A comparative assessment of plant flammability through a functional approach: The case of woody species from Argentine Chaco region

Recent changes to fire regimes in many regions of the world have led to renewed interest in plant flammability experiments to understand and predict the consequences of such changes. These experiments require the development of practical and standardised flammability testing protocols. The research aims were (i) to compare plant flammability assessments carried out using two different approaches, namely functional trait analysis and testing with a shoot‐level device; and (ii) to evaluate the effect of disturbances and seasonal variability on flammability. The study area was located in the Western Chaco region, Argentina, and 11 species were selected based on their representativeness in forests. We studied six functional traits related to flammability, growth habit and foliar persistence, in forests without disturbances over the three last decades as well as in disturbed forests. The seasonal variation of these functional traits was evaluated over two consecutive years. Functional trait flammability index (FI) and shoot‐level measurements followed standard protocols. Sixty per cent of the species measured presented a high to very high FI. The results of both assessment methods were significantly correlated. Both methods identified the same species as having medium flammability, but differed in regards to the most flammable species. Senegalia gilliesii was identified as the most flammable species when using functional trait analysis, whereas shoot‐level assessments found Larrea divaricata and Schinus johnstonii to be the most flammable. There were no disturbance effects on the FI but there was seasonal variation. Our results validate the use of functional traits as a predictive method of flammability testing and represent the first global effort comparing flammability obtained through functional trait analysis with empirical measurements. The significant correlation between both methods allows the selection of the one that is more appropriate for the size of the area to be evaluated and for the availability of technical resources. Abstract in Spanish is available with online material.     

Advanced fertility diagnosis in stallion semen using transmission electron microscopy

Routine semen analysis of stallions is based on light microscopy (LM). However, there are still a number of animals that are subfertile or even infertile not being identified with conventional semen analysis. The objective of this study was to investigate the suitability of transmission electron microscopy (TEM) for advanced fertility diagnosis in stallion. We examined ejaculates of 46 stallions with known fertility. Animals were divided into three different groups: group 1, fertile stallions (pregnant mares> or =70%, n=29); group 2, subfertile stallions (pregnant mares 10-69%, n=14); group 3, infertile stallions (pregnant mares<10%, n=3). Ejaculates were collected in spring 2002. Conventional semen analysis (volume, sperm concentration, motility, live:dead ratio and percentage of morphologically normal sperm) was immediately performed after semen collection. Ultrastructural analysis included the evaluation of 200 acrosomes, heads, midpieces and cross-sections of tails as well as 100 longitudinal sections of tails from every ejaculate. Using LM, we found a significant increase of morphological deviations from 24.5% (x ) in group 1 to 34.5% in group 2 and 73.5% in group 3. Using TEM, we found a significant increase of detached acrosomes from 6.1% in group 1 to 7.6% in group 2 and 21.4% in group 3. Deviations in tubule pattern were also increased (but not significant) from 2.7% in fertile and 2.8% in subfertile to 11.4% in infertile stallions as well as multiple tails from 1.9% in fertile to 2.0% in subfertile and 8.9% in infertile. Our data indicate that TEM is suitable for advanced fertility diagnostic in stallions, giving a connection between fertility and morphology. It suggests that the most likely reason for sub- and infertility in stallion in case of increased LM pathomorphology of semen are acrosomal alterations, especially detached acrosomes.     

LCA of energetic biomass utilization: actual projects and new developments—April 23, 2012, Berne, Switzerland

Introduction

In the last years, the use of biomass for energy purposes has been seen as a promising option to reduce the use of nonrenewable energy sources and the emissions of fossil carbon. However, LCA studies have shown that the energetic use of biomass also causes impacts on climate change and, furthermore, that different environmental issues arise, such as land use and agricultural emissions. While biomass is renewable, it is not an unlimited resource. Its use, to whatever purpose, must therefore be well studied to promote the most efficient option with the least environmental impacts. The 47th LCA Discussion Forum gathered several national and international speakers who provided a broad and qualified view on the topic.

Summary of the topics presented in DF 47

Several aspects of energetic biomass use from a range of projects financed by the Swiss Federal Office of Energy (SFOE) were presented in this Discussion Forum. The first session focused on important aspects of the agricultural biogas production like the use of high energy crops or catch crops as well as the influence of plant size on the environmental performance of biogas. In the second session, other possibilities of biomass treatment like direct combustion, composting, and incineration with municipal waste were presented. Topic of the first afternoon session was the update and harmonization of biomass inventories and the resulting new assessment of biofuels. The short presentations investigated some further aspects of the LCA of bioenergy like the assessment of spatial variation of greenhouse gas (GHG) emissions from bioenergy production in a country, the importance of indirect land use change emissions on the overall results, the assessment of alternative technologies to direct spreading of digestate or the updates of the car operation datasets in ecoinvent.

