Antiviral Sulfoquinovosyldiacylglycerols (SQDGs) from the Brazilian Brown Seaweed Sargassum vulgare

Total lipids from the Brazilian brown seaweed Sargassum vulgare were extracted with chloroform/methanol 2:1 and 1:2 (v/v) at room temperature. After performing Folch partition of the crude lipid extract, the lipids recovered from the Folch lower layer were fractionated on a silica gel column eluted with chloroform, acetone and methanol. The fraction eluted with methanol, presented a strong orcinol-positive band characteristic of the presence of sulfatides when examined by TLC. This fraction was then purified by two successive silica gel column chromatography giving rise to fractions F4I86 and F4II90 that exhibited strong activity against herpes simplex virus type 1 and 2. The chemical structures present in both fractions were elucidated by ESI-MS and ¹H/¹³C NMR analysis HSQC fingerprints based on their tandem–MS behavior as sulfoquinovosildiacylglycerols (SQDGs). The main SQDG present in both fractions and responsible for the anti-herpes activity observed was identified as 1,2-di-O-palmitoyl-3-O-(6-sulfo-α-d-quinovopyranosyl)-glycerol.     

Getting to the root of the matter: landscape implications of plant-fungal interactions for tree migration in Alaska

Context

Forecasting the expansion of forest into Alaska tundra is critical to predicting regional ecosystem services, including climate feedbacks such as carbon storage. Controls over seedling establishment govern forest development and migration potential. Ectomycorrhizal fungi (EMF), obligate symbionts of all Alaskan tree species, are particularly important to seedling establishment, yet their significance to landscape vegetation change is largely unknown.

Objective

We used ALFRESCO, a landscape model of wildfire and vegetation dynamics, to explore whether EMF inoculum potential influences patterns of tundra afforestation and associated flammability.

Methods

Using two downscaled CMIP3 general circulation models (ECHAM5 and CCCMA) and a mid-range emissions scenario (A1B) at a 1 km² resolution, we compared simulated tundra afforestation rates and flammability from four parameterizations of EMF effects on seedling establishment and growth from 2000 to 2100.

Results

Modeling predicted an 8.8–18.2 % increase in forest cover from 2000 to 2100. Simulations that explicitly represented landscape variability in EMF inoculum potential showed a reduced percent change afforestation of up to a 2.8 % due to low inoculum potential limiting seedling growth. This reduction limited fuel availability and thus, cumulative area burned. Regardless of inclusion of EMF effects in simulations, landscape flammability was lower for simulations driven by the wetter and cooler CCCMA model than the warmer and drier ECHAM5 model, while tundra afforestation was greater.

Conclusions

Results suggest abiotic factors are the primary driver of tree migration. Simulations including EMF effects, a biotic factor, yielded more conservative estimates of land cover change across Alaska that better-matched empirical estimates from the previous century.

     

Comparison of culture versus quantitative real-time polymerase chain reaction for the detection of Taylorella equigenitalis in field samples from naturally infected horses in Canada and Germany

A quantitative real-time polymerase chain reaction method (qPCR) was developed and tested for the detection of Taylorella equigenitalis. It was shown to have an analytical sensitivity of 5 colony-forming units (CFU) of T. equigenitalis when applied to the testing of culture swabs that mimicked field samples, and a high analytical specificity in not reacting to 8 other commensal bacterial species associated with horses. As designed, it could also differentiate specifically between T. equigenitalis and T. asinigenitalis. The qPCR was compared to standard culture in a study that included 45 swab samples from 6 horses (1 stallion, 5 mares) naturally infected with T. equigenitalis in Canada, 39 swab samples from 5 naturally infected stallions in Germany, and 311 swab samples from 87 culture negative horses in Canada. When the comparison was conducted on an individual sample swab basis, the qPCR had a statistical sensitivity and specificity of 100% and 96.4%, respectively, and 100% and 99.1% when the comparison was conducted on a sample set basis. A comparison was also made on 203 sample swabs from the 5 German stallions taken over a span of 4 to 9 mo following antibiotic treatment. The qPCR was found to be highly sensitive and at least as good as culture in detecting the presence of T. equigenitalis in post-treatment samples. The work demonstrates that the qPCR assay described here can potentially be used to detect the presence of T. equigenitalis directly from submitted sample swabs taken from infected horses and also for determining T. equigenitalis freedom following treatment.     

