Developing semiochemical repellents for protecting Picea from Dendroctonus rufipennis (Coleoptera: Curculionidae) in Alaska and Utah, USA.

Spruce beetle, Dendroctonus rufipennis (Kirby) (Coleoptera: Curculionidae), is the most destructive pest of mature spruce (Picea) in western North America. Recent outbreaks in Alaska and other western US states highlight the need for tools to protect Picea from D. rufipennis. The primary antiaggregation pheromone of D. rufipennis (3-methylcyclohex-2-en-1-one, MCH) and various combinations of potential repellents (1-octen-3-ol, exo-brevicomin, endo-brevicomin, ipsdienol, ipsenol, limonene, and verbenone) were tested for their ability to disrupt the response of D. rufipennis to attractant-baited multiple-funnel traps. Two assays were conducted on the Kenai Peninsula, Alaska, in June and July 2021. All treatments significantly reduced the mean number of D. rufipennis caught compared to the baited control. No other significant differences were observed among treatments. Informed by these and other data, tree protection studies were established in Lutz spruce, Picea × lutzii, on the Kenai Peninsula in 2022 and in Engelmann spruce, Pi. engelmannii, in the Uinta Mountains, Utah, in 2021. All experimental trees were baited with frontalin. Repellent treatments included MCH (SPLAT MCH, ISCA Inc., Riverside, CA, USA) and at least 1 additional repellent combination. In Alaska, all treatments significantly reduced colonization (strip attacks + mass attacks) and mortality of individually treated Pi. × lutzii and all Picea within 11.3-m radius of each treated Pi. × lutzii compared to the control. In Utah, all treatments except for SPLAT MCH + octenol significantly reduced colonization compared to the control. Only SPLAT MCH + Acer kairomone blend (AKB) and SPLAT MCH + octenol reduced Pi. engelmannii mortality compared to the control. SPLAT MCH + AKB and SPLAT MCH + acetophenone and green leaf volatiles (PLUS) were the most effective across both studies. The implications of these and other results to the development of an effective semiochemical repellent for D. rufipennis are discussed.

MCH-Based Semiochemical Repellents for Protecting Engelmann Spruce Trees From Dendroctonus rufipennis (Coleoptera: Curculionidae).

Spruce beetle, Dendroctonus rufipennis Kirby (Coleoptera: Curculionidae), is a lethal pest of spruce trees in North America. Despite decades of research, a semiochemical repellent that consistently and effectively protects spruce trees remains elusive. We evaluated the efficacy of 3-methyl-2-cyclohexen-1-one (MCH) in a proprietary, volatile compound release technology (SPLAT) alone and with two adjuvants, Acer kairomone blend (AKB) and acetophenone + green leaf volatiles (PLUS) to protect individually treated Picea engelmannii Parry ex. Engelm. (Pinales: Pinaceae), and Pi. engelmannii within 11.3-m radius of the individually treated trees from colonization and mortality attributed to D. rufipennis in western Wyoming. Ninety-one Pi. engelmannii were baited with frontalin and randomly assigned to one of seven treatments (n = 13): 3.5 g of MCH applied as SPLAT MCH (SPLAT3.5), 3.5AKB, 3.5PLUS, 7 g of MCH applied as SPLAT MCH (SPLAT7), 7AKB, 7PLUS, and baited control (bait only). All repellents except SPLAT3.5 and SPLAT7 significantly reduced colonization of individually treated Pi. engelmannii compared to the baited control. 3.5PLUS, 7AKB, and 7PLUS reduced colonization most effectively, and all repellents significantly reduced mortality of individually treated Pi. engelmannii compared to the baited control. All repellents also significantly reduced colonization and mortality of neighboring Pi. engelmannii.

Comparisons of Efficiency of Two Formulations of Verbenone (4, 6, 6-trimethylbicyclo [3.1.1] hept-3-en-2-one) for Protecting Whitebark Pine, Pinus albicaulis (Pinales: Pinaceae) From Mountain Pine Beetle (Colopetera: Curculionidae).

