Selection of Beet Necrotic Yellow Vein Virus Specific Single-chain Fv Antibodies from a Semi-synthetic Combinatorial Antibody Library

Methods for the generation of monoclonal antibodies against plant viruses are limited because current hybridoma techniques do not allow efficient exploitation of the immune repertoire. Moreover, the immunization procedures often lead to a bias towards an immunodominant contaminant in the immunogen preparation and not to the plant virus itself. The selection of six different single-chain antibody variable fragments (scFv) against beet necrotic yellow vein virus from a semi-synthetic human combinatorial antibody library showed the feasibility of the phage display system. No bias towards minor contaminants in the purified virus preparation was observed in ELISA, as all the selected scFvs reacted only with beet necrotic yellow vein virus infected plant homogenates. In addition, two of the isolated beet necrotic yellow vein virus-specific scFvs could be produced in E. coli as a scFv fusion protein with alkaline phosphatase, and were applied in ELISA as specific ready to use antibody-enzyme conjugates. Because of their specificity, these antibodies have potential to be used as reagents in sensitive diagnostic assays for routine testing for beet necrotic yellow vein virus in sugar beets.     

Computed tomographic morphometry of the biceps brachii muscle tendon of dogs affected by the medial coronoid disease

The objective of this study was to measure the biceps brachii muscle (BBM) attachment to the medial coronoid process (MCP) using computed tomography images and to compare these data between clinically healthy and dogs affected by medial coronoid disease (MCD). Computed tomography was performed in MCD-affected and healthy dogs. Multiplanar reconstruction views were obtained to measure distance (cm) of the BBM attachment point to the MCP. Results showed that the mean BBM attachment point-to-MCP was 1.42 ± 0.23 cm in MCD-affected and 1.27 ± 0.20 cm in healthy dogs, respectively. In dogs with MCD, the BBM attachment point-to-MCP was significantly more abaxial than in clinically healthy dogs (p < .001). The results of this study support the hypothesis that the BBM attachment site is more abaxial to the MCP and therefore may contribute to supraphysiological overload leading to MCD. Albeit more research is necessary, this study proves a relationship between the BBM attachment point and the development of MCD.     

Different cultures,different values: The role of cultural variation in public’s WTP for marine species conservation

Understanding the cultural variation in public preference for marine species is a necessary pre-requisite if conservation objectives are to include societal preferences in addition to scientific considerations. We report the results of a contingent study undertaken at three case-study sites: Azores islands (Portugal), Gulf of Gdansk (Poland) and Isles of Scilly (UK). The study considered species richness of five specific marine taxa (mammals, birds, fish, invertebrates and algae) as proxies of marine biodiversity and the aim of analysis was to estimate from a multi-site perspective public’s willingness to pay (WTP) to avoid increased levels of species loss (reduction of species richness) for different marine taxa. Results, based on 1502 face-to-face interviews, showed that income, education and environmental awareness of the respondents were significant predictors of WTP for marine species conservation. Results also indicated that respondents in each of the European locations had different preferences for marine taxa. In the Azores, although mammals and fish were valued highly, small differences occurred in the WTP among different taxa. Respondents in the Isles of Scilly put a relatively low value on fish while algae and marine mammals were highly valued. In Gdansk, respondents defined a clear order of preference for marine mammals > fish > birds > invertebrates and algae. These findings suggested that cultural differences may be important drivers of valuation and undermines the commonly held premise that charismatic/likeable taxa consistently have a disproportionately strong influence on WTP for biodiversity conservation. We conclude that conservation policy must take account of cultural diversity alongside biological diversity.     

Digested Ara h 1 loses sensitizing capacity when separated into fractions

The major peanut allergen Ara h 1 is an easily digestible protein under physiological conditions. The present study revealed that pepsin digestion products of Ara h 1 retained the sensitizing potential in a Brown Norway rat model, while this sensitizing capacity was lost by separating the digest into fractions by gel permeation chromatography. Protein chemical analysis showed that the peptide composition as well as the aggregation profiles of the fractions of Ara h 1 digest differed from that of the whole pool. These results indicate that the sensitizing capacity of digested Ara h 1 is a consequence of the peptides being in an aggregated state resembling the intact molecule or that most peptides of the digests need to be present in the same solution, having a synergistic or adjuvant effect and thereby augmenting the immune response against other peptides.     

Can agricultural cultivation methods influence the healthfulness of crops for foods?

The aim of the current study was to investigate if there are any health effects of long-term consumption of organically grown crops using a rat model. Crops were retrieved over two years from a long-term field trial at three different locations in Denmark, using three different cultivation systems (OA, organic based on livestock manure; OB, organic based on green manure; and C, conventional with mineral fertilizers and pesticides) with two field replicates. The cultivation system had an impact on the nutritional quality, affecting γ-tocopherol, some amino acids, and fatty acid composition. Additionally, the nutritional quality was affected by harvest year and location. However, harvest year and location rather than cultivation system affected the measured health biomarkers. In conclusion, the differences in dietary treatments composed of ingredients from different cultivation systems did not lead to significant differences in the measured health biomarkers, except for a significant difference in plasma IgG levels.     