Identification of grouper (Epinephelus guaza), wreck fish (Polyprion americanus), and Nile perch (Lates niloticus) fillets by polyclonal antibody-based enzyme-linked immunosorbent assay

An indirect enzyme-linked immunosorbent assay (ELISA) has been developed for the species identification of grouper (Epinephelus guaza), wreck fish (Polyprion americanus), and Nile perch (Lates niloticus) fillets. The assay was performed in two different formats, microtiter plates and immunostick tubes, and uses polyclonal antibodies raised in rabbits against muscle-soluble proteins of grouper (anti-GSP), wreck fish (anti-WSP), and Nile perch (anti-PSP). The antibodies were made species-specific by blocking them with the heterologous soluble muscle proteins. Immunorecognition of polyclonal antibodies adsorbed to their specific fish samples was made with swine antirabbit immunoglobulins conjugated to the enzyme horseradish peroxidase. Subsequent enzymatic conversion of the substrate allowed unequivocal identification of the species studied.     

Foliar response of wild and domesticated Phaseolus vulgaris L. to water stress

It is surmised that domestication has reduced geneticvariability in the common bean (Phaseolusvulgaris L.). However, little is known about leaf anatomy responses to water stress in wild or domesticated forms ofP. vulgaris. In this study,anatomical variation was evaluated in watered and water stressed plants of wild and domesticated genotypes of P.vulgaris from Mexico, growing under greenhouse conditions. Five terminal leaflets of completely expanded leaves wereused to evaluate leaf area (LA) and theirmiddle-regions to evaluate leaf thickness (LT), and various anatomical features. Domesticated genotypes showed bigger LAthan that of the wild ones; there was an effect due to the origin of materials, LA being bigger in genotypes from warm and humid lands that those from dry and temperate ones. No differences were detected in LT among materials; however, in domesticated plants the palisadeparenchyma predominated over the spongy one, contrasting with thewild genotypes. Under drought conditions, stomatal abortion was observed in wild and domesticated genotypes, especially on theabaxial surface. We conclude that, in fact, there are contrasting differences in leaf anatomy among wild and domesticated forms of Mexican Phaseolus vulgaris. Results suggest that domestication not only favours the enlargement of the LA, but also an increase in palisade parenchyma, suggesting a divergence toward assimilation in domesticated forms, in contrast with a better internal gas diffusion in wild forms. Domestication seems to have accentuated stomatal abortion as means of a transpirational control mechanism to counteract the effects of drought.     

The effect of solvent interactions on alpha-, beta-, and gamma-cyclodextrin/flavor molecular inclusion complexes

Three commonly used flavor industry solvents (propylene glycol, triacetin, and triethyl citrate) were tested for their capacity to interfere with the ability of alpha-, beta-, and gamma-cyclodextrin to form molecular inclusion complexes with flavors. Six flavor compounds (ethyl butyrate, ethyl heptanoate, l-menthol, methyl anthranilate, neral, and geranial) were measured by headspace gas chromatography above 2:1 water/ethanol containing appropriate additions of cyclodextrin and flavor solvent. The smallest and most polar solvent molecule represented by propylene glycol had the least effect on cyclodextrin/flavorant complex formation. In contrast, triacetin, intermediate in size among the three flavor diluents studied, had the greatest effect, even though, based on at least some computed molecular parameters, it appears to be more polar than triethyl citrate. The explanation for this apparent anomaly may lie in differences in the extent to which triacetin and triethyl citrate are able to interact with cyclodextrins by means of partial interaction with the hydrophobic cavities of the latter.     

