing us to use their site. We would also like to thank the United States Department of Agriculture Forest Service for funding this work (15-CA-11420004-0260). Public domain – authored by a U.S. government employee

Laurel wilt (LW) is a vascular disease caused by the fungus

Background: Laurel wilt disease has caused the extensive mortality of lauraceous species in the southeastern United States. The causal agent is an invasive fungus, Raffaelea lauricola, which is a symbiont of the beetle Xyleborus glabratus and causes a rapid, fatal vascular wilt. Early diagnosis of laurel wilt is imperative for efficient disease management. The current diagnostic process, however, is slow due to the lengthy laboratory procedures required to confirm pathogen presence. Methods: We tested the robustness and field-portability of a recently developed, species-specific, loop-mediated isothermal amplification (LAMP) assay for R. lauricola, with the overall goal of eliminating the need for a laboratory confirmation of the diagnosis. We tested the robustness of the assay using benchtop equipment with naturally infected samples. We then tested the assay directly in the field using a portable device. Results: The assay successfully detected R. lauricola directly from symptomatic wood tissue using crude DNA extracts. Furthermore, the assay readily allowed users to distinguish between symptoms caused by R. lauricola infection and similar symptoms caused by other agents. In-field, we assayed wood samples from symptomatic redbay (Persea borbonia [L.] Spreng) and sassafras (Sassafras albidum [Nutt.] Nees) across the Southeast and successfully detected R. lauricola-infected trees in less than an hour. Conclusion: Results of this study confirmed that the field-deployable LAMP assay is robust and can rapidly and accurately detect R. lauricola in infected trees directly on-site. LAMP technology is well suited for in-field implementation, and these results serve as an incentive for further development and use of this technology in the field of forest pathology.

Phytosanitary treatments for logs and barked wood products are needed to mitigate the spread of thousand cankers disease through the movement of these commodities. The disease threatens eastern black walnut (Juglans nigra L.) populations in the United States. It is caused by repeated attacks by the walnut twig beetle (Pityophthorus juglandis Blackman) and subsequent canker development caused by the fungal pathogen Geosmithia morbida M. Kolařík et al. Methyl bromide (MB) fumigations were evaluated for efficacy against P. juglandis and G. morbida in J. nigra bolts. Fumigation with 82 mg/L MB for 24 h at 4.5° C eliminated P. juglandis in J. nigra, but was ineffective against G. morbida. Subsequent experiments focused on eliminating G. morbida, but results were inconclusive because of low rates of pathogen recovery from naturally infested control bolts. Final experiments used J. nigra bolts artificially inoculated with G. morbida. Fumigations with 240 and 320 mg/L MB for 72 h at 10° C were effective in eliminating G. morbida from J. nigra bolts. Results confirm that the USDA fumigation treatment schedule for logs with the oak wilt pathogen will also mitigate the risk of spreading the thousand cankers disease vector and pathogen by movement of walnut bolts and wood products.

Efficient separation of insects from plant material for quantification and collection is an important component of entomological research. This paper reports on a novel, easily replicable container designed to efficiently collect two different biological control agents dispersing from hemlock (Tsuga spp.) foliage infested with the invasive hemlock woolly adelgid (HWA), Adelges tsugae Annand (Hemiptera: Adelgidae). The container utilizes a simplified Berlese-style funnel design to collect Laricobius spp. (Coleoptera: Derodontidae) larvae dropping from the foliage into a removable bottom jar, a central jar to house the foliage sample, and a removable top jar to collect adult silver flies (Leucopis spp., Diptera: Chamaemyiidae) emerging from puparia on the twigs. The efficacy of two designs (with and without a funnel leading to the top collection jar) was evaluated using western hemlock [Tsuga heterophylla (Raf.) Sarg.] foliage naturally colonized with HWA and the two predator genera. All Laricobius larvae were effectively collected in the bottom jar, and the addition of an inverted funnel leading to the top collection jar increased the proportion of Leucopis flies reaching the target jar from 60% to 94%. This ‘Lari-Leuco’ container is presented as a research and motoring tool to benefit the integrated pest management program for HWA in eastern North America and for potential use in simultaneously separating ascending and descending life stages in other insect-plant or predator-prey systems.

The introduction, establishment, and spread of invasive species in terrestrial and aquatic environments is widely recognized as one of the most serious threats to the health, sustainability, and productivity of native ecosystems (Holmes et al. 2009; Mack et al. 2000; Pyšek et al. 2012; USDA Forest Service 2013). In the United States, invasive species are the second leading cause of native species endangerment and extinction, and their costs to society have been estimated at $120 billion annually (Crowl et al. 2008; Pimentel et al. 2000, 2005). These costs include lost production and revenue from agricultural and forest products, compromised use of waterways and terrestrial habitats, harm to human and animal health, reduced property values and recreational opportunities, and diverse costs associated with managing (e.g., monitoring, preventing, controlling, and regulating) invasive species (Aukema et al. 2011; Pimentel et al. 2005). The national significance of these economic, ecological, and social impacts in the United States has prompted various actions by both legislative and executive branches of the Federal Government (e.g., the Nonindigenous Aquatic Nuisance Prevention and Control Act of 1990; the Noxious Weed Control and Eradication Act of 2002; Executive Order 13112 of 1999, amended in 2016).