Whitebark pine, Pinus albicaulis Engelm., is a subalpine tree endemic to western North America. This species provides multiple ecosystem services and is suffering widespread mortality from mountain pine beetle, Dendroctonus ponderosae Hopkins. Verbenone is a pheromone produced as D. ponderosae feed, and high air concentrations of verbenone deter D. ponderosae from colonizing trees. Synthetic verbenone has been formulated into products used to prevent D. ponderosae from colonizing trees. We compared the ability of verbenone pouches and SPLAT Verb to protect individuals and small stands of P. albicaulis. With individual trees in Montana, all treated trees survived regardless of verbenone formulation and rate, whereas untreated trees suffered 70 and 90% mortality in 2015 and 2016. In plot experiments in California from 2015 to 2017, and Oregon from 2015 to 2018, verbenone was applied to trees spaced ~10 m apart, and survival of small (12.7-23 cm DBH = diameter at 1.37 m height), medium (23.1-33 cm DBH) and large (>33 cm DBH) trees was compared. In California, where >80% of untreated trees survived, pouches increased survival ~2 to 3% and SPLAT Verb increased survival ~4 to 7% regardless of tree size. In Oregon, verbenone pouches and SPLAT Verb performed similarly on medium and small trees, but large trees had greater survival when treated with SPLAT Verb (~93%) than pouches (~82%). Compared to verbenone pouches, SPLAT Verb appears to better protect P. albicaulis from D. ponderosae.

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.

Efficacy of Spring and Fall Treatments of Carbaryl for Protecting Ponderosa Pine From Mortality Attributed to Mountain Pine Beetle (Coleoptera: Curculionidae).

High-value trees, such as those growing in residential, recreational, or administrative sites, are often susceptible to colonization by bark beetles (Coleoptera: Curculionidae: Scolytinae) as a result of increased amounts of stress associated with off-site plantings, drought, soil compaction, and/or mechanical injury. The value of these trees, cost of removing dead trees, and loss of aesthetics often justify the use of insecticides to protect trees from mortality attributed to bark beetles. Carbaryl (1-naphthyl methylcarbamate) is among the most effective, economically-viable, and ecologically-compatible insecticides available for protecting conifers from several species of bark beetles in the western United States. Treatments are usually applied in spring prior to initiation of flight of the target species. We evaluated the efficacy of spring and fall applications of carbaryl for protecting individual ponderosa pine, Pinus ponderosa Dougl. ex Laws. (Pinales: Pinaceae), from mortality attributed to mountain pine beetle, Dendroctonus ponderosae Hopkins (Coleoptera: Curculionidae), in Idaho. Both spring and fall treatments of 2.0% a.i. carbaryl (maximum label rate; Sevin SL, Bayer Environmental Science, Montvale, NJ 07645) provided one field season of protection, and thus should be applied annually if tree protection is desired for multiple years. Our research also provides some insight on the efficacy of carbaryl treatments after wildfire. We found no evidence that a mixed-severity wildfire negatively affected the efficacy of carbaryl treatments.

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.

Lethal Trap Trees and Semiochemical Repellents as Area Host Protection Strategies for Spruce Beetle (Coleoptera: Curculionidae, Scolytinae) in Utah.

We tested lethal trap trees and repellent semiochemicals as area treatments to protect host trees from spruce beetle (Dendroctonus rufipennis Kirby) attacks. Lethal trap tree treatments ("spray treatment") combined a spruce beetle bait with carbaryl treatment of the baited spruce. Repellent treatments ("spray-repellent") combined a baited lethal trap tree within a 16-m grid of MCH (3-methylcyclohex-2-en-1-one) and two novel spruce beetle repellents. After beetle flight, we surveyed all trees within 50 m of plot center, stratified by 10-m radius subplots, and compared attack rates to those from baited and unbaited control plots. Compared to the baited controls, spruce in the spray treatment had significantly reduced likelihood of a more severe attack classification (e.g., mass-attacked over strip-attacked or unsuccessful-attacked over unattacked). Because spruce in the spray treatment also had significantly heightened probability of more severe attack classification than those in the unbaited controls, however, we do not recommend lethal trap trees as a stand-alone beetle suppression strategy for epidemic beetle populations. Spruce in the spray-repellent treatment were slightly more likely to be classified as more severely attacked within 30 m of plot center compared to unbaited controls but, overall, had reduced probabilities of beetle attack over the entire 50-m radius plots. The semiochemical repellents deployed in this study were effective at reducing attacks on spruce within treated plots despite the presence of a centrally located spruce beetle bait. Further testing will be required to clarify operational protocols such as dose, elution rate, and release device spacing.

