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Effects of long-term storage on potency of TM Biocontrol-1, the registered viral insecticide of Orgyia pseudotsugata (Lepidoptera: Lymantriidae).

The article presents the results of bioassaying 39 samples of TM Biocontrol-1, a viral insecticide, from 10 different lots and various sizes of vacuum-sealed packages that were stored at -10 degrees C for 5-15 yr. This is the first study to present potency data for a registered virus product stored for this length of time. Laboratory bioassays in insects from the same colony from which the TM Biocontrol-1 was produced showed that the stored viral insecticide is still effective, although it lost approximately 30% of its effectiveness during storage. A direct correlation of this loss with the length of time in storage is not apparent. Bioassays also showed that there are significant differences in the susceptibility of Douglas-fir tussock moth, Orgyia pseudotsugata (McDunnough), larvae from different geographic regions to OpMNPV (family Baculoviridae, genus Nucleopolyhedrovirus) infection. Package size did not affect the potency of stored TM Biocontrol-1. There were no clear, significant differences in the effectiveness among lots of TM Biocontrol-1 processed by different organizations.

Animals↗

Frequent fire alters nitrogen transformations in ponderosa pine stands of the inland northwest.

Recurrent, low-severity fire in ponderosa pine (Pinus ponderosa)/interior Douglas-fir (Pseudotsuga menziesii var. glauca) forests is thought to have directly influenced nitrogen (N) cycling and availability. However, no studies to date have investigated the influence of natural fire intervals on soil processes in undisturbed forests, thereby limiting our ability to understand ecological processes and successional dynamics in this important ecosystem of the Rocky Mountain West. Here, we tested the standing hypothesis that recurrent fire in ponderosa pine/Douglas-fir forests of the Inland Northwest decreases total soil N, but increases N turnover and nutrient availability. We compared soils in stands unburned over the past 69-130 years vs. stands exposed to two or more fires over the last 130 years at seven distinct locations in two wilderness areas. Mineral soil samples were collected from each of the seven sites in June and July of 2003 and analyzed for pH, total C and N, potentially mineralizable N (PMN), and extractable NH4+, NO3-, PO4(-3), Ca+2, Mg+2, and K+. Nitrogen transformations were assessed at five sites by installing ionic resin capsules in the mineral soil in August of 2003 and by conducting laboratory assays of nitrification potential and net nitrification in aerobic incubations. Total N and PMN decreased in stands subjected to multiple fires. This loss of total N and labile N was not reflected in concentrations of extractable NH4+ and NO3-. Rather, multiple fires caused an increase in NO3 sorbed on ionic resins, nitrification potential, and net nitrification in spite of the burned stands not having been exposed to fire for at least 12-17 years. Charcoal collected from a recent fire site and added to unburned soils increased nitrification potential, suggesting that the decrease of charcoal in the absence of fire may play an important role in N transformations in fire-dependent ecosystems in the long term. Interestingly, we found no consistent effect of fire frequency on extractable P or alkaline metal concentrations. Our results corroborate the largely untested hypothesis that frequent fire in ponderosa pine forests increases inorganic N availability in the long term and emphasize the need to study natural, unmanaged sites in far greater detail.

Ecosystem↗

Species richness and stand stability in conifer forests of the Sierra Nevada.

