DNA Vaccination Partially Protects Muskellunge against Viral Hemorrhagic Septicemia Virus (VHSV-IVb)

Abstract

A DNA vaccine containing the glycoprotein (G) gene of the North American viral hemorrhagic septicemia virus (VHSV) genotype IVb was developed to evaluate the immune response of fish following vaccination and evaluate its efficacy in protecting a susceptible species, the Muskellunge Esox masquinongy, against VHSV-IVb challenge. Seven weeks (539 degree-days) following vaccination with 10 μg of either pVHSivb-G or a control plasmid, Muskellunge were challenged by immersion with 10⁵ plaque-forming units (pfu)/mL of VHSV-IVb. Fish vaccinated with pVHSivb-G had a relative percent survival (RPS) of 45%. Vaccinated fish also had significantly lower mean viral titers in tissues (4.2 × 10² pfu/g) and viral prevalence (4%) than fish receiving the plasmid control vaccine (3.3 × 10⁵ pfu/g; 82%). Neutralizing antibodies were detected 28 d (308 degree-days) postchallenge (11 weeks postvaccination) in 100% of Muskellunge vaccinated with pVHSivb-G compared with only 12% of plasmid-control-vaccinated Muskellunge, suggesting robust induction of a secondary, adaptive immune response. In addition, pVHSivb-G–vaccinated Rainbow Trout Oncorhynchus mykiss challenged 7 d (100 degree-days) postvaccination with the heterologous novirhabdovirus, infectious hematopoietic necrosis virus (IHNV), experienced an RPS of 61%, compared to control fish, suggesting induction of an early and transient nonspecific antiviral immune response. This study provides an important starting point for VHSV-IVb vaccine development and useful information about the antiviral immune response elicited by DNA vaccination in a nondomesticated fish species.

Received May 1, 2016; accepted September 1, 2016     

Effect of the addition of bran fractions on bread properties

High fibre breads were produced adding durum wheat bran fractions of different composition and particle size. Fresh products were characterized for texture, crumb grain, volume, colour, water status (water activity, moisture content, frozen water content, ¹H molecular mobility).     

Smenamides A and B,Chlorinated Peptide/Polyketide Hybrids Containing a Dolapyrrolidinone Unit from the Caribbean Sponge Smenospongia aurea. Evaluation of Their Role as Leads in Antitumor Drug Research

An in-depth study of the secondary metabolites contained in the Caribbean sponge Smenospongia aurea led to the isolation of smenamide A (1) and B (2), hybrid peptide/polyketide compounds containing a dolapyrrolidinone unit. Their structures were elucidated using high-resolution ESI-MS/MS and homo- and heteronuclear 2D NMR experiments. Structures of smenamides suggested that they are products of the cyanobacterial metabolism, and 16S rRNA metagenomic analysis detected Synechococcus spongiarum as the only cyanobacterium present in S. aurea. Smenamides showed potent cytotoxic activity at nanomolar levels on lung cancer Calu-1 cells, which for compound 1 is exerted through a clear pro-apoptotic mechanism. This makes smenamides promising leads for antitumor drug design.     

Simultaneous Extraction and Depolymerization of Fucoidan from Sargassum muticum in Aqueous Media

The biomass components of the invasive seaweed Sargassum muticum were fractionated to allow their separate valorization. S. muticum (Sm) and the solid residue remaining after alginate extraction of this seaweed (AESm) were processed with hot, compressed water (hydrothermal processing) to assess the effects of temperature on fucoidan solubilization. Fucose-containing oligosaccharides were identified as reaction products. Operating under optimal conditions (170 °C), up to 62 and 85 wt% of the dry mass of Sm and AESm were solubilized, respectively. The reaction media were subjected to precipitation, nanofiltration and freeze-drying. The dried products contained 50% and 85% of the fucoidan present in Sm and AESm, respectively; together with other components such as phenolics and inorganic components. The saccharidic fraction, accounting for up to 35% of the dried extracts, contained fucose as the main sugar, and also galactose, xylose, glucose and mannose. The concentrates were characterized for antioxidant activity using the TEAC assay.     

Mobilization of Cu and Pb from multi-metal contaminated soils by dissolved humic substances extracted from leonardite and factors affecting the process

Journal of Soils and Sediments - The aim of the research was to determine the potential of dissolved humic substances extracted from leonardite to mobilize Cu and Pb from multi-metal contaminated...     

Biodegradation of humic substances by microscopic filamentous fungi: chromatographic and spectroscopic proxies

Purpose

The study of interactions between humic substances (HSs) and soil filamentous fungi is the key to understanding the sustainable soil functioning. The present work aims to examine the decomposition of HSs by filamentous dark-pigmented fungus Alternaria alternatа under the laboratory conditions and to determine the effect of easily assimilable organic carbon on this process. Analyzing such polydisperse substances like HSs by a complex integrated methodology makes it possible to explore the data on their decomposition by microorganisms.

