Morphological traits for allometric scaling of the European Sea Bass Dicentrarchus labrax (Linnaeus, 1758) from Southern Portugal population.

The present study aimed to determine the allometric scaling among a selection of morphological traits in European sea bass (Dicentrarchus labrax) to estimate fish body weight. A set of morphological traits (fish body weight, length, height, and width) were directly measured in 146 fish of a recirculating aquaculture system, with body weights ranging from 17.11 to 652.21 g. In addition, a collection of digital imagery of each anesthetized fish from the side and top views were used to estimate other traits (indirect measures). Multiple regression analysis and regression coefficients were calculated using all possible combinations of biometric data (predictors) to estimate fish body weight, applying different numerical fitting models (linear, log-linear, quadratic, exponential). The results showed that the best combination of traits for estimating fish body weight were fish body width, length and height, collected from direct measure (R2 = 0.995), for a log-linear model fitting, which revealed more accurate determinations than the most commonly used length-weight relationship. Nevertheless, other combinations of morphological traits and fitting models were also found to be suitable in successfully predict fish body weight, with variability ranging between 92.5% and 98.5%. For indirect measures, the best predictor was a combination of traits from top view (width, eye distance and area without fins) fitted with a log-linear function. These results comprise a relevant baseline in supporting the high potential of noninvasive methods to accurately follow the growth of European sea bass juveniles, recurring to imagery analysis of anesthetized fish. It has major potential applications in feeding consumption trials and fish growth models, as it allows for continuously following up fish growth under different experimental conditions without therein distress derived from manipulation.

Influence of some parameters on the germination assessment of mycopesticides.

The substantial negative impact of some parameters on the germination of low-quality conidia (high proportion of slow-germinating propagules) was demonstrated, whereas for high-quality batches their effect was small or even absent. Germination was increased as the initial hydration status of conidia immediately prior to suspension preparation was increased, being ca. 33% and 80% for dehydrated Metarhizium anisopliae propagules (water activity ≤0.314) from low- or high-quality batches after an 18 h incubation period, respectively, and 63% and 95% for hydrated propagules (water activity = 0.933). Germination of low-quality propagules also increased as the time dry conidia were kept in aqueous suspension prior to inoculation onto culture media (15 min, 3 or 24 h) or the incubation time at 25°C before counts (18, 48 or 72 h) was increased. Depending on treatment conditions, average germination of low-quality conidia varied from 53% to 98%. On the other hand, germination for high-quality conidia was always ≥94%. Regarding the relative humidity (RH) of the incubation atmosphere, the average germination rates for low-quality conidia on Potato Dextrose Agar (PDA) in Petri plates was 49%, while germination of these conidia on PDA blocks kept under lower RH inside plastic boxes was ≤23%. Use of lactophenol-staining and/or use of coverslips had a negative effect when germination assessment was performed for low-quality conidia, resulting in distorted counts or increased standard deviations compared to high-quality conidial batches. The occurrence of dislodged conidia (ungerminated conidia outside the inoculation zone due to hydraulic pressure exercised by addition of stains and/or coverslips added to the substrate by the time germination is assessed) was common place, whereas dislodged conidia were not seen in treatments with high-quality batches. This work underscores the importance of a number of parameters that anyone working with low-quality fungi needs to be cognizant of in their research.

Stressors of emerging concern in deep-sea environments: microplastics, pharmaceuticals, personal care products and deep-sea mining.

Although most deep-sea areas are remote in comparison to coastal zones, a growing body of literature indicates that many sensitive ecosystems could be under increased stress from anthropogenic sources. Among the multiple potential stressors, microplastics (MPs), pharmaceuticals and personal care products (PPCPs/PCPs) and the imminent start of commercial deep-sea mining have received increased attention. Here we review recent literature on these emerging stressors in deep-sea environments and discuss cumulative effects with climate change associated variables. Importantly, MPs and PPCPs have been detected in deep-sea waters, organisms and sediments, in some locations in comparable levels to coastal areas. The Atlantic Ocean and the Mediterranean Sea are the most studied areas and where higher levels of MPs and PPCPs have been detected. The paucity of data for most other deep-sea ecosystems indicates that many more locations are likely to be contaminated by these emerging stressors, but the absence of studies hampers a better assessment of the potential risk. The main knowledge gaps in the field are identified and discussed, and future research priorities are highlighted to improve hazard and risk assessment.