Conclusions

One main outcome of this Discussion Forum is that bioenergy is not environmentally friendly per se. In many cases, energetic use of biomass allows a reduction of GHG and fossil energy use. However, there is often a tradeoff with other environmental impacts linked to agricultural production like eutrophication or ecotoxicity. Methodological challenges still exist, like the assessment of direct and indirect land use change emissions and their attribution to the bioenergy production, or the influence of heavy metal flows on the bioenergy assessment. Another challenge is the implementation of a life cycle approach in certification or legislation schemes, as shown by the example of the Renewable Energy Directive of the European Union.     

Thermofluor-based optimization strategy for the stabilization and crystallization of Campylobacter jejuni desulforubrerythrin

Desulforubrerythrin from Campylobacter jejuni has recently been biochemical and spectroscopically characterized. It is a member of the rubrerythrin family, and it is composed of three structural domains: the N-terminal desulforedoxin domain with a non-heme iron center, followed by a four-helix bundle domain harboring a binuclear iron center and finally a C-terminal rubredoxin domain. To date, this is the first example of a protein presenting this kind of structural domain organization, and therefore the determination of its crystal structure may unveil unexpected structural features. Several attempts were made in order to obtain protein crystals, but always without success. As part of our strategy the thermofluor method was used to increase protein stability and its propensity to crystallize. This approach has been recently used to optimize protein buffer formulation, thus yielding more stable and homogenous protein samples. Thermofluor has also been used to identify cofactors/ligands or small molecules that may help stabilize native protein states. A successful thermofluor approach was used to select a pH buffer condition that allowed the crystallization of Campylobacter jejuni desulforubrerythrin, by screening both buffer pH and salt concentration. A buffer formulation was obtained which increased the protein melting temperature by 7°C relatively to the initial purification buffer. Desulforubrerythrin was seen to be stabilized by lower pH and high salt concentration, and was dialyzed into the new selected buffer, 100mM MES pH 6.2, 500mM NaCl. This stability study was complemented with a second thermofluor assay in which different additives were screened. A crystallization screening was carried out and protein crystals were rapidly obtained in one condition. Protein crystal optimization was done using the same additive screening. Interestingly, a correlation between the stability studies and crystallization experiments using the additive screening could be established. The work presented here shows an elegant example where thermofluor was shown to be a key biophysical method that allowed the identification of an improved buffer formulation and the applicability of this technique to increase the propensity of a protein to crystallize is discussed.     

Availability and toxicity of cadmium to forage grasses grown in contaminated soil

It is important to know the mechanisms for forage development, especially those related to the tolerance of potentially toxic elements, when considering their use in phytoremediation in heavy metal contaminated areas. In this study, we evaluated plant growth, concentration, and the availability of cadmium (Cd) for forage grasses (Panicum maximum Jacq. cv. Aruana and cv. Tanzânia; Brachiaria decumbens cv. Basilisk; Brachiaria brizantha cv. Xaraés and cv. Marandu) cultivated in Cd contaminated soils. The experiments were performed under greenhouse conditions over a 90-day evaluation period, and the Cd rates were 2, 4, and 12 mg/kg of soil. The relative growth rate of the forage grasses decreased as Cd rates increased, and the following descending order of susceptibility was observed: Marandu > Xaraés > Aruana > Tanzânia > Basilisk, with regard to phytotoxicity in these plants. The forage Cd concentration increased in line with increases in the Cd rates. Cd contents extracted by Mehlich-1 and by diethylenetriaminepentaacetic acid presented high positive correlation with forage relative growth. The forage plants did not block Cd entry into the food chain because they were not capable of limiting Cd absorption.     

Genotoxic damage in Solea senegalensis exposed to sediments from the Sado Estuary (Portugal): Effects of metallic and organic contaminants

Juvenile Solea senegalensis (Senegalese sole) were exposed to freshly collected sediments from three sites of the Sado Estuary (West-Portuguese coast) in 28-day laboratory assays in order to assess the ecological risk from sediment contaminants, by measuring two genotoxicity biomarkers in peripheral blood: the percentage of Erythrocyte Nuclear Abnormalities (ENA) by use of an adaptation of the micronucleus test, and the percentage of DNA strand-breakage (DNA-SB) with the Comet assay. Sediments were surveyed for metallic (Cr, Ni, Cu, Zn, As, Cd and Pb) and organic (PAHs (polycyclic aromatic hydrocarbons), PCBs (polychlorinated biphenyls) and DDTs (dichloro-diphenyl-trichloroethane)) contaminants. Sediments from site A (farthest from hotspots of contamination) were found to be the least contaminated and weaker inducers of genotoxic damage, whereas sediments from sites B (urban influence) and C (affected by industrial effluents and agricultural runoffs) were responsible for a very significant increase in both ENA and DNA-SB, site B being most contaminated with metals and site C mainly with organic pollutants, especially PAHs and PCBs . Analysis of genotoxic effects showed a strong correlation between the concentrations of PAHs and PCBs and both biomarkers at sampling times T₁₄ and T₂₈, while the amounts of Cu, As, Cd and Pb were less strongly correlated, and at T₂₈ only, with ENA and DNA-SB. These results show that organic contaminants in sediment are stronger and faster acting genotoxic stressors. The results also suggest that metals may have an inhibitory effect on genotoxicity when interacting with organic contaminants, at least during early exposure. ENA and DNA-SB do not show a linear relationship, but a strong correlation exists between the overall increase in genotoxicity caused by exposure to sediment, confirming that they are different, and possibly non-linked effects that respond similarly to exposure. Although the Comet assay showed enhanced sensitivity, the two analyses are complementary and suitable for the biomonitoring of sediment contaminants in a benthic species like S. senegalensis.     

