Identification of sperm proteins as candidate biomarkers for the analysis of reproductive isolation in Mytilus: a case study for the enkurin locus

Sperm proteins of the marine sessile mussels of the Mytilus edulis species complex are models to investigate reproductive isolation and speciation. This study aimed at identifying sperm proteins and their corresponding genes. This was aided by the use of monoclonal antibodies that preferentially bind to yet unknown sperm molecules. By identifying their target molecules, this approach identified proteins with relevance to Mytilus sperm function. This procedure identified 16 proteins, for example, enkurin, laminin, porin and heat shock proteins. The potential use of these proteins as genetic markers to study reproductive isolation is exemplified by analysing the enkurin locus. Enkurin evolution is driven by purifying selection, the locus displays high levels of intraspecific variation and species-specific alleles group in distinct phylogenetic clusters. These findings characterize enkurin as informative candidate biomarker for analyses of clinal variation and differential introgression in hybrid zones, for example, to understand determinants of reproductive isolation in Baltic Mytilus populations.     

DNA damage in oral epithelial cells of individuals chronically exposed to indoor radon (<Superscript>222</Superscript>Rn) in a hydrothermal area

Hydrothermal areas are potentially hazardous to humans as volcanic gases such as radon (²²²Rn) are continuously released from soil diffuse degassing. Exposure to radon is estimated to be the second leading cause of lung cancer, but little is known about radon health-associated risks in hydrothermal regions. This cross-sectional study was designed to evaluate the DNA damage in the buccal epithelial cells of individuals chronically exposed to indoor radon in a volcanic area (Furnas volcano, Azores, Portugal) with a hydrothermal system. Buccal epithelial cells were collected from 33 individuals inhabiting the hydrothermal area (Ribeira Quente village) and from 49 individuals inhabiting a non-hydrothermal area (Ponta Delgada city). Indoor radon was measured with Ramon 2.2 detectors. Chromosome damage was measured by micronucleus cytome assay, and RAPD-PCR was used as a complementary tool to evaluate DNA damage, using three 10-mer primers (D11, F1 and F12). Indoor radon concentration correlated positively with the frequency of micronucleated cells (r _s = 0.325, p = 0.003). Exposure to radon is a risk factor for the occurrence micronucleated cells in the inhabitants of the hydrothermal area (RR = 1.71; 95% CI, 1.2–2.4; p = 0.003). One RAPD-PCR primer (F12) produced differences in the banding pattern, a fact that can indicate its potential for detecting radon-induced specific genomic alterations. The observed association between chronic exposure to indoor radon and the occurrence of chromosome damage in human oral epithelial cells evidences the usefulness of biological surveillance to assess mutations involved in pre-carcinogenesis in hydrothermal areas, reinforcing the need for further studies with human populations living in these areas.     

Trade-offs between maternal foraging and fawn predation risk in an income breeder

The choice of neonatal hiding place is critical for ungulates adopting hiding anti-predator strategies, but the consequences of different decisions have rarely been evaluated with respect to offspring survival. First, we investigated how landscape-scale choices made by roe deer fawns and their mothers affected predation risk by red foxes in a forest–farmland mosaic in southeastern Norway. After, we examined the effect of site-specific characteristics and behaviour (i.e. visibility, mother–fawn distance and abundance of the predator’s main prey item—small rodents) on predation risk. The study of habitat use, selection and habitat-specific mortality revealed that roe deer utilised the landscape matrix in a functional way, with different habitats used for feeding, providing maternal care and as refugia from predation. Mothers faced a trade-off between foraging and offspring survival. At the landscape-scale decisions were primarily determined by maternal energetic constraints and only secondarily by risk avoidance. Indeed, forage-rich habitats were strongly selected notwithstanding the exceptionally high densities of rodents which increased fawn predation. At fine spatial scales, a high visibility of the mother was the major factor determining predation risk; however, mothers adjusted their behaviour to the level of risk at the bed site to minimise predation. Fawns selected both landscape-scale refugia and concealed bed sites, but failure to segregate from the main prey of red foxes led to higher predation. This study provides evidence for the occurrence of spatial heterogeneity in predation risk and shows that energetically stressed individuals can tackle the foraging-safety trade-off by adopting scale-dependent anti-predator responses.     