Pine Sawyers (Coleoptera: Cerambycidae) Attracted to α-Pinene, Monochamol, and Ipsenol in North America.

Detection tools are needed for Monochamus species (Coleoptera: Cerambycidae) because they are known to introduce pine wilt disease by vectoring nematodes in Asia, Europe, and North America. In 2012-2014, we examined the effects of the semiochemicals monochamol and ipsenol on the flight responses of the sawyer beetles Monochamus carolinensis (Olivier), Monochamus clamator (LeConte), Monochamus mutator LeConte, Monochamus notatus (Drury), Monochamus obtusus Casey, Monochamus scutellatus (Say), and Monochamus titillator (F.) complex (Coleoptera: Cerambycidae) to traps baited with α-pinene. Experiments were set in pine forests in New Brunswick and Ontario (Canada), and Arizona, Georgia, Michigan, Montana, Oregon, South Carolina, Utah, and Washington (United States). In brief, 40 traps were placed in 10 blocks of 4 traps per block per location. Traps were baited with: 1) α-pinene; 2) α-pinene + monochamol; 3) α-pinene + ipsenol; and 4) α-pinene + monochamol + ipsenol. Monochamol increased catches of six species and one species complex of Monochamus with an additive effect of ipsenol for five species and one species complex. There was no evidence of synergy between monochamol and ipsenol on beetle catches. Monochamol had no effect on catches of other Cerambycidae or on any associated species of bark beetles, weevils, or bark beetle predators. We present a robust data set suggesting that the combination of α-pinene, ipsenol, and monochamol may be a useful lure for detecting Monochamus species.

A Novel Semiochemical Tool for Protecting Pinus contorta From Mortality Attributed to Dendroctonus ponderosae (Coleoptera: Curculionidae).

Verbenone (4,6,6-trimethylbicyclo[3.1.1]hept-3-en-2-one) is an antiaggregant of the mountain pine beetle, Dendroctonus ponderosae Hopkins (Coleoptera: Curculionidae), a notable forest insect capable of causing extensive levels of tree mortality in western North America. Several formulations of verbenone are registered for tree protection, but failures in efficacy are not uncommon, particularly when applied during large infestations. A formulation of (-)-verbenone was developed (Specialized Pheromone & Lure Application Technology [SPLAT] Verb, ISCA Technologies Inc., Riverside, CA) and evaluated for protecting individual lodgepole pine, Pinus contorta Douglas ex Loudon, and small stands of P. contorta from mortality attributed to D. ponderosae. SPLAT Verb applied to individual P. contorta resulted in complete tree protection, while 93.3% mortality occurred in the untreated controls. Significantly fewer P. contorta were killed by D. ponderosae within 0.041-ha circular plots surrounding P. contorta treated with SPLAT Verb compared with the untreated control. In a second study, a smaller percentage of P. contorta were colonized and killed on 0.4-ha square plots treated with SPLAT Verb compared with the untreated control. No significant differences in levels of tree mortality were observed between the untreated control and another formulation of verbenone (7-g pouch) or between the 7-g pouch and SPLAT Verb. In a trapping bioassay, no significant differences were observed among captures in multiple-funnel traps at 1, 2, or 4 m from the point of release of SPLAT Verb. Significantly fewer D. ponderosae were collected at 1 and 2 m compared with 8 m. Significantly more D. ponderosae were captured at the farthest distance evaluated (16 m) than at any other distance. Our data indicate that SPLAT Verb is effective for protecting individual P. contorta and small stands of P. contorta from mortality attributed to D. ponderosae at moderate doses. The high levels of tree protection observed are attributed to the ability of applying release points (dollops) at high densities, and a larger zone of inhibition than reported for other formulations of verbenone. SPLAT Verb was registered by the U.S. Environmental Protection Agency for use on pines, Pinus spp., in 2013.

