Thesis

Intraspecific variation in the sensitivity of aquatic macrophytes to chemical contamination: the case of copper

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  • Environment and Climate Change Canada
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Abstract

Intraspecific variability plays a pivotal role in short and long term responses of species to environmental fluctuations. This variability, expressed through different traits of individuals, can potentially influence species sensitivity to chemical contamination. This intraspecific variability is currently not taken into account in ecotoxicological risk assessment, whereas it can mislead its results. To examine this hypothesis, the importance of intraspecific variability in the response to copper (Cu) was quantified in controlled conditions for three aquatic macrophyte species, Lemna minor, Myriophyllum spicatum and Ceratophyllum demersum. Variations among genotypes of each of these 3 species were compared to interspecific variability. Results have highlighted a significant genotypic variability, whose importance depends on the species considered. Indeed, L. minor demonstrated a low variability, contrarily to M. spicatum whose variability in growth inhibition by Cu was higher than interspecific differences. In order to specify the extent and the mechanisms of genotypic variability in M. spicatum, other experiments involving measurements of life-history traits have been conducted on 7 genotypes exposed to Cu. Results showed that some genotypes were up to eightfold more sensitive to Cu than others (at concentrations ranging between 0.15 and 0.5 mg/L). These differences in sensitivity were partly explained by the traits measured, but physiological or transcriptomic endpoints may explain more precisely the source of these differences in sensitivity. Finally, 3 experiments with fluctuations in nutrient concentrations, light intensity and Cu pre-exposure have demonstrated that phenotypic plasticity plays an important role in L. minor sensitivity to Cu. Indeed, the weakening of individuals, as a result of unfavorable environmental conditions, can lead to a two-fold increase in sensitivity to Cu.[...]

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... The variation in qualitative and quantitative characters among M. crenata populations in different habitats reflects intraspecific variability, which is defined as the differences occurring between various individuals of the same species, recognized through morphological characteristics (Dumont 2018). One of the M. crenata's characteristics that varies and shows significant differences between the aquatic and terrestrial samples is the petioles' length. ...
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... To investigate whether different genotypes of D. pulex vary in their response to acute copper exposure, we analyzed patterns of gene expression and splicing separately in each clone. Interclonal variation in heavy metal sensitivity has been shown in plants [59,60] and previous studies on Daphnia have reported that molecular response to heavy metals varied across genotypes [49,53,61]. Gene expression response to acute Cu stress was more similar among the three clones compared to splicing. ...
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Changes in plant community traits along an environmental gradient are caused by interspecific and intraspecific trait variation. However, little is known about the role of interspecific and intraspecific trait variation in plant community responses to the restoration of a sandy grassland ecosystem. We measured five functional traits of 34 species along a restoration gradient of sandy grassland (mobile dune, semi-fixed dune, fixed dune, and grassland) in Horqin Sand Land, northern China. We examined how community-level traits varied with habitat changes and soil gradients using both abundance-weighted and non-weighted averages of trait values. We quantified the relative contribution of inter- and intraspecific trait variation in specific leaf area (SLA), leaf dry matter content (LDMC), leaf carbon content (LCC), leaf nitrogen content (LNC), and plant height to the community response to habitat changes in the restoration of sandy grassland. We found that five weighted community-average traits varied significantly with habitat changes. Along the soil gradient in the restoration of sandy grassland, plant height, SLA, LDMC, and LCC increased, while LNC decreased. For all traits, there was a greater contribution of interspecific variation to community response in regard to habitat changes relative to that of intraspecific variation. The relative contribution of the interspecific variation effect of an abundance-weighted trait was greater than that of a non-weighted trait with regard to all traits except LDMC. A community-level trait response to habitat changes was due largely to species turnover. Though the intraspecific shift plays a small role in community trait response to habitat changes, it has an effect on plant coexistence and the maintenance of herbaceous plants in sandy grassland habitats. The context dependency of positive and negative covariation between inter- and intraspecific variation further suggests that both effects of inter- and intraspecific variation on a community trait should be considered when understanding a plant community response to environmental changes in sandy grassland ecosystems.