Effect of the nitrification inhibitor dicyandiamide on microbial communities and N<Subscript>2</Subscript>O from an arable soil fertilized with ammonium sulphate

Nitrous oxide (N₂O) affects climate change as a greenhouse gas and indirectly contributes to stratospheric ozone depletion. The main source of N₂O in soils is denitrification which requires high soil moisture, carbon and nitrate. Nitrification inhibitors can be used to mitigate emissions of N₂O from soils. In Portugal, fertilisers are often applied when soils are still relatively warm and moist conditions conducive to denitrification. A Portuguese arable soil was inhibited with dicyandiamide, a nitrification inhibitor and the effect on soil microbiological activity and composition was determined after 46 days. Soils were then incubated and received carbon and ammonium under high soil water conditions and mineral N and N₂O fluxes were measured during 22 days. We found that dicyandiamide decreased microbial populations and activity, but did not alter composition. Pre-conditioning of the soil with dicyandiamide was 80% more effective in reducing fluxes of N₂O than simultaneous application with fertiliser.     

Beurteilung des Ozonrisikos für die Waldregionen Bayerns am Beispiel des Jahres 2002 und des Extremtrockenjahres 2003 auf der Basis der externen Ozonexposition und der internen Ozonaufnahme

Background, aim, and scope Increasing background concentrations of ground-level tropospheric ozone and more frequent and prolonged summer drought incidences due to climate change are supposed to increase the stress on Bavarian forests. For such scenarios growth reduction and yield losses are predicted. Sustainable forest management in Bavaria aims to significantly increase the proportion of beech (Fagus sylvatica L.) because of its broad ecological amplitude. In our regional study different approaches for calculating ozone impact were used to estimate the risks for Bavarian forests in the average climatic, rather moist year 2002 and the extremely dry year 2003.Materials and methods Measurements were conducted for eleven forest ecosystem sites and two forest research sites representing typical Bavarian forest stands under different climatic conditions and situated in different altitudes. For risk assessment currently used approaches were applied either based on the calculation of the cumulative ozone exposure (external dose; MPOC maximal permitted ozone concentration; critical level AOT40_phen? accumulated ozone exposure over a threshold of 40 nl [O₃] l^–1, for the effective phenolgy of beech) or based on the calculation of the phytomedically relevant ozone flux into the stomata (internal dose, critical level AF_st>1,6, accumulated stomatal flux above a flux threshold of 1.6 nmol O₃?m^–2 PLA; PLA = projected leaf area). For calculations continuously recorded ozone concentrations and meteorological and phenological data from nearby rural open field background measuring stations from the national air pollution control and from forested sites were used. Additionally ozone induced leaf symptoms were assessed.Results The exposure-based indices AOT40_phen and MPOC as well as the flux-based index AF_st>1.6suggest that Bavarian forests are at risk from O₃ during a rather moist average year concerning climate conditions (2002) as well as in an extreme dry year (2003). Thus, growth reductions of 5?% are predicted when thresholds are exceeded. Threshold exceedance occurred in both years at all plots, mostly already at the beginning of the growing season and often even many times over. Ozone induced leaf symptoms could be detected only on a few plots in a very slight occurrence.Discussion The results for the applied critical level indices differed depending on climatic conditions during the growing seasons: Regarding exposure-based indices, the highest degree of threshold exceedance occurred in the dry year of 2003 at all plots; the flux-based approach indicated the highest stomatal ozone uptake and thus an increased risk at moist sites or during humid years, whereas the risk was decreasing at dry sites with prolonged water limitation. Hence, soil and accordingly plant water availability was the decisive factor for the flux-modelled internal ozone uptake via stomata. Drought and increased ozone impact can generate synergistic, but also antagonistic effects for forest trees. At water limited rather dry forest sites restricted transpiration and thus production, but concurrently lower ozone uptake and reduced risk for damage can be expected.Conclusions, recommendations, and perspectives For realistic site-specific risk assessment in forest stands the determination of the internal ozone dose via modeling flux based internal stomatal ozone uptake is more appropriate than the calculation of the external ozone dose. The predicted 5?% growth reductions are in discrepancy with the frequently observed increment increase during the last decades in forest stands. Comprehensive and significant statistical verification for ozone induced forest growth reduction as well as the systematic validation of thresholds for ozone in the field is still lacking. However, a multiplicity of different specific new and retrospective growth analysis data should allow closing the gap. Moreover, the determination of canopy transpiration with sap flow measurements is a novel approach to provide cause-effect related, site specific results for the effective internal ozone dose as well as for canopy water supply and consecutively for regional risk estimation. A further future objective is the refinement of O₃ flux modelling by further consideration of soil/water budget characteristics and the above mentioned improved estimations of crown and canopy transpiration. Further, the introduction of threshold ranges for forest trees in view of their specific regional climatic conditions and their validation in real forest stands is necessary for developing meaningful ozone risk predictions for forests.     