Hemlock woolly adelgid (HWA), Adelges tsugae Annand (Hemiptera: Adelgidae), has devastated eastern hemlock (Tsuga canadensis [L.] Carriere) in a major portion of its native range in eastern North America. Population dynamics of HWA in the absence of predators have been studied for decades. After many years and much effort directed towards rearing and releasing biological control agents to manage HWA, one of these agents, Laricobius nigrinus Fender (Coleoptera: Derodontidae), is now successfully established at significant densities at sites from the southern to the mid-Atlantic states of the eastern U.S. However, high densities of HWA still persist at many locations throughout the region and spread of HWA and associated damage to hemlock continues. Population models for HWA have suggested that even upwards of 90% predation on eggs laid by the overwintering sistens generation will have minimal effect in reducing the population densities of HWA, if HWA are at high density. In this study, we tested the ability of L. nigrinus to reduce HWA densities, and experimentally tested these model predictions to better understand what impact, if any, L. nigrinus has on HWA densities. By using predator exclusion cages at field sites with well-established populations of L. nigrinus, we were able to record HWA densities, fecundity, overwintering mortality, and predation by L. nigrinus, as well as the proportion of branch tips producing new growth on study trees. Using our field-collected data, we refitted the model in ways that allowed us to predict what population densities we could expect for the following summertime progrediens generation given previous HWA density and levels of L. nigrinus. In both years, we found that despite high rates (greater than 80% ovisac predation) of predation by L. nigrinus on uncaged branches compared to caged branches, there were no significant differences in subsequent densities of the HWA spring generation between caged and uncaged treatments, as predicted by our model. In 2018, our field-collected densities of the summer progrediens generation were lower than what was predicted by the model in both predator exclusion treatments, possibly due to the model not incorporating tree health and climatic factors. Simulation models of pest insect populations based on field-collected data such as fecundity, density, overwintering mortality, and predation, could prove to be important in informing researchers and managers about the role of the biological control agent in the population dynamics of the target host. USDA Forest Service, USA [14-CA-11420004-181] This work was funded by a cooperative agreement with the USDA Forest Service, USA (No. 14-CA-11420004-181). We thank R., Reardon, N. Schneeburger, R. Rhea of the Forest Service for arranging and administrating our funding, M. Meyer for helping us find field sites in NJ and R. Mays, A. Dechaine (Virginia Tech) and B. Mudder (USDA Forest Service) for helping collect samples from the field. We thank A. Roehrig, A. Langevin for processing samples. We thank H. Broadley, J. Anderson, and B. Padilla for help with statistical analysis and graphics. We thank J. Chandler, R. Van Driesche, and D. Orwig for reviewing earlier drafts of this MS. Public domain – authored by a U.S. government employee

In the eastern United States, hemlock woolly adelgid (HWA), Adelges tsugae Annand (Hemiptera: Adelgidae) is considered an invasive pest of eastern hemlocks; an ecologically foundational tree species. Current management of HWA focuses on chemical and biological controls, with recent research suggesting that these two tactics could be integrated successfully. The approach is to protect a subset of hemlocks with systemic insecticides while releasing predatory insects onto adjacent, unprotected trees. The goal of this study was to assess the effects of chemical and biological control tactics, alone and in combination, on hemlock health and HWA densities at three southern Appalachian sites (KY, WV, and TN) from 2010 to 2016. Although insecticide applications were effective at protecting individual trees, none of the overall treatments (chemical, biological, or combined) had a significant effect on tree health or HWA population index values relative to untreated plots. Tree health generally declined at all sites over time. HWA populations were highly variable over time and were likely more strongly influenced by extremely low, winter temperatures than by the treatments. Cross-correlation analysis of tree health and HWA population indicated a time-lag effect. At two of the three sites, recovery of tree health lagged 0 – 3 years behind decline in HWA population, and decline in HWA populations lagged approximately 0 – 1 years behind decline in tree health. The predatory beetle, Laricobius nigrinus, was recovered two-years, post-release at the KY and WV sites in 2012 and 2013, but was not recovered from the TN site. The lack of sustained recovery of L. nigrinus may be attributable to the occurrence of extremely low, winter temperatures in 2014 and 2015, which produced subsequent crashes in the HWA populations. In TN, the L. nigrinus population may have been unrecoverable due to a decline in the HWA population shortly after initial release.

The walnut twig beetle (Pityophthorus juglandis Blackman) (Coleoptera: Curculionidae) is a regulated pest in the United States due to its causal role in thousand cankers disease of walnut trees, including the commercially valuable eastern black walnut (Juglans nigra L.). Several state quarantines designed to limit spread of P. juglandis regulate movement of kiln-dried walnut lumber that contains bark. Previous research demonstrated that P. juglandis will enter and re-emerge from bark of kiln-dried, J. nigra slabs subjected to extreme beetle pressure (baited with a pheromone lure and hung in infested J. nigra trees). This study evaluated P. juglandis bark colonization of both kiln-dried and fresh J. nigra slabs, varying the presence of aggregation pheromone and relative proximity to a beetle source. Wood treatment, slab location, and pheromone presence all significantly affected P. juglandis colonization, as assessed by subsequent beetle emergence. When placed on the ground directly beneath infested trees, kiln-dried slabs were not colonized, and fresh slabs were colonized only when baited with the pheromone lure (6/14 replicates). When placed in crowns of infested trees, kiln-dried slabs were colonized only when baited with pheromone (3/14 replicates), whereas fresh slabs were colonized with and without pheromone (14/14 and 1/13 replicates, respectively). Timing of emergence indicated that beetles did not reproduce in kiln-dried bark. Results suggest that the risk of kiln-dried walnut bark becoming colonized by the P. juglandis during movement of commercial wood products is very low. This information may be useful to government agencies that administer quarantines regulating the transport of walnut lumber.

Thousand cankers disease (TCD), caused by the walnut twig beetle (WTB), Pityophthorus juglandis , and its associated fungal symbiont, Geosmithia morbida , is a disease of economic and ecological concern on eastern black walnut, Juglans nigra . Numerous attacks and gallery formation by the WTB and subsequent development of cankers caused by the fungus result in progressive crown dieback. The disease can kill affected trees in a few to several years following initial infestation and very few management options are available for preventing or reducing impact of TCD on black walnut trees. Since advanced development of TCD requires multiple WTB generations and numerous beetle attacks, control strategies that reduce beetle attacks and brood production, without completely eliminating infestation, could still significantly benefit tree health and survival. We evaluated the use of entomopathogenic fungi Beauveria bassiana and Metarhizium brunneum against the WTB. Laboratory and field studies showed that WTB adults are susceptible to commercial strains B. bassiana GHA and M. brunneum F52. Exposure of beetles to sprayed walnut bolts resulted in reduced brood production, primarily due to the death of parental adults from fungal infection prior to egg laying. Spraying walnut bolts with B. bassiana GHA and M. brunneum F52 prior to field exposure to natural WTB populations reduced emergence of next generation adults by up to 98% and 96%, respectively, due to a combination of fewer beetle attacks and mortality among those that tunneled. These results demonstrate the potential use of entomopathogenic fungi in the integrated management of TCD in walnut trees.