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Interspecific variation in plant functional traits is the basis of species coexistence in natural ecosystems. However, intraspecific variation is extremely important to community assemblage as well. Here, we quantify leaf trait variation within and across 32 dominant shrub species within two different leaf forms (16 evergreen species and 16 deciduous species) in subtropical evergreen and deciduous broadleaf mixed forest in the Karst topography of Guilin, southwest China. Results showed that leaf area (LA) and leaf thickness (LT) of evergreen species were significantly lower than those of deciduous species, whereas specific leaf areas (SLA) and leaf dry matter content (LDMC) showed the opposite pattern. For SLA and LA, the majority variance was found among species (50.82% vs. 65.01%) and little was found within species (22.98% vs. 27.89%), whereas the largest variation was within species (44.87% vs. 48.2%) with little among species (40.73% vs. 43%) for LDMC and LT. The large variability of LDMC and LT within species may reflect niche differentiation and the importance of intraspecific variation on species coexistence. Our results will help design conservation strategies for this unique subtropical evergreen-deciduous broadleaved forest ecosystem.
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Deteriorating environments in shallow waters place significant pressure on aquatic plants. To estimate the morphological and genetic diversity of submerged macrophytes at 5 sites characterized by varying degrees of human impact, Myriophyllum spicatum L. was studied. Sediment nitrogen concentrations were found to be significantly correlated with the morphological properties of M. spicatum: the site with high sediment nitrogen concentration had 5 to 10 times larger, but rather uniform (low morphological variability) plants, while the site with the least nitrogen in the sediment had smaller M. spicatum plants that were 4 times more variable than the larger plants. Nevertheless, genetic variability of the studied plants showed no response to nutrient loading in the shallow lakes. The adverse environmental conditions therefore seem not to have affected the genetic material of the sampled plants, thus the large morphological variability observed in association with the nitrogen poor sites was predominantly a result of plastic reactions of M. spicatum to environmental conditions.
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Biogeographic barriers for freshwater biota can be effective at various spatial scales. At the largest spatial scale, freshwater organisms can become genetically isolated by their high mountain ranges, vast deserts, and inability to cross oceans. Isolation by distance of aquatic plants is expected to be stronger across than alongside mountain ridges whereas the heterogeneity of habitats among populations and temporary droughts may influence connectivity and hamper dispersal. Suitable aquatic plant habitats became reduced, even for the widespread submerged Potamogeton pectinatus L. (also named Stuckenia pectinata) giving structure to various aquatic habitats. We compared the level of genetic diversity in a heterogeneous series of aquatic habitats across Iran and tested their differentiation over distances and across mountain ranges (Alborz and Zagros) and desert zones (Kavir), with values obtained from temperate region populations. The diversity of aquatic ecosystems across and along large geographic barriers provided a unique ecological situation within Iran. P. pectinatus were considered from thirty-six sites across Iran at direct flight distances ranging from 20 to 1,200 km. Nine microsatellite loci revealed a very high number of alleles over all sites. A PCoA, NJT clustering and STRUCTURE analysis revealed a separate grouping of individuals of southeastern Iranian sites and was confirmed by their different nuclear ITS and cpDNA haplotypes thereby indicating an evolutionary significant unit (ESU). At the level of populations, a positive correlation between allelic differentiation Dest with geographic distance was found. Individual-based STRUCTURE analysis over 36 sites showed 7 genetic clusters. FST and RST values for ten populations reached 0.343 and 0.521, respectively thereby indicating that allele length differences are more important and contain evolutionary information. Overall, higher levels of diversity and a stronger differentiation was revealed among Iranian P. pectinatus than previously observed for temperate European regions, due to regional differences across mountain ranges over long distances.
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The goal of protecting the aquatic environment through testing thousands of chemicals against hundreds of aquatic species with thousands of endpoints while also considering mixtures is impossible given the present resources. Much of the impetus for studies on micropollutants, such as pharmaceuticals, came from the topic of endocrine disruption in wild fish. But despite concern over reductions in fish fertility, there is little evidence that fish populations are in peril. Indeed, fish biologists suggest that many cyprinid populations have been recovering for the past 30 to 40 yr. The central assumption, key to current risk assessment, that effects observed in the laboratory or predicted by models are readily transferrable to the population level, is therefore questionable. The neglect in monitoring wildlife populations is the key weakness in environmental protection strategies. If we do not know whether aquatic wildlife species are declining or increasing, how valuable are our other ecotoxicological activities? Environ Toxicol Chem 2016;35:1609-1616. © 2016 SETAC.
