Publications by authors named "Julián Blasco"

99 Publications

Meta-omic evaluation of bacterial microbial community structure and activity for the environmental assessment of soils: overcoming protein extraction pitfalls.

Environ Microbiol 2021 Aug 30;23(8):4706-4725. Epub 2021 Jul 30.

Departamento de Bioquímica y Biología Molecular, Campus de Excelencia Internacional Agroalimentario CeiA3, Universidad de Córdoba, Campus de Rabanales, Edificio Severo Ochoa, Córdoba, E-14071, Spain.

Microorganisms play unique, essential and integral roles in the biosphere. This work aims to assess the utility of soil's metaomics for environmental diagnosis. Doñana National Park (DNP) was selected as a natural lab since it contains a strictly protected core that is surrounded by numerous threats of pollution. Culture-independent high-throughput molecular tools were used to evaluate the alterations of the global structure and metabolic activities of the microbiome. 16S rRNA sequencing shows lower bacterial abundance and diversity in areas historically exposed to contamination that surround DNP. For metaproteomics, an innovative post-alkaline protein extraction protocol was developed. After NaOH treatment, successive washing with Tris-HCl buffer supplemented with glycerol was essential to eliminate interferences. Starting from soils with different physicochemical characteristics, the method renders proteins with a remarkable resolution on SDS-PAGE gels. The proteins extracted were analysed by using an in-house database constructed from the rRNA data. LC-MS/MS analysis identified 2182 non-redundant proteins with 135 showing significant differences in relative abundance in the soils around DNP. Relevant global biological processes were altered in response to the environmental changes, such as protective and antioxidant mechanisms, translation, folding and homeostasis of proteins, membrane transport and aerobic respiratory metabolism.
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http://dx.doi.org/10.1111/1462-2920.15673DOI Listing
August 2021

Assessment of pharmaceutical mixture (ibuprofen, ciprofloxacin and flumequine) effects to the crayfish Procambarus clarkii: A multilevel analysis (biochemical, transcriptional and proteomic approaches).

Environ Res 2021 09 29;200:111396. Epub 2021 May 29.

Department of Ecology and Coastal Management, Instituto de Ciencias Marinas de Andalucía (CSIC), Campus Rio San Pedro, 11510, Puerto Real, Cádiz, Spain. Electronic address:

The knowledge about the effects of pharmaceuticals on aquatic organisms has been increasing in the last decade. However, due to the variety of compounds presents in the aquatic medium, exposure scenarios and exposed organisms, there are still many gaps in the knowledge on how mixtures of such bioactive compounds affect exposed non target organisms. The crayfish Procambarus clarkii was used to analyze the toxicity effects of mixtures of ciprofloxacin, flumequine and ibuprofen at low and high concentrations (10 and 100 μg/L) over 21 days of exposure and to assess the recovery capacity of the organism after a depuration phase following exposure during additional 7 days in clean water. The crayfish accumulated the three compounds throughout the entire exposure in the hepatopancreas. The exposure to the mixture altered the abundance of proteins associated with different cells functions such as biotransformation and detoxification processes (i.e. catalase and glutathione transferase), carbohydrate metabolism and immune responses. Additionally changes in expression of genes encoding antioxidant enzymes and in activity of the corresponding enzymes (i.e. superoxide dismutase, glutathione peroxidase and glutathione transferase) were reported. Alterations at different levels of biological organization did not run in parallel under all circumstances and can be related to changes in the redox status of the target tissue. No differences were observed between control and exposed organisms for most of selected endpoints after a week of depuration, indicating that exposure to the drug mixture did not produce permanent damage in the hepatopancreas of P. clarkii.
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http://dx.doi.org/10.1016/j.envres.2021.111396DOI Listing
September 2021

Constructing a de novo transcriptome and a reference proteome for the bivalve Scrobicularia plana: Comparative analysis of different assembly strategies and proteomic analysis.

Genomics 2021 May 25;113(3):1543-1553. Epub 2021 Mar 25.

Departamento de Bioquímica y Biología Molecular, Campus de Excelencia Internacional Agroalimentario CeiA3, Universidad de Córdoba, Campus de Rabanales, Edificio Severo Ochoa, E-14071 Córdoba, Spain. Electronic address:

Scrobicularia plana is a coastal and estuarine bivalve widely used in ecotoxicological studies. However, the underlying molecular mechanisms for S. plana pollutant responses are hardly known due to the lack of molecular databases. Thus, in this study we present a holistic approach to assess a robust reference transcriptome and proteome of this clam. A mixture of control and metal-exposed individuals was used for mRNA isolation. Four sets of high quality filtered preprocessed reads were generated (two quality scores and two sequenced lengths) and assembled with Mira, Ray and Trinity algorithms. The sixty-four generated assemblies were refined, filtered and evaluated for their proteomic quality. Eight assemblies presented top Detonate scores but one was selected due to its compactness and biological representation, which was generated: (i) from the highest quality dataset (Q20L100), (ii) using Trinity algorithm with all k-mers (AtKa), (iii) removing redundancy by CD-HIT (RR80), and (iv) filtering out poor contigs (F), that was subsequently named Q20L100AtKaRR80F. S. plana proteomic analysis revealed 10,017 peptide groups that corresponded to 2066 proteins with a wide coverage of molecular functions and biological processes, confirming the strength of the database generated.
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http://dx.doi.org/10.1016/j.ygeno.2021.03.025DOI Listing
May 2021

High-throughput molecular analyses of microbiomes as a tool to monitor the wellbeing of aquatic environments.

Microb Biotechnol 2021 05 9;14(3):870-885. Epub 2021 Feb 9.

Departamento de Bioquímica y Biología Molecular, Campus de Excelencia Internacional Agroalimentario CeiA3, Universidad de Córdoba, Campus de Rabanales, Edificio Severo Ochoa, Córdoba, E-14071, Spain.