The rapid loss of eastern hemlock (Tsuga canadensis) due to infestation with hemlock woolly adelgid (Adelges tsugae, HWA) has greatly altered structure and function of eastern forests. Numerous control strategies including local pesticide use and biocontrol with predator beetles have been implemented with considerable cost and varying success. Silviculture treatments that increase incident light on surviving hemlock trees to reduce infestation and ameliorate carbon starvation have been proposed as another possible conservation strategy, yet no controlled studies have evaluated the coupled plant-insect responses to increased light. We conducted a nursery experiment on artificially infested eastern hemlock seedlings under varying levels of ambient light, ranging from 0 to 90% shade. We measured HWA infestation (ovisac density), short- and long-term indicators of carbon balance (leaf chlorophyll fluorescence, net photosynthesis, total nonstructural carbohydrate content, and shoot growth), and nutrition (leaf N content). We hypothesized that higher light would result in reduced HWA densities, higher C assimilation rates, and improved tissue non-structural carbohydrate balance; and these effects would ameliorate the effects of infestation on C balance and lead to improved seedling growth. HWA density decreased with increasing light, and was highest in the 90% shade treatments. However, photosystem II efficiency and net assimilation were also lower under the higher light treatment. Despite tradeoffs between reduced infestation and reduced leaf function from higher light, and little variation in sugar content among treatments, both leaf and root starch content and seedling growth were higher under the higher light treatment. Increasing light levels improves long-term carbon balance for hemlock seedlings in the presence of HWA. Although hemlock typically occurs in deeply shaded forests, our results suggest that silvicultural treatments such as forest thinning that increase light exposure may reduce HWA abundance and lead to better tree C balance, and may be an effective component of large-scale conservation and restoration strategies.

Laurel wilt is a vascular disease of redbay (Persea borbonia (L.) Spreng.) and other plants in the family Lauraceae in the southeastern United States. It is caused by a fungus (Raffaelea sp.) that is vectored by a non-native insect of Asian origin, the redbay ambrosia beetle (Xyleborus glabratus Eichhoff) (1). Since the initial detection of the redbay ambrosia beetle near Savannah, GA in 2002, laurel wilt has caused widespread mortality of redbay in Georgia, South Carolina, and Florida (1). In September 2007, an avocado (Persea americana Mill.) tree planted approximately 10 years earlier in a residential neighborhood in Jacksonville, FL was discovered to be infected with laurel wilt. The crown was in various stages of decline, including upper branches that were dead and leafless, those with wilted and drooping foliage, and those with healthy foliage. Removal of bark from wilted branch sections revealed black-to-brown streaks of discoloration in the sapwood and a few ambrosia beetle holes from which the discoloration extended into the adjacent wood. A Raffaelea sp. was isolated from discolored wood samples by surface sterilizing wood chips by submersion in a 5% sodium hypochlorite solution for 30 s and plating them on cycloheximide streptomycin malt agar (2). Small subunit (18S) sequences from the rDNA were amplified by PCR and sequenced with primers NS1 and NS4 (3). BLASTn searches revealed homology to Raffaelea sp. C2203 (GenBank Accession No. EU123076, 100% similarity, e-value of 0.0, and a total score of 1,886), which is known to be the causal agent of laurel wilt (1). The small-subunit rDNA sequence for this isolate has been deposited into GenBank and has been assigned accession No. EU257806. Pathogenicity of the laurel wilt pathogen on Persea spp. in growth chamber trials has been previously demonstrated (1). Laurel wilt is of concern to the commercial avocado industry and is a potential threat to the Lauraceae elsewhere in the Americas. References: (1) S. W. Fraedrich et al. Plant Dis. 92:215, 2008. (2) T. C. Harrington. Mycologia 73:1123, 1981. (3) 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.

Laurel wilt is a lethal, nonnative vascular wilt disease of redbay (Persea borbonia), sassafras (Sassafras albidum), and other trees in the Lauraceae (1,4). It is caused by a fungus (Raffaelea lauricola) and transmitted by the redbay ambrosia beetle (Xyleborus glabratus), a nonnative insect first detected in Georgia in 2002 (1,2). Since introduction of the pathogen and vector (presumably from Asia), laurel wilt has caused extensive mortality to redbays in Georgia, Florida, and South Carolina (1). In June 2009, a landowner in Gautier, MS reported dead redbay trees. Signs and symptoms were identical to those reported for laurel wilt along the Atlantic Coast (wilted, bronze red foliage, and dark gray-to-black vascular discoloration) (1). Infected trees have subsequently been confirmed in and near the Pascagoula River Basin. Size of infected redbays ranged from 5 to 20 cm (diameter at breast height). No heavily decomposed or fallen redbays were noted. Many individual specimens exhibited extensive drying of stem wood and dry, wilted, light brown foliage. This indicates that introduction to the area may have occurred within the last 3 years. X. glabratus adults were collected (30°26′44.45″N, 88°39′41.83″W) in a Lindgren funnel trap baited with phoebe and manuka oil lures. Beetle identification was confirmed by USDA-APHIS, and voucher specimens were submitted to the Smithsonian National Museum of Natural History and the Mississippi Entomological Museum. Symptomatic redbay wood chips from the same location were surface sterilized and plated on cycloheximide-streptomycin malt agar and R. lauricola was isolated. A 1,026-bp portion of 18S rDNA (GenBank No. GQ996063) was amplified by PCR and sequenced using primers NS1 and NS4. BLASTn searches revealed perfect homology to R. lauricola isolate PL 697 (GQ329704). Two isolates of R. lauricola were recovered and prepared into separate spore suspensions (1 × 108 CFU/ml). Each isolate was inoculated into two healthy redbays. The inoculated redbays were placed in a growth chamber with two water-only controls. All inoculated plants, and none of the controls, exhibited wilt symptoms and died within 20 days. R. lauricola was recovered from the discolored sapwood of the inoculated plants, completing Koch's postulates. A model prediction for the natural dispersion of X. glabratus and R. lauricola estimated that these organisms may not reach Mississippi for 10 to 15 years (3). The current detection of laurel wilt in Mississippi is substantially ahead of this estimate. Currently, no records of laurel wilt have been reported from western Georgia, all of Alabama, or the panhandle of Florida. Confirmed locations in Mississippi are in Jackson County, along the Interstate 10 corridor and the Pascagoula River drainage. Due to the relatively large extent of the infestation (~64 km2, including hundreds of infected trees) eradication is not being attempted. Surveys, remote sensing, and phylogeographic analysis are underway to delineate the extent of infestation and discover the mode of introduction. The current outbreak of laurel wilt in Mississippi is likely the result of human transport of infested wood, either from Asia as a separate, new introduction or from previously infested areas in the southeastern United States. References: (1) S. W Fraedrich et al. Plant Dis. 92:215, 2008. (2) T. C. Harrington et al. Mycotaxon 104:399, 2008. (3) F. Koch and W. Smith. Environ. Entomol. 37:442, 2008. (4) J. A. Smith et al. Plant Dis. 93:198, 2009.