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Highly purified, intact chloroplasts were prepared from pea (Pisum sativum L.) and spinach (Spinacia oleracea L.) following an identical procedure, and were used to investigate the cupric cation inhibition on the photosynthetic activity. In both species, copper inhibition showed a similar inhibitor concentration that decreases the enzyme activity by 50% (IC(50) approximately 1.8 microM) and did not depend on the internal or external phosphate (Pi) concentration, indicating that copper did not interact with the Pi translocator. Fluorescence analysis suggested that the presence of copper did not facilitate photoinhibition, because there were no changes in maximal fluorescence (F(m)) nor in basal fluorescence (F(o)) of copper-treated samples. The electron transport through the photosystem II (PSII) was also not affected (operating efficiency of PSII-F'v/F'm similar in all conditions). Yet, under Cu(2+) stress, the proportion of open PSII reaction centers was dramatically decreased, and the first quinone acceptor (Q(A)) reoxidation was fully inhibited, as demonstrated by the constant photochemical quenching (q(P)) along experiment time. The quantum yield of PSII electron transport (Phi(PSII)) was also clearly affected by copper, and therefore reduced the photochemistry efficiency. Manganese, when added simultaneously with copper, delayed the inhibition, as measured by oxygen evolution and chlorophyll fluorescence, but neither reversed the copper effect when added to copper-inhibited plastids, nor prevented the inhibition of the Hill activity of isolated copper-treated thylakoids. Our results suggest that manganese competed with copper to penetrate the chloroplast envelope. This competition seems to be specific because other divalent cations e.g. magnesium and calcium, did not interfere with the copper action in intact chloroplasts. All results do suggest that, under these conditions, the stroma proteins, such as the Calvin-Benson cycle enzymes or others are the most probable first target for the Cu(2+) action, resulting in the total inhibition of chloroplast photosynthesis and in the consequent unbalanced rate of production and consumption of the reducing power.
Chapter
Description Discusses approaches for determining the impact of pollution on aquatic ecosystems through toxicological and chemical fate assessments. Five sections cover: methods development, refinement, and evaluation; chemicals and chemical wastes; impact of wastes on aquatic ecosystems; bioavailability; and national water quality criteria.
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The paper presents a new index for assessing water trophy and organic pollution. It is based on only true aquatic macrophytes being calculated on species score, coefficient of ecological amplitude and degree of cover. The method was tested in an acidic lowland river and an alkaline mountain river, and is shown to be validated by bio-indication scales based on macrophyte communities. The practical interest is discussed regarding the Water Framework Directive.
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This study investigated the ability of Potamogeton pectinatus L. to accumulate copper and its effects on plants. In accumulation tests, macrophytes were exposed (96 h) to different copper concentrations (0-1000 µM) and the metal was measured in media and plant tissues (roots, stems and leaves) to determine the bioconcentration factor (BCF). Plants accumulated high concentrations of copper in a dose-dependent manner and roots was the main organ for copper accumulation. However, the more copper increased in water, the more BCF values decreased. It may be due to either saturation of copper uptake or down-regulation of metal uptake by plants. In the physiological and morphological analyses, plants were kept (96 h) in Hoagland nutrient solution without copper, in full Hoagland solution (0.5 µM Cu) and in Hoagland medium with copper from 1 to 100 µM. The absence and the presence of copper above to 1 µM inhibited photosynthesis. Chlorophylls and carotenoid levels also decreased with the excess of copper, a fact that may have affected the photosystem II-dependent of chlorophyll and caused photosynthesis suppression. Only macrophytes at 10 µM Cu showed decrease in length and number of leaves on the 10th day of the test, when they died. Chlorosis and necrosis were observed in control groups and groups with extra copper, but not in Hoalgand group. Overall, the macrophyte P. pectinatus can be considered a suitable plant for monitoring environments contaminated by copper, based on results of copper accumulation in the plant, decrease in pigment concentration and presence of chlorosis and necrosis. However, values of BCF based on fresh water tissues was not proper to indicate the use of P. pectinatus for cleaning environments contaminated by copper.