Soil enzyme activity in an old-growth northern hardwood forest: Interactions between soil environment,ectomycorrhizal fungi and plant distribution

Plants and soil microbes produce extracellular enzymes (EE) that catalyze the hydrolysis of nitrogen (N) and phosphorus (P) containing compounds in soil and other enzymes involved in degradation of lignin and cellulose. We explored whether soil enzyme activity involved in carbon (C), N and P cycling were correlated with plant distribution, soil chemical conditions and the identity of fungi colonizing tree roots in an old growth forest remnant. Terminal restriction fragment length polymorphism (TRFLP) was used to determine the presence of root fungi and standard fluorometric analysis was used to determine soil enzyme activities. Soil enzymes were consistently positively correlated with soil C and N, but not CN ratio. Soil P was also correlated with enzyme activity during both June and September sampling. We saw no significant relationships between herbaceous plant cover and enzyme activity in June, but there were significant positive correlations between α-glucosidase and herbaceous plant coverage in September. We also found that some enzymes were significantly correlated with the identity of fungi colonizing tree roots separated from the soil cores. Chitinase and β-glucosidase were positively correlated with the genera Russula and Piloderma while chitinase was negatively correlated with Amanita and Entoloma. In addition, phosphatase was positively correlated with Russula, Meliniomyces and Solenopezia. Our results suggest that enzyme activity in old growth forest soils are affected by a variety of environmental factors, and that herbaceous plants and some root fungi may be associated with sites of elevated or decreased decomposition potential and nutrient cycling.     

Stormwater impact on urban waterways in a cold climate: variations in sediment metal concentrations due to untreated snowmelt discharge

Purpose

Stormwater discharges include contaminated sediments that accumulate in the receiving water body. It is thus important to investigate sediment and pollutant processes and pathways from the catchment to, and within, the receiving water. These processes may be influenced by seasonal changes. The objective of this study was to investigate the stormwater impact on receiving waters in the Luleå area, Northern Sweden; seasonal changes in contamination loads in the receiving waters due to snowmelt; and factors influencing the pollutant pathways in the receiving waters.

Materials and methods

In front of three storm sewer outlets in Luleå, samples of bottom sediment (surface layer 0–2 cm) were collected from the connecting ditches and the downstream water body in autumn and spring (before and after the snow season 2009/2010). The characteristics of the receiving waters differed in geomorphology and vegetation. The sediment was analyzed for loss-on-ignition (LOI), grain size, and bulk concentrations of SiO₂, Al₂O₃, CaO, Fe₂O₃, MnO, Na₂O, P₂O₅, TiO₂, As, Cd, Co, Cr, Cu, Hg, Ni, Pb, S, V, and Zn. The sediment contamination was compared to concentrations at a reference point in Luleå where the bottom sediment was not affected by stormwater discharges and with Swedish environmental quality guidelines. Pearson’s correlation and a principal component analysis were used to further evaluate the results.

Results and discussion

Relative to the reference point, elevated trace metal concentrations were detected in sediments at all three sampling stations. At two of the stations, seasonal variations in ditch sediment grain size, LOI, and contaminant concentrations were observed, originating from stormwater sediment. Snowmelt runoff caused an increased proportion of fine-grained sediment fractions (<0.063 mm) in spring, mainly due to changes in runoff intensity and high sediment loads in the snowmelt runoff. The retention of metals appeared to be due to low turbulence in the water and the presence of organic material.

Conclusions

Stormwater discharge affected the contaminant concentrations in the bottom sediments. The observed seasonal variation of contaminants indicated that relatively high amounts of contaminants are discharged during snowmelt and then reallocated within the receiving water body, either directly or after some temporal retention, depending on the characteristics of the receiving water. A calm water column and the presence of organic material in the receiving water body were crucial for the retention of metals.

     

Zinc hyperaccumulation in Thlaspi caerulescens. I. Influence on growth and mineral nutrition

Thlaspi caerulescens, a metallophyte that is able to accumulate up to 4% zinc (Zn) in leaf dry matter, has attracted much attention for its possible use in phytoremediation of metal contaminated soils. In the present study, the influence of Zn supply on mineral nutrition in T. caerulescens was investigated, in order to establish the extent to which growth stimulation by high Zn supply is related to changes in the levels of other essential nutrients. The plants were exposed to nutrient solutions containing 1.5, 100, 500, 750, 1000, or 1500 μM Zn. Zinc supply significantly influenced root and shoot concentrations of essential nutrients, but excepting Zn, the concentrations stayed within the range considered adequate for optimum growth in Brassicaceae crops. Best performance was achieved with the supply of 500 μM Zn. Growth stimulation by this treatment was accompanied by increased translocation of iron (Fe) from root to shoot and a significant correlation between shoot dry weight and Fe concentrations in shoots was found.     

Identification of Four New agr Quorum Sensing-Interfering Cyclodepsipeptides from a Marine Photobacterium

During our search for new natural products from the marine environment, we discovered a wide range of cyclic peptides from a marine Photobacterium, closely related to P. halotolerans. The chemical fingerprint of the bacterium showed primarily non-ribosomal peptide synthetase (NRPS)-like compounds, including the known pyrrothine antibiotic holomycin and a wide range of peptides, from diketopiperazines to cyclodepsipeptides of 500–900 Da. Purification of components from the pellet fraction led to the isolation and structure elucidation of four new cyclodepsipeptides, ngercheumicin F, G, H, and I. The ngercheumicins interfered with expression of virulence genes known to be controlled by the agr quorum sensing system of Staphylococcus aureus, although to a lesser extent than the previously described solonamides from the same strain of Photobacterium.