Aquatic environments are the recipients of many sources of environmental stress that trigger both local and global changes. To evaluate the associated risks to organisms and ecosystems more sensitive and accurate strategies are required. The analysis of the microbiome is one of the most promising candidates for environmental diagnosis of aquatic systems. Culture-independent interconnected meta-omic approaches are being increasing used to fill the gaps that classical microbial approaches cannot resolve. Here, we provide a prospective view of the increasing application of these high-throughput molecular technologies to evaluate the structure and functional activity of microbial communities in response to changes and disturbances in the environment, mostly of anthropogenic origin. Some relevant topics are reviewed, such as: (i) the use of microorganisms for water quality assessment, highlighting the incidence of antimicrobial resistance as an increasingly serious threat to global public health; (ii) the crucial role of microorganisms and their complex relationships with the ongoing climate change, and other stress threats; (iii) the responses of the environmental microbiome to extreme pollution conditions, such as acid mine drainage or oil spills. Moreover, protists and viruses, due to their huge impacts on the structure of microbial communities, are emerging candidates for the assessment of aquatic environmental health.
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http://dx.doi.org/10.1111/1751-7915.13763DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8085945PMC
May 2021

"Nanosize effect" in the metal-handling strategy of the bivalve Scrobicularia plana exposed to CuO nanoparticles and copper ions in whole-sediment toxicity tests.

Sci Total Environ 2021 Mar 3;760:143886. Epub 2020 Dec 3.

Universidad de Cádiz, INMAR, Campus Rio San Pedro, 11510 Puerto Real, Spain. Electronic address:

To date, the occurrence, fate and toxicity of metal-based NPs in the environment is under investigated. Their unique physicochemical, biological and optical properties, responsible for their advantageous application, make them intrinsically different from their bulk counterpart, raising the issue of their potential toxic specificity or "nanosize effect". The aim of this study was to investigate copper bioaccumulation, subcellular distribution and toxic effect in the marine benthic species Scrobicularia plana exposed to two forms of sediment-associated copper, as nanoparticles (CuO NPs) and as soluble ions (CuCl). Results showed that the exposure to different copper forms activated specific organism's metal handling strategies. Clams bioaccumulated soluble copper at higher concentrations than those exposed to sediment spiked with CuO NPs. Moreover, CuO NPs exposure elicited a stronger detoxification response mediated by a prompt mobilization of CuO NPs to metal-containing granules as well as a delayed induction of MT-like proteins, which conversely, sequestered soluble copper since the beginning of the exposure at levels significantly different from the control. Eventually, exposure to high concentrations of either copper form led to the same acute toxic effect (100% mortality) but the outcome was delayed in bivalves exposed to CuO NPs suggesting that the mechanisms underlying toxicity were copper form-specific. Indeed, while most of soluble copper was associated to the mitochondrial fraction suggesting an impairment of the ATP synthesis capacity at mitochondrial level, CuO NPs toxicity was most likely caused by the oxidative stress mediated by their bioaccumulation in the enzymatic and mitochondrial metabolically available fractions.
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http://dx.doi.org/10.1016/j.scitotenv.2020.143886DOI Listing
March 2021

Not Only Toxic but Repellent: What Can Organisms' Responses Tell Us about Contamination and What Are the Ecological Consequences When They Flee from an Environment?

Toxics 2020 Dec 12;8(4). Epub 2020 Dec 12.

Department of Ecology and Coastal Management, Institute of Marine Sciences of Andalusia (CSIC), Puerto Real, 11519 Cadiz, Spain.

The ability of aquatic organisms to sense the surrounding environment chemically and interpret such signals correctly is crucial for their ecological niche and survival. Although it is an oversimplification of the ecological interactions, we could consider that a significant part of the decisions taken by organisms are, to some extent, chemically driven. Accordingly, chemical contamination might interfere in the way organisms behave and interact with the environment. Just as any environmental factor, contamination can make a habitat less attractive or even unsuitable to accommodate life, conditioning to some degree the decision of organisms to stay in, or move from, an ecosystem. If we consider that contamination is not always spatially homogeneous and that many organisms can avoid it, the ability of contaminants to repel organisms should also be of concern. Thus, in this critical review, we have discussed the dual role of contamination: toxicity (disruption of the physiological and behavioral homeostasis) vs. repellency (contamination-driven changes in spatial distribution/habitat selection). The discussion is centered on methodologies (forced exposure against non-forced multi-compartmented exposure systems) and conceptual improvements (individual stress due to the toxic effects caused by a continuous exposure against contamination-driven spatial distribution). Finally, we propose an approach in which Stress and Landscape Ecology could be integrated with each other to improve our understanding of the threat contaminants represent to aquatic ecosystems.
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http://dx.doi.org/10.3390/toxics8040118DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7768353PMC
December 2020

Metal body burden and tissue oxidative status in the bivalve Venerupis decussata from Tunisian coastal lagoons.

Mar Environ Res 2020 Jul 14;159:105000. Epub 2020 May 14.

Departamento de Bioquímica y Biología Molecular, Campus de Excelencia Internacional Agroalimentario CeiA3, Universidad de Córdoba, Campus de Rabanales, Edificio Severo Ochoa, E-14071, Córdoba, Spain. Electronic address:

Coastal transitional waters are exposed to many anthropogenic threats. This study aims to assess the trace metals' pollution status of transitional waters by evaluating its biological effects in the clam Venerupis decussata. Among the studied sites along the Tunisian littoral, South Tunis and Boughrara were the most impacted, since clams from these two lagoons presented significant differences in: (i) trace metal contents, (ii) in-cell hydrogen peroxide, (iii) enzymatic and non-enzymatic defenses, (iv) damage to lipids and proteins, and (v) protein post-translational modifications. These changes related to evident histopathological traits. PCA showed a clear separation between the digestive gland and gills tissues and illustrated an impact gradient in Tunisian coastal lagoons. Water temperature was revealed as an added natural stressor that, when concurring with high pollution, may jeopardize an ecosystem's health and contribute to the accumulation of hazardous metals in organisms.
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http://dx.doi.org/10.1016/j.marenvres.2020.105000DOI Listing
July 2020

Immunotoxicity of polystyrene nanoplastics in different hemocyte subpopulations of Mytilus galloprovincialis.