Deep-sea mining: using hyperbaric conditions to study the impact of sediment plumes in the subtidal clam Spisula solida.

With the growing interest to exploit mineral resources in the deep-sea, there is the need to establish guidelines and frameworks to support hazard and risk assessment schemes. The present study used a subtidal species of filter-feeding bivalve, the clam Spisula solida, as a proxy to better understand the impacts of sediment plumes in marine organisms under hyperbaric conditions. Four concentrations of suspended sediments (0 g/L, 1 g/L, 2 g/L, and 4 g/L) were used in a mixture with different grain sizes at 4 Bar for 96 h. Functional (filtration rate-FR) and biochemical endpoints (catalase-CAT, glutathione s-transferase-GST, and lipid peroxidation-LPO) were analyzed in the gonads, digestive gland, and gills of S. solida after a 96-h exposure at 4 Bar (the natural limit of the species vertical distribution). The FR showed a decreasing trend with the increasing sediment concentrations (significant effects at 2 and 4 g/L). Additionally, significant changes were observed for some of the tested oxidative stress biomarkers, which were concentration and tissue-dependent, i.e., CAT activity was significantly elevated in gills (1 g/L treatment), and GST was decreased in digestive gland (1 g/L treatment). Overall, the results show that suspended sediments, at 2 and 4 g/L, have negative functional impacts in the bivalve S. solida providing additional insights to improve hazard assessment of deep-sea mining. These findings represent a step forward to ensure the mitigation of the potential negative effects of deep-sea resource exploitation.

A modelling framework to assess multiple metals impacts on marine food webs: Relevance for assessing the ecological implications of deep-sea mining based on a systematic review.

Industrial deep-sea mining will release plumes containing metals that may disperse over long distances; however, there is no general understanding of metal effects on marine ecosystems. Thus, we conducted a systematic review in search of models of metal effects on aquatic biota with the future perspective to support Environmental Risk Assessment (ERA) of deep-sea mining. According to results, the use of models to study metal effects is strongly biased towards freshwater species (83% freshwater versus 14% marine); Cu, Hg, Al, Ni, Pb, Cd and Zn are the best-studied metals, and most studies target few species rather than entire food webs. We argue that these limitations restrain ERA on marine ecosystems. To overcome this gap of knowledge, we suggest future research directions and propose a modelling framework to predict the effects of metals on marine food webs, which in our view is relevant for ERA of deep-sea mining.

Prioritizing the Effects of Emerging Contaminants on Estuarine Production under Global Warming Scenarios.

Due to non-linear interactions, the effects of contaminant mixtures on aquatic ecosystems are difficult to assess, especially under temperature rise that will likely exacerbate the complexity of the responses. Yet, under the current climatic crisis, assessing the effects of water contaminants and temperature is paramount to understanding the biological impacts of mixtures of stressors on aquatic ecosystems. Here, we use an ecosystem model followed by global sensitivity analysis (GSA) to prioritize the effects of four single emerging contaminants (ECs) and their mixture, combined with two temperature rise scenarios, on the biomass production of a NE Atlantic estuary. Scenarios ran for 10 years with a time-step of 0.1 days. The results indicate that macroinvertebrate biomass was significantly explained by the effect of each single EC and by their mixture but not by temperature. Globally, the most adverse effects were induced by two ECs and by the mixture of the four ECs, although the sensitivity of macroinvertebrates to the tested scenarios differed. Overall, the present approach is useful to prioritize the effects of stressors and assess the sensitivity of the different trophic groups within food webs, which may be of relevance to support decision making linked to the sustainable management of estuaries and other aquatic systems.

Boom and bust: Simulating the effects of climate change on the population dynamics of a global invader near the edge of its native range.