Laurel wilt disease, caused by Raffaelea lauricola (T.C. Harr., Fraedrich & Aghayeva sp. nov.), which is a fungal symbiont of the nonnative redbay ambrosia beetle (Xyleborus glabratus Eichhoff), has caused widespread mortality of native redbay (Persea borbonia (L.) Spreng) in Georgia, South Carolina, and Florida since 2002. The disease has been noted on other species in the Lauraceae including sassafras in Georgia (1), and more recently, on avocado and camphor in Florida (4). Since 2005, wilted shoots, branch dieback, and tree death have been observed in sassafras trees (Sassafras albidum (L.)) in Liberty, McIntosh, Chatham, Effingham, Bulloch, Evans, and Screven counties in Georgia; Bamberg, Beaufort, Charleston, Colleton, Hampton, and Orangeburg counties in South Carolina; and Putnam County in Florida. Symptomatic sassafras trees ranged from 1 to 12 m high and 2.5 to 25 cm in diameter at breast height. In contrast to red bay trees that retain wilted foliage, symptomatic sassafras defoliate rapidly as trees wilt and die. Multiple symptomatic ramets originating from a common root system have been observed. Removal of bark from stem and root sections from wilted trees revealed black-to-brownish staining in the sapwood, characteristic of laurel wilt. Wood chips from symptomatic areas of branches and roots were surface sterilized and plated on cycloheximide-streptomycin malt agar as previously described (1) and R. lauricola was routinely isolated. Small subunit (18S) sequences from rDNA were amplified by PCR and sequenced using primers NS1 and NS4 (3) for isolates from sassafras from Florida and South Carolina. BLASTn searches revealed homology to Raffaelea sp. C2203 (GenBank Accession No. EU123076, 100% similarity) described by Fraedrich et al. (1) from redbay and later named R. lauricola (2). The small subunit rDNA sequences for these isolates have been deposited into GenBank ( http://www.ncbi.nlm.nih.gov/Genbank/index.html ) and assigned Accession Nos. EU980448 (Florida) and GQ329704 (South Carolina). Koch's postulates have been completed with R. lauricola on this host previously (1). Laurel wilt on sassafras often was geographically isolated from other symptomatic hosts in Georgia and South Carolina and appears to occur on this host independently of proximity to redbay. Further studies to determine the epidemiology of laurel wilt on sassafras, potential resistance, and impact on sassafras life history and distribution are needed. Given the clonal nature of sassafras, the disease would appear to have the potential to move through roots of trees once established in a stand. References: (1) S. W Fraedrich et al. Plant Dis. 92:215, 2008. (2) T. C. Harrington et al. Mycotaxon 104:399, 2008. (3) M. A. Innis et al. PCR Protocols, A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990. (4) J. A. Smith et al. Plant Dis. 93:198, 2009.

Formulations of entomopathogenic (insect-killing) fungi represent alternatives to synthetic insecticides in the management of forest and shade tree insects. We evaluated bark spray applications of the entomopathogen Beauveria bassiana (Balsamo) Vuillemin (Hypocreales: Cordycipitaceae) strain GHA (BotaniGardES), permethrin insecticide (Astro), and water (control) on colonization of black walnut (Juglans nigra L.) (Fagales: Juglandaceae) bolts by the walnut twig beetle (Pityophthorus juglandis Blackman) (Coleoptera: Curculionidae), vector of the fungus that causes thousand cankers disease. Treated bolts were baited with a P. juglandis aggregation pheromone lure and deployed in infested walnut trees. Bark application of permethrin prevented P. juglandis colonization of the phloem. Although treatment of bolts with the B. bassiana suspension did not reduce P. juglandis colonization or short-term emergence relative to the control treatment, it increased the B. bassiana infection rate from 25 to 62% of emerged adults. Results suggest that commercial applications of B. bassiana strain GHA may help augment natural levels of infection by this entomopathogen in the eastern United States, and support continued exploration of entomopathogens for biological control of the walnut twig beetle.

Laurel wilt is a fungal vascular disease of redbay (Persea borbonia (L.) Spreng) and other plants in the family Lauraceae in the southeastern United States (1). The disease is caused by Raffaelea lauricola T. C. Harr., Fraedrich & Aghayeva, which is vectored by the exotic redbay ambrosia beetle (Xyleborus glabratus Eichhoff) (2). Pondspice (Litsea aestivalis (L.) Fern.) is an obligate wetland shrub listed as endangered in Florida and Maryland and threatened in Georgia (4). On 29 August 2008, 369 of 430 (85%) pondspice trees observed at St. Marks Pond in St. John's County, Florida were dead and/or dying (4). Stem samples were collected from plants with wilted and reddened foliage, entrance holes with boring dust characteristic of ambrosia beetle attack, and dark discoloration in the outer sapwood. Discolored stem sections were surface disinfested for 30 s in a 5% sodium hypochlorite solution and then plated onto cycloheximide streptomycin malt extract agar (1). Smooth, cream-buff, submerge hyphae with uneven margins resembling R. lauricola (2) was observed growing from all sapwood pieces. DNA was extracted from a single isolate (PL 392) and the 18s small subunit rDNA was PCR amplified and sequenced with primers NS1 and NS4 (3), resulting in a 1,026-bp amplicon. A BLASTn search showed identical homology to R. lauricola strain PL 159 (GenBank Accession No. EU257806). The 18s small subunit rDNA sequence was deposited into GenBank (FJ514097). In May 2011, a spore suspension was made by flooding a single-spore culture plate of isolate PL 392 with 2 ml of sterile water, collecting the spores by pipette, and quantification by hemacyometer to 1.5 × 106 spores/ml. Pathogenicity tests were conducted on 1 to 1.5 m tall pondspice plants. Six saplings were wounded by a 3/32-inch drill bit, with four receiving 50 μl of the spore suspension and two serving as water-inoculated controls. All plants were kept in a greenhouse under ambient temperature. Within 21 days, all fungal-inoculated saplings displayed complete canopy wilt, typical of laurel wilt. R. lauricola was later recovered from all four infected plants, completing Koch's postulates. To determine if the vector can reproduce in pondspice, infected stem sections were placed in a plastic rearing box indoors at room temperature, and both callow and mature adult female X. glabratus emerged in October and November 2008. Although laurel wilt has been previously observed on pondspice in South Carolina and Georgia (1), this is the first confirmation of the disease on pondspice in Florida and the first confirmation of the vector from stem material of this host. References: (1) S. W. Fraedrich et al. Plant Dis. 92:215, 2008. (2) T. C. Harrington et al. Mycotaxon 104:399, 2008. (3) M. A. Innis et al. PCR Protocols, A Guide to Methods and Applications. Academic Press. San Diego, CA, 1990. (4) J. A. Surdick and A. M. Jenkins. Pondspice (Litsea aestivalis) Population Status and Response to Laurel Wilt Disease in Northeast Florida. Florida Natural Areas Inventory, Tallahassee, FL, 2009.