Theoretical and empirical studies have long suggested that stability and complexity are intimately related, but evidence from long-lived systems at large scales is lacking. Stability can either be driven by complex species interactions, or it can be driven by the presence/absence and abundance of a species best able to perform a specific ecosystem function. We use 64 years of stand productivity measures in forest systems composed of four dominant conifer tree species to contrast the effect of species richness and abundance on three stability measures. To perform this contrast, we measured the annual growth increments of > 900 trees in mixed and pure forest stands to test three hypotheses: increased species richness will (1) decrease stand variance, (2) increase stand resistance to drought events, and (3) increase stand resilience to drought events. In each case, the alternate hypothesis was that species richness had no effect, but that species composition and abundance within a stand drove variance, resistance, and resilience. In pure stands, the four species demonstrated significant differences in productivity, and in their resistance and resilience to drought events. The two pine species were the most drought resistant and resilient, whereas mountain hemlock was the least resistant and resilient, and red fir was intermediate. For community measures we found a moderately significant (P = 0.08) increase in the community coefficient of variation and a significant (P = 0.03) increase in resilience with increased species richness, but no significant relationship between species richness and community resistance, though the variance in community resistance to drought decreased with species richness. Community resistance to drought was significantly (P = 0.001) correlated to the relative abundance of lodgepole pine, the most resistant species. We propose that resistance is driven by competition for a single limiting resource, with negative diversity effects. In contrast resilience measures the capacity of communities to partition resources in the absence of a single limiting resource, demonstrating positive diversity effects.

Analysis of Variance↗

Influence of overstory removal on growth of epiphytic mosses and lichens in western Oregon.

Will old-growth-associated epiphytes survive if the forest canopy is opened around them by thinning or partial harvest? If old-growth association is due to a species' environmental tolerances, it may not survive in the relatively open stands that result from such treatments. If, however, old-growth association is due to dispersal limitations rather than environmental tolerances, retention of host trees as refugia and sources of inoculum might carry populations of old-growth-associated epiphytes into young stands. We studied growth rates of lichen and moss transplants in a Douglas-fir (Pseudotsuga menziesii) forest (tree ages approximately 55 yr) in western Oregon for nine months before and 27 months after moderate thinning, creation of 0.4-ha patch cuts, and in control areas. We also assessed moss sporophyte production. We contrasted responses of one moss species, Isothecium myosuroides sensu lato, which is ubiquitous in forests of varying ages, with those of another moss, Antitrichia curtipendula, and a lichen, Lobaria oregana, which are both associated with old-growth forests. Both old-growth associates grew faster in thinned areas and patch cuts than in controls, while Isothecuim grew most slowly and produced fewest sporophytes in patch cuts. These species are likely to survive in remnants, assuming they can remain attached, and may be successful in young stands if they can disperse and establish there. Our results suggest that logging with green-tree retention and other silvicultural practices that preserve trees or shrubs hosting the species studied here are likely to encourage these species' development in managed forests.

Bryophyta↗

Immortalization of rat spleen and thymus T cells by human T-cell leukemia virus type I.

Co-cultivation of thymus and spleen cells of Fisher and Lewis rats with lethally irradiated MT-2 cells harboring human T-cell leukemia virus type I (HTLV-I) resulted in the establishment of lymphoid cell lines, FIRT-1, FIRS-1, LERT-1, and LERS-1, respectively. Cells of these cell lines had rat T-cell characters as demonstrated by the positive reaction to monoclonal antibodies (MAbs) to rat T cell antigens (Thy 1 and pan T). They lacked surface immunoglobulins and strongly expressed rat interleukin-2 receptor antigen (Tac) and Ia antigen. Karyotypic analysis revealed that they had the normal rat karyotype in early cultures, but showed marked aneuploidy after long cultivation. None of them expressed HTLV gag proteins (p19 and p24) or virus particles, but they contained HTLV-I proviral DNA monoclonally and weakly expressed pX gene products (p40x). They were not transplantable into syngeneic newborn rats.

Animals↗

Constituents within pulp mill effluent deplete retinoid stores in white sucker and bind to rainbow trout retinoic acid receptors and retinoid X receptors.