Despite the increasing awareness of climate change, few studies have used the Intergovernmental Panel on Climate Change (IPCC) scenarios to simulate the effects of climate change on estuarine populations of crustaceans. The objective of this study was to investigate the effects of temperature and salinity fluctuations on the population dynamics of the shore crab Carcinus maenas at the southern edge of its native range. To this end, a population dynamics model was developed based on experimental and literature data on the biology, ecology and physiology of the species. Results showed that the shore crab will be more affected by changes in temperature than in salinity. The parameter sensitivity analysis revealed that the larval phase of the species is the most sensitive stage of the shore crab life cycle. Three IPCC scenarios (SSP1-2.6, SSP2-4.5, and SSP3-8.5) were used to simulate the effects of temperature increase on the population of C. maenas in the near- (2040), mid- (2060), and long-term (2100). Two scenarios of drought conditions accompanied by the estimated salinity change were also simulated (10 % and 40 % drought). Results suggested that slight increases in temperature (up to 2 °C) lead to a strong increase on the density of C. maenas in the mid-term, while further temperature increases lead to a decline or local extinction of the shore crab population at the southern edge of its distribution range. Results indicated that a salinity increase in the estuary had a negative effect on the shore crab population. Given the importance of the species to temperate coastal ecosystems, both population increase and local extinction are likely to have significant impacts on estuarine communities and food webs, with unknown ecological and socioeconomic consequences.

Trichoderma from Brazilian garlic and onion crop soils and description of two new species: Trichoderma azevedoi and Trichoderma peberdyi.

Fifty four Trichoderma strains were isolated from soil samples collected from garlic and onion crops in eight different sites in Brazil and were identified using phylogenetic analysis based on combined ITS region, tef1-α, cal, act and rpb2 sequences. The genetic variability of the recovered Trichoderma species was analysed by AFLP and their phenotypic variability determined using MALDI-TOF. The strain clusters from both typing techniques coincided with the taxonomic determinations made from phylogenetic analysis. The phylogenetic analysis showed the occurrence of Trichoderma asperellum, Trichoderma asperelloides, Trichoderma afroharzianum, Trichoderma hamatum, Trichoderma lentiforme, Trichoderma koningiopsis, Trichoderma longibrachiatum and Trichoderma erinaceum, in the soil samples. We also identified and describe two new Trichoderma species, both in the harzianum clade of section Pachybasium, which we have named Trichoderma azevedoi sp. nov. and Trichoderma peberdyi sp. nov. The examined strains of both T. azevedoi (three strains) and T. peberdyi (12 strains) display significant genotypic and phenotypic variability, but form monophyletic clades with strong bootstrap and posterior probability support and are morphologically distinct from their respective most closely related species.

Abundance and composition of floating marine macro litter on the eastern sector of the Mediterranean Sea.

Floating marine litter is known to be currently a widespread pollution problem, especially in the Mediterranean basin, but records on the levels of floating debris in the eastern part are less known. By using an observation platform (32 days, 137 transects, 1784 km), this study provides evidence of a high amount of floating debris in the Levantine sub-basin, within the eastern Mediterranean Sea. The density of floating litter varied between 18 and 1593 items km-2 (average 232 ±â€¯325 items km-2), and small plastic debris accounted for >90% of the items surveyed. These values tend to be higher than densities reported for the central and western Mediterranean areas, which may be related to the circulation patterns and inputs from coastal sources. Significant correlations of floating macro litter density with wind force and sea state were found, thus strengthening the need to standardise protocols (including oceanographic variables) of visual surveys of floating debris.

MODELPlastics workshop - Modelling Ocean Plastic Litter in a Changing Climate: Gaps and future directions.

Here we summarize the overarching issues that emerged from a workshop held to discuss scientific challenges and future directions on the use of numerical models to predict the amount, distribution and effects of plastic pollution on marine ecosystems. The need for multi-disciplinary approaches, standardized protocols for plastic quantification and analyses, using realistic contaminant concentrations in laboratorial experiments and targeting early-life stages of marine organisms were pointed out as needs to improve data accuracy. Participants also enumerated a list of gaps that include, identification of indicator organisms of plastic contamination, selection of biomarkers and the role of extreme events on plastic dynamics. Responding to these gaps will contribute to improve data quantity and quality and, thus, allow developing more reliable models. A crucial role is foreseen for modelling tools as they can incorporate the cumulative impacts of multiple stressors, including the individual-level effects of plastics, to the population- and ecosystem level.

Development of physical modelling tools in support of risk scenarios: A new framework focused on deep-sea mining.