Evaluations of emamectin benzoate and propiconazole for protecting individual Pinus contorta from mortality attributed to colonization by Dendroctonus ponderosae and associated fungi.

BACKGROUND: Protection of conifers from bark beetle colonization typically involves applications of liquid formulations of contact insecticides to the tree bole. An evaluation was made of the efficacy of bole injections of emamectin benzoate alone and combined with the fungicide propiconazole for protecting individual lodgepole pine, Pinus contorta Dougl. ex Loud., from mortality attributed to colonization by mountain pine beetle, Dendroctonus ponderosae Hopkins, and progression of associated blue stain fungi. RESULTS: Injections of emamectin benzoate applied in mid-June did not provide adequate levels of tree protection; however, injections of emamectin benzoate + propiconazole applied at the same time were effective for two field seasons. Injections of emamectin benzoate and emamectin benzoate + propiconazole in mid-September provided tree protection the following field season, but unfortunately efficacy could not be determined during a second field season owing to insufficient levels of tree mortality observed in the untreated control, indicative of low D. ponderosae populations. CONCLUSION: Previous evaluations of emamectin benzoate for protecting P. contorta from mortality attributed to D. ponderosae have failed to demonstrate efficacy, which was later attributed to inadequate distribution of emamectin benzoate following injections applied several weeks before D. ponderosae colonization. The present data indicate that injections of emamectin benzoate applied in late summer or early fall will provide adequate levels of tree protection the following summer, and that, when emamectin benzoate is combined with propiconazole, tree protection is afforded the year that injections are implemented.

Trap lure blend of pine volatiles and bark beetle pheromones for Monochamus spp. (Coleoptera: Cerambycidae) in pine forests of Canada and the United States.

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.

Population densities and tree diameter effects associated with verbenone treatments to reduce mountain pine beetle-caused mortality of lodgepole pine.

Mountain pine beetle, Dendroctonus ponderosae Hopkins (Coleoptera: Curculionidae: Scolytinae), is among the primary causes of mature lodgepole pine, Pinus contorta variety latifolia mortality. Verbenone is the only antiaggregant semiochemical commercially available for reducing mountain pine beetle infestation of lodgepole pine. The success of verbenone treatments has varied greatly in previous studies because of differences in study duration, beetle population size, tree size, or other factors. To determine the ability of verbenone to protect lodgepole pine over long-term mountain pine beetle outbreaks, we applied verbenone treatments annually for 3 to 7 yr at five western United States sites. At one site, an outbreak did not develop; at two sites, verbenone reduced lodgepole pine mortality in medium and large diameter at breast height trees, and at the remaining two sites verbenone was ineffective at reducing beetle infestation. Verbenone reduced mountain pine beetle infestation of lodgepole pine trees in treated areas when populations built gradually or when outbreaks in surrounding untreated forests were of moderate severity. Verbenone did not protect trees when mountain pine beetle populations rapidly increase.

Spray deposition from ground-based applications of carbaryl to protect individual trees from bark beetle attack.