The impact of global change on economic values of water for Public Irrigation Schemes at the São Francisco River Basin in Brazil

Economic values of water for the main Public Irrigation Schemes in the sub-middle region of the São Francisco River Basin, in northeastern Brazil, are determined in this study using an integration of a global agro-economic land and water use (MAgPIE) with a local economic model (Positive Mathematical Programming). As in the latter, the water values depend on the crops grown, and as Brazilian agriculture is strongly influenced by the global market, we used a regionalized version of the global model adapted to the region in order to simulate the crop land use, which is in turn determined by changes in global demand, trade barriers, and climate. The allocation of sugarcane and fruit crops projected with climate change by the global model, showed an impact on the average yields and on the water costs in the main schemes resulting in changes in the water values locally. The economic values for all schemes in the baseline year were higher than the water prices established for agricultural use in the basin. In the future, these water values will be higher in all the schemes. The highest water values currently and in the future were identified in municipalities with a significant proportion of area growing irrigated sugarcane. Being aware of current water values of each user in a baseline year and in a projected future under global climate and socioeconomic changes, decision makers should improve water allocation policies at local scale, in order to avoid conflicts and unsustainable development in the future.     

Wastewater reuse: modeling chloroform formation

The chloroform is a substance that presents a significant risk to or via the aquatic environment. Thus, the emissions, discharges and losses of this substance need to be controlled during wastewater disinfection for reclamation and reuse purposes. Due to its carcinogenetic potential, multiple studies have been carried out on drinking and surface/natural waters but less consideration has been directed to the wastewater disinfection. The focus of this work studied the formation of chloroform during chlorination in prepared waters or artificial matrices that intended to simulate wastewaters stored in landscape ponds for green areas irrigation. The relation between reaction time, chlorine dose, and chloroform formation and the variation of the dissolved organic carbon (DOC) content during the reaction was assessed. A two-variant model was proposed to simulate breakpoint chlorination practices (when chlorine dose is equal or lower than chlorine demand) and super chlorination techniques (when chlorine dose tends to surpass chlorine demand). The model was validated by the application of actual data from working conditions of six wastewater treatment plants located in Algarve, Portugal, including other data obtained in previous research studies that were not used in the model development, and by comparing the predicted values with real measured ones.     

Economic assessment of managing processionary moth in pine forests: a case-study in Portugal

This paper assesses the private and social profitability of current strategies for managing processionary moth (Thaumetopoea pityocampa) in Portuguese pine forests, looking at economic and environmental costs and benefits. Costs include the expenses for forest treatment and the social costs of threats to human health (dermatitis amongst others); benefits are assessed in terms of both revenue and social benefits such as carbon fixation and recreation. The evaluation was done using Cost Benefit Analysis (CBA) as an analytical framework. While this tool is currently applied to forest and environmental assessment and specific applications to pest management strategies are to be found in agricultural economics, rather few attempts have been made in the field of forest pest management. In order to assess and compare with--without options, a case-study was analysed for the Setúbal Peninsula, south of Lisbon, an area where extensive stands of maritime pine (Pinus pinaster) grow. The exercise has shown that CBA can be a valuable tool for assessing the economic and social profitability of pest management. The results demonstrate that the loss of revenues in the no-management option is not sufficient to make pest management profitable for private forest owners in the short-term. Conversely, a social profit is gained as pest management minimizes health risks for humans and avoids possible recreational losses.     

Numerical simulations on the influence of matrix diffusion to carbon sequestration in double porosity fissured aquifers

The double porosity model for fissured rocks, such as limestones and dolomites, has some features that may be relevant for carbon sequestration. Numerical simulations were conducted to study the influence of matrix diffusion on the trapping mechanisms relevant for the long-term fate of CO₂ injected in fissured rocks. The simulations show that, due to molecular diffusion of CO₂ into the rock matrix, dissolution trapping and hydrodynamic trapping are more effective in double porosity aquifers than in an equivalent porous media. Mineral trapping, although assessed indirectly, is also probably more relevant in double porosity aquifers due to the larger contact surface and longer contact time between dissolved CO₂ and rock minerals. However, stratigraphic/structural trapping is less efficient in double porosity media, because at short times CO₂ is stored only in the fissures, requiring large aquifer volumes and increasing the risk associated to the occurrence of imperfections in the cap-rock through which leakage can occur. This increased risk is also a reality when considering storage in aquifers with a regional flow gradient, since the CO₂ free-phase will move faster due to the higher flow velocities in fissured media and discharge zones may be reached sooner.