A rapid and simple method for processing wood to crude cellulose for analysis of stable carbon isotopes in tree rings

For analysis of carbon isotope discrimination in wood, cellulose or holocellulose is often preferred to whole tissue because of the variability in isotopic composition of different wood components and the relative immobility of cellulose. Most currently used methods for the preparation of wood components for stable isotope analysis (e.g., the Jayme-Wise method) produce a residue of holocellulose. The Jayme-Wise method was initially developed to extract holocellulose from small (~1 g) samples of wood, and, despite subsequent modifications, the method requires specialized glassware, considerable time and entails the risk of sample loss. For carbon isotope analysis, we adapted an acid-catalyzed solvolytic method for preparing crude cellulose by treating wood meal with acidified di-glycol methyl ether (diglyme). The one-step process requires no special glassware, is complete within 24 hours and enables over 100 samples to be processed in a day. This method gives similar delta(13)C values to the Jayme-Wise method for wood of Eucalyptus globulus Labill., Pinus radiata D. Don and Pinus pinaster Ait. The relationship between delta(13)C of wood and crude cellulose is as strong as that observed between wood and alpha-cellulose and stronger than that observed between wood and holocellulose in other species. These relationships suggest that variation in delta(13)C of wood may result from hemicellulose and that analysis of stable carbon isotopes in crude cellulose is preferable. If the consistent -0.3 bias in the value of delta(13)C of cellulose resulting from residual lignin is corrected for, then the relationship between delta(13)C of wood and crude cellulose may be used to predict delta(13)C of cellulose from a small sub-sample. The method is well suited to species with low concentrations of extractives, but further testing is needed to assess its general applicability.     

Improving social acceptability of marine protected area networks: A method for estimating opportunity costs to multiple gear types in both fished and currently unfished areas

We present a novel method for calculating the opportunity costs to fishers from their displacement by the establishment of marine protected areas (MPAs). We used a fishing community in Kubulau District, Fiji to demonstrate this method. We modelled opportunity costs as a function of food fish abundance and probability of catch, based on gear type and market value of species. Count models (including Poisson, negative binomial and two zero-inflated models) were used to predict spatial abundance of preferred target fish species and were validated against field surveys. A profit model was used to investigate the effect of restricted access to transport on costs to fishers. Spatial distributions of fish within the three most frequently sighted food fish families (Acanthuridae, Lutjanidae, Scaridae) varied, with greatest densities of Lutjanidae and Acanthuridae on barrier forereefs and greatest densities of Scaridae on submerged reefs. Modelled opportunity cost indicated that highest costs to fishers arise from restricting access to the barrier forereefs. We included our opportunity cost model in Marxan, a decision support tool used for MPA design, to examine potential MPA configurations for Kubulau District, Fiji Islands. We identified optimum areas for protection in Kubulau with: (a) the current MPA network locked in place; and (b) a clean-slate approach. Our method of modelling opportunity cost gives an unbiased estimate for multiple gear types in a marine environment and can be applied to other regions using existing species data.     

Clinical and imaging findings in five dogs with intracranial blastomycosis (Blastomyces dermatiditis)

Fungal infections affecting the central nervous system are rare. The purpose of this study was to describe clinical and imaging findings in dogs with intracranial blastomycosis (Blastomyces dermatiditis). The radiology database was searched retrospectively for patients with a diagnosis of intracranial blastomycosis which had computed tomography performed as part of their diagnostic work-up. Medical records and imaging studies were reviewed. Five dogs met the inclusion criteria. Major presenting complaints were stertor/nasal discharge (n=2), exophthalmos (n=1), and seizures (n=2). Clinical and laboratory findings were variable. Computed tomographic examination revealed a single contrast-enhancing intra-axial mass (n=1), a nasal mass disrupting the cribriform plate (n=3), and an intracranial mass extending into the orbit and nasal cavity (n=1). Findings in intracranial blastomycosis in dogs are variable, and the disease may mimic other inflammatory disorders or neoplasia.     