Sci Rep 2020 05 25;10(1):8637. Epub 2020 May 25.

Department of Ecology and Coastal Management, Institute of Marine Sciences of Andalusia (CSIC), Campus Río San Pedro, 11510, Puerto Real, Cádiz, Spain.

Plastic represents 60-80% of litter in the ocean. Degradation of plastic to small fragments leads to the formation of microplastics (MPs <5 mm) and nanoplastics (NPs <1 µm). One of the most widely used and representative plastics found in the ocean is polystyrene (PS). Among marine organisms, the immune system of bivalves is recognized as suitable to assess nanomaterial toxicity. Hemocyte subpopulations [R1 (large granular cells), R2 (small semi-granular cells) and R3 (small agranular or hyaline cells)] of Mytilus galloprovincialis are specialized in particular tasks and functions. The authors propose to examine the effects of different sizes (50 nm, 100 nm and 1 μm) PS NPs on the different immune cells of mussels when they were exposed to (1 and 10 mg·L-1) of PS NPs. The most noteworthy results found in this work are: (i) 1 µm PS NPs provoked higher immunological responses with respect to 50 and 100 nm PS NPs, possibly related to the higher stability in size and shape in hemolymph serum, (ii) the R1 subpopulation was the most affected with respect to R2 and R3 concerning immunological responses and (iii) an increase in the release of toxic radicals, apoptotic signals, tracking of lysosomes and a decrease in phagocytic activity was found in R1.
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http://dx.doi.org/10.1038/s41598-020-65596-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7248110PMC
May 2020

Lead toxicity on a sentinel species subpopulation inhabiting mangroves with different status conservation.

Chemosphere 2020 Jul 3;251:126394. Epub 2020 Mar 3.

Universidade Federal de São Paulo, Departamento de Ciências do Mar, Campus Baixada Santista, Rua Maria Máximo, 168, Ponta da Praia, Santos, SP, 11030-100, Brazil; Universidade Santa Cecília, Laboratório de Ecotoxicologia, Rua Oswaldo Cruz, 266, Santos, SP, 11045-907, Brazil. Electronic address:

Lead is a priority pollutant introduced in the aquatic environment by different sources commonly located in estuarine regions, such as ports, marinas and industries. Environmental agencies around the world set the maximum allowable concentration of lead in effluents, surface water and sediment, but few studies reported its accumulation and chronic toxicity in mangrove benthic invertebrates using concentrations believed to be safe. In the case of Brazilian mangrove environments, Ucides cordatus is a crab species of choice to be used in bioaccumulation studies. We have assessed biomarkers' responses (DNA strand breaks, micronucleated cells, metallothioneins, enzymatic activity of aminolevulinic acid dehydratase and neutral red retention time) and the total bioaccumulation in six tissues of U. cordatus crabs resident to mangrove areas under different conservation status during a 28-day period bioassay. We also investigated Pb subcellular partition and biomarkers' responses using a supposedly safe concentration (10 μg L). During the Pb exposure, the highest concentration of Pb was observed in crab gills. Crabs also showed a high ability to allocate Pb in detoxified forms. Multivariate analysis pointed out that bioaccumulation (total, active and detoxified) is linked to biomarkers. Even in supposedly safe dosage, U. cordatus triggered its defense mechanisms expressing more metallothioneins and presented relevant cyto-genotoxic damage. Our data suggest the development of biological tolerance to Pb in crabs from polluted areas. Our results provided a new insight about lead toxicity even at concentrations considered environmentally safe, which could support new strategies to manage estuarine areas considering their respective conservation status.
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http://dx.doi.org/10.1016/j.chemosphere.2020.126394DOI Listing
July 2020

Stress under the sun: Effects of exposure to low concentrations of UV-filter 4- methylbenzylidene camphor (4-MBC) in a marine bivalve filter feeder, the Manila clam Ruditapes philippinarum.

Aquat Toxicol 2020 Apr 3;221:105418. Epub 2020 Feb 3.

Instituto Universitario de Investigación Marina (INMAR), Campus de Excelencia Internacional del Mar (CEI•MAR), Universidad de Cádiz, Av. República Saharaui s/n, 11510, Puerto Real, Cádiz, Spain. Electronic address:

UV filters are a class of emerging contaminants with an annual estimated production of 10,000 tons worldwide that continuously enter aquatic environments. Among UV filters, 4-methylbenzylidenecamphor (4-MBC) is an organic camphor derivative used in the cosmetic industry for its ability to protect the skin against UV, specifically UV B radiation. Individuals of the Japanese clam, Ruditapes philippinarum, were exposed to 4-MBC at environmentally relevant and slightly higher concentrations (nominal: 0, 1, 10, 100 μg L) using a semi-static exposure system over a 7-days period followed by a 3-days depuration period (total 10 days) where no 4-MBC was added to the tanks. Assessed mortality reached up to 100 % at the highest exposure concentration and a LC50 value of 7.71 μg·L4-MBC was derived. Environmental risk assessment carried out in a site specific environment, the Cadiz bay in the south of Spain, revealed a potential risk produced by the presence of 4-MBC. Digestive glands tissues were analysed for differential expression of genes encoding proteins involved in the stress response (SOD, MT, GST, EIF1, BCL2, TP53, CAT, 18S, GADPH, GPX, GADD45, THIO9) by RT-qPCR for relative quantification. Results showed that the presence of 4-MBC at environmentally relevant concentrations induced the expression of genes that encode for antioxidant enzymes (GST) and for proteins related to the inhibition of apoptosis (BCL2) and cellular stress (GADD), suggesting a physiological stress response.
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http://dx.doi.org/10.1016/j.aquatox.2020.105418DOI Listing
April 2020

Do Contaminants Influence the Spatial Distribution of Aquatic Species? How New Perspectives on Ecotoxicological Assays Might Answer This Question.

Environ Toxicol Chem 2020 01;39(1):7-8

Center for Functional Ecology, Department of Life Sciences, University of Coimbra, Coimbra, Portugal.