Materials and methods

To achieve the aforementioned goals, we used chromatographic and spectroscopic approaches: low-pressure size-exclusion and hydrophobic interaction chromatography accompanied by absorption and fluorescence spectroscopy. To determine the effect cometabolism conditions produced on HS decomposition, two types of carbon substrates were added to the nutrient media: easily assimilable organic carbon (standard 0.3% or reduced 0.03% sucrose content) and hardly assimilable organic carbon (HSs), as well as their combinations. Five HS samples of different organic matter origin have been inspected: potassium humates (HPs) and humic acids (HAs) from coal, peat, and lignosulfonate. Correlation matrix and principal component analysis (PCA) were calculated for comprehensive data analysis.

Results and discussion

Transformations of the investigated HSs under fungal cultivation lead to the increase in the low molecular weight fraction, rise of hydrophilic fraction, enlargement of absorbance ratio A₂₅₀/A₃₆₅, shortening of the emission wavelength of the humic-type fluorescence, and growth in the fluorescence quantum yield measured with excitation at 355 nm. A positive correlation was observed between the accumulation of fungal biomass and the degree of HS decomposition. PCA analysis confirms that the difference in the results of HS decomposition largely depends on the sucrose content and the nature of HSs. We divided all the HS samples into four groups according to the degree of HS decomposition: original HS solutions, HPs altered using fungal cultivation at 0.03% sucrose, HAs after fungal cultivation at 0.03% sucrose, and finally, HSs (both HPs and HAs) after fungal cultivation at 0.3% sucrose.

Conclusions

In the laboratory experiments, we showed that (1) the isolated HAs were more effectively degraded than the parent HPs, and this process was more pronounced at a reduced sucrose content, and (2) the decomposition of stable organic compounds (HSs) was activated by the easily assimilable carbon sources (especially 0.3% sucrose) being present. We assume that it is the easily assimilable organic carbon that most likely triggers the HS degradation working as the priming effect in natural environments.

     

Glyphosate Dissipation in Different Soils Under No-Till and Conventional Tillage

Glyphosate is the most used herbicide in Argentina, accounting for 62% of the commercialized pesticides on the market. It is used as a weed controller in no-till systems, and it is also applied to various genetically modified crops (e.g., soybean, corn, and cotton). Although it has a high solubility in water, it tends to adsorb and accumulate in agricultural soils. The main objectives of this work were to compare the dissipation of glyphosate and the accumulation of its metabolite aminomethylphosphonic acid (AMPA) over time in three soils from agricultural areas of Argentina under long-term management with no-till (NT) and conventional tillage (CT) practices. There were no differences in dissipation between NT and CT, indicating that the glyphosate-degrading microflora was not modified by the different tillage managements. Moreover, tillage practices did not alter the general soil properties; therefore, glyphosate bioavailability was not affected by NT or CT practice. Forty percent of the applied glyphosate was degraded within the first three days in all soils, indicating a fast initial dissipation rate. However, the dissipation rate considerably decreased over time, and the degradation kinetics followed a bi-exponential (or two-compartment) kinetic model. No differences were found between tillage practices. Dissipation was not related to the microbial activity measured as soil respiration. The fast decrease in the concentration of glyphosate at the beginning of the dissipation study was not reflected in an increase in the concentration of AMPA. The estimated half-lives for glyphosate ranged between 9 and 38 d. However, glyphosate bioavailability decreases over time, as it is strongly adsorbed to the soil matrix. This increases its residence time, which may lead to its accumulation in agricultural soils.     

Organic acid contents in onion cultivars (Allium cepa L.)

The following organic acids (glutamic, oxalic, pyruvic, malic, tartaric, citric, and fumaric), pungency, Brix degree, acidity, and pH were determined in onion cultivars (Texas, Guayonje, San Juan de la Rambla, Carrizal Alto, Carrizal Bajo, and Masca) harvested in the same agroclimatic conditions. Glutamic acid was the most abundant organic acid (325 +/- 133 mg/100 g) followed by citric acid (48.5 +/- 24.1 mg/100 g) and malic acid (43.6 +/- 10.4 mg/100 g). There were significant differences between the onion cultivars in the mean concentrations of all of the analyzed parameters. The San Juan de la Rambla and Masca cultivars presented, in general, higher concentrations of the organic acids than the other cultivars. Significant differences in most of the analyzed parameters were observed between the two seed origins for the Masca and San Juan de la Rambla cultivars. The onion samples tended to be classified according to the cultivar and, in the case of San Juan de la Rambla cultivar, according to the precedence of the seeds after applying discriminant analysis.