Field Of Ecotoxicology Research Articles

The genome of the harpacticoid copepod Tigriopus japonicus: Potential for its use in marine molecular ecotoxicology

The copepod Tigriopus japonicus has been widely used as an experimental species in the field of ecotoxicology. We have sequenced and assembled the whole genome of T. japonicus with comparative analysis of gene families that represent detoxification phases in two additional public genomes of Tigriopus spp., namely, T. californicus and T. kingsejongensis. The total length of the T. japonicus assembled genome was 196.6 Mb with an N50 value of 10.65 Mb and consisted of 339 scaffolds and 25,143 annotated genes. The detoxification gene families encoding cytochrome P450s (CYP450s), glutathione S-transferases (GSTs), and ATP-binding cassette (ABC) proteins in Tigriopus spp. have shown species-dependent diversity in several gene sets, suggesting that these genes have undergone a species-specific expansion to increase their fitness to different marine habitats and environmental pressures. Our study will provide a better understanding of the detoxification system in Tigriopus spp. and will contribute to various areas of research, including ecotoxicology.

ANTHROPOGENIC POLLUTANTS: 10 YEARS OF PROGRESS IN ECOTOXICOLOGICAL STUDIES AND AQUATIC RISK ASSESSMENT

The market expansion due to globalization generated more anthropogenic contaminants, enhancing their negative impact on the environment. In order to prevent and balanced their negative effect, more rapid, sensitive and eco-friendly ecotoxicological studies as risk assessments were developed. Each country developed national programs to ensure the research infrastructure for implementation and development of new testing methodologies, generating new experimental data transferable to governmental authorities, economic agents and academia. In this review, we intend to highlighted the progress of Romanian researchers in the last 10 years in the field of ecotoxicology and aquatic risk assessment. The studies focused on aquatic systems as a non-targeted system to the anthropogenic pollutants impact. The implemented methodologies according with international requirements for chemical control like REACH Regulation, Detergents Regulation, Biocide Products Regulation or the Aquatic Risk Assessment Guidelines were presented. The ecotoxicological effects of various contaminants such as detergents, pharmaceuticals, pesticides, biocides or heavy metals and environmental polluted samples (surface water, sediment or sludge) were provided. Overall, the studies presented in our studies have a nationally novelty through the variety of test substances, the complexity of laboratory experiments and data processing. The research work has a continue applicability in industry and for control authorities. New accredited testing services that providing useful information about the chemicals and their impact on the environmental were introduced. In addition, the ecotoxicology studies and their challenges were a great source for professional training like PhD and master studies and for impact published papers.

Milli-channel array for direct and quick reading of root elongation bioassays

A novel platform to perform systematic analysis and direct reading of root elongation bioassays is presented. The device was designed to include multiplexed microenvironments for the germination and growth of individual seeds, which allows observation by the naked eye or by optical systems, notably cellphone cameras. Prototypes were fabricated by laser micromachining on a highly transparent material that is fully compatible with biological systems. The effectiveness of the milli-channel array was verified against the conventional system (Petri dish). Lactuca sativa was chosen as a model species and glyphosate as a typical toxic agent. All tests were run according to standardized procedures and data analysis was carried out through different statistical indicators such as the root elongation and germination indexes. Results attained in the milli-channel array were identical to those in Petri dish, with the remarkable benefit that several steps required in the conventional system were avoided, which enormously decreases the operation time and the possibility of experimental errors. Further advantages of the milli-channel array are also reported, such as the capability to achieve live imaging of plant organs growth through a simple experiment. The developed device has been proven to be effective, versatile, easy-to-use, and integrable to cellphones, which naturally provide facilities for data recording, analysis, and networking. These improvements open the route to novel applications of bioassays in the wide field of ecotoxicology and environmental studies.

