Western Spruce Budworm Effects on Forest Resilience.

Western spruce budworm (Choristoneura freemani Razowski) is the most destructive defoliator of forests in the western US. Forests in northern New Mexico experienced high levels of WSBW-caused defoliation and subsequent mortality between the 1980s and 2010s. The effects of severe western spruce budworm outbreaks on stand dynamics in the US Southwest are still relatively unknown, but understanding the impacts is important to the management and resilience of these forests. To begin addressing this knowledge gap, we conducted a study along two gradients: an elevational gradient from mixed-conifer to spruce-fir forests and a gradient of WSBW-caused defoliation intensity. We recorded overstory and understory stand conditions (size structure, species composition, damaging agents). Western spruce budworm was the primary damaging agent of host trees in all stands andcaused host tree mortality across all size classes, particularly in spruce-fir stands. Results indicate an unsustainable level of mortality in spruce-fir stands and a transition towards non-host species in mixed-conifer stands. Low levels of regeneration coupled with high overstory mortality rates indicate a potential lack of resilience in spruce-fir stands, whereas resilience to future western spruce budworm defoliation events may have increased in mixed-conifer stands affected by these outbreaks.

A Biodegradable Formulation of MCH (3-Methylcyclohex-2-en-1-one) for Protecting Pseudotsuga menziesii from Dendroctonus pseudotsugae (Coleoptera: Curculionidae) Colonization.

Douglas-fir, Pseudotsuga menziesii (Mirb.) Franco, trees and stands can be protected from Douglas-fir beetle, Dendroctonus pseudotsugae Hopkins (DFB)-caused mortality by application of synthetic formulations of the beetle's antiaggregation pheromone, 3-methylcyclohex-2-en-1-one (MCH). A biodegradable formulation of MCH, SPLAT MCH, was developed and evaluated for protecting individual Douglas-fir trees and small stands from colonization and mortality by DFB. In an individual-tree experiment in Idaho, both MCH bubble capsules and SPLAT MCH significantly reduced the proportion of treated trees colonized and killed by DFB compared to untreated controls. SPLAT MCH was as effective as MCH bubble capsules for protecting individual trees. Both MCH bubble capsules and SPLAT MCH significantly reduced the proportion of trees colonized and killed by DFB within 0.04-ha circular plots surrounding each treated tree compared to untreated controls. In 0.41 ha stands in New Mexico, both MCH bubble capsules and SPLAT MCH significantly reduced the proportion of trees colonized and killed by DFB compared to untreated controls, again with no differences observed between MCH treatments. In a similar stand level trial in Idaho, neither MCH treatment significantly reduced the proportion of trees colonized by DFB, and only MCH bubble capsules significantly reduced levels of tree mortality compared to untreated controls, but no significant difference was observed between SPLAT MCH and MCH bubble capsules. Overall, the results indicate that SPLAT MCH is as effective as MCH bubble capsules for protecting individual trees and small stands of Douglas-fir from DFB-caused mortality.

Advances in Semiochemical Repellents to Mitigate Host Mortality From the Spruce Beetle (Coleoptera: Curculionidae).

We tested 3-methyl-2-cyclohexen-1-one (MCH) and novel semiochemicals as potential spruce beetle (Dendroctonus rufipennis Kirby) (Coleoptera: Curculionidae, Scolytinae) repellents over multiple years in Utah and Colorado trapping bioassays. MCH is a known spruce beetle repellent and our testing revealed Acer kairomone blend (AKB) and isophorone plus sulcatone as repellents. We subsequently tested these semiochemicals for area and single tree protection to prevent spruce beetle attacks at locations in Utah, Colorado, Wyoming, New Mexico, and Alaska. Individual tree protection trials found MCH-AKB provided significant protection against spruce beetle attacks in the southern Rocky Mountains but not in Alaska. Adding sulcatone or doubling MCH-AKB pouches did not further enhance protection. A degree of protection was extended to spruce at least 10 m distant from the repellents, including in Alaska. Tree diameter was not a significant covariate among treated trees but was positively correlated with the probability of infestation for surrounding spruce. In area protection trials, spruce in control plots were 2.4 times more likely to be in a higher severity attack class compared with spruce in plots treated with MCH-AKB pouches deployed at 30 sets per hectare. Tree diameter had a significant, positive relationship to the probability of infestation. We found MCH-AKB to offer a high degree of protection against beetle attack in Engelmann spruce (Picea engelmannii Parry ex Engelm.) (Pinales: Pinaceae) (Picea engelmannii Parry ex Engelm.) (Pinales: Pinaceae), especially for single tree protection (66% of control trees were strip- or mass-attacked compared with 6% of repellent-treated trees). AKB requires registration and labeling, however, before this economical and environmentally benign semiochemical can be used operationally.