In recent years, outbreaks of nonnative invasive insects and pathogens have caused significant levels of tree mortality and disturbance in various forest ecosystems throughout the United States. Laurel wilt, caused by the pathogen Raffaelea lauricola (T.C. Harr., Fraedrich and Aghayeva) and the primary vector, the redbay ambrosia beetle (Xyleborus glabratus Eichhoff), is a nonnative pest-disease complex first reported in the southeastern United States in 2002. Since then, it has spread across eleven southeastern states to date, killing hundreds of millions of trees in the plant family Lauraceae. Here, we examine the impacts of laurel wilt on selected vulnerable Lauraceae in the United States and discuss management methods for limiting geographic expansion and reducing impact. Although about 13 species belonging to the Lauraceae are indigenous to the United States, the highly susceptible members of the family to laurel wilt are the large tree species including redbay (Persea borbonia (L.) Spreng) and sassafras (Sassafras albidum (Nutt.) Nees), with a significant economic impact on the commercial production of avocado (Persea americana Mill.), an important species native to Central America grown in the United States. Preventing new introductions and mitigating the impact of previously introduced nonnative species are critically important to decelerate losses of forest habitat, genetic diversity, and overall ecosystem value.

Laurel wilt kills American members of the Lauraceae plant family, including avocado (Persea americana). The disease threatens commercial production in the United States and other countries, and currently impacts the avocado industry in Florida. As laurel wilt spreads, the National Germplasm Repository for avocado in Miami (USDA-ARS) and commercial and residential production in other states (e.g., California and Hawaii), U.S. protectorates (Puerto Rico), and other countries are at risk. In the United States, value-added production of avocado of more than $1.3 billion/year is threatened. This recovery plan was produced as part of the National Plant Disease Recovery System (NPDRS), called for in Homeland Security Presidential Directive Number 9 (HSPD-9) to insure that the tools, infrastructure, communication networks, and capacity required to mitigate the impact of high-consequence plant disease outbreaks are such that a reasonable level of crop production is maintained. It is intended to provide a brief primer on the disease, assess the status of critical recovery components, and identify disease management research, extension, and education needs.

Hemlock woolly adelgid (HWA), Adelges tsugae Annand (Hemiptera: Adelgidae), is an invasive pest causing significant mortality to eastern and Carolina hemlock in eastern North America. Since 2003, management of HWA has included targeted release of the HWA predator Laricobius nigrinus Fender (Coleoptera: Derodontidae), native to western North America. Establishment of L. nigrinus at release sites is well documented, but investigations of its impact on HWA populations have been limited. A four-year (2014-2018), two-phase study using predator exclusion cages to assess the impact of L. nigrinus on HWA was conducted at nine previous release sites in the eastern United States. Significantly more HWA sistens ovisacs were disturbed on no-cage and open-cage branches than on caged branches where predators were excluded. Mean disturbance levels on cage, no-cage and open-cage branches was 8, 38, and 27 percent, respectively. Seven of nine sites had a mean HWA ovisac disturbance greater than 50% for at least one year. Winter temperatures were also a significant factor in overall mortality of the sistens generation with a mean of 46% on study branches. Six of nine sites had a mean overall mortality (winter mortality and predation) greater than 80% for at least one year. Larvae of Laricobius spp. were recovered at all sites during this study. Sequencing of the COI gene from recoveries in Phase One (2015 and 2016) indicated that 88% were L. nigrinus and 12% were L. rubidus LeConte. Microsatellite analysis performed during Phase Two (2017 and 2018) indicated that approximately 97% of larval recoveries were L. nigrinus, 2% were hybrids of L. nigrinus and L. rubidus, and 1% were L. rubidus. Results of this study suggest that L. nigrinus can significantly impact the HWA sistens generation ovisacs and continued investment in the use of this species as a biological control is recommended. USDA Forest Service cooperative agreementUnited States Department of Agriculture (USDA)United States Forest Service [14-CA-11420004-028] We gratefully acknowledge the assistance of Dr. Thomas Kuhar and Dr. Douglas Pfieffer (Virginia Tech) for review of this project, as well as Dr. Carlyle Brewster (Virginia Tech) for assistance with data analysis. We thank Dr. Nathan Havill (USDA Forest Service), Kari Stanley (Virginia Tech), and Melissa Fischer (Washington State Department of Natural Resources) for assistance with molecular protocols, as well as Mark Mayer (New Jersey Department of Agriculture), Scott Passwaters (James River State Park), Biff Thompson (Maryland Department of Agriculture), Jesse Webster (Great Smoky Mountains National Park), and the staff at Blackberry Farm (Tennessee) for site assistance. Thank you to Arthur Morgan School, Celo Community, and the Chattahoochee National Forest for permission to use sites in NC and GA. We are grateful to Natalie Morris, Kara Jeffries, Andy Dechaine, Ryan Mays, Ashley Toland, and James Wahls (Virginia Tech), Bryan Mudder and Andrew Tait (USDA Forest Service), David Bechtel, Philip Hensley, Elizabeth McCarty, Forest Palmer, Pat Parkman, and David Paulsen (University of Tennessee), and Peter Menzies, Marika Lapham, and Ivy Haas (University of North Carolina Asheville), for laboratory and field assistance. This work was funded under USDA Forest Service cooperative agreement 14-CA-11420004-028. Public domain – authored by a U.S. government employee