Long-term preservation of echinoderm sperm under non-cryo condition for ecotoxicological bioassay

The preservation of sea urchin gametes was examined to make them more convenient material for research and education, especially on the embryology and ecotoxicology field. The possibility of egg preservation for enough period, two weeks to one month, were reported (Epel et al., 2004; Kiyomoto et al., 2014). The sperm storage is usually done without seawater (dry sperm) for several days. This storage period is lengthened by the addition of antibiotics up to 10 days (Hata, 1998). To maximize the preserved period, we examined the dilution or replacement of seminal plasma with seawater containing antibiotics. Because the activation of sperm is induced by dilution, the condition to inhibit flagella motility was also investigated. Neither high potassium nor low pH improved the period of sperm preservation. The dilution of dry sperm around 100 times with normal seawater containing antibiotics was enough to keep the motility and fertilization ability for longer period, where sperm motility was prevented possibly by the effect of carbon dioxide.

Heavy Metal Threats to Plants and Soil Life in Southern Africa: Present Knowledge and Consequences for Ecological Risk Assessment.

In recent times there has been remarkable development in the field of soil ecotoxicology and risk assessment (RA) models. It is, however, debatable if these RA models are robust representatives for worldwide relevance. In order to investigate this, the current overview aims to address heavy metal threats to soil life in southern Africa by investigating present knowledge and consequences for RA using research in southern Africa as a case. To this end, the focus is on southern African soils, soil life and living conditions. To critically discuss these issues, we report on extensive research conducted in the southern African context and looked how comparable these findings are to RA models employed in the western world. This is done by providing an inventory of selected studies focused on the ecotoxicity of metals towards soil life. It is concluded that there is a dearth of information on southern African soil life, most of which are laboratory-based studies carried out by a handful of researchers. Future research incorporating the available information into a soil ecosystem assessment procedure is paramount. It is recommended that a starting point to tackle this might be the development of holistic sight-specific guidelines for ecological risk assessment at larger spatial scales (km2) which takes into cognizance landscapes, vegetation and faunal characteristics.

Current Status and Future Prognosis of Malaria Vector Control Pesticide Ecotoxicology and Xenopus sp.

Anurans from the genus Xenopus have long been used as standard testing organisms and occur naturally in tropical and sub-tropical areas where malaria vector control pesticides are actively used. However, literature on the toxic effects of these pesticides is limited. This review analyses the available data pertaining to both Xenopus and the pesticides used for malaria vector control in order to determine the pesticides that have the greatest potential to influence amphibian health while also identifying gaps in literature that need to be addressed. Amphibian diversity has shown the fastest decline of any group, yet there are still voids in our understanding of how this is happening. The lack of basic toxicity data on amphibians with regard to pesticides is an issue that needs to be addressed in order to improve effectiveness of amphibian conservation strategies. Meta-analyses performed in this review show that, at current usage, with the available acute toxicity literature, the pyrethroid pesticide group could hold the highest potential to cause acute toxicity to Xenopus sp. in relation to the other MVCPs discussed, but the lack of data cripples the efficacy with which meta-analyses can be performed and conclusions made from such analyses. Several studies have shown that DDT accumulates in Xenopus sp. from malaria vector control areas, but accumulation of other MVCPs in frogs is still largely unknown. Through this review we hope to encourage future research into the field of amphibian ecotoxicology and to promote the use of the Xenopus standard model in order to build comprehensive datasets that may be used in amphibian conservation.

Interactions of Bromocarbazoles with Human Serum Albumin Using Spectroscopic Methods

The 1,3,6,8-tetrabromocarbazole and 3-bromocarbazole have attracted great attention in the ecotoxicology field recently as hazardous environmental contaminants. In this study, the quenching mechanism of these two substances binding with human serum albumin (HSA) has been investigated with spectroscopic methods. Through fluorescence quenching and binding site experiments with steady-state fluorescence and UV-Vis spectra, the intrinsic fluorescence of HSA quenched by 1,3,6,8-tetrabromocarbazole and 3-bromocarbazole both in static process, are activated by binding to site II (subdomain IIIA) of the HSA. In addition, it was not only found that the conformation and secondary structure of the proteins changes, but also that their spontaneous binding processes were driven by electrostatic interactions as well as hydrophobic forces for HSA-1,3,6,8-tetrabromocarbazole, and by typical hydrophobic forces for HSA-3-bromocarbazole. The above studies are beneficial to enhance our understanding of the ecotoxicology and environmental behaviors of halogenated carbazoles.