Thawing permafrost alters nematode populations and soil habitat characteristics in an Antarctic polar desert ecosystem

Spatial distribution of soil nematode populations in Antarctic terrestrial ecosystems is tightly controlled by environmental factors and thus highly sensitive to changes in soil properties. Increases in the magnitude and frequency of episodic warming events as well as eventual warming trends are likely to result in increased water availability due to glacial melting and permafrost thaw, and may also incite changes in soil physical and chemical characteristics that determine nematode habitat suitability. We hypothesized that climate warming would result in new suitable soil habitats leading to heightened diversity and activity in nematode communities. In order to test this hypothesis, we compared nematode populations in patches of soil wetted by naturally enhanced permafrost thaw versus adjacent soils unaffected by thaw. We found that thaw sites had significantly lower nematode abundances and living to dead ratios, contradicting our hypothesis. We also observed significantly altered soil texture (finer particle size), lower pH and higher salinity in permafrost seeps. These observations suggest that current and future changes in climate may alter soil properties and result in significant changes in nematode population structure, distribution and function.     

High throughput real‐time RT‐PCR assays for specific detection of cassava brown streak disease causal viruses,and their application to testing of planting material

Cassava brown streak disease (CBSD) caused by Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV) is causing severe losses in cassava production in Kenya, Tanzania and Uganda. Two real‐time RT‐PCR assays based on TaqMan chemistry capable of detecting and distinguishing these two viruses are described. These assays were used to screen 493 cassava samples collected from western and coastal Kenya, the main cassava regions of Uganda and inland Tanzania. Both viruses were found in all three countries and across regions therein. Association of CBSD leaf symptom status with CBSV and UCBSV assay results was weak, confirming the need for a diagnostic assay. For leaf samples that were observed with CBSD‐like leaf symptoms but shown as CBSV and UCBSV negative by the RT‐PCR assay, deep sequencing using a Roche 454 GS‐FLX was used to provide additional evidence for the absence of the viruses. The probability of the CBSD associated diagnostics detecting a single CBSV or UCBSV positive sample amongst other non‐CBSD samples was modelled. The results of this study are discussed in the context of the application of diagnostics of CBSD‐associated viruses under the Great Lakes Cassava Initiative and the need to minimize the risk of further spread of the viruses with cassava multiplication material. It is shown that high throughput testing undertaken at Fera of 300 cassava leaves taken from fields for seed multiplication, when analysed in pools of 10, has given a 95% probability of detecting 1% infected plants in the field.     

Comparative Study on the Performance of Three Detection Methods for the Quantification of Pacific Ciguatoxins in French Polynesian Strains of Gambierdiscus polynesiensis

Gambierdiscus and Fukuyoa dinoflagellates produce a suite of secondary metabolites, including ciguatoxins (CTXs), which bioaccumulate and are further biotransformed in fish and marine invertebrates, causing ciguatera poisoning when consumed by humans. This study is the first to compare the performance of the fluorescent receptor binding assay (fRBA), neuroblastoma cell-based assay (CBA-N2a), and liquid chromatography tandem mass spectrometry (LC-MS/MS) for the quantitative estimation of CTX contents in 30 samples, obtained from four French Polynesian strains of Gambierdiscus polynesiensis. fRBA was applied to Gambierdiscus matrix for the first time, and several parameters of the fRBA protocol were refined. Following liquid/liquid partitioning to separate CTXs from other algal compounds, the variability of CTX contents was estimated using these three methods in three independent experiments. All three assays were significantly correlated with each other, with the highest correlation coefficient (r² = 0.841) found between fRBA and LC-MS/MS. The CBA-N2a was more sensitive than LC-MS/MS and fRBA, with all assays showing good repeatability. The combined use of fRBA and/or CBA-N2a for screening purposes and LC-MS/MS for confirmation purposes allows for efficient CTX evaluation in Gambierdiscus. These findings, which support future collaborative studies for the inter-laboratory validation of CTX detection methods, will help improve ciguatera risk assessment and management.