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http://dx.doi.org/10.1002/etc.4594DOI Listing
January 2020

Summary of the special issue.

Sci Total Environ 2020 03 29;706:134934. Epub 2019 Nov 29.

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http://dx.doi.org/10.1016/j.scitotenv.2019.134934DOI Listing
March 2020

Ingestion and bioaccumulation of polystyrene nanoplastics and their effects on the microalgal feeding of Artemia franciscana.

Ecotoxicol Environ Saf 2020 Jan 5;188:109853. Epub 2019 Nov 5.

Department of Ecology and Coastal Management, Institute of Marine Sciences of Andalusia (ICMAN), National Research Council (CSIC), Campus Río San Pedro, 11510 Puerto Real, Cádiz, Spain.

Nanoplastics (NPs) have become one of the most serious environmental problems nowadays. The environmental issues linked to NPs are attributed to the effects after ingestion in marine organisms. Due to the incipient and controversial information about the effects of PS NPs on the feeding of organisms, the aim of this work is to assess (i) digestion dynamics of Artemia franciscana when exposed to PS NPs as the lowest concentration of PS NPs reported in toxicity test [0 (control), 0.006 and 0.6 mg·L-1] and possible interferences in the ingestion of microalgae and (ii) the accumulation and depuration of PS NPs by A. franciscana. Artemia were subjected to ingestion experiments [24 h and 3.5 h], in which the organisms were exposed to PS NPs or to PS NPs + microalgae. Post-exposure feeding (24 h exposure and 2 h feeding) and depuration (24 h exposure and 24 h of depuration) were also carried out. More than 90% of the PS NPs were ingested by Artemia and bioaccumulated in the mandible, stomach, gut, tail gut and appendages after 24 h. The ingestion of microalgae was not affected by the presence of the PS NPs. Data of post-exposure feeding indicated that Artemia previously exposed to plastic and/or microalgae presented similar microalgal ingestion (around 70%); the highest microalgal consumption (around 90%) was recorded in the treatment in which Artemia were previously starved (no plastic and no microalgae). The presence of PS NPs in the gut after the depuration experiments indicates that 24 h was not enough to eliminate the PS NPs.
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http://dx.doi.org/10.1016/j.ecoenv.2019.109853DOI Listing
January 2020

Might the interspecies interaction between fish and shrimps change the pattern of their avoidance response to contamination?

Ecotoxicol Environ Saf 2019 Dec 10;186:109757. Epub 2019 Oct 10.

Department of Ecology and Coastal Management, Institute of Marine Sciences of Andalusia (CSIC), 11510, Puerto Real, Cádiz, Spain.

Contamination seems to exert a crucial role in the spatial distribution of some organisms, such as shrimps and fish. Both, especially the freshwater fish Danio rerio and the shrimp Atyaephyra desmarestii, have been tested experimentally for their avoidance response and have showed the ability to escape from toxic effects. As the behavior of avoiding or not the contamination might be altered in the presence of other factors, the aim of the current study was to verify whether the avoidance response of both species, when exposed jointly (multispecies tests), to a copper gradient is different from the avoidance response observed in monospecies tests. The avoidance was assessed in a multi-compartmented exposure system, in which a copper gradient was simulated. Organisms were tested individually and together. Both species avoided potentially toxic copper concentrations; however, shrimps were slightly more sensitive in the monospecies tests: AC (avoidance concentration for 50% of the population) of 60 (53-68) μg/L for the zebrafish and 50 (45-56) μg/L for the shrimp. In the multispecies tests, the sensitivity pattern changed: the avoidance response by the fish [AC: 30 (14-46) μg/L] was greater than by the shrimps [AC: 70 (22-141) μg/L]. Although the AC values are in the same order of magnitude, a slight trend to change the avoidance pattern was observed in the shrimps during multispecies test: the avoidance was lower and time-delayed. This behavioral change could be linked to the stress caused by the zebrafish sharing the space with the shrimps, perhaps increasing the territorialism of the fish, or a delay in the shrimps detecting the risk of contamination.
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http://dx.doi.org/10.1016/j.ecoenv.2019.109757DOI Listing
December 2019

Biochemical response of the clam Ruditapes philippinarum to silver (AgD and AgNPs) exposure and application of an integrated biomarker response approach.

Mar Environ Res 2019 Dec 4;152:104783. Epub 2019 Sep 4.

Department of Ecology and Coastal Management, Institute of Marine Sciences of Andalusia (CSIC), Campus Río S. Pedro, 11510, Puerto Real, Cádiz, Spain.

Silver is a ubiquitous metal in the marine environment which can be accumulated by marine organisms. In order to assess the effect of dissolved silver (AgD) and AgNPs in R. philippinarum, the organisms were exposed to 20 μg L of AgD and AgNPs (15 nm) over 7 days. Bioaccumulation of the metal and oxidative and detoxification biomarkers were studied in control and exposed clams. Ag was accumulated in gills and digestive glands. Results for biochemical biomarkers (superoxide dismutase, catalase and glutathione reductase activity, lipid peroxidation and metallothionein provoked a general increase in the integrated biomarker response index (IBR) values) indicating the induction of oxidative stress in the clams exposed to both Ag treatments. Therefore, the presence of Ag forms at the tested concentration in the aquatic medium represent a risk for R. philippinarum.
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http://dx.doi.org/10.1016/j.marenvres.2019.104783DOI Listing
December 2019

Does the previous exposure to copper alter the pattern of avoidance by zebrafish in a copper gradient scenario? Hypothesis of time-delayed avoidance due to pre-acclimation.

Sci Total Environ 2019 Dec 31;694:133703. Epub 2019 Jul 31.

Department of Ecology and Coastal Management, Institute of Marine Sciences of Andalusia (CSIC), 11510 Puerto Real, Cádiz, Spain.

