Combination of On-Line Solid-Phase Extraction with LC-MS for the Determination of Potentially Hazardous Glycoalkaloids in Potato Products

A simple and rapid method for the determination of naturally occurring, potentially hazardous glycoalkaloids (GAs) in potatoes and their products has been developed. The procedure is based on the on-line solid-phase extraction of the acetic acid extracts from potato products and combined with liquid chromatography (LC)-mass spectrometry (MS) in a fully automated system (Symbiosis™, Spark Holland Instruments, Emmen, The Netherlands). As sorbent material HySphere™ 18HD was used for alkaloid enrichment. GAs were eluted with the LC gradient and directly analysed by MS. Detection of the analytes was achieved in the sensitive multiple reaction monitoring mode using two characteristic ions (m/z 98 as a qualifier for GAs and m/z 868.3 as a quantifier for α-solanine or m/z 852.4 for α-chaconine). Typical validation data for method precision (v _k α-solanine = 5.3–6.5, v _k α-chaconine = 3.4–15.4), accuracy (average recovery of α-solanine = 84%, average recovery of α-chaconine = 87%) and linearity over the range from 1 to 1,000 ng ml⁻¹ (R ² = 0.9915 for α-solanine, R ² = 0.9939 for α-chaconine) with detection limits of 0.3 ng ml⁻¹ for α-chaconine and 0.5 ng ml⁻¹ for α-solanine were obtained. GA contents of commercial potato products were determined by the new on-line method and afterwards compared with those obtained with an established high-performance LC routine procedure. Better performance of the on-line procedure was obvious from the standard deviations of both methods. Other advantages included a strong reduction of overall analysis time, human intervention and solvent consumption as well as waste production. The time required for the on-line analysis was 5 min, which would allow processing of almost 100 samples in 8 h.     

Use of Dried Fermented Biomass as a Fish Meal Replacement in Practical Diets of Florida Pompano,Trachinotus carolinus

Substitution of fish meal (FM) as a protein source in aquaculture diets is required to reduce cost and promote a sustainable industry. Potential protein sources include a range of traditional products from agricultural production such as plant or animal proteins. However, there are also a number of manufacturing coproducts, including bacterial dried fermented biomass (DFB,) for which there is an increased interest. Hence, the objective of this study was to evaluate DFB as a replacement for FM in practical diets for Florida pompano, Trachinotus carolinus. Four diets were prepared in which different levels of FM (15%, 10%, 5%, and 0% of the diet) were replaced with DFB (0%, 4.27%, 8.54%, and 12.82%) on an iso-nitrogenous and iso-lipidic basis, to contain 40% crude protein and 10% lipid. A 10-week growth trial was conducted in a recirculating system consisting of 12 indoor 1.0 m³ tanks, stocked with 20 juvenile pompano (mean initial weight 8.1 g) per tank. There were no significant differences in final weight (64.5 g?79.5 g), survival (81.7%–96.7%), percent weight gain (711.6%–879.0%), FCR (2.1–2.0), or thermal-unit growth coefficient (0.1013?0.1149), regardless of the levels of dietary FM or DFB tested. Based on these results, it is recommended that DFB can be included up to 12.8% in practical diets for Florida pompano without significantly reducing growth performance. The response to higher levels of DFB inclusion should be evaluated.     

Ectomycorrhizal communities in a Mediterranean forest ecosystem dominated by <Emphasis Type="Italic">Quercus ilex</Emphasis>: seasonal dynamics and response to drought in the surface organic horizon

• Introduction   

Millions of hectares of Quercus ilex forests dominate disturbed landscapes in the western part of the Mediterranean basin. Although these forests are very widespread, little is known about the composition and structure of their associated ectomycorrhizal fungal communities.     