Book
This book is the 2nd edition of a book published in 1995. The first book was written parallel to major developments in the science of risk assessment following the introduction of EU-legislation on industrial chemicals in the period 1970-1995. The present edition reflects the progress and experience since 1995 and again aims at providing background and training material for a new generation of risk assessors, specifically for those who will be involved in implementing legislation in the EU (REACH, the new legislative framework for industrial chemicals) and, in addition, the USA, Japan and Canada. The book is an introduction to risk assessment of chemicals and contains basic background information on sources, emissions, distribution and fate processes for the estimation of exposure of plant and animal species in the environment and humans exposed via the environment, consumer products, and at the workplace. This book includes chapters on environmental chemistry, toxicology and ecotoxicology as well as information on data requirements, data estimation methodologies and intelligent testing strategies. It describes the basic principles and methods of risk assessment in the legislative frameworks of the EU, USA, Japan, and Canada. It also provides an overview of the OECD Chemicals Program. The book is intended to be used by those who are involved in risk assessment of chemicals in government, research institutes, academia and industry as well as by students in technology, health and environmental sciences.
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Dwarf eelgrasses (Zostera noltei) populations have decreased since 2005 in Arcachon Bay (southwest France). Various stressors have been pointed out, however the role of xenobiotics like pesticides or copper (Cu) and of parameters like water temperature warming have not yet been explored. To determine their impact, Z. noltei individuals were collected in a pollution-free site and transferred to the laboratory in seawater microcosms. This dwarf eelgrass was exposed to a pesticide cocktail and copper, alone or simultaneously, at temperatures (10°C, 20°C, 28°C) representative of different seasons. After a two-week contamination, leaf growth, leaf bioaccumulation of Cu, and differential expression of target genes were studied. Eelgrasses bioaccumulated Cu regardless of the temperature, with reduced efficiency in the presence of the Cu and pesticide cocktail at the two higher temperatures. High temperature also exacerbated the effect of contaminants, leading to growth inhibition and differential gene expression. Mitochondrial activity was strongly impacted and higher mortality rates occurred. Experimental results have been confirmed during field survey. This is the first report on the impacts on Z. noltei of pesticides and Cu associate to temperature.
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"An important, controversial account ... of the way in which man's use of poisons to control insect pests and unwanted vegetation is changing the balance of nature." Booklist.
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Chlorophyll fluorescence analysis has become one of the most powerful and widely used techniques available to plant physiologists and ecophysiologists. This review aims to provide an introduction for the novice into the methodology and applications of chlorophyll fluorescence. After a brief introduction into the theoretical background of the technique, the methodology and some of the technical pitfalls that can be encountered are explained. A selection of examples is then used to illustrate the types of information that fluorescence can provide.
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Reciprocal transplantations of Spartina patens genotypes from adjacent salt marsh, swale, and dune habitats provided evidence for genetic differentiation among subpopulations, due at least in part to contrasting selection regimes. Genet survival in the different habitats was related to the amount of genetic divergence. In the dune habitat, marsh ramets showed the lowest survival, swale ramets showed intermediate survival, and dune ramets showed the highest survival. This relationship was not reciprocal, however. The marsh habitat afforded an environment where survival was maximal for all genotypes. Thus, by comparison, the dune environment appeared to impose a more intense selection pressure, and the swale an intermediate selection pressure on Spartina patens. In each site resident genotypes tended to show greater relative fitness than aliens. This evidence for genetic divergence corroborates that previously reported on morphometric (Silander and Antonovics, 1979) and allozymic traits (Silander, 1984).
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An understanding of the mineral nutrition of plants is of fundamental importance in both basic and applied plant sciences. The Second Edition of this book retains the aim of the first in presenting the principles of mineral nutrition in the light of current advances. This volume retains the structure of the first edition, being divided into two parts: Nutritional Physiology and Soil-Plant Relationships. In Part I, more emphasis has been placed on root-shoot interactions, stress physiology, water relations, and functions of micronutrients. In view of the worldwide increasing interest in plant-soil interactions, Part II has been considerably altered and extended, particularly on the effects of external and interal factors on root growth and chapter 15 on the root-soil interface. The second edition will be invaluable to both advanced students and researchers.