Bark beetles (Coleoptera: Curculionidae, Scolytinae) are recognized as the most important tree mortality agent in western coniferous forests. A common method of protecting trees from bark beetle attack is to saturate the tree bole with carbaryl (1-naphthyl methylcarbamate) using a hydraulic sprayer. In this study, we evaluate the amount of carbaryl drift (ground deposition) occurring at four distances from the tree bole (7.6, 15.2, 22.9, and 38.1 m) during conventional spray applications for protecting individual lodgepole pine (Pinus contorta Dougl. ex Loud.) from mountain pine beetle (Dendroctonus ponderosae Hopkins) attack and Engelmann spruce (Picea engelmannii Parry ex Engelm.) from spruce beetle (D. rufipennis [Kirby]) attack. Mean deposition (carbaryl + alpha-naphthol) did not differ significantly among treatments (nozzle orifices) at any distance from the tree bole. Values ranged from 0.04 +/- 0.02 mg carbaryl m(-2) at 38.1 m to 13.30 +/- 2.54 mg carbaryl m(-2) at 7.6 m. Overall, distance from the tree bole significantly affected the amount of deposition. Deposition was greatest 7.6 m from the tree bole and quickly declined as distance from the tree bole increased. Approximately 97% of total spray deposition occurred within 15.2 m of the tree bole. Application efficiency (i.e., percentage of insecticide applied that is retained on trees) ranged from 80.9 to 87.2%. Based on review of the literature, this amount of drift poses little threat to adjacent aquatic environments. No-spray buffers of 7.6 m should be sufficient to protect freshwater fish, amphibians, crustaceans, bivalves, and most aquatic insects. Buffers >22.9 m appear sufficient to protect the most sensitive aquatic insects (Plecoptera).

Soil enzyme inhibition by condensed litter tannins may drive ecosystem structure and processes: the case of Kalmia angustifolia.

Kalmia angustifolia is an ericaceous shrub that can rapidly spread on recently harvested boreal forest sites, causing a slow-down in soil nutrient cycling and reduced growth of spruce seedlings. It has been hypothesized that tannins released from Kalmia litter suppress soil enzyme activity, and are thus important in controlling ecosystem structure and processes. Here the effects of different concentrations of tannins extracted from both Kalmia and black spruce (Picea mariana) foliage were tested on enzyme activities of soil extracts. Then the effects of various Kalmia-black spruce litter mixtures on soil enzyme activity were investigated. Lastly, the correlation between Kalmia cover in the field and soil enzyme activity was measured. Both tannin types suppressed beta-glucosidase and acid phosphatase activities, and the magnitude of these effects was concentration-dependent. beta-glucosidase and amidase activity decreased linearly with an increasing Kalmia : spruce litter ratio added to soil. A field survey of 24 sites revealed a negative relationship between percentage Kalmia cover and beta-glucosidase activity. Collectively, results of the three experiments converge to support the claim that enzyme inhibition by litter tannins has evolved as an important mechanism controlling ecosystem processes and structure following Kalmia invasion on recently disturbed forest sites.

Risk assessment in the face of a changing environment: gypsy moth and climate change in Utah.

The importance of efficaciously assessing the risk for introduction and establishment of pest species is an increasingly important ecological and economic issue. Evaluation of climate is fundamental to determining the potential success of an introduced or invasive insect pest. However, evaluating climatic suitability poses substantial difficulties; climate can be measured and assessed in a bewildering array of ways. Some physiological filter, in essence a lens that focuses climate through the requirements and constraints of a potential pest introduction, is required. Difficulties in assessing climate suitability are further exacerbated by the effects of climate change. Gypsy moth (Lymantria dispar L.) is an exotic, tree-defoliating insect that is frequently introduced into the western United States. In spite of an abundance of potential host species, these introductions have yet to result in established populations. The success of eradication efforts and the unsuccessful establishment of many detected and undetected introductions may be related to an inhospitable climate. Climatic suitability for gypsy moth in the western United States, however, is potentially improving, perhaps rapidly, due to a general warming trend that began in the mid 1970s and continues today. In this work, we describe the application of a physiologically based climate suitability model for evaluating risk of gypsy moth establishment on a landscape level. Development of this risk assessment system first required amassing databases that integrated the gypsy moth climatic assessment model, with host species distributions, and climate (historical, present, and future). This integrated system was then used to evaluate climate change scenarios for native host species in Utah, with the result that risk of establishment will dramatically increase during the remainder of the 21st century under reasonable climate change scenarios. We then applied the risk assessment system to several case histories of detected gypsy moth introductions in Utah. These applications demonstrated the general utility of the system for predicting risk of establishment and for designing improved risk detection strategies.