Epigenetic profiling to environmental stressors in model and non-model organisms: Ecotoxicology perspective.

Epigenetics, potentially heritable changes in genome function that occur without alterations to DNA sequence, is an important but understudied component of ecotoxicology studies. A wide spectrum of environmental challenge, such as temperature, stress, diet, toxic chemicals, are known to impact on epigenetic regulatory mechanisms. Although the role of epigenetic factors in certain biological processes, such as tumourigenesis, has been heavily investigated, in ecotoxicology field, epigenetics still have attracted little attention. In ecotoxicology, potential role of epigenetics in multi- and transgenerational phenomenon to environmental stressors needs to be unrevealed. Natural variation in the epigenetic profiles of species in responses to environmental stressors, nature of dose-response relationships for epigenetic effects, and how to incorporate this information into ecological risk assessment should also require attentions. In this review, we presented the available information on epigenetics in ecotoxicological context. For this, we have conducted a systemic review on epigenetic profiling in response to environmental stressors, mostly chemical exposure, in model organisms, as well as, in ecotoxicologically relevant wildlife species.

Use of integrated biomarker indexes for assessing the impact of receiving waters on a native neotropical teleost fish

In the field of aquatic ecotoxicology, indexes obtained from a battery of biomarkers have proved to be a useful tool for assessing quantifiable and integrated health responses of organisms exposed to pollutants. The objective of this work was to evaluate the effects of exposure to the Reconquista River water (RR) on adults of Cnesterodon decemmaculatus using different integrated indexes. We conducted a 12-d laboratory assay involving the exposure of fish to RR, a negative control (moderately hard water - MHW medium), and a positive control (for genotoxicity with MHW + Cyclophosphamide, CP). There were measured metabolic (food intake and specific assimilation, specific metabolic rate, oxygen extraction efficiency, ammonia excretion, and ammonia quotient), genotoxic (comet assay, micronucleus test, and nuclear abnormalities), morphological variables (total length, body and liver weight) and biochemical variables (Electron Transport System - ETS, Acetylcholinesterase activity - AChE, Catalase - CAT, Glutathione-S-transferase - GST, Glutathione content - GSH and tissue proteins). These variables were grouped into different indexes: morphological (Condition Factor - K and Liver Somatic Index-LSI), metabolic (Scope for Growth-SFG), genetic damage (GDI) and integrated biomarker response - IBR (AChE brain, CAT, GST and GSH liver, GSH gills, ETS muscle) indexes. Results indicated that RR water induced metabolic, biochemical and genetic damages. The SFG, GDI and IBR were suitable to assess the effects of exposure to an environmental sample in an integrated approach, reducing uncertainty due to inherent biomarker variability. These indexes have emerged as promising tools for environmental monitoring studies.

Selection of reference genes for reverse transcription-qPCR analysis in the biomonitor macrophyte Bidens laevis L.

The RT-qPCR has been the method used to analyze gene expression in plants but its benefits have not been completely exploited in the field of plants ecotoxicology when used as molecular biomarkers. The correct use of RT-qPCR demands to establish a certain number of reference genes (RG) which are expected to be invariable in their expression although it does not always happen. The main goals of this work were to: (1) analyze the stability of six potential RG, (2) establish the optimum number of RG, (3) select the most suitable RG to be applied in Bidens laevis under different test conditions and tissues and (4) confirm its convenience by normalizing the expression of one gene of interest under three different challenges. When all data were pooled together, the geNorm algorithm pointed out beta-actin and beta-tubulin (TUB) as the optimal RG pair while NormFinder algorithm selected nicotinamide adenine dinucleotide dehydrogenase (NADHD) and histone 3 (H3) as possessing the most invariable levels of expression. On the other hand, when data were grouped by tissues, ANOVA test selected H3 and TUB, while data grouped by conditions indicated that H3 and NADHD were the most stable RG under this analysis. Therefore, for a general-purpose set of RG, the overall analysis showed that a set of three RG would be optimum, and H3, TUB and NADHD were the selected ones. On the other hand, as RG can vary depending on the tissues or conditions, results achieved with ANOVA would be more reliable. Thus, appropriate normalization process would clearly need more than one RG.