Wild female and male white sucker (Catostomus commersoni) inhabiting an area receiving pulp mill effluent had reduced hepatic levels of retinol, didehydroretinol, retinyl esters, and didehydroretinyl esters, while vitamin E levels were unaffected. This disruption of the retinoid system led us to test methanol and dichloromethane extracts from the effluent of 11 pulp mills from across Canada for their ability to bind to rainbow trout (Oncorhynchus mykiss) retinoic acid receptors (RARs) from the gill and retinoid X receptors (RXRs) from the liver. Concentrated extracts of the final effluent from 6 of the 11 pulp mills were able to displace greater than 25% of the receptor-bound [3H]all-trans retinoic acid (RA) or [3H]9-cis RA from trout RARs and RXRs, respectively. The ability of the extracts to displace retinoic acid did not appear to be linked to the pulping or treatment processes. Moreover, extracts with the greatest activity came from thermomechanical mills, suggesting the compounds may originate from the wood furnish. In addition, extracts prepared from wood furnish (wood chips: white spruce [50%], lodgepole pine [47%], and balsam fir [3%]) from one mill were able to displace [3H]RA from the RARs and RXRs. The 4-hydroxy RA, a metabolite of RA that has been shown to be generated in greater quantities in fish exposed to P450-inducing xenobiotics, was able to displace [3H]all-trans RA from trout RARs as effectively as unlabeled all-trans RA. These results suggest that pulp mill effluent may impact the retinoid system of fish at multiple sites, either by decreasing hepatic retinoid stores or through contributing additional ligands (from the wood furnish) that can bind to RA receptors.

Animals↗

Decomposition and nitrogen mineralization from biosolids and other organic materials: relationship with initial chemistry.

Biosolids are effective forest fertilizers. In order to facilitate their use it is important that one be able to predict the amount and rate of mineralization of nutrients, particularly nitrogen, and the relationship between substrate chemistry and N release. We examined the relationships between substrate quality and nitrogen release in a variety of organic materials. Rates of decomposition and net N mineralization from four biosolids, wheat straw, paper fines, and Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco] needle litter were measured during 391-d incubations in a greenhouse, and at two field sites in wet coastal and dry interior forests. Decomposition rates were best predicted by a model incorporating the ratio of carbon to organic matter. The decomposition model extrapolated well to the field when site-specific correction factors were applied. There was a weak relationship between rates of decomposition and net N mineralization. Rates of net N mineralization were best predicted by a model incorporating the initial organic N concentration and the proportion of phenolic C determined from solid-state 13C nuclear magnetic resonance (NMR) spectroscopy. The mineralization model extrapolated less well to the field, but the effect of substrate chemistry was still apparent. Among the four biosolids there was a strong correlation between organic N concentration and indices or protein determined from 13C NMR, suggesting that these protein indices may be useful for predicting N mineralization from biosolids. There was some evidence that the protein content and N mineralization in biosolids may be predictable from the sewage treatment process employed.

Chemical Phenomena↗

Greenhouse gas emissions from forestry operations: a life cycle assessment.

Most forest carbon assessments focus only on biomass carbon and assume that greenhouse gas (GHG) emissions from forestry activities are minimal. This study took an in-depth look at the direct and indirect emissions from Pacific Northwest (PNW) Douglas-fir [Pseudotsuga menziesii (Mirbel) Franco] forestry activities to support or deny this claim. Greenhouse gas budgets for 408 "management regimes" were calculated using Life Cycle Assessment (LCA) methodology. These management regimes were comprised of different combinations of three types of seedlings (P + 1, 1 + 1, and large plug), two types of site preparation (pile and burn, and chemical), 17 combinations of management intensity including fertilization, herbicide treatment, pre-commercial thinning (PCT), commercial thinning (CT), and nothing, and four different rotation ages (30, 40, 50, and 60 yr). Normalized to 50 yr, average direct GHG emissions were 8.6 megagrams (Mg) carbon dioxide equivalents (CO2e) ha(-1), which accounted for 84% of total GHG emissions from the average of 408 management regimes. Harvesting (PCT, CT, and clear cutting) contributed the most to total GHG emissions (5.9 Mg CO2e per 700 m3 harvested timber), followed by pile and burn site preparation (4.0 Mg CO2e ha(-1) or 32% of total GHG emissions) and then fertilization (1.9 Mg CO2e ha(-1) or 15% of total GHG emissions). Seedling production, seedling transportation, chemical site preparation, and herbicide treatment each contributed less than 1% of total GHG emissions when assessed per hectare of planted timberland. Total emissions per 100 m3 averaged 1.6 Mg CO2e ha(-1) over all 408 management regimes. An uncertainty analysis using Monte Carlo simulations revealed that there are significant differences between most alternative management regimes.