The rapid loss of eastern hemlock (Tsuga canadensis) in the southern Appalachian Mountains due to hemlock woolly adelgid (Adelges tsugae, HWA) infestation has resulted in substantial changes to ecosystem structure and function. Several restoration strategies have been proposed, including silvicultural treatments that increase incident light in forest understories. We conducted a four-year manipulative field experiment on surviving mid-story hemlock trees to investigate the effects of release from light limitation on HWA infestation and physiological parameters, expecting that higher light levels would improve tree carbon balance. Mixed hardwood forest sites were either previously uninfested with HWA (UN), infested with HWA (I), or infested with HWA and had a history of predatory beetle releases for biological control (I+P). At each site, we identified ten eastern hemlock trees in the mid-story and cut ~15 m radius canopy gaps around half of them while leaving the canopy intact over the other half. We compared short- and long-term indices of carbon gain and stress: leaf net photosynthesis (Anet); leaf fluorescence (Fv/Fm); leaf total non-structural carbohydrate concentration ([TNC]); new shoot growth; HWA density; and basal area growth. We found that trees experienced greater leaf-level stress in gaps and when HWA was actively feeding. Despite being more stressed, trees in gaps fixed 2x more carbon than those in reference conditions. High Anet in the spring translated into high leaf [TNC] in the spring, coinciding with when HWA was actively feeding. Although infested and uninfested trees had similar leaf [TNC] maxima, infestation prevented trees from allocating this carbon to shoot and basal area growth; this was particularly true for reference trees. Greater shoot growth in gap trees translated to greater annual basal area growth—by the end of the study, trees in gaps were growing 9x more than trees in reference conditions, and this was generally regardless of infestation status. In terms of growth and carbon balance, eastern hemlock consistently benefited from the increased light and soil moisture found in gaps; there was inconsistent and rather weak evidence that predator beetles conferred an additional advantage. Our results indicate that silvicultural treatments may improve long-term health and survival of infested trees and that integration of such treatments with existing strategies is worthy of continued exploration.

Laricobius nigrinus Fender (Coleoptera: Derodontidae) is a specialist predator of the hemlock woolly adelgid, Adelges tsugae Annand (Hemiptera: Adelgidae), native to the Pacific Northwest. It has been introduced into the eastern United States for biological control of exotic hemlock woolly adelgid populations that threaten native hemlock. The possible role of olfactory cues in host finding by this predator has received little study. We used gas chromatography–electroantennographic detection (GC-EAD) to test adult L. nigrinus olfactory sensitivity to volatiles from foliage of both adelgid-infested and uninfested eastern hemlock [Tsuga canadensis (L.) Carriere] and western hemlock [Tsuga heterophylla (Raf.) Sarg.], and two hemlock woolly adelgid nonhost species (eastern white pine, Pinus strobus L., and western white pine, Pinus monticola Douglas ex D. Don). Adelgid infestation did not alter L. nigrinus EAD response profiles to volatiles from either hemlock species. In total, antennal preparations...

Several North American walnut species (Juglans spp.) are threatened by thousand cankers disease which is caused by the walnut twig beetle (Pityophthorus juglandis Blackman) and its associated fungal plant pathogen, Geosmithia morbida M. Kolarík, E. Freeland, C. Utley and N. Tisserat sp. nov. Spread of this disease may occur via movement of infested black walnut (Juglans nigra L.) wood. This study evaluated the ability of P. juglandis to colonize J. nigra wood previously treated with various phytosanitation methods. Steam-heated and methyl bromide-fumigated J. nigra logs, as well as kiln-dried natural wane J. nigra lumber (with and without bark) were subsequently exposed to P. juglandis colonization pressure in two exposure scenarios. Following a pheromone-mediated, high-pressure scenario in the canopy of infested trees, beetles readily colonized the bark of steam-heated and methyl bromide-fumigated logs, and were also recovered from kiln-dried lumber on which a thin strip of bark was retained. In the simulated lumberyard exposure experiment, during which samples were exposed to lower P. juglandis populations, beetles were again recovered from bark-on steam-heated logs, but were not recovered from kiln-dried bark-on lumber. These data suggest logs and bark-on lumber treated with phytosanitation methods should not be subsequently exposed to P. juglandis populations. Further beetle exclusion efforts for phytosanitized, bark-on walnut wood products transported out of quarantined areas may be necessary to ensure that these products do not serve as a pathway for the spread of P. juglandis and thousand cankers disease.

An integrated management approach is needed to maintain eastern hemlock (Tsuga canadensis (L.) Carriere) in eastern North America and to minimize tree damage and mortality caused by the invasive hemlock woolly adelgid (Adelges tsugae Annand). This study examined the hypothesis that chemical control with low rates of insecticide and biological control can be combined in the same stand to impact adelgid populations, prolong crown health, and allow predator proliferation. Sixty T. canadensis trees in northern Georgia were individually treated via soil injection with 0%, 10%, or 25% of the label rate of imidacloprid insecticide, and the biological control predator Laricobius nigrinus Fender was released in the stand, two and four years later. By year seven, hemlocks treated with the 25% imidacloprid rate lost their insecticide protection, had significantly better crown health and higher adelgid densities than untreated trees, and supported as many Laricobius predator larvae as untreated trees. In year seven, no residues of imidacloprid were detected in Laricobius larvae feeding on previously-treated hemlocks. Most (77%) of the predators collected on study trees were identified as L. nigrinus, 12% were the native congener Laricobius rubidus LeConte, and 11% were hybrids between the introduced and native species. The hybridization rate remained stable over time. The density of undisturbed A. tsugae ovisacs was twice as high on branches protected from predators as compared with branches exposed to predators. Results suggest that chemical and biological control of A. tsugae can be successfully integrated to help prolong hemlock health, although additional predators may be necessary to protect hemlock trees in the southern Appalachians.

Thousand cankers disease, caused by the invasive bark beetle Pityophthorus juglandis Blackman and an associated fungal pathogen Geosmithia morbida M.Kolařík, E. Freeland, C. Utley, N. Tisserat, currently threatens the health of eastern black walnut (Juglans nigra L.) in North America. Both the beetle and pathogen have expanded beyond their native range via transport of infested walnut wood. Geosmithia morbida can develop in seedlings following inoculation, but the ability of P. juglandis to colonize young, small diameter trees has not been investigated. This study assessed the beetle's colonization behavior on J. nigra nursery trees. Beetles were caged directly onto the stems of walnut seedlings from five nursery sources representing a range of basal stem diameter classes. Seedlings were also exposed to P. juglandis in a limited choice, field-based experiment comparing pheromone-baited and unbaited stems. When beetles were caged directly onto stems, they probed and attempted to colonize seedlings across the range of diameters and across sources tested, including stems as small as 0.5 cm in diameter. In the field experiment, beetles only attempted to colonize seedlings that were baited with a pheromone lure and appeared to prefer (though not statistically significant) the larger diameter trees. Despite several successful penetrations into the phloem, there was no evidence of successful progeny development within the young trees in either experiment. Further investigation is recommended to better elucidate the risk nursery stock poses as a pathway for thousand cankers disease causal organisms.