Effectiveness of bifenthrin (Onyx) and carbaryl (Sevin SL) for protecting individual, high-value conifers from bark beetle attack (Coleoptera: Curculionidae: Scolytinae) in the Western United States.

High-value trees, such as those located in residential, recreational, or administrative sites, are particularly susceptible to bark beetle (Coleoptera: Curculionidae: Scolytinae) attack as a result of increased amounts of stress associated with drought, soil compaction, mechanical injury, or vandalism. Tree losses in these unique environments generally have a substantial impact. The value of these individual trees, cost of removal, and loss of esthetics may justify protection until the main thrust of a bark beetle infestation subsides. This situation emphasizes the need for ensuring that effective insecticides are available for individual tree protection. In this study, we assess the efficacy of bifenthrin (Onyx) and carbaryl (Sevin SL) for protecting: ponderosa pine, Pinus ponderosa Dougl. ex. Laws., from western pine beetle, Dendroctonus brevicomis LeConte, in California; mountain pine beetle, Dendroctonus ponderosae Hopkins in South Dakota; and Ips spp. in Arizona; lodgepole pine, Pinus contorta Dougl. ex Loud., from D. ponderosae in Montana; pinyon, Pinus edulis Engelm. in Colorado and Pinus monophylla Torr. and Frem. in Nevada from pinyon ips, Ips confusus (LeConte); and Engelmann spruce, Picea engelmannii Parry ex. Engelm. from spruce beetle, Dendroctonus rufipennis (Kirby) in Utah. Few trees were attacked by Ips spp. in Arizona and that study was discontinued. Sevin SL (2.0%) was effective for protecting P. ponderosa, P. contorta, and P. monophylla for two field seasons. Estimates of efficacy could not be made during the second field season in P. edulis and P. engelmannii due to insufficient mortality in untreated, baited control trees. Two field seasons of efficacy was demonstrated in P. ponderosa/D. brevicomis and P. monophylla for 0.06% Onyx. We conclude that Onyx is an effective individual tree protection tool, but repeated annual applications may be required in some systems if multiyear control is desired.

Respiratory exposure to diesel exhaust particles decreases the spleen IgM response to a T cell-dependent antigen in female B6C3F1 mice.

We investigated the systemic immunotoxic potential of respiratory exposure to diesel exhaust particles (DEP) in this study. Female B6C3F1 mice (approximately 8 weeks old) were exposed to increasing concentrations of DEP intratracheally, 3 times every two weeks, and sacrificed 2 or 4 weeks after the first exposure. The systemic toxicity and immune status in mice were evaluated. Mice exposed to DEP (1 to 15 mg/kg) showed no significant changes in body, spleen, or liver weights. Lung weights were increased in the mice exposed to 15 mg/kg DEP for 2 or 4 weeks. Except for a decreased platelet count, no significant alterations occurred in hematological parameters following DEP exposure. The number of splenic anti-sheep red blood cell (sRBC) IgM antibody-forming cells (AFC) decreased following DEP exposure for 2 weeks. This effect was less severe following 4 weeks of exposure and was only evident in the high dose group. Exposure to DEP also resulted in a significant decrease in the absolute numbers and the percentages of total spleen cells for total, CD4(+), and CD8(+) T cells, while the numbers of B cells and total nucleated cells in spleen were not significantly changed. The proliferative response of splenocytes to the T-cell mitogen, concanavalin A (ConA), as well as their production of IL-2 and IFN-gamma, was decreased dose-dependently following exposure of mice to DEP for 2 weeks, whereas proliferation was not changed in response to anti-CD3 monoclonal antibody. In summary, short-term respiratory exposure of mice to DEP resulted in systemic immunosuppression with evidence of T cell-mediated and possibly macrophage-mediated mechanisms.