Agriculture↗

Auditory brainstem evoked potentials peak identification by finite impulse response digital filters.

Linear phase finite impulse response (FIR) filtering can be used to differentiate auditory brainstem evoked potentials (ABEP) components. The power spectrum of ABEP at high intensities indicates that they contain 3 frequency bands that can be distinguished by applying appropriate digital filters with the following characteristics: up to 240 Hz (revealing slow components), 240-483 Hz (resulting in medium components) and above 500 Hz (leaving only fast components). The results using these filters, indicate that the medium components coincide with peaks I, III and V and that the slow filter results in a 'pedestal' whose peak coincides with peak V. These findings were used for automatic identification of ABEP peaks. A coincidence of the 'pedestal' peak with a medium component was sought and labelled peak V. The preceding medium peaks were labelled, in order of decreasing latency, III and I. Validation of this procedure was conducted on ABEP from normal subjects, using different stimulus rates and intensities, as well as from selected neurological patients with lesions affecting the brainstem. Provided the waveform included a 'pedestal', the results proved this procedure to be reliable and in very good agreement with manual identification and measurement of ABEP peaks.

Adult↗

Use of pectinase to dissociate plant nuclei for squash preparations: effect of hydration procedures.

Scale tissues taken from cones of pendent Douglas-fir Pseudotsuga menziesii (Mirb.) Franco were treated with pectinase to dissociate nuclei for Feulgen staining. Hydration of fixed tissues with water or a graded series of ethyl alcohol before pectinase treatment resulted in distention and faint staining of nuclei and inadequate disintegration of cell walls. These problems were overcome by adopting citrate buffer hydration. Analysis of digitized scans of nuclei provided information about the distribution of the stained material in density classes and nuclear size which is usually unavailable through the conventional double wavelength DNA measuring method.

Buffers↗

Cytophotometric differentiation of high elevation spruces: physiological and ecological implications.

Red and black spruce and their hybrids can be determined by morphological indices; however, the criteria are somewhat subjective and increasingly difficult to use at higher elevations. Although the chromosome number is identical (2n = 24), red spruce has twice as much nuclear DNA (48 pg) than black spruce (24 pg) and thus the species and their hybrids can also be separated by cytophotometry. This is relevant to spruce decline studies because black spruce is much more resistant to high elevation environmental stresses, both natural and anthropogenic. It also has implications for the effect of climatic changes on the composition of high elevation spruce-fir forests because red spruce can outcompete black spruce under more mesic conditions. Four elevation transects sampling spruce on the east and west sides of Mount Washington (New Hampshire) and Camels Hump (Vermont) and a single transect on the southwest side of Whiteface Mountain (New York) were made to investigate the degree of hybridization and introgression between these two species. A positive correlation was found between increased elevation and increased black spruce genes on Mount Washington and Camels Hump. Pure black spruce was found on Mount Washington from 1356 m to 1582 m. No pure black or red spruce was found on Camels Hump although the proportion of red spruce alleles was significantly greater on Camels Hump. All trees sampled at all elevations on Whiteface Mountain were pure red spruce. Thus the proportion of black spruce alleles in high elevation spruce populations decreases from east to west. This closely parallels the increase in spruce decline which increases from east to west.

Cell Differentiation↗

Spatial analysis of within-population microsatellite variability reveals restricted gene flow in the Pacific golden chanterelle (Cantharellus formosus).