Mycobiota associated with insect galleries in walnut with thousand cankers disease reveals a potential natural enemy against Geosmithia morbida.

Thousand Cankers Disease (TCD) affects Juglans and Pterocarya species. This disease poses not only a major threat to the nut and timber industries but also to native stands of walnut trees. Galleries created by Pityophthorus juglandis (vector) are colonized by the fungus Geosmithia morbida (causal agent of necrosis). It is unknown if other fungi colonizing these galleries might act antagonistically towards G. morbida. The objectives of this study were to: (1) characterize the fungal community associated with TCD-infected trees and (2) develop a pilot study addressing their potential antagonism towards G. morbida. We collected non-Geosmithia fungi from ten TCD-infected walnut trees from California and Tennessee. Four hundred and fifty-seven isolates, representing sixty-five Operational Taxonomic Units (99 % ITS similarity) were obtained. Fungal communities were found to be highly diverse. Ophiostoma dominated the communities associated with TCD-compromised trees from California, whereas Trichoderma dominated TCD-compromised trees in Tennessee. Six Trichoderma isolates showed varying levels of antagonism against three isolates of G. morbida, suggesting potential applications for the biological control of TCD. Furthermore, results from this study contribute to the growing knowledge about the observed differential disease development between the western and eastern USA and could overall impact our understanding of TCD etiology.

Efficacy of Systemic Insecticides for Control of the Invasive Goldspotted Oak Borer (Coleoptera: Buprestidae) in California.

From 2009 to 2013, we tested four systemic insecticide formulations and five application methods against the invasive goldspotted oak borer, Agrilus auroguttatus Schaeffer (Coleoptera: Buprestidae), in California. The insecticides were evaluated in three experiments: 1) 2009 remedial applications of emamectin benzoate (stem-injection) and imidacloprid (stem-injection and soil-injection); 2) 2009-2012 emamectin benzoate and imidacloprid initially applied at different times during the dormant season with varying injection technologies; and 3) 2013 dinotefuran applied to several tree diameter size classes. Adult leaf-feeding bioassays were used to assess the impact of systemic treatments against A. auroguttatus, whereas enzyme-linked immunosorbent assays determined the quantity of the active ingredient of insecticide residues in foliage. Imidacloprid (experiment 1) persisted at elevated levels in foliage of coast live oak, Quercus agrifolia Née, for 1.5 yr following stem injections. Stem injections of emamectin benzoate (experiment 2) sometimes significantly decreased survival in adults fed foliage from treated Q. agrifolia, and both the emamectin benzoate and imidacloprid treatments reduced adult feeding in some trials. Imidacloprid residues in Q. agrifolia and California black oak, Quercus kelloggii Newb., foliage remained at elevated levels (>10 µg/g) ∼2 yr postapplication. In 2013 (experiment 3), dinotefuran residues were highest in foliage collections 2 wk postapplication and greatest in smaller diameter oaks, but insecticide treatment had no effect on survival or frass production by adults fed foliage from treated trees. Systemic injections of emamectin benzoate and imidacloprid applied during the dormant season to uninfested or lightly infested oaks can reduce adult A. auroguttatus survival and maturation feeding.