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Genetic diversity and population structure reflect complex interactions among a diverse set of processes that may vary temporally, limiting their potential to predict ecological and evolutionary outcomes. Yet, the stability of these patterns is rarely tested. We resampled eelgrass (Zostera marina) meadows from published studies to determine variability in genetic diversity and structure within and between meadows over 5-12 years. The meadows sampled (San Francisco, Tomales and Bodega Bays in California and the Virginia coastal bays) represent a range of life histories (annual vs perennial), age (well-established vs restored) and environments (rural vs urbanized). In all of these systems, neither diversity nor differentiation (FST) changed over time. Differences among tidal heights within Bodega Bay were also remarkably consistent, with the high intertidal being more diverse than the subtidal, and tidal height differentiation being modest but significant at both time points. Historical studies used only a few microsatellite loci; therefore, our temporal comparisons were based on 4-5 loci. However, analysis of the current data using a set of 12 loci show that 4-5 loci are sufficient to describe diversity and differentiation patterns in this system. This temporal consistency was not because of the resampling of large clones, underscoring the feasibility and relevance of understanding drivers of the differences. Because seagrasses are declining at rapid rates, restoration and conservation are increasingly a coastal management priority. Our results argue that surveys of eelgrass genetic structure and diversity at decadal scales can provide accurate depictions of populations, increasing the utility of published genetic data for restoration and designing networks of reserves.Heredity advance online publication, 28 December 2016; doi:10.1038/hdy.2016.114.
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Copper at high concentrations is an ionoregulatory toxicant in fish and its toxicity is known to be strongly modulated by the water chemistry. The toxicity of Cu to the tropical fish cardinal tetra (Paracheirodon axelrodi) was investigated in waters from two major rivers of the Amazon watershed: the Rio Negro (filtered <0.45 μm, pH 5.6, DOC = 8.4 mg L⁻¹, Na = 33 μM, Ca = 8 μM) and the Rio Solimões (filtered <0.45 μm, pH 6.7, DOC = 2.8 mg L⁻¹, Na = 185 μM, Ca = 340 μM), as well as in a natural “reference water” (groundwater) which was almost DOC-free (pH 6.0, DOC = 0.34 mg L⁻¹, Na = 53 μM, Ca = 5 μM). Acute 96-h mortality, Cu bioaccumulation and net flux rates of Na⁺, Cl⁻, K⁺ and total ammonia were determined in P. axelrodi exposed in each water. Copper speciation in each water was determined by two thermodynamic models and by potentiometry, and its toxicity was predicted based on the biotic ligand model (BLM) framework. Our results indicate that high Na⁺ loss is the main mode of toxic action of Cu in P. axelrodi, in accordance with general theory. Cardinal tetra showed a particularly high ability to tolerate Cu and to maintain Na⁺ balance, similar to the ability of this and other endemic Rio Negro species to tolerate low pH and ion-poor conditions. Cu toxicity was lower in Rio Negro than in the other two waters tested, and the free [Cu²⁺] at the LC50, as determined by any of the three speciation methods tested, was approximately 10-fold higher. This variation could not be captured by a realistic set of BLM parameters. At least in part, this observation may be due to gill physiological alterations induced by the abundant dissolved organic matter of the Rio Negro. The implication of this observation is that, for metals risk assessment in tropical waters, similar to the Rio Negro, care must be used in applying BLM models developed using temperate DOC and temperate species.
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The environmental impact of mining and metallurgy is an issue that has affected societies in the ancient Near East over the past 8000 years. We present the results of a multidisciplinary project using agricultural sediments from ancient terraces as a cultural archive of environmental pollution and land use in the copper ore-rich Faynan valley of southern Jordan. Due to the simultaneous production of agricultural goods and copper metallurgy throughout the last 6000 years in the valley, environmental pollution and its consequences for human health have been considered as a factor in settlement abatement. Sediments from two farming terrace systems adjacent to the major mining and smelting locales were analyzed. The sediment analyses included metal concentrations, lead-isotopes and phytolith analysis, and OSL dating. Although measurable concentrations of lead and other heavy metals persist in ancient metallurgical waste piles, our investigations found minimal evidence for contamination in the adjacent terrace systems. Based on these results, we argue that the occurrence of environmental pollution in the Faynan valley is highly variable, and that the distribution of heavy metals resulted from a combination of natural and cultural factors, including persistent landscape features that helped contain the most polluted metallurgical deposits. These findings are significant for understanding the processes of landscape change and human impacts on desert environments, including the ways in which past human actions have negatively affected the environment, as well as preserved and protected the environment from further degradation.