We examined the within-population genetic structure of the Pacific golden chanterelle (Cantharellus formosus) in a 50 y old forest stand dominated by Douglas-fir (Pseudotsuga menziesii) and western hemlock (Tsuga heterophylla) with spatial autocorrelation analysis. We tested the null hypothesis that multilocus genotypes possessed by chanterelle genets were randomly distributed within the study area. Fruit bodies from 203 C. formosus genets were collected from a 50 ha study plot. One hundred six unique multilocus genotypes were identified after scoring these collections at five microsatellite loci. Statistically significant positive spatial autocorrelation was detected indicating the presence of fine-scale genetic structure within the area. Repeated autocorrelation analyses with varied minimum distance classes (50-500 m) detected positive spatial genetic structure up to 400 m. Therefore nonrandom evolutionary processes (e.g., isolation by distance) can cause fine-scale genetic structure in C. formosus. The implications of this research for future broad-scale population studies of this species are that population samples should be separated by at least 400 m to be considered statistically independent. Sampling designs that account for fine-scale genetic structure will better characterize heterogeneity distributed across the landscape by avoiding the effects of pseudo replication.

Basidiomycota↗

Comparison of taxonomic, colony morphotype and PCR-RFLP methods to characterize microfungal diversity.

We compared three methods for estimating fungal species diversity in soil samples. A rapid screening method based on gross colony morphological features and color reference standards was compared with traditional fungal taxonomic methods and PCR-RFLP for estimation of ecological indices of soil microfungal community composition. Normalized counts of colony morphotypes on dichloran rose bengal medium were used to estimate species richness (S) and evenness (J) and to calculate Shannon's diversity (H) and Simpson's (SI) dominance indices. Isolates were obtained by dilution plating techniques from litter and soil layer samples taken from Douglas-fir forest and clear-cut areas at two locations in the Cascade Mountains. The highest correspondence (97%) was observed between taxonomic identification and RFLP patterns (32:33). Cladistic analyses of PCR-RFLP patterns indicated an 81% correspondence between RFLP patterns:colony morphotypes (33:41). A correspondence of 78% was observed between traditional taxonomic identification:colony morphotypes (32:41). Statistical analyses of ecological indices based on quantitative application of the colony morphotyping method indicated significant differences (P < 0.05) in fungal community composition between forested and clear-cut areas at the Toad Road site but not at the Falls Creek site. Comparisons of ecological indices based on traditional identification of taxa by microscopic characterization on defined culture media resulted in identical findings of statistical significance. The colony morphotyping approach is proposed as a screening method to identify potential effects of land management practices, edaphic factors and pollutants on microfungal diversity.

Culture Media↗

Characteristics of the inactivation of Escherichia coli by infrared irradiative heating.

The inactivation of Escherichia coil by infrared (IR) irradiation was quantitatively evaluated and compared to that by thermal conductive heating. After the bulk temperature of the sterile saline irradiated by IR reached a steady given temperature, the E. coil culture was inoculated and IR heating was conducted. A mullite cylinder FIR heater, of which main wavelength is 4-7 microm, was used for IR heating. The killing of the E. coil by IR heating and thermal conductive heating both followed first-order reaction kinetics, and the apparent death rate constants (k) under different conditions were obtained. At the same temperatures, pasteurization by IR heating was more effective than that by thermal conductive heating, and its effectiveness increased with a decrease in liquid layer thickness. The values of k due to IR heating were approximately 2-3 times and one order larger than those due to thermal conductive heating at the liquid layer thickness of 10 mm and 1.5 mm, respectively. The activation energy (Ea) for the killing of E. coli by IR irradiation was slightly lower than that by thermal conductive heating, indicating that there are some differences in the action mechanisms.

Disinfection↗

A case-referent study on nasal cancer and exposure to wood dust in the province of Siena, Italy.