3-Methylcyclohex-2-en-1-one for area and individual tree protection against spruce beetle (Coleoptera: Curculionidae: Scolytinae) attack in the southern Rocky Mountains.

We tested 3-methylcyclohex-2-en-1-one (MCH) and an Acer kairomone blend (AKB) as repellent semiochemicals for area and single tree protection to prevent spruce beetle (Dendroctonus rufipennis Kirby) attacks at locations in Utah and New Mexico. In the area protection study, we compared host infestation rates of MCH applications at three densities (20, 40, and 80 g MCH ha-1) against a control treatment over 0.64 ha plots centered within ~1.25 ha treatment blocks. All treatments included two baited funnel traps within the plot to assure spruce beetle pressure. Following beetle attack, plots were surveyed for new spruce beetle attacks and to quantify stand characteristics. The probability of more severe spruce beetle attacks was significantly reduced, by ~50%, in each of the MCH area treatments compared with the control treatment but there was no significant treatment difference among the MCH deployment densities. For the single tree protection study, we compared attack rates of MCH, Acer kairomone blend (AKB), and MCH plus AKB on spruce beetle-baited trees against bait-only trees. Each treatment was applied over a range of host diameters to test for host size effects. Seventy-five percent of control trees were mass-attacked, about one-third of MCH- and AKB-alone spruce was mass-attacked, and no MCH plus AKB spruce were mass-attacked. These results suggest that MCH alone is a marginal area and single tree protectant against spruce beetle but that deployment with other repellents can significantly increase treatment efficacy.

Effect of Contact Insecticides Against the Invasive Goldspotted Oak Borer (Coleoptera: Buprestidae) in California.

The goldspotted oak borer, Agrilus auroguttatus Schaeffer (Coleoptera: Buprestidae), was linked in 2008 to ongoing tree mortality in oak woodlands of southern California. Mortality of coast live oak, Quercus agrifolia Née, and California black oak, Q. kelloggii Newb., continues as this exotic phloem borer spreads in southern California. Management options are needed to preserve high-value oaks and maintain management objectives. From 2009 to 2012, we tested four contact insecticide formulations in four experiments against A auroguttatus in California. The impact of contact insecticides was evaluated ∼<1, 8, and 12 mo postapplication against A auroguttatus adults in no-choice leaf-feeding or walking bioassays. At <1 mo postapplication, bifenthrin, carbaryl, lambda-cyhalothrin, and permethrin all reduced adult survival and feeding in leaf-feeding and walking bioassays. At 8 mo postapplication, only bifenthrin reduced adult feeding, but had no effect on survivorship. At 12 mo postapplication, adult A auroguttatus survived fewer days and fed less in leaf-feeding bioassays with bifenthrin, carbaryl, and permerthin. These results support the annual application of contact insecticides prior to A auroguttatus' flight period to reduce adult leaf maturation feeding and activity on the bark surface (e.g., oviposition), but additional studies are needed to show these contact treatments can prevent tree mortality from this invasive species.

Phylogeography of the walnut twig beetle, Pityophthorus juglandis, the vector of thousand cankers disease in North American walnut trees.

Thousand cankers disease (TCD) of walnut trees (Juglans spp.) results from aggressive feeding in the phloem by the walnut twig beetle (WTB), Pityophthorus juglandis, accompanied by inoculation of its galleries with a pathogenic fungus, Geosmithia morbida. In 1960, WTB was only known from four U.S. counties (in Arizona, California, and New Mexico), but the species has now (2014) invaded over 115 counties, representing much of the western USA, and at least six states in the eastern USA. The eastern expansion places TCD in direct proximity to highly valuable (> $500 billion) native timber stands of eastern black walnut, Juglans nigra. Using mitochondrial DNA sequences, from nearly 1100 individuals, we examined variation among 77 samples of WTB populations across its extended range in the USA, revealing high levels of polymorphism and evidence of two divergent lineages. The highest level of genetic diversity for the different lineages was found in the neighboring Madrean Sky Island and Western New Mexico regions, respectively. Despite their proximity, there was little evidence of mixing between these regions, with only a single migrant detected among 179 beetles tested. Indeed, geographic overlap of the two lineages was only common in parts of Colorado and Utah. Just two haplotypes, from the same lineage, predominated over the vast majority of the recently expanded range. Tests for Wolbachia proved negative suggesting it plays no role in "driving" the spread of particular haplotypes, or in maintaining deep levels of intraspecific divergence in WTB. Genotyping of ribosomal RNA corroborated the mitochondrial lineages, but also revealed evidence of hybridization between them. Hybridization was particularly prevalent in the sympatric areas, also apparent in all invaded areas, but absent from the most haplotype-rich area of each mitochondrial lineage. Hypotheses about the specific status of WTB, its recent expansion, and potential evolutionary origins of TCD are discussed.