Mouse heart valve structure and function: echocardiographic and morphometric analyses from the fetus through the aged adult

The purpose of this study is to provide standard echocardiographic and morphometric data for normal mouse valve structure and function from late fetal to aged adult stages. Cross-sectional, two-dimensional and Doppler transthoracic echocardiography was performed in C57BL6 mice anesthetized with 1% to 2% isoflurane at embryonic day 18.5 (late fetal), 10 days (neonate), 1 mo (juvenile), 2 mo (young adult), 9 mo (old adult), and 16 mo (aged adult). Normal annulus dimensions indexed to age or weight, and selected flow velocities, were established by echocardiography. After echocardiographic imaging, hearts were harvested and histological and morphometric analyses were performed. Morphometric analysis demonstrated a progressive valve thinning and elongation during the fetal and juvenile stages that plateaued during adult stages (ANOVA, P < 0.01); however, there was increased thickening of the hinge of the aortic valve with advanced age, reminiscent of human aortic valve sclerosis. There was no age-related calcification. The results of this study provide comprehensive echocardiographic and morphometric data for normal mouse valve structure and function from late fetal to aged adult stages and should prove useful as a reference standard for future studies using mouse models of progressive valve disease.     

Nuclear localization of TRK-A in liver cells

The liver represents a site of expression of neurotrophins and their receptors. We have characterized the expression and intracellular localization of the nerve growth factor (NGF) receptor, Trk-A, in liver cells in vivo and in vitro. In both normal and fibrotic liver tissue, Trk-A immunostaining was present in different cell types, including parenchymal cells and cells of the inflammatory infiltrate. In hepatocytes and activated stellate cells (HSC), Trk-A showed a predominant nuclear localization, both in the presence and absence of injury. In cultured HSC, Trk-A was found to be functional, because exposure of the cells to recombinant NGF resulted in stimulation of cell migration and activation of intracellular signaling pathways, including Ras-ERK and PI3K/Akt. Remarkably, in cultured HSC, Trk-A staining was found constitutively in the nucleus. In these cells, Trk-A could be stained only by antibodies directed against the intracellular domain but not by those recognizing the extracellular portion of Trk-A suggesting that the intracellular portion of the receptor is the major determinant of nuclear Trk-A staining. In contrast to HSC, freshly isolated hepatocytes did not show any nuclear localization of the intracellular portion of Trk-A. In pheocromocytoma cells, nuclear staining for Trk-A was not present in conditions of serum deprivation, but could be induced by exposure to NGF or to a mixture of soluble mediators. We conclude that nuclear localization of the intracellular domain of Trk-A is observed constitutively in liver cells such as HSC, while in other cell types it could be induced in response to soluble factors.     

A novel meta‐analytical approach to improve systematic review of rates and patterns of microevolution

A classic topic in ecology and evolution, phenotypic microevolution of quantitative traits has received renewed attention in the face of rapid global environmental change. However, for plants, synthesis has been hampered by the limited use of standard metrics, which makes it difficult to systematize empirical information. Here we demonstrate the advantages of incorporating meta‐analysis tools to the review of microevolutionary rates. We perform a systematic survey of the plant literature on microevolution of quantitative traits over known periods of time, based on the scopus database. We quantify the amount of change by standard mean difference and develop a set of effect sizes to analyze such data. We show that applying meta‐analysis tools to a systematic literature review allows the extraction of a much larger volume of information than directly calculating microevolutionary rates. We also propose derived meta‐analysis effect sizes (h, LG and LR) which are appropriate for the study of evolutionary patterns, the first being similar to haldanes, the second and third allowing the application of a preexisting analytical framework for the inference of evolutionary mechanisms. This novel methodological development is applicable to the study of microevolution in any taxa. To pilot test it, we built an open‐access database of 1,711 microevolutionary rates of 152 angiosperm species from 128 studies documenting population changes in quantitative traits following an environmental novelty with a known elapsed time (<260 years). The performance of the metrics proposed (h, LG and LR) is similar to that of preexisting ones, and at the same time they bring the advantages of lower estimation bias and higher number of usable observations typical of meta‐analysis.