Investigation of the molecular mechanisms of hepatic injury upon naphthalene exposure in zebrafish (Danio rerio).

Naphthalene has been used worldwide as a commercial insecticide for decades, which when detected in the environment can have various negative effects on non-target organism, such as hepatotoxicity. However, the molecular mechanisms of how naphthalene acts to affect the liver in zebrafish (Danio rerio) remains unknown. In this study, we evaluated the potential toxic effects of naphthalene on livers in female adult zebrafish over a 21-day subacute exposure. Global hepatic gene expression was examined by microarrays and the results indicated the regulated genes were associated significantly with vital hepatic injury pathways and GO categories upon naphthalene exposure, such as disruptions in lipid metabolism, inflammatory response, and the carcinogenic processes. According to our observations of liver histology, nuclear enlargement as a potential indicator of cancers and hepatic lipometabolic disorder precisely were supported. The 96 h acute naphthalene tests on Tg(lysC:DsRed) and LiPan lines larvae revealed recruitment of neutrophils by the liver, as well as decreased liver size, which further confirmed hepatic inflammation response to naphthalene exposure. Thus, these findings advance the field of ecotoxicology by unveiling a new role of naphthalene as a leading cause of liver damage and provide potential biomarker-genes for environmental naphthalene monitoring.

Protocol for Acute and Chronic Ecotoxicity Testing of the Turquoise Killifish Nothobranchius furzeri.

The killifish Nothobranchius furzeri is an emerging model organism in the field of ecotoxicology and its applicability in acute and chronic ecotoxicity testing has been demonstrated. Overall, the sensitivity of the species to toxic compounds is in the range with, or higher than, that of other model species. This work describes protocols for acute, chronic, and multigenerational bioassays of single and combined stressor effects on N. furzeri. Due to its short maturation time and life-cycle, this vertebrate model allows the study of endpoints such as maturation time and fecundity within four months. Transgenerational full life-cycle exposure trials can be performed in as little as 8 months. Since this species produces eggs that are drought-resistant and remain viable for years, the on-site culture of the species is not needed but individuals can be recruited when required. The protocols are designed to measure life-history traits (mortality, growth, fecundity, weight) and critical thermal maximum.

Metacommunities, metaecosystems and the environmental fate of chemical contaminants

Abstract Although pollution is a major driver of ecosystem change, models predicting the environmental fate of contaminants suffer from critical uncertainties related to oversimplifying the dynamics of the biological compartment. It is increasingly recognized that contaminant processing is an outcome of ecosystem functioning, that ecosystem functioning is contingent on community structure and that community structure is influenced by organismal dispersal. We propose a conceptual organization of the contribution of organismal dispersal to local contaminant fate. Direct dispersal effects occur when the dispersing organism directly couples contaminant stocks in spatially separate ecosystems by transporting contaminants in its biomass. Indirect dispersal effects occur when the dispersing organism indirectly influences contaminant fate via community assembly. This can occur either when the dispersing organism is a contaminant processor or when the dispersing organism alters, via species interactions, the abundance of contaminant biotransporters or processors already established in the ecosystem. The magnitude of direct and indirect dispersal effects is modulated by many factors, including other contaminants. These will influence population growth rates of the dispersing species in the donor ecosystem, or the probability that a dispersing individual reaches the recipient ecosystem. We provide a review of pertinent literature demonstrating that these two mechanisms, and their chemical modulation, are well supported or likely to occur in many natural and human‐modified landscapes. The literature also demonstrates that they can operate in concert with each other. Synthesis and applications. Managed ecosystems thought to be important contaminant and nutrient sinks, such as artificial ponds and constructed wetlands, should be monitored and controlled for in‐and‐out animal movement if contaminant export is found to be relevant. Uncontaminated fishing grounds linked to contaminated sites via movement of dispersing species should be monitored and resident species evaluated for health consumption advisories. Assessing the success of contaminated site remediation can be improved by better matching the spatial extent of site remediation and the home range of monitored species. Finally, interagency research fund programmes should be developed that narrow the current gap between the fields of ecology and ecotoxicology.