Population structure of Geosmithia morbida, the causal agent of thousand cankers disease of walnut trees in the United States.

The ascomycete Geosmithia morbida and the walnut twig beetle Pityophthorus juglandis are associated with thousand cankers disease of Juglans (walnut) and Pterocarya (wingnut). The disease was first reported in the western United States (USA) on several Juglans species, but has been found more recently in the eastern USA in the native range of the highly susceptible Juglans nigra. We performed a comprehensive population genetic study of 209 G. morbida isolates collected from Juglans and Pterocarya from 17 geographic regions distributed across 12 U.S. states. The study was based on sequence typing of 27 single nucleotide polymorphisms from three genomic regions and genotyping with ten microsatellite primer pairs. Using multilocus sequence-typing data, 197 G. morbida isolates were placed into one of 57 haplotypes. In some instances, multiple haplotypes were recovered from isolates collected on the same tree. Twenty-four of the haplotypes (42%) were recovered from more than one isolate; the two most frequently occurring haplotypes (H02 and H03) represented 36% of all isolates. These two haplotypes were abundant in California, but were not recovered from Arizona or New Mexico. G. morbida population structure was best explained by four genetically distinct groups that clustered into three geographic regions. Most of the haplotypes isolated from the native range of J. major (Arizona and New Mexico) were found in those states only or present in distinct genetic clusters. There was no evidence of sexual reproduction or genetic recombination in any population. The scattered distribution of the genetic clusters indicated that G. morbida was likely disseminated to different regions at several times and from several sources. The large number of haplotypes observed and the genetic complexity of G. morbida indicate that it evolved in association with at least one Juglans spp. and the walnut twig beetle long before the first reports of the disease.

Developing monitoring techniques for the invasive goldspotted oak borer (Coleoptera: Buprestidae) in California.

The goldspotted oak borer, Agrilus auroguttatus Schaeffer (Coleoptera: Buprestidae), is an invasive species that has colonized oak woodlands in southern California. To better define its seasonal flight activity, assist with forest and integrated pest management activities, and define the current distribution in California, an effective monitoring technique for A. auroguttatus is necessary. We assessed the efficacy of two colors of flight-intercept prism traps, the placement of these traps at three heights, and several commercially available lures [Manuka oil, Phoebe oil, and a green leaf volatile, (3Z)-hexenol] for monitoring the flight of adult A. auroguttatus. Landing rates and the densities of D-shaped emergence holes of A. auroguttatus adults were assessed on the lower stems of coast live oak, Quercus agrifolia Née, of varying size and crown health classes. Purple flight-intercept prism traps placed at heights of 3 m and 4.5 m caught significantly more female A. auroguttatus than green prism traps. In one experiment, males also responded at a significantly higher level to purple than to green prism traps placed at 3 m height. The addition of commercially available lures significantly enhanced male, but not female, A. auroguttatus trap catch when compared with unbaited control traps. There were no differences among male flight responses to the three lures. A. auroguttatus landing rates and emergence hole densities were significantly greater on the largest-diameter trees (>76.2 cm diameter at breast height) and on trees with severe crown thinning or complete crown collapse. The annual increment in emergence hole densities was also significantly greater on trees with severe crown thinning or complete crown collapse. In three trapping studies over multiple years in southern California, the adult flight period began as early as mid-May, peaked in mid-June to early July, and ended in early- to mid-September. To demonstrate the efficacy of the detection method for A. auroguttatus (unbaited purple traps at 3 m height), a delimitation survey conducted from 2009 to 2012 confirmed that the species was only present in San Diego Co., but that the distribution was expanding northward.