Leucopis argenticollis (Zetterstedt) and Leucopis piniperda (Malloch) are known to feed on the lineage of Adelges tsugae Annand that is native to western North America, but it is not known if they will survive on the lineage that was introduced from Japan to the eastern USA. In 2014, western Leucopis spp. larvae were brought to the laboratory and placed on A. tsugae collected in either Washington (North American A. tsugae lineage) or Connecticut (Japanese lineage). There were no significant differences in survival or developmental times between flies reared on the two different adelgid lineages. In 2015 and 2016, western Leucopis spp. adults were released at two different densities onto enclosed branches of A. tsugae infested eastern hemlock (Tsuga canadensis (L.) Carr.) in Tennessee and New York. Cages were recovered and their contents examined 4 weeks after release at each location. Leucopis spp. larvae and puparia of the F1 generation were recovered at both release locations and adults of the F1 generation were collected at the Tennessee location. The number of Leucopis spp. offspring collected increased with increasing adelgid density, but did not differ by the number of adult flies released. Flies recovered from cages and flies collected from the source colony were identified as L.argenticollis and L. piniperda using DNA barcoding. These results demonstrate that Leucopis spp. from the Pacific Northwest are capable of feeding and developing to the adult stage on A. tsugae in the eastern USA and they are able to tolerate environmental conditions during late spring and early summer at the southern and northern extent of the area invaded by A. tsugae in the eastern USA.

The hemlock woolly adelgid Adelges tsugae Annand (Hemiptera: Adelgidae), is a non-native invasive pest that has caused widespread decline and mortality of eastern hemlock (Tsuga canadensis (L.) Carr. (Pinales: Pinaceae)) and Carolina hemlock (T. caroliniana Engelm.) in the eastern United States. Our preliminary experiments evaluated the utility of a rain-down technique to induce artificial infestations of A. tsugae on hemlock seedlings en masse. Experiments were conducted in PVC (1 m3) cages topped with poultry wire for placement of A. tsugae-infested branches, and with 1 m2 gridded glue sheets and/or hemlock seedlings placed below to capture adelgid abundance, distribution, and infestation rate data. In the March 2011 experiment, the density of progrediens crawlers (adelgid nymphs, first instars) that rained down inside the PVC cages was significantly higher in the high ovisac treatment compared to the low ovisac treatment, with an estimated 513,000 and 289,000 crawlers per m2 falling beneath each treatment, respectively. Resulting A. tsugae infestation rates on Carolina hemlock seedlings placed inside the cages did not differ between the treatments but were at or above established damage threshold densities for the adelgid. Infestation rates on eastern hemlock seedlings that were placed in cages nine days after the experiment started were below damage threshold levels and did not differ between the treatments. In the May 2011 experiment, the density of sistens crawlers raining down was substantially lower, with 17,000 and 33,000 falling per m2 in the low and high ovisac treatments, respectively. Resulting infestation rates on Carolina hemlock seedlings were extremely low and well below damage threshold levels. Although A. tsugae crawlers were well distributed across the 1 m2 gridded glue sheets placed at the bottom of each cage, hot spots of unusually high crawler density did occur in both experiments. This rain-down technique shows potential for use in an operational tree-breeding program where screening large numbers of hemlock seedlings for resistance to A. tsugae is required.

A method for determining the sex of live adult Laricobius nigrinus Fender (Coleoptera: Derodontidae) is described. Beetles were briefly chilled and positioned ventral-side-up under a dissecting microscope. Two forceps with blunted ends were used to gently brace the beetle and press on the centre of the abdomen to extrude its terminal segments. Male beetles were distinguished by a sclerotised, reticulate ninth abdominal sternite. In females, the distinct ovipositor (tergite, valvifers (ninth sternite), and laterotergites of the ninth abdominal segment) was visible. The procedure was rapid and harmed only a small number of individuals (fewer than 5%). Resume—Nous decrivons une methode pour determiner le sexe chez des adultes vivants de Laricobius nigrinus Fender (Coleoptera: Derodontidae). Les coleopteres sont brievement refroidis et places sur le dos sous une loupe microscopique. Deux brucelles a bouts emousses permettent de delicatement immobiliser l'insecte et appuyer sur le centre de l'abdomen pour faire sortir les segments terminaux. Les coleopteres mâles se reconnaissent par leur neuvieme sternite abdominal sclerifi ee t reticule. Chez les femelles, l'ovipositeur est bien visible; il se compose du tergite, des valviferes (neuvieme sternite) et des laterotergites du neuvieme segment abdominal. La procedure est rapide et ne blesse qu'un petit nombre d'individus (moins de 5%).

Thousand cankers disease, caused by the walnut twig beetle (Pityophthorus juglandis Blackman) and an associated fungal pathogen (Geosmithia morbida M. Kolarík, E. Freeland, C. Utley, and N. Tisserat), threatens the health and commercial use of eastern black walnut (Juglans nigra L.), one of the most economically valuable tree species in the United States. Effective phytosanitary measures are needed to reduce the possibility of spreading this insect and pathogen through wood movement. This study evaluated the efficacy of heat treatments and debarking to eliminate P. juglandis and C. morbida in J. nigra logs 4-18 cm in diameter and 30 cm in length. Infested logs were steam heated until various outer sapwood temperatures (60, 65, and 70 degrees C in 2011; 36, 42, 48, 52, and 56 degrees C in 2012) were maintained or exceeded for 30-40 min. In 2011, all heat treatments eliminated G. morbida from the bark, but logs were insufficiently colonized by P. juglandis to draw conclusions about treatment effects on the beetle. Debarking did not ensure elimination of the pathogen from the sapwood surface. In 2012, there was a negative effect of increasing temperature on P. juglandis emergence and G. morbida recovery. G. morbida did not survive in logs exposed to treatments in which minimum temperatures were 48 degrees C or higher, and mean P. juglandis emergence decreased steadily to zero as treatment minimum temperature increased from 36 to 52 degrees C. A minimum outer sapwood temperature of 56 degrees C maintained for 40 min is effective for eliminating the thousand cankers disease vector and pathogen from walnut logs, and the current heat treatment schedule for the emerald ash borer (60 degrees C core temperature for 60 min) is more than adequate for treating P. juglandis and G. morbida in walnut firewood.