Antioxidant responses in the earthworm Aporrectodea caliginosa of eastern Slovakia: application of principal component analysis as a tool to identify metal contaminated areas.

The soil of Slovak Republic is severely contaminated with heavy metals, creating hazards to soil health. In order to assess the current status with the prospect of selecting the appropriate treatment methods and land use, this investigation aimed to determine a panel of complementary and ecologically relevant biomarkers that reflect adverse biological responses towards terrestrial pollutants. To attain this objective, the concentration of reduced glutathione and enzymes of glutathione antioxidant system were assessed in clitellate earthworm, Aporrectodea caliginosa sampled from selected sites of eastern Slovakia along with the pH and total metal concentration (As, Cd, Pb, Cr, Hg, Mn, Fe, Co, Ni, Cu, Zn) of soils. Positive, significant (p < 0.05) induction of glutathione peroxidase, glutathione reductase, and glutathione-S-transferase activities and depletion of reduced glutathione level (negative correlation) were associated with the increased soil metal concentrations. Metal interference was found in the detoxification process and antioxidant defense mechanism does not efficiently counteract the oxidative stress induced by chronic metal exposure. The tested biomarkers confirmed sensitive and affective response to the pollution of soil contaminants, in this case metals. This has a potential use in ecotoxicological field monitoring. The proposed principal component analysis is a multivariate model of data analysis that represents a cost-effective approach to differentiate metalliferous soils of eastern Slovakia with different health status.

Aquatic live animal radiotracing studies for ecotoxicological applications: Addressing fundamental methodological deficiencies

The use of live animal gamma radioisotope tracer techniques in the field of ecotoxicology allows laboratory studies to accurately monitor contaminant biokinetics in real time for an individual organism. However, methods used in published studies for aquatic organisms are rarely described in sufficient detail to allow for study replication or an assessment of the errors associated with live animal radioanalysis to be identified. We evaluate the influence of some important methodological deficiencies through an overview of the literature on live aquatic animal radiotracer techniques and through the results obtained from our radiotracer studies on four aquatic invertebrate species. The main factors discussed are animal rinsing, radioanalysis and geometry corrections. We provide examples of three main techniques in live aquatic animal radiotracer studies to improve data quality control and demonstrate why each technique is crucial in interpreting the data from such studies. The animal rinsing technique is also relevant to non-radioisotope tracer studies, especially those involving nanoparticles. We present clear guidance on how to perform each technique and explain the importance of proper reporting of the validation of each technique for individual studies. In this paper we describe methods that are often used in lab-based radioecology studies but are rarely described in great detail. We hope that this paper will act as the basis for standard operating procedures for future radioecology studies to improve study replication and data quality control.

Mapping of the spatial distribution of silver nanoparticles in root tissues of Vicia faba by laser-induced breakdown spectroscopy (LIBS)

The manuscript presents a procedure for optimal sample preparation and the mapping of the spatial distribution of metal ions and nanoparticles in plant roots using laser-induced breakdown spectroscopy (LIBS) in a double-pulse configuration (DP LIBS) in orthogonal reheating mode. Two Nd:YAG lasers were used; the first one was an ablation laser (UP-266 MACRO, New Wave, USA) with a wavelength of 266nm, and the second one (Brilliant, Quantel, France), with a fundamental wavelength of 1064nm, was used to reheat the microplasma. Seedlings of Vicia faba were cultivated for 7 days in CuSO4 or AgNO3 solutions with a concentration of 10µmoll−1 or in a solution of silver nanoparticles (AgNPs) with a concentration of 10µmoll−1 of total Ag, and in distilled water as a control. The total contents of the examined metals in the roots after sample mineralization as well as changes in the concentrations of the metals in the cultivation solutions were monitored by ICP-OES. Root samples embedded in the TissueTek medium and cut into 40µm thick cross sections using the Cryo-Cut Microtome proved to be best suited for an accurate LIBS analysis with a 50µm spatial resolution. 2D raster maps of elemental distribution were created for the emission lines of Cu(I) at 324.754nm and Ag(I) at 328.068nm. The limits of detection of DP LIBS for the root cross sections were estimated to be 4pg for Cu, 18pg for Ag, and 3pg for AgNPs. The results of Ag spatial distribution mapping indicated that unlike Ag+ ions, AgNPs do not penetrate into the inner tissues of Vicia faba roots but stay in their outermost layers. The content of Ag in roots cultivated in the AgNP solution was one order of magnitude lower compared to roots cultivated in the metal ion solutions. The significantly smaller concentration of Ag in root tissues cultivated in the AgNP solution also supports the conclusion that the absorption and uptake of AgNPs by roots of Vicia faba is very slow. LIBS mapping of root sections represents a fast analytical method with sufficient precision and spatial resolution that can provide very important information for researchers, particularly in the fields of plant science and ecotoxicology.