Deep-sea mining has gained international interest to provide materials for the worldwide industry. European oceans and, particularly, the Portuguese Exclusive Economic Zone present a recognized number of areas with polymetallic sulphides rich in metals used in high technology developments. A large part of these resources are in the vicinity of sensitive ecosystems, where the mineral extraction can potentially damage deep-ocean life services. In this context, technological research must be intensified, towards the implementation of environmental friendly solutions that mitigate the associated impacts. To reproduce deep-sea dynamics and evaluate the effects of the mining activities, reliable numerical modelling tools should be developed. The present work highlights the usefulness of a new framework for risk and impact assessment based on oceanographic numerical models to support the adoption of good management practices for deep-sea sustainable exploitation. This tool integrates the oceanic circulation model ROMS-Agrif with the semi-Lagrangian model ICHTHYOP, allowing the representation of deep-sea dynamics and particles trajectories considering the sediments physical properties. Numerical simulations for the North Mid-Atlantic Ridge region, revealed the ability of ROMS-Agrif to simulate real deep-sea dynamics through validation with in situ data. Results showed a strong diversity in the particle residence time, with a dependency on their density and size but also on local ocean conditions and bottom topography. The highest distances are obtained for the smaller and less dense particles, although they tend to be confined by bathymetric constrains and deposited in deepest regions. This work highlights the potential of this modelling tool to forecast laden plume trajectories, allowing the definition of risk assessment scenarios for deep-sea mining activities and the implementation of sustainable exploitation plans. Furthermore, the coupling of this numerical solution with models of biota inhabiting deep-sea vent fields into ecosystem models is discussed and outlined as cost-effective tools for the management of these remote ecosystems.

Impacts of macroalgal spores on the dynamics of adult macroalgae in a eutrophic estuary: high versus low hydrodynamic seasons and long-term simulations for global warming scenarios.

A model describing macroalgal early life stages and adult dynamics was used to quantify the effects of non-adult forms on the productivity of adult macroalgae in a eutrophic estuary. Predictions indicate that during years with mild winters and low rainfall, spring blooms will occur at the expense of the growth and reproduction of overwintering adults and without the contribution of the spore bank. In these circumstances, there is a positive correlation between the intensity of the blooms and the biomass of overwintering adults until a maximum threshold value. On the contrary, in years with high rainfall and low or inexistent biomass of overwintering adults, the onset of adult's growth depends on the biomass of viable dormant spores, the growing season occurs later and adult productivity is very limited. Long-term predictions for climate change scenarios suggest that, in general, global warming will have adverse affects on Ulva intestinalis productivity, with the adults being more affected than the early life stages.

Assessing the effects of temperature and salinity oscillations on a key mesopredator fish from European coastal systems.

A population dynamics model was developed to assess the short and long-term effects of temperature and salinity variations in the common goby Pomatoschistus microps in a Portuguese estuary (Minho estuary, NW Portugal). The population was divided into juveniles, females and males, which constituted the model's state variables. Linear regressions between the observed and the predicted density of juveniles, females and the total population were significant. Parameter's sensitivity and uncertainty analysis were estimated. The model was able to satisfactory describe the P. microps population dynamics, and thus was used to simulate the effects of climatic changes on the fish population. Simulations indicated that the common goby population is sensitive to both temperature and salinity changes. Overall, scenarios of more than 3 °C increase caused significant population decreases. Similarly, increased salinities led to a population shrinkage, whereas scenarios of salinity decrease generated an opposite variation on the population. According to the IPCC predictions for climatic tendencies, the population of the common goby will tend to decrease in the near future, experiencing marked oscillations (decrease or increase) during climatic extremes, namely droughts and floods, respectively. These results may be a useful for future planning and management of estuarine systems given that the common goby is an important species of estuarine food webs in many temperate ecosystems.

Population level effects of intersexuality in the marine environment.