The Ecological Dose of Nickel in a Semiarid Soil Amended with Sewage Sludge Related to the Unamended Soil

Nickel pollution may affect microbial communitydevelopment and its activity in soil and, therefore, itsfertility, while organic amendment may affect Ni mobilityand bioavailability. The aim of this research was toascertain the extent to which the addition of sewage sludgeto soil may affect Ni toxicity and to establish theecological dose of this heavy metal. Unamended and amendedsoils with sewage sludge were spiked with differentconcentrations of Ni (0–8000 mg kg^-1) and then incubatedfor 3 hours, 12 days and 40 days. Soil dehydrogenaseactivity, ATP content, microbial respiration and microbialbiomass C were measured in the samples containing differentNi concentrations. A mathematical model which describe theinhibition of these soil parameters were used to calculatethe ecological dose of Ni: ED₅₀. Soil microbialbiomass C measurements of all treatments better fitted themodel (r² = 0.95–0.69 with p < 0.05) than the otherparameters studied; but the ED₅₀ values predicted by the modelhad a high coefficient of variation. For both ATP contentand microbial biomass C, the ED₅₀ values calculatedfor the amended soil were higher than for the unamendedsoil and so that the effect of Ni toxicity on theseparameters of microbial activity may be considered lower inthe former. In this study, an increase in ED₅₀ valueswith the time elapsed was also observed.     

Sublethal infection of different pine species by the pinewood nematode

Bursaphelenchus xylophilus, the pinewood nematode and causal agent of pine wilt disease (PWD), is a globally destructive pine pest. PWD is favoured by high temperatures and drought, but B. xylophilus can also persist in symptomless hosts in areas not prone to PWD. We compared seedlings of different Pinus species that were inoculated with B. xylophilus when well-watered and under moderate temperatures, conditions that favour sublethal infections. We compared isolates of B. xylophilus in south-west Europe (Portugal) and south-east USA (Louisiana). P. pinea, P. pinaster, and P. radiata were challenged in both areas with inoculations of local B. xylophilus. P. sylvestris in Portugal and P. palustris in Louisiana were also tested. Seedling mortality was low, as expected, but nematode establishment was common. There were higher nematode densities and more damage in seedlings of P. sylvestris and P. radiata, followed by P. pinaster, with P. pinea and P. palustris proving to be more tolerant. Life history of hosts was a better predictor of resistance to B. xylophilus than geographical origin, even under sublethal conditions. Fast-growing species (P. radiata and P. pinaster) appeared most likely to be symptomless carriers in areas not prone to the development of PWD. Cold-adapted species (P. sylvestris) may still be suitable hosts for the pathogen, even at the sublethal conditions of their distribution areas. There were more nematodes per gram of host using nematode isolates from Portugal. Nonetheless, our results opposed the hypothesis that North American pinewood nematodes have such low virulence that they are obligatorily saprophytic.     

Characterization of whitebait (Galaxias maculatus) respiratory rates to optimize intensive culture carrying capacities

Galaxias maculatus is an osmeriform native fish of the Southern Hemisphere, in which the crystalline larvae is considered as a luxury delicacy, for this reason, it has been commercially exploited in Chile, Argentina and New Zealand. However, the fisheries have been rapidly decreasing due to the overexploitation and the predation of introduced species. Because of these events, there is a need to determine a carrying capacity for an intensive fish culture. In order to optimize stocking densities for fish culture, this paper proposes objectives to determine oxygen consumption (OC) rates, dissolved oxygen concentrations that produce signs of hypoxia and the average time elapsed between food intake and peak OC in G. maculatus. In the oxygen experiments under routine metabolism conditions, we found that G. maculatus adults and whitebait showed signs of asphyxia at dissolved oxygen concentrations between 1.3 and 2.2 mg L^?1 and that adults tolerated dissolved oxygen levels as low as 1.3 mg L^?1. The results showed that G. maculatus individuals with an average weight of 0.04 g consumed 0.048 mg O₂ h^?1, whereas individuals with an average weight of 1.4 g consumed 0.345 mg O₂ h^?1. Galaxias maculatus increased the OC rate by 31%, from 0.39 to 0.51 mg O₂ h^?1 g^?1, occurring 14 min after food intake. The carrying capacities for industrial cultures of G. maculatus, were estimated using an allometric equation (OC=0.2363 ×W ^0.612 ), a water flow rate of 1 m³ h^?1 and an input oxygen concentration of 10 mg L^?1 at 12 °C. The density culture of whitebait (4 g) can be allowed to reach 8–11 kg m^?3; therefore, these stocking densities reduce the risk of hypoxia and mortality, ensuring the appropriate growth and feed conversion rates.