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Sucrose as carbon source in axenic tests affects plant growth and physiology. We postulate that the high sucrose concentration in the OECD TG238 for the submerged growing aquatic plant Myriophyllum spicatum may modify pollutant effects, thus impairing environmental risk assessment. In a factorial design experiment with axenic M. spicatum exposed to three sucrose concentrations (no, low and high) with or without cadmium, we measured growth, dry matter content (DMC), content in pigments or phenolic compounds, and elemental stoichiometry of carbon, nitrogen and phosphorus. We show that sucrose is crucial for growth, but can be used at lower concentrations than currently considered. Sucrose-treated plants had higher DMC and carbon content, but lower contents in chlorophyll and nitrogen. Cadmium affected the content in chlorophyll, phenolic compounds, and elemental stoichiometry. Interactive effects were observed on length growth, the carbon and nitrogen content and the C:N and N:P molar ratios. Remarkably, cadmium led to increased shoot length at low but not at high sucrose concentration. This contrasting effect might result from differences in osmotic potential caused by sucrose. Overall, we found a strong effect of sucrose concentration on the growth and physiology of M. spicatum, and modifications of the response to cadmium. Further studies should establish the lowest sucrose level needed to account for realistic environmental risk assessment based on the axenic OECD TG238. This article is protected by copyright. All rights reserved.
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La profesion –formacion- docente es un tema crucial en los actuales debates educativos. La existencia de dos decretos y el desplazamiento del verdadero sentido del ser maestro reclaman de los analisis un ejercicio de comprension del orden discursivo oficial. La calidad es el sustrato de la sociedad de control. En este marco se agencia nuevas practicas de subjetivacion del maestro los cuales podriamos situar en la calidad, flexibilidad, adaptabilidad, eficiencia, eficacia. En cualquier caso, el esfuerzo por hacer del maestro un intelectual de la educacion fue borrado. La gran cuestion consiste en saber que discursos regula el saber del docente a la luz de la sociedad de control.
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A species sensitivity distribution (SSD) is a probability model of the variation of species sensitivities to a stressor, in particular chemical exposure. The SSD approach has been used as a decision support tool in environmental protection and management since the 1980 s, and the ecotoxicological, statistical and regulatory basis and applications continue to evolve. This article summarizes the findings of a 2014 workshop held by ECETOC (the European Center for Toxicology and Ecotoxicology of Chemicals) and the UK Environment Agency in Amsterdam, the Netherlands on the ecological relevance, statistical basis, and regulatory applications of SSDs. An array of research recommendations categorized under the topical areas of Use of SSDs, Ecological Considerations, Guideline Considerations, Method Development and Validation, Toxicity Data, Mechanistic Understanding and Uncertainty were identified and prioritized. A rationale for the most critical research needs identified in the workshop is provided. The workshop reviewed the technical basis and historical development and application of SSDs, described approaches to estimating generic and scenario specific SSD-based thresholds, evaluated utility and application of SSDs as diagnostic tools, and presented new statistical approaches to formulate SSDs. Collectively, these address many of the research needs to expand and improve their application. The highest priority work, from a pragmatic regulatory point of view, is to develop a guidance of best practices that could act as a basis for global harmonization and discussions regarding the SSD methodology and tools. This article is protected by copyright. All rights reserved
Article
In the past few decades, copper based biocides have been extensively used in food crop protection including citrus, small fruits and in all garden vegetable production facilities. Continuous and rampant use of copper based biocides over decades has led to accumulation of this metal in the soil and the surrounding ecosystem. Toxic levels of copper and its derivatives in both the soil and in the run off pose serious environmental and public health concerns. Alternatives to copper are in great need for the agriculture industry to produce food crops with minimal environmental risks. A combination of copper and zinc metal containing biocide such as Nordox 30/30 or an improved version of zinc-only containing biocide would be a good alternative to copper-only products if the efficacy can be maintained. As of yet there is no published literature on the comparative study of the materials characteristics and phyto-compatibility properties of copper and zinc-based commercial products that would allow us to evaluate the advantages and disadvantages of both versions of pesticides. In this report, we compared copper hydroxide and zinc oxide based commercially available biocides along with suitable control materials to assess their efficacy as biocides. We present a detailed material characterization of the biocides including morphological studies involving electron microscopy, molecular structure studies involving X-ray diffraction, phytotoxicity studies in model plant (tomato) and antimicrobial studies involving surrogate plant pathogens (Xanthomonas alfalfae subsp. citrumelonis, Pseudomonas syringae pv. syringae and Clavibacter michiganensis subsp. michiganensis). Zinc based compounds were found to possess comparable to superior antimicrobial properties while exhibiting significantly lower phytotoxicity when compared to copper based products thus suggesting their potential as an alternative.