Whilst there have been many studies indicating links between anthropogenic contaminants and reproductive abnormalities in wildlife, very few studies have addressed the effects that might be occurring at the population level. In this study we aimed to assess the mid and long-term population effects of different levels of intersexuality in an amphipod species, Echinogammarus marinus. This aim was achieved by modelling the dynamics of a population of E. marinus with possible contaminant or parasite induced intersexuality over a ten-year period. The model considers five population groups: juveniles, males, immature females, adult normal females and adult intersex females. The costs associated with intersexuality in E. marinus were incorporated into the model using data from previous studies. The basic run of the model considers a population with 50% normal males and 50% normal females, which are prevalent in some field populations of E. marinus. Results of increasing proportions of intersex females were compared with the basic run and suggest that if intersex females occur at the expense of normal females, the population collapses after approximately 6 and 2.3 years when intersex females account for 5 and 10% of the population, respectively. Conversely, the population density increases exponentially if intersex females occur at the expense of males. However, if the number of intersex females reaches approximately (1/2) of the number of normal females, even if the percent of males in the population is as low as 27% (e.g. 27% males, 45% normal females, 28% intersex females), the population will be extinguished within 10 years. These results suggest a selective advantage in female-biased sex ratios in populations with significant levels of intersexuality, up to a certain threshold, where the increase in the total number of females seems to compensate the lower recruitment rates of intersex individuals, namely, intersex females. Furthermore, even in this scenario, if the recruitment rate of normal females is negatively affected, the population survivorship may be compromised. Modelling the effects of intersexuality in this manner should aid both environmental toxicologists and ecologists in predicting the population level effects of endocrine disrupting chemicals and/or parasites.

Significant variations in the productivity of green macroalgae in a mesotidal estuary: implications to the nutrient loading of the system and the adjacent coastal area.

A spatially dynamic model for the productivity of spores and adults of green macroalgae (Enteromorpha sp.) was developed for a mesotidal estuary (Mondego estuary, Portugal). Many of the algal processes and parameters included in the model were experimentally obtained. Model predictions were compared to a real time series (1993-1997) of macroalgal biomass variation and the two sets show a good agreement (ANOVA, P<0.001). Results suggest that algal growth is highly sensitive to small changes in depth and exhibits different patterns of variation in different seasons. On a yearly basis, global calculations for the south channel of the estuary (137 ha) suggest that during bloom years, macroalgal biomass may reach about 21,205 ton DW compared to 240 ton DW in regular years. On a seasonal basis, the difference may be even more significant. The consequences of such variations on the nitrogen and phosphorus loading of the system and the adjacent coastal area are discussed.

Assessing estuarine quality: A cost-effective in situ assay with amphipods.

In situ assays based on feeding depression can be powerful ecotoxicological tools that can link physiological organism-level responses to population and/or community-level effects. Amphipods are traditional target species for toxicity tests due to their high sensitivity to contaminants, availability in the field and ease of handling. However, cost-effective in situ assays based on feeding depression are not yet available for amphipods that inhabit estuarine ecosystems. The aim of this work was to assess a short-term in situ assay based on postexposure feeding rates on easily quantifiable food items with an estuarine amphipod. Experiments were carried out under laboratory conditions using juvenile Echinogammarus marinus as the target individual. When 60 Artemia franciscana nauplii (as prey) were provided per individual for a period of 30 min in dark conditions, feeding rates could be easily quantified. As an endpoint, postexposure feeding inhibition in E. marinus was more sensitive to cadmium contamination than mortality. Assay calibration under field conditions demonstrated the relevance of sediment particle size in explaining individual feeding rates in uncontaminated water bodies. An evaluation of the 48-h in situ bioassay based on postexposure feeding rates indicated that it is able to discriminate between unpolluted and polluted estuarine sites. Using the harmonized protocol described here, the in situ postexposure feeding assay with E. marinus was found to be a potentially useful, cost-effective tool for assessing estuarine sediment and water quality.

Ecological modelling and toxicity data coupled to assess population recovery of marine amphipod Gammarus locusta: Application to disturbance by chronic exposure to aniline.

A population agent-based model of marine amphipod Gammarus locusta was designed and implemented as a basis for ecological risk assessment of chemical pollutants impairing life-history traits at the individual level. We further used the model to assess the toxic effects of aniline (a priority hazardous and noxious substance, HNS) on amphipod populations using empirically-built dose-response functions derived from a chronic bioassay that we previously performed with this species. We observed a significant toxicant-induced mortality and adverse effects in reproductive performance (reduction of newborn production) in G. locusta at the individual level. Coupling the population model with the toxicological data from the chronic bioassay allowed the projection of the ecological costs associated with exposure to aniline that might occur in wild populations. Model simulations with different scenarios indicated that even low level prolonged exposure to the HNS aniline can have significant long-term impacts on G. locusta population abundance, until the impacted population returns to undisturbed levels. This approach may be a useful complement in ecotoxicological studies of chemical pollution to transfer individual-collected data to ecological-relevant levels.