The traditional ecotoxicity assays (forced exposure) tend to use organisms that are cultured under controlled conditions or that come from undisturbed ecosystems, with no (or negligible) previous contact with contamination. The same occurs in the non-forced approach, in which organisms are exposed to a contamination gradient and can move between different concentrations choosing the less toxic one. Considering that organisms inhabiting contaminated ecosystems tend to be gradually exposed to contamination, an abrupt exposure from uncontaminated conditions to a contaminated environment might present two problems: lack of ecological relevance to a scenario where the contamination occurs gradually and a magnification of the toxicity due to the sudden change in the environmental conditions. Therefore, a key question should be addressed: might a previous exposure to contamination reduce the organisms' perception of the danger of a contaminant (hypothesis of time-delayed avoidance due to pre-acclimation-TDADP), altering their avoidance response pattern? We tested the avoidance of zebrafish (Danio rerio: ±2 months old) populations when exposed to a copper gradient (0-400 μg/L). The populations differed according to the period (24 h and 7 and 30 days) in which they were acclimated to copper (ca. 400 μg/L). The avoidance in the 2 h experiments changed as a consequence of the acclimation period. In the population that was not previously acclimated, 40% of the fish moved to the less contaminated compartment and only 6.7% stayed in the most contaminated one; for the other populations those values were, respectively, 31 and 11% (24 h-acclimation), 28 and 26% (7 day-acclimation) and 19 and 27% (30 day-acclimation). An abrupt exposure to a contaminant might overestimate the response if this is analyzed in the short-term. When the avoidance tests were prolonged to 24 h, the avoidance tended to reach similar values to those of the non-acclimated population, thus supporting our TDADP hypothesis.
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http://dx.doi.org/10.1016/j.scitotenv.2019.133703DOI Listing
December 2019

Risk of triclosan based on avoidance by the shrimp Palaemon varians in a heterogeneous contamination scenario: How sensitive is this approach?

Chemosphere 2019 Nov 21;235:126-135. Epub 2019 Jun 21.

Department of Ecology and Coastal Management, Institute of Marine Sciences of Andalusia (CSIC), 11510, Puerto Real, Cádiz, Spain.

As the exposure of organisms to contaminants can provoke harmful effects, some organisms try to avoid a continuous exposure by using different strategies. The aim of the current study was to assess the ability of the shrimp Palaemon varians to detect a triclosan gradient and escape to less contaminated areas. Two multi-compartmented exposure systems (the linear system and the HeMHAS-Heterogeneous Multi-Habitat Assay System) were used and then results were compared. Finally, it was aimed how sensitive the avoidance response is by comparing it with other endpoints through a sensitivity profile by biological groups and the species sensitive distribution. The distribution of the shrimps along the triclosan gradient was dependent on the concentrations, not exceeding 3% for 54 μg/L in the linear system and 7% for 81 μg/L in the HeMHAS; 25% of organisms preferred the compartment with the lowest concentrations in both systems. Half of the population seems to avoid concentrations around 40-50 μg/L. The triclosan concentration that might start (threshold) to trigger an important avoidance (around 20%) was estimated to be of 18 μg/L. The profile of sensitivity to triclosan showed that avoidance by shrimps was less sensitive than microalgae growth and avoidance by guppy; however, it might occur even at concentrations considered safe for more than 95% of the species. In summary, (i) the HeMHAS proved to be a suitable system to simulate heterogeneous contamination scenarios, (ii) triclosan triggered the avoidance response in P. varians, and (iii) the avoidance was very sensitive compared to other ecotoxicological responses.
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http://dx.doi.org/10.1016/j.chemosphere.2019.06.139DOI Listing
November 2019

Addition to Metals in the European Marine Strategies Legislation: A Challenge for the Managers and Decision-Makers.

Environ Sci Technol 2019 May 10;53(9):5528. Epub 2019 Apr 10.

ICMAN-Instituto de Ciencias Marinas de Andalucía (CSIC) . Campus Universitario Río San Pedro , 11510 Puerto Real , Cádiz . Spain.

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http://dx.doi.org/10.1021/acs.est.9b01851DOI Listing
May 2019

Are the primary characteristics of polystyrene nanoplastics responsible for toxicity and ad/absorption in the marine diatom Phaeodactylum tricornutum?

Environ Pollut 2019 Jun 21;249:610-619. Epub 2019 Mar 21.

Department of Ecology and Coastal Management, Institute of Marine Sciences of Andalusia (CSIC), Campus Río San Pedro, 11510, Puerto Real, Cádiz, Spain.

Nowadays, the occurrence of a large volume of plastic litter in oceanic and coastal zones has increased concern about its impacts on marine organisms. The degradation of plastic polymers leads to the formation of smaller fragments at both micro and nano scale (<5 mm and <1 μm respectively). Nanoplastics (NPs), due to their smaller size and high specific surface area can establish colloidal interactions with marine microalgae, therefore potential toxicity can be led. . To assess this hypothesis, the aim of the present study is to examine the behaviour of polystyrene nanoparticles (PS NPs) of different sizes (50 and 100 nm) in marine water and their possible effects at different physiological and cellular levels in the marine diatom Phaeodactylum tricornutum. Different biomarkers and stress responses in P. tricornutum were analysed when organisms were exposed to environmentally relevant PS NPs concentrations between 0.1 and 50 mg L. Our results showed significant differences between controls and exposure microalgae, indicating toxicity. After 24 h, an increase in oxidative stress biomarkers, damage to the photosynthetic apparatus, DNA damage and depolarization of mitochondrial and cell membrane from 5 mg L were observed. Further after 72 h the inhibition of population growth and chlorophyll content were observed. Examining effects the effects related to PS NPs size, the smallest (50 nm) induced greater effects at 24 h while bigger PS NPs (100 nm) at72 h. This bigger particles (100 nm) showed more stability (in size distribution and spherical form) in the different culture media assayed, when compared with the rest of particles used. Strong adsorption and/or internalization of PS NPs was confirmed through changes in cell complexity and cell size as well as the fluorescence of 100 nm fluoresbrite PS NPs after washing cell surface.
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http://dx.doi.org/10.1016/j.envpol.2019.03.047DOI Listing
June 2019

Habitat selection response of the freshwater shrimp Atyaephyra desmarestii experimentally exposed to heterogeneous copper contamination scenarios.