Quantifying uncertainty in the wild‐caught fisheries goal of the Ocean Health Index

Sustainability indices are proliferating, both to help synthesize scientific understanding and inform policy. However, it remains poorly understood how such indices are affected by underlying assumptions of the data and modelling approaches used to compute indicator values. Here, we focus on one such indicator, the fisheries goal within the Ocean Health Index (OHI), which evaluates the sustainable provision of food from wild fisheries. We quantify uncertainty in the fisheries goal status arising from the (a) approach for estimating missing data (i.e., fish stocks with no status) and (b) reliance on a data‐limited method (catch‐MSY) to estimate stock status (i.e., B/B_MSY). We also compare several other models to estimate B/B_MSY, including an ensemble approach, to determine whether alternative models might reduce uncertainty and bias. We find that the current OHI fisheries goal model results in overly optimistic fisheries goal statuses. Uncertainty and bias can be reduced by (a) using a mean (vs. median) gap‐filling approach to estimate missing stock scores and (b) estimating fisheries status using the central tendency from a simulated distribution of status scores generated by a bootstrap approach that incorporates error in B/B_MSY. This multitiered approach to measure and describe uncertainty improves the transparency and interpretation of the indicator and allows us to better understand uncertainty around our OHI fisheries model and outputs for country‐level interpretation and use.     

Host resistance to lung infection mediated by major vault protein in epithelial cells

The airway epithelium plays an essential role in innate immunity to lung pathogens. Ribonucleoprotein particles primarily composed of major vault protein (MVP) are highly expressed in cells that encounter xenobiotics. However, a clear biologic function for MVP is not established. We report here that MVP is rapidly recruited to lipid rafts when human lung epithelial cells are infected with Pseudomonas aeruginosa, and maximal recruitment is dependent on bacterial binding to the cystic fibrosis transmembrane conductance regulator. MVP was also essential for optimal epithelial cell internalization and clearance of P. aeruginosa. These results suggest that MVP makes a substantial contribution to epithelial cell-mediated resistance to infection.     

Amplification of the <Emphasis Type="Italic">Phytophthora infestans</Emphasis> RG57 loci Facilitates <Emphasis Type="Italic">in planta</Emphasis> T-RFLP Identification of Late Blight Genotypes

Late blight, caused by the oomycete Phytophthora infestans (Mont.) de Bary, is a devastating disease in potato and tomato and causes yield and quality losses worldwide. The disease first emerged in central America and has since spread in North America including the United States and Canada. Several new genotypes of P. infestans have recently emerged, including US-22, US-23 and US-24. Due to significant economic and environmental impacts, there has been an increasing interest in the rapid identification of P. infestans genotypes. In addition to providing details regarding the various phenotypic characteristics such as fungicide resistance, host preference, and pathogenicity associated with various P. infestans genotypes, information related to pathogen movement and potential recombination may also be determined from the genetic analyses. Restriction fragment length polymorphism (RFLP) analysis with the RG57 loci is one of the most reliable procedures used to genotype P. infestans. However, the RFLP procedure requires propagation and isolation of the pathogen and relatively large amounts of DNA. Isolation of the late blight pathogen is sometimes impossible due to the poor condition of the infected tissues or the presence of fungicide residues. In this study, we describe a procedure to identify P. infestans at the molecular level in planta using terminal restriction fragment length polymorphism (T-RFLP) of the RG57 loci. This T-RFLP assay is sufficiently sensitive to detect and differentiate P. infestans genotypes directly in planta without propagation and isolation of the pathogen, to facilitate the timely implementation of best management practices.     

Calcium oxalate accumulation and soil weathering in mats of the hypogeous fungus Hysterangium crassum

Fungal mats of Hysterangium crassum (Tul. and Tul.) Fischer occupied a mean of 9.6% of the upper 10 cm of soil developed under a 40–65 yr old stand of Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) in Oregon. This hypogeous basidiomycete exudes large amounts of oxalic acid, some of which precipitates with Ca in microscopic crystals of calcium oxalate, resulting in a mean CaC₂O₄ content of 82g m^?2 for the entire soil. Soil oxalate concentration was significantly greater within fungal mats (P < 0.01) and soil pH was significantly lower (P < 0.01) than in soil adjacent to mats. The quantity of Ca present as CaC₂O₄ is 0.5 the amount of exchangeable Ca in the soil and exceeds the mass of Ca lost annually in runoff. Scanning electron micrographs show intense chemical weathering, attributable to oxalate attack, in the immediate vicinity of hyphae. X-ray diffraction patterns of clay indicate bulk weathering is more intense within the fungal mats than in adjacent uncolonized soil.