Potential nitrification and denitrification on different surfaces in a constructed treatment wetland.

Improved understanding of the importance of different surfaces in supporting attached nitrifying and denitrifying bacteria is essential if we are to optimize the N removal capacity of treatment wetlands. The aim of this study was therefore to examine the nitrifying and denitrifying capacity of different surfaces in a constructed treatment wetland and to assess the relative importance of these surfaces for overall N removal in the wetland. Intact sediment cores, old pine and spruce twigs, shoots of Eurasian watermilfoil (Myriophyllum spicatum L.), and filamentous macro-algae were collected in July and November 1999 in two basins of the wetland system. One of the basins had been constructed on land that contained lots of wood debris, particularly twigs of coniferous trees. Potential nitrification was measured using the isotope-dilution technique, and potential denitrification was determined using the acetylene-inhibition technique in laboratory microcosm incubations. Nitrification rates were highest on the twigs. These rates were three and 100 times higher than in the sediment and on Eurasian watermilfoil, respectively. Potential denitrification rates were highest in the sediment. These rates were three times higher than on the twigs and 40 times higher than on Eurasian watermilfoil. The distribution of denitrifying bacteria was most likely due to the availability of organic material, with higher denitrification rates in the sediment than on surfaces in the water column. Our results indicate that denitrification, and particularly nitrification, in treatment wetlands could be significantly increased by addition of surfaces such as twigs.

Predicting the variation in Echinogammarus marinus at its southernmost limits under global warming scenarios: can the sex-ratio make a difference?

Understanding the environmental parameters that constrain the distribution of a species at its latitudinal extremes is critical for predicting how ecosystems react to climate change. Our first aim was to predict the variation in the amphipod populations of Echinogammarus marinus from the southernmost limit of its distribution under global warming scenarios. Our second aim was to test whether sex-ratio fluctuations - a mechanism frequently displayed by amphipods - respond to the variations in populations under altered climate conditions. To achieve these aims, scenarios were run with a validated model of E. marinus populations. Simulations were divided into: phase I - simulation of the effect of climate change on amphipod populations, and phase II - simulation of the effect of climate change on populations with male and female proportions. In both phases, temperature (T), salinity (S) and temperature and salinity (T-S) were tested. Results showed that E. marinus populations are highly sensitive to increases in temperature (>2 °C), which has adverse effects on amphipod recruitment and growth. Results from the climate change scenarios coupled with the sex-ratio fluctuations depended largely on the degree of female bias within population. Temperature increase of 2 °C had less impact on female-biased populations, particularly when conjugated with increases in salinity. Male-biased populations were highly sensitive to any variation in temperature and/or salinity; these populations exhibited a long-term decline in density. Simulations in which temperature increased more than 4 °C led to a continuous decline in the E. marinus population. According to this work, E. marinus populations at their southernmost limit are vulnerable to global warming. We anticipate that in Europe, temperature increases of 2 °C will incite a withdrawal of the population of 5°N from the amphipod species located at southernmost geographical borders. This effect is discussed in relation to the distribution of E. marinus along the Atlantic coast.

Hypocholesterolaemic pharmaceutical simvastatin disrupts reproduction and population growth of the amphipod Gammarus locusta at the ng/L range.

Simvastatin (SIM), a hypocholesterolaemic drug, is among the most widely used pharmaceuticals worldwide and is therefore of emerging environmental concern. Despite the ubiquitous nature of SIM in the aquatic ecosystems, significant uncertainties exist about sublethal effects of the drug in aquatic organisms. Therefore, here we aimed at investigating a multi-level biological response in the model amphipod Gammarus locusta, following chronic exposures to low levels of SIM (64 ng/L to 8 µg/L). The work integrated a battery of key endpoints at individual-level (survival, growth and reproduction) with histopathological biomarkers in hepatopancreas and gonads. Additionally, an individual-based population modelling was used to project the ecological costs associated with long-term exposure to SIM at the population level. SIM severely impacted growth, reproduction and gonad maturation of G. locusta, concomitantly to changes at the histological level. Among all analysed endpoints, reproduction was particularly sensitive to SIM with significant impact at 320 ng/L. These findings have important implications for environmental risk assessment and disclose new concerns about the effects of SIM in aquatic ecosystems.