Sanitation options for managing oak wood infested with the invasive goldspotted oak borer (Coleoptera: Buprestidae) in southern California.

Movement of invasive wood-boring insects in wood products presents a threat to forest health and a management challenge for public and private land managers. The goldspotted oak borer, Agrilus auroguttatus Schaeffer (Coleoptera: Buprestidae), is a new pest in San Diego and Riverside Cos., CA, believed to have been introduced on firewood. This beetle has caused elevated levels of oak mortality since 2002. From 2009-2011, we tested several sanitation methods, applicable to large and small land parcels, to reduce or prevent goldspotted oak borer emergence from infested oak wood. In most experiments, emergence of goldspotted oak borer adults from the positive controls demonstrated that the beetle could complete development in firewood-sized pieces of cut oak wood. In 2009, adult emergence from sun-exposed oak wood began and peaked 2- to 4-wks earlier at a low elevation site than at a high elevation site (late May to late June). However, there were no significant effects of elevation or host species on the emergence response of goldspotted oak borer by solarization treatment in this study. Solarization of infested wood with thick (6 mil) and thin (1 mil) plastic tarpaulins (tarps) did not significantly reduce emergence of adults despite recordings of greater mean and maximum daily temperatures in both tarped treatments and greater relative humidity in the thick-tarped treatment (all compared with nontarped controls). Grinding wood with a 3"-minus screen (< or = 7.6 cm) significantly reduced goldspotted oak borer emergence compared with control treatments, and this was the best method for reducing adult emergence among those tested. In a separate grinding study, no adults emerged when wood was ground to 9"-minus (22.9 cm), 2"-minus (5.1 cm), or 1"-minus (2.5 cm) screen sizes, but a low level of adult emergence from the positive controls limited any inferences from this experiment. Debarking cut wood pieces eliminated goldspotted oak borer emergence from the wood fraction, but adults emerged from the shaved bark and phloem.

Susceptibility of Walnut and Hickory Species to Geosmithia morbida.

Thousand cankers disease (TCD) of walnut is a result of feeding in the phloem by the walnut twig beetle (WTB), Pityophthorus juglandis, and subsequent canker formation caused by Geosmithia morbida around galleries. TCD has caused extensive morbidity and mortality to Juglans nigra in the western United States and, in 2010, was discovered in the eastern United States, where the tree is a highly valuable timber resource. WTB and G. morbida also have been found in J. regia orchards throughout major production areas in California, and the numbers of damaged trees are increasing. We tested the susceptibility of walnut and hickory species to G. morbida in greenhouse and field studies. Carya illinoinensis, C. aquatica, and C. ovata were immune. All walnut species tested, including J. ailantifolia, J. californica, J. cinerea, J. hindsii, J. major, J. mandshurica, J. microcarpa, J. nigra, and J. regia, developed cankers following inoculation with G. morbida. J. nigra was the most susceptible, whereas J. major, a native host of the WTB and, presumably, G. morbida, had smaller and more superficial cankers. Canker formation differed among maternal half-sibling families of J. nigra and J. cinerea, indicating genetic variability in resistance to G. morbida. Our inoculation studies with G. morbida have corroborated many of the field observations on susceptibility of walnut and hickory species to TCD, although the ability of the WTB to successfully attack and breed in walnut is also an important component in TCD resistance.

Management of a patient with a clot-filled pulmonary arteriovenous malformation.

Transcatheter coil embolization is an established treatment for pulmonary arteriovenous malformations (PAVMs). Encountering a patient with a symptomatic PAVM partially occluded with a prominent clot is rare. Successful and safe coil embolization of such a PAVM is described in this case report.