Sci Total Environ 2019 Apr 24;662:816-823. Epub 2019 Jan 24.

Department of Ecology and Coastal Management, Institute of Marine Sciences of Andalusia, CSIC, 11510 Puerto Real, Cádiz, Spain. Electronic address:

In contaminated aquatic ecosystems, it is expected that organisms suffer some effects caused by the contaminants. However, for mobile organisms inhabiting heterogeneously contaminated ecosystems, the continuous exposure to contaminants can be avoided by moving to less contaminated habitats. The present study evaluated the habitat selection of the freshwater shrimp Atyaephyra desmarestii experimentally exposed to different copper concentrations to verify whether the heterogeneous contamination distribution and the connectivity between habitats with different copper levels could generate a random population distribution similar to metapopulation. The experiments were performed in the HeMHAS (Heterogeneous Multi-Habitat Assay System), a non-forced multi-compartmented exposure system, in which it is possible to simulate the distribution of contaminants in a linear gradient or as patches of contamination. Copper was used to simulate a linear contamination gradient (26 to 105 μg/L Cu) and two patchy scenarios with three contamination levels [reference zone (R: 26 ± 7 μg/L Cu), mixing zone (M: 61 ± 2 μg/L Cu) and disturbed zone (D: 101 ± 12 μg/L Cu)], with two mixing zones or one central mixing zone in a heterogeneous scenario. In the copper gradient scenario, a clear trend of shrimps (59.6 ± 8.0% of the population) moving to the reference zones and an avoidance of 66.7 ± 11.1% of the most contaminated zone were observed. For the patchy scenarios, a random distribution of organisms (34, 36 and 30% for R, M and D zones, respectively) was observed in the scenario with one mixing zone; on the other hand, a slight preference for the reference zones (44.9 ± 4.8%) was evidenced in the scenario with two mixing zones. As shrimps are able to select less contaminated areas, it is highly important to preserve clean zones in heterogeneously contaminated environments, such as the arrangement in meta-ecosystems, as the less- or uncontaminated zones might represent less stressful areas to protect populations against continuous contamination exposure.
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http://dx.doi.org/10.1016/j.scitotenv.2019.01.304DOI Listing
April 2019

Assessing the effect of human pharmaceuticals (carbamazepine, diclofenac and ibuprofen) on the marine clam Ruditapes philippinarum: An integrative and multibiomarker approach.

Aquat Toxicol 2019 Mar 9;208:146-156. Epub 2019 Jan 9.

Instituto de Ciencias Marinas de Andalucía (CSIC), Campus Rio San Pedro, 11510 Puerto Real, Cádiz, Spain. Electronic address:

The presence of pharmaceuticals in the aquatic ecosystem has become a topic of growing interest in recent years. In this study, the marine clam Ruditapes philippinarum was exposed during 14 days to concentrations close to those found in the environment: (15 μg L) of carbamazepine (CBZ), diclofenac (DCF) and ibuprofen (IBU), three pharmaceuticals widely used worldwide and commonly found within the aquatic environment. Additionally, exposure was followed by a depuration phase (7 days). A battery of biomarkers (superoxide dismutase SOD, catalase CAT, glutathione reductase GR, total glutathione peroxidase T-GPx, glutathione transferase GST, lipid peroxidation LPO, acetylcholinesterase AChE and metallothionein MT) was evaluated throughout the exposure and depuration. The Integrated Biomarker Response index was calculated with all selected biomarkers and used as a complementary tool in the evaluation of the organisms' health status. Exposure induced changes in the clams' biochemical responses that led to the hypothesis of the harmful role of the pharmaceuticals resulting in negative effects (changes in enzyme activities, LPO and MT levels, related to ROS production) particularly after short-term exposure. However, the clams showed the ability to cope with these imbalances by recovering their general oxidative status by the end of exposure.
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http://dx.doi.org/10.1016/j.aquatox.2019.01.004DOI Listing
March 2019

Disturbance of ecological habitat distribution driven by a chemical barrier of domestic and agricultural discharges: An experimental approach to test habitat fragmentation.

Sci Total Environ 2019 Feb 16;651(Pt 2):2820-2829. Epub 2018 Oct 16.

Department of Ecology and Coastal Management, Institute of Marine Sciences of Andalusia (CSIC), 11510 Puerto Real, Cadiz, Spain.

Contamination is an important factor for determining the pattern of habitat selection by organisms. Since many organisms are able to move from contaminated to more favorable habitats, we aimed to: (i) verify if the contamination along the river Guadalete (Spain) could generate a chemical barrier, restricting the displacement of freshwater shrimps (Atyaephyra desmarestii) and (ii) discriminate the role of the contaminants concerning the preference response by the shrimps. A. desmarestii was experimentally tested in a multi-compartmented, non-forced exposure system, simulating the spatial arrangement of the samples just like their distribution in the environment. Water and sediment samples were chemically characterized by analyses of 98 chemical compounds and 19 inorganic elements. Shrimps selected the less contaminated water and sediment samples, with two marked preference patterns: (i) upstream displacement avoiding the sample located at the point of pollutant discharges and those samples downstream from this point and (ii) fragmentation of the population with spatial isolation of the upstream and downstream populations. The preference was related to the avoidance of artificial sweeteners, flame retardants, fragrances, PAHs, PCBs, pesticides, UV filters and some inorganic elements. The threat of contamination was related to its potential to isolate populations due to the chemical fragmentation of their habitat.
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http://dx.doi.org/10.1016/j.scitotenv.2018.10.200DOI Listing
February 2019

Spatial avoidance as a response to contamination by aquatic organisms in nonforced, multicompartmented exposure systems: A complementary approach to the behavioral response.

Environ Toxicol Chem 2019 02 19;38(2):312-320. Epub 2018 Dec 19.

Department of Ecology and Coastal Management, Institute of Marine Sciences of Andalusia, Cádiz, Spain.