Correlation of suppressed natural killer cell activity with altered host resistance models in B6C3F1 mice.

A number of methods have been developed to assess the impact of a xenobiotic on the various components of the immune system. For risk analysis, it is necessary to determine what degree of chemically induced immune perturbation translates into altered host resistance. Natural killer (NK) cells play a pivotal role in the innate immune system with the ability to lyse cells infected with intracellular pathogens and certain tumors without previous exposure to the antigen. Spontaneous NK activity in B6C3F1 mice could be incrementally and consistently decreased by 20 to > or =80% by the intravenous administration of a range of dilutions of anti-asialo GM1 (AAGM1) antibody. The decrease in spontaneous NK activity following a single iv administration of AAGM1 antibody persisted for up to approximately 3 weeks when the initial suppression (e.g., 24 h after AAGM1 antibody injection) was almost 100%. Treatment with AAGM1, however, did not appear to perturb the function of other immune cells, based on results of the plaque assay, the mixed lymphocyte response, the cytotoxic T lymphocyte assay, the reticuloendothelial system clearance of sRBC assay, and the Streptococcus pneumoniae host resistance assay. Following a > or =80% decrease in spontaneous NK activity in mice, challenge with > or =1 x 10(3) B16F10 melanoma cells resulted in an increase in tumor burden based on the number of lung nodules. However, following challenge with 1 x 10(5) melanoma cells, a significant increase in tumor burden in mice was not observed until spontaneous NK activity had been decreased by > or =50-60%. Altered host resistance is a function not only of the magnitude of the decrease in NK activity but also of the magnitude of the challenge to the host.

The eyeless mouse mutation (ey1) removes an alternative start codon from the Rx/rax homeobox gene.

The eyeless inbred mouse strain ZRDCT has long served as a spontaneous model for human anophthalmia and the evolutionary reduction of eyes that has occurred in some naturally blind mammals. ZRDCT mice have orbits but lack eyes and optic tracts and have hypothalamic abnormalities. Segregation data suggest that a small number of interacting genes are responsible, including at least one major recessive locus, ey1. Although predicted since the 1940s, these loci were never identified. We mapped ey1 to chromosome 18 using an F2 genome scan and there found a Met10-->Leu mutation in Rx/rax, a homeobox gene that is expressed in the anterior headfold, developing retina, pineal, and hypothalamus and is translated via a leaky scanning mechanism. The mutation affects a conserved AUG codon that functions as an alternative translation initiation site and consequently reduces the abundance of Rx protein. In contrast to a targeted Rx null allele, which causes anophthalmia, central nervous system defects, and neonatal death, the hypomorphic M10L allele is fully viable.

Dynamic forces in the cell cycle affecting fibroblasts in pressure ulcers.

Utilizing specific cell cycle markers of gene activity, temporal changes in the equilibrium of proliferating and non proliferating fibroblasts were shown in pressure ulcers after 36 days of quality care. Average cell counts from multiple tissue sections showed that fibroblast nuclei were stained in decreasing order by antibodies to p21, p21/proliferating cell nuclear antigen (PCNA) and PCNA. P21 labeling suggested that the majority of ulcer fibroblasts were senescent. Fibroblast nuclei showing PCNA staining identified those fibroblasts that were capable of synthesizing DNA and contributing to pressure ulcer repair. Increased rates of wound closure were correlated with a decreasing number of p21 positive cells and an increasing portion of PCNA labeled cells. While the proportion of antigens appeared to correlate with the status of wound closure after 36 days of quality care, they did not always appear to reflect the final outcome of the pressure ulcer. No significant differences were observed in ulcer fibroblasts labeled with p21 at 0 and 10 days, however, the differences were significant after 36 days of quality care (p = 0.05, analysis of variance, post hoc Tukey test). The cellular contribution to pressure ulcer repair appeared to occur from ulcer fibroblasts that were capable of division, of emerging from quiescence, and that were successful in repairing their DNA.