The redbay ambrosia beetle (Xyleborus glabratus Eichhoff) is an invasive pest and vector of the pathogen that causes laurel wilt disease in Lauraceous tree species in the eastern United States. This insect uses olfactory cues during host finding, but use of visual cues by X. glabratus has not been previously investigated and may help explain diameter-related patterns in host tree mortality. The objective of this study was to determine whether X. glabratus females visually detect silhouettes of tree stems during host finding and are more likely to land on large diameter stems than smaller ones. Three field experiments were conducted in which stem silhouettes (black cylinders or standing nonhost pines) of varying diameters and identical capture surface areas were baited with essential oil lures. The Log10-transformed number of X. glabratus trapped per week increased as a function of silhouette diameter in 2011 and 2012, using artificial silhouette diameters ranging 2-18 and 3-41 cm, respectively. When lures and capture surfaces were attached to standing pines ranging 4-37 cm in diameter, a positive relationship between Log10(X. glabratus trap catch) and stem diameter was modeled using nonlinear quadratic plateau regression and indicated a diameter above which visual attraction was not enhanced; however, there was not a maximum diameter for enhanced X. glabratus attraction that was generally consistent across all experiments. These results 1) indicate that X. glabratus incorporates visual information during host finding, 2) help explain diameter-related patterns of redbay (Persea borbonia (L.) Sprengel) mortality observed during laurel wilt epidemics, and 3) are applicable to the management of this forest pest.

In 2007-2008, we examined the flight responses of Monochamus titillator (F.) complex [M. titillator, Monochamus carolinensis (Olivier), and any possible hybrids], Monochamus scutellatus (Say), Monochamus clamator (LeConte), Monochamus obtusus Casey, and Monochamus mutator LeConte (Coleoptera: Cerambycidae) to multiple-funnel traps baited with and without host volatiles and bark beetle pheromones. Experiments were conducted in mature pine (Pinus) stands in Alberta (Canada), and Arkansas, Arizona, California, Florida, Idaho, Michigan, Montana, New Hampshire, North Carolina, Ohio, Oregon, Tennessee, Utah, and Wisconsin (United States). At each location, traps were deployed in 10 replicate blocks of four traps per block. The trap treatments were: 1) blank control; 2) ipsenol and ipsdienol; 3) ethanol and alpha-pinene; and 4) a quaternary blend of ipsenol, ipsdienol, ethanol, and alpha-pinene. All five species or species complex of Monochamus preferred traps baited with the quaternary blend over all other treatments. The consistency of these results across such a large geographic area suggests that similar selection pressures may be acting on Monochamus spp. in pine forests, regardless of variation in stand composition and climatic conditions. Our results suggest that multiple-funnel traps baited with the quaternary blend ofipsenol, ipsdienol, ethanol, and alpha-pinene may be highly effective for monitoring various Monochamus spp. in pine forests of North America, and may have utility in trapping and detection programs in North America and overseas.

The redbay ambrosia beetle Xyleborus glabratus Eichhoff is a non-native vector of the pathogen that causes laurel wilt, a deadly disease of trees in the family Lauraceae in the southeastern U.S.A. 2 Concern exists that X. glabratus and its fungal symbiont could be transported to the western U.S.A. and cause damage to California bay laurel Umbellularia californica (Hook. & Arn.) Nutt. in California and Washington. 3 The present study evaluated in-flight attraction, attack density and emergence of X. glabratus and another invasive ambrosia beetle Xylosandrus crassiusculus (Motschulsky) on cut bolts of California bay laurel and eight related tree species in an infested forest in South Carolina. Xylosandrus crassiusculus is not a vector of the laurel wilt pathogen but is a pest of nursery and ornamental trees. 4 Mean catch of X. glabratus on California bay laurel bolts was not significantly different from catches on bolts of known X. glabratus hosts sassafras Sassafras albidum (Nutt.) Nees and swampbay Persea palustris (Raf.) Sarg. Mean attack density and adult emergence of both beetle species from California bay laurel was equal to or greater than all other tree species tested. Both beetle species readily produced brood in California bay laurel bolts. 5 The results obtained in the present study suggest that California bay laurel may be negatively impacted by both of these invasive ambrosia beetles if they become established in the tree's native range.

Hemlock woolly adelgid (Adelges tsugae Annad; or HWA) is a non-native invasive pest that attacks and kills eastern hemlock (Tsuga canadensis (L.) Carriere) and Carolina hemlock (Tsuga caroliniana Engelm.). Hemlock is a ‘‘foundation species’’ due to its strong influence on ecosystem structure and function, especially in riparian areas. HWA management involves the integrated use of multiple approaches including chemical control, biological control, cultural treatments, host resistance, and host gene conservation. Despite extensive control efforts, large areas in the eastern US, but especially in the southern Appalachian region, have experienced extensive hemlock mortality. Most of the short-term impacts of HWA induced mortality on ecosystem structure and function are localized and small; however, long-term impacts such as large pulses of woody debris and changes in species composition that impact structure and function could be significant. Using a decision analysis framework, land managers should begin to strategically implement land management decisions to address observed short-term impacts and plan and manage for projected longer-term impacts. In order to maintain ecosystem services in response to long-term impacts, restoration efforts may require novel approaches, such as the introduction of non-native species, facilitated movement of native species to new habitats (e.g., white pine), and aggressive management of existing species (e.g., Rhododendron) with mechanical removal, fire, or chemicals.

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.

Understanding the seasonal phenology of an insect pest in a specific geographic region is essential for optimizing the timing of management actions or research activities. We examined the phenology of hemlock woolly adelgid, Adelges tsugae Annand, near the southern limit of the range of eastern hemlock, Tsuga canadensis (L.) Carriere, in the Appalachians of northern Georgia, where adelgid phenology has not been previously reported. Adelges tsugae- infested hemlock trees were visited at various sites from 2004 - 2007. Two hemlock twigs were collected from each of 3 hemlock trees per site, except during the final 3 months of sampling when 1 twig was collected from each of 3 trees per site. Progrediens adults initiated oviposition by midMay, 2 - 4 weeks earlier than has been reported for more northern parts of the adelgid range. Sistens eggs were present until late-June (2006) or early-July (2004 - 2005). After aestivation, sistens nymphs resumed development by early October. Sistens adults were first found ...