The idea that the hazard of contaminants is exclusively related to their toxic effects does not consider the fact that some organisms can avoid contamination, preventing toxicity. Although inferences about avoidance are made in most behavioral ecotoxicology studies, assessment of the real spatial displacement (organisms moving toward another habitat to escape contamination) is difficult due to the type of exposure (confined and mandatory) used in the bioassays: a forced exposure approach. A complementary approach using nonforced exposure systems to assess how contaminants affect the spatial distribution of organisms in a bicompartmented (toxic or nontoxic) environment has long been described. Recently, this nonforced approach has been developed to include a multi compartmented system in which different samples can be simultaneously tested. The aim of the present review was to describe the importance of the nonforced, multicompartmented exposure approach to simulate a gradient or patches of contamination, to describe the 2 main exposure systems, and to highlight the ecological relevance of including spatial avoidance and habitat preference in ecotoxicological studies. The multicompartmentalization of the system makes it possible to simulate more complex scenarios and therefore include new ecological concepts in bioassays. We also contrasted spatial avoidance in the nonforced exposure systems with the behavioral endpoints measured under other exposure systems. Finally, we showed that the nonforced, multicompartmented exposure approach makes it possible 1) to improve environmental risk assessments by adding the dispersion pattern of organisms in a multihabitat scenario, and 2) to integrate ecological concepts such as recolonization of recovering habitats, loss of habitat connectivity, habitat fragmentation, and contamination-driven metapopulation, which have received limited attention in ecotoxicological studies. Environ Toxicol Chem 2019;38:312-320. © 2018 SETAC.
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http://dx.doi.org/10.1002/etc.4310DOI Listing
February 2019

Spatial avoidance, inhibition of recolonization and population isolation in zebrafish (Danio rerio) caused by copper exposure under a non-forced approach.

Sci Total Environ 2019 Feb 29;653:504-511. Epub 2018 Oct 29.

Department of Ecology and Coastal Management, Institute of Marine Sciences of Andalusia (CSIC), Campus Río S. Pedro, 11510 Puerto Real, Cadiz, Spain.

Aquatic ecosystems receive run-off and discharges from different sources that lead to the accumulation of contaminants such as copper. Besides producing lethal and sub-lethal effects, copper has shown to be aversive to zebrafish (Danio rerio) by triggering avoidance response. The primary aim of the present study was to evaluate how a copper gradient could affect the spatial distribution of D. rerio by triggering avoidance, preventing recolonization and isolating populations. Secondly, to what extent the food availability in a previously avoided environment could make it a less aversive environment was assessed. A non-forced, multi-compartmented exposure system with a copper gradient (0-300 μg·L), through which fish could move, was used for the avoidance and recolonization assays. To test the effect of copper on population isolation, two uncontaminated connected zones were separated by a chemical barrier with a copper concentration of 90 μg·L (a concentration producing an avoidance of 50% - AC). Zebrafish avoided copper and the 2 h-AC was 90.8 μg·L. The recolonization was in accordance with avoidance and the relationship AC/RC (RC: recolonization concentration) was around 2.5. When food was provided in the highest copper concentration, the recolonization pattern was altered, although the distribution of the fish was not statistically different from the scenario without food. The chemical barrier formed by copper (90 μg·L) impaired the migratory potential of the fish population by 41.3%; when food was provided in the last compartment, no statistically significant trend of fish moving towards that concentration was observed. Copper might act as an environmental disruptor by triggering spatial avoidance, preventing recolonization and isolating populations in zebrafish. The present study allows simultaneously including three ecological concepts to ecotoxicological studies that have received little attention: habitat selection, recolonization and habitat chemical fragmentation.
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http://dx.doi.org/10.1016/j.scitotenv.2018.10.375DOI Listing
February 2019

Erythromycin sensitivity across different taxa of marine phytoplankton. A novel approach to sensitivity of microalgae and the evolutionary history of the 23S gene.

Aquat Toxicol 2018 Nov 27;204:190-196. Epub 2018 Sep 27.

Department of Ecology and Coastal Management, Institute of Marine Sciences of Andalusia (CSIC), Campus Río S. Pedro, 11510, Puerto Real, Cádiz, Spain.

Erythromycin has been recorded in coastal waters and could pose a severe threat to marine microbial life. Macrolides such as erythromycin may affect microalgae by inhibiting the pathways involved in protein synthesis. Toxicological testing of microalgae has proven to be a useful tool for the risk assessment of a substance affecting phytoplankton. Due to the controversial results concerning the sensitivity of microalgal species to erythromycin found in the literature, the goals of this work were, initially, to assess the erythromycin sensitivity of different species of marine microalgae from different and representative taxonomic groups; and, secondly, to examine whether the sensitivity to erythromycin could be explained by the differences in the phylogenetic evolution. We chose eight species: two green algae, four heterokonts, one haptophyte and one dinoflagellate, which were then exposed to erythromycin (0.1 to 10 mg L). Our results showed a wide range of sensitivities indicating that the biology of each species was primarily responsible for the variation observed. To test the second objective, we contrasted different ecotoxicological endpoints (growth, cellular properties and status of the photosynthetic apparatus) with the phylogenetic distribution [eukaryotic host (concatenated nuclear tree), evolutionary history of the chloroplast (16S tree), efficiency and repair of photosystem II (psbA tree), and the binding site of erythromycin (23S tree)] of the species. We found that the growth inhibition of microalgae as a toxicological endpoint was the endpoint best explained by the topology of the 23S rRNA gene tree when it was modelled following a non-stationary evolutionary process.
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http://dx.doi.org/10.1016/j.aquatox.2018.09.008DOI Listing
November 2018

Avoidance response by shrimps to a copper gradient: Does high population density prevent avoidance of contamination?

Environ Toxicol Chem 2018 12 15;37(12):3095-3101. Epub 2018 Nov 15.

Department of Ecology and Coastal Management, Institute of Marine Sciences of Andalusia, Cádiz, Spain.