The aim of the present study was to assess the risk of nasal cancer associated with exposure to dust in the wood and furniture industry in the province of Siena, Italy. Four to seven percent of the active male population is employed in this sector. A case-referent approach was used. The cases included male subjects seen at the Ear, Nose and Throat Clinic and the Radiotherapy Unit of Siena and diagnosed as having cancer of the nasal cavities or paranasal sinuses between 1963 and 1981. The referents were male patients admitted to the Medical Clinic of Siena for all causes except nasal neoplasia; they were matched 5:1 to the cancer patients for age and time of admission. Information was gathered (by postal questionnaires) on the occupational histories of all 36 of the cancer patients and 164 of the 180 referents. The odds ratio associated with exposure to wood dust was 5.4 (1.7-17.2) for all carcinomas, and 87.7 (19.8-407.3) for mucinous adenocarcinoma. The woods used by the exposed cancer patients were mainly oak, chestnut, poplar, and fir. The median duration of exposure was 40 years; no exposed cancer patient reported the presence of exhaust systems in their work environment.

Adult↗

Prevalence of meningeal worm (Parelaphostrongylus tenuis) in white-tailed deer from New Hampshire.

Parelaphostrongylus tenuis was found in 79 (62.2%) of 127 white-tailed deer heads (Odocoileus virginianus) collected in New Hampshire. Prevalence of infection was higher in fawns and male deer, but older female deer (5.0 + years) had a higher intensity of infection (3.1) than any other age or sex class. Male deer in forest cover type 2 (northern hardwoods-hemlock-white pine) had a significantly greater prevalence of infection than the females in the same habitat (P less than .05). Adult females had a higher prevalence of infection (67.0%) than female fawns (20%) in forest cover type 1 (spruce-fir-northern hardwoods). In the craniums examined, P. tenuis was found most frequently in the tentorium cerebelli (23.6%) while the falx cerebri, the most prevalent site of infection in fawns, was a significantly less frequent site of infection in older deer (P less than .02). Male deer of age class 3.5 years had a significantly higher rate of infection in the dura mater-olfactory lobe than any other age or sex group (P less than .02). The cavernous sinus was the most frequent site of parasitism in male fawns (25.0%) and was the second most common location in all deer (13.4%).

Animals↗

Differences in urinary chemistry profiles of moose on Isle Royale during winter.

During winters 1987-1988 (I) and 1988-1989 (II), we assessed the physiological status of moose (Alces alces) residing on the west and east ends of Isle Royale (Michigan, USA) by collecting and chemically analyzing urine deposited in snow (snow-urine) from January to early March. Samples were assayed for urea nitrogen (U), sodium (Na), potassium (K), calcium (Ca), phosphorus (P) and creatinine (C). Throughout both winters, elevated urinary U:C ratios in snow-urine samples collected from east-end moose compared to west-end moose indicated greater dietary energy deprivation and accelerated net catabolism of endogenous protein. Sodium: C ratios were low throughout the study and were similar between moose from both ends of the island, except during the middle of winter I. Greater K:C, P:C and Ca:C ratios in east-end moose compared to west-end moose throughout winter I, and increases in these ratios and U:C in east-end moose from middle to late winter during the second year provided additional evidence of a greater deterioration in condition in east-end moose. The superior nutrition provided to moose on the west end of the island was associated with more developed soils and diverse vegetation and a lower stem density of balsam fir compared to the east end.

Animal Nutritional Physiological Phenomena↗

Cryptococcus gattii in wildlife of Vancouver Island, British Columbia, Canada.

Although Cryptococcus gattii has emerged as an important pathogen of humans and domestic animals on Vancouver Island, Canada since 1999; its distribution in regional wildlife species is largely unknown. Opportunistic sampling methods were employed to obtain nasal swabs for fungal culture from wild mammal species residing within the coastal Douglas fir biogeoclimatic zone on the southeast coast of the island. Samples were collected from 91 animals representing 14 species. Cryptococcus gattii was isolated from the nasal swabs of two eastern gray squirrels (Sciurus carolinensis) trapped in Duncan, British Columbia. The relative proportion of nasal colonization in wild mammal species is consistent with findings in domestic animals, suggesting that animals may be good indicators of environmental organisms.

Animals↗