How we can make ecotoxicology more valuable to environmental protection

There is increasing awareness that the value of peer-reviewed scientific literature is not consistent, resulting in a growing desire to improve the practice and reporting of studies. This is especially important in the field of ecotoxicology, where regulatory decisions can be partly based on data from the peer-reviewed literature, with wide-reaching implications for environmental protection. Our objective is to improve the reporting of ecotoxicology studies so that they can be appropriately utilized in a fair and transparent fashion, based on their reliability and relevance. We propose a series of nine reporting requirements, followed by a set of recommendations for adoption by the ecotoxicology community. These reporting requirements will provide clarity on the the test chemical, experimental design and conditions, chemical identification, test organisms, exposure confirmation, measurable endpoints, how data are presented, data availability and statistical analysis. Providing these specific details will allow for a fuller assessment of the reliability and relevance of the studies, including limitations. Recommendations for the implementation of these reporting requirements are provided herein for practitioners, journals, reviewers, regulators, stakeholders, funders, and professional societies. If applied, our recommendations will improve the quality of ecotoxicology studies and their value to environmental protection.

ChemInform Abstract: An Overview of Detection Techniques for Monitoring Dioxin‐Like Compounds: Latest Technique Trends and Their Applications

AbstractReview: 117 refs.

Endocrine Disruption in the Omics Era: New Views, New Hazards, New Approaches

The genome revolution has brought about a complete change on our view of biological systems. The quantitative determination of changes in all the major molecular components of the living cells, the "omics" approach, opened whole new fields for all health sciences, including toxicology. Endocrine disruption,i.e., the capacity of anthropogenic pollutants to alter the hormonal balance of the organisms, is one of the fields of Ecotoxicology in which omics has a relevant role. In the first place, the discovery of scores of potential targets in the genome of almost any Metazoan species studied so far, each of them being a putative candidate for interaction with endocrine disruptors. In addition, the understanding that ligands, receptors, and their physiological functions suffered fundamental variations during animal evolution makes it necessary to assess disruption effects separately for each major taxon. Fortunately, the same deal of knowledge on genes and genomes powered the development of new high-throughput techniques and holistic approaches. Genomics, transcriptomics, proteomics, metabolomics, and others, together with appropriate prediction and modeling tools, will mark the future of endocrine disruption assessment both for wildlife and humans.

Time Matters in Ecotoxicological Sampling Due to Circadian Rhythm.

As in general, technological inventions also drive the development in the field of toxicology and ecotoxicology. In the past decade, gene expression analysis has become a universally applied technology allowing many insights into toxicological pathways of environmental contaminants. Due to the novel technologies, including quantitative determination of mRNA by quantitative reverse transcription analysis (qRT-PCR), and semiquantitative methods, such as microarrays and RNA-sequencing technologies, toxicological profiles of contaminants could be identified. For instance, gene expression analysis of genes associated with the hypothalamic-pituitary-gonadal axis (HPG axis) in fish had become a conventional end point for endocrine disrupting chemicals. While these gene expression data provide novel insights into identifying potential toxicological end points and molecular mechanisms, often not enough attention is given to the question of mRNA stabilities and reliabilities of transcriptional data, in particular when links to physiological effects are difficult to make. A crucial factor in this issue is the endogenous circadian oscillations of genes during sampling.