Bioassays using the nonforced exposure approach have been shown to be a relevant tool that might complement the traditional ecotoxicological risk assessment. Because the nonforced exposure approach is based on spatial displacement of organisms and the consequent habitat selection processes, the population density might play an important role in the decision to avoid or prefer an ecosystem. Therefore, the aim of the present study was to assess if the avoidance response to contamination, measured in a nonforced exposure system, is density-dependent and how determinant contamination could be for the habitat-selection process in comparison with the population density. The freshwater shrimp Atyaephyra desmarestii was exposed to a copper gradient in a nonforced exposure system formed by 7 interconnected compartments (total volume 600 mL), which contained different copper concentrations. The density treatments used were 3, 5, and 10 organisms per compartment, corresponding to 0.5, 0.8, and 1.7 organisms per 100 mL, respectively. Clearly, the avoidance response to copper was more intense in the population with the lower density: the highest population density showed the lowest avoidance. The concentrations that triggered an avoidance of 50% of the population were 47, 134, and 163 μg L . In summary, it was observed that shrimps were able to detect and avoid potentially toxic copper concentrations but that the avoidance response was affected by population density. Environ Toxicol Chem 2018;37:3095-3101. © 2018 SETAC.
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http://dx.doi.org/10.1002/etc.4277DOI Listing
December 2018

Effect of erythromycin and modulating effect of CeO NPs on the toxicity exerted by the antibiotic on the microalgae Chlamydomonas reinhardtii and Phaeodactylum tricornutum.

Environ Pollut 2018 Nov 3;242(Pt A):357-366. Epub 2018 Jul 3.

Department of Ecology and Coastal Management, Institute of Marine Sciences of Andalusia (CSIC), Campus Río S. Pedro, 11510, Puerto Real, Cádiz, Spain.

Erythromycin is an antibiotic employed in the treatment of infections caused by Gram positive microorganisms and the increasing use has made it a contaminant of emerging concern in aqueous ecosystems. Cerium oxide nanoparticles (CeO NPs), which are known to have catalytic and antioxidant properties, have also become contaminants of emerging concern. Due to the high reactivity of CeO NPs, they can interact with erythromycin magnifying their effects or on the other hand, considering the redox potential of CeO NPs, it can alleviate the toxicity of erythromycin. The present study was carried out to assess the toxicity of both single compounds as well as mixed on Chlamydomonas reinhardtii and Phaeodactylum tricornutum (freshwater and marine microalgae respectively) employed as target species in ecotoxicological tests. Mechanisms of oxidative damage and those harmful to the photosynthetic apparatus were studied in order to know the toxic mechanisms of erythromycin and the joint effects with CeO NPs. Results showed that erythromycin inhibited the microalgae population growth and effective quantum yield of PSII (E.Q.Y.) in both microalgae. However, the freshwater microalgae Chlamydomonas reinhardtii was more sensitive than the marine diatom Phaeodactylum tricornutum. Responses related to the photosynthetic apparatus such as E.Q.Y. was affected by the exposure to erythromycin of both microalgae, as chloroplasts are target organelle for this antibiotic. Mixed experiments (CeO NPs + erythromycin) showed the protective role of CeO NPs in both microalgae preventing erythromycin toxicity in toxicological responses such as the growth of the microalgae population and E.Q.Y.
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http://dx.doi.org/10.1016/j.envpol.2018.07.009DOI Listing
November 2018

Stress-driven emigration in complex field scenarios of habitat disturbance: The heterogeneous multi-habitat assay system (HeMHAS).

Sci Total Environ 2018 Dec 3;644:31-36. Epub 2018 Jul 3.

Aquatic BioTechnology, El Puerto de Santa María, Spain.

Lines of evidence used in ecological risk assessment (ERA) are essentially three: chemistry, biology and ecotoxicology. Until now, the fundamental assumption made when measuring ecological risks is that organisms are forcedly exposed to stressors. However, when organisms can avoid disturbed habitats by escaping to less stressful areas, the assumption that exposure is mandatory to pose risk may not match field disturbance scenarios. A non-forced exposure approach using a linear free-choice multi-compartmented system has been proposed previously as a complementary tool to assess the effects on organisms' spatial avoidance/preference responses. Yet, the linearity of the latter system limits avoidance measurements to one spatial dimension. A novel, heterogeneous multi-habitat assay system (HeMHAS) consisting of 18 connected circular compartments (3 compartments on a vertical axis in each one of 6 zones on a longitudinal axis; a 2D system) is put forward here to be used in heterogeneous-habitat selection studies, as it makes it possible to assess the ability of organisms to detect contamination and other stressors and select more favorable habitats. In the present study, the avoidance to copper by zebrafish (Danio rerio) was tested after exposing organisms to a copper gradient in the HeMHAS and compared with that in the linear system. Avoidance occurred for all copper concentrations: 43% in the lowest (21 μg·L) to 72% in the highest (221 μg·L). Results obtained within the HeMHAS (AC: 60 μg·L) were statistically (p = 0.72) similar to avoidance of copper by D. rerio in the linear non-forced system (AC: 89 μg·L). In summary, the simulation of a copper gradient in the HeMHAS (2D system) allowed to assess the potential repellency of copper to zebrafish and to corroborate the ability of organisms to detect and avoid potentially toxic concentrations.
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http://dx.doi.org/10.1016/j.scitotenv.2018.06.336DOI Listing
December 2018

Preface.

Sci Total Environ 2018 Jul 13;628-629:441-442. Epub 2018 Feb 13.

Institute of Marine Sciences of Andalusia (ICMAN-CSIC), Spain.

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http://dx.doi.org/10.1016/j.scitotenv.2018.01.329DOI Listing
July 2018

Metals in the European Marine Strategies Legislation: A Challenge for the Managers and Decision-Makers.

Environ Sci Technol 2018 07 25;52(14):7601-7603. Epub 2018 Jun 25.

ICMAN-Instituto de Ciencias Marinas de Andalucía (CSIC) , Campus Universitario Río San Pedro , 11510 Puerto Real, Cádiz , Spain.

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http://dx.doi.org/10.1021/acs.est.8b02906DOI Listing
July 2018
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