Chemicals sorbed to environmental microplastics are toxic to early life stages of aquatic organisms.

Microplastics are ubiquitous in aquatic ecosystems, but little information is currently available on the dangers and risks to living organisms. In order to assess the ecotoxicity of environmental microplastics (MPs), samples were collected from the beaches of two islands in the Guadeloupe archipelago, Petit-Bourg (PB) located on the main island of Guadeloupe and Marie-Galante (MG) on the second island of the archipelago. These samples have a similar polymer composition with mainly polyethylene (PE) and polypropylene (PP). However, these two samples are very dissimilar with regard to their contamination profile and their toxicity. MPs from MG contain more lead, cadmium and organochlorine compounds while those from PB have higher levels of copper, zinc and hydrocarbons. The leachates of these two samples of MPs induced sublethal effects on the growth of sea urchins and on the pulsation frequency of jellyfish ephyrae but not on the development of zebrafish embryos. The toxic effects are much more marked for samples from the PB site than those from the MG site. This work demonstrates that MPs can contain high levels of potentially bioavailable toxic substances that may represent a significant ecotoxicological risk, particularly for the early life stages of aquatic animals.

Proteomic response of Macrobrachium rosenbergii hepatopancreas exposed to chlordecone: Identification of endocrine disruption biomarkers?

The present work is the first study investigating the impacts of chlordecone, an organochlorine insecticide, on the proteome of the decapod crustacean Macrobrachium rosenbergii, by gel-free proteomic analysis. The hepatopancreas protein expression variations were analysed in organisms exposed to three environmental relevant concentrations of chlordecone (i.e. 0.2, 2 and 20µg/L). Results revealed that 62 proteins were significantly up- or down-regulated in exposed prawns compared to controls. Most of these proteins are involved in important physiological processes such as ion transport, defense mechanisms and immune system, cytoskeleton dynamics, or protein synthesis and degradation. Moreover, it appears that 6% of the deregulated protein are involved in the endocrine system and in the hormonal control of reproduction or development processes of M. rosenbergii (e.g. vitellogenin, farnesoic acid o-methyltransferase). These results indicate that chlordecone is potentially an endocrine disruptor compound for decapods, as already observed in vertebrates. These protein modifications could lead to disruptions of M. rosenbergii growth and reproduction, and therefore of the fitness population on the long-term. Besides, these disrupted proteins could be suggested as biomarkers of exposure for endocrine disruptions in invertebrates. However, further investigations are needed to complete understanding of action mechanisms of chlordecone on proteome and endocrine system of crustaceans.

Polycyclic aromatic hydrocarbons (PAHs) in surface sediment and oysters (Crassostrea rhizophorae) from mangrove of Guadeloupe: levels, bioavailability, and effects.

Surface sediment and oysters (Crassostrea rhizophorae) from the coastlines of Guadeloupe were analysed for polycyclic aromatic hydrocarbons (PAHs) using GC/MS. Biomarkers of oxidative stress were used to assess the response of these oysters to hydrocarbons exposure. The total concentration of PAHs in the sediment ranged from 49 to 1065 ng/g dw, while concentrations in oyster ranged from 66 to 961 ng/g dw. Molecular indices based on isomeric PAHs ratios characterize the pollution sources and show that most of the contaminations in sediment originate from pyrolytic inputs. Bioaccumulation factors (BAFs) have been related to isomeric ratio calculated for oysters in order to refine PAHs sources. The variations of BAFs observed in the different compounds resulted from different uptake pathways in the mangrove oysters according to the type of inputs. Response of biomarkers showed inhibition of catalase and an increase of lipid peroxidation at the station where PAHs concentrations were the highest. Taken together, data obtained point to the relevance of considering environmental conditions as factors influencing biomarker responses in environmental monitoring programs. These data also indicate the need for regular environmental follow-up studies in Guadeloupe.

Environmental fate of chlordecone in coastal habitats: recent studies conducted in Guadeloupe and Martinique (Lesser Antilles).

The organochlorine pollution by chlordecone, an insecticide spread in the past in banana plantations, is now recognized as a major ecological, economic, and social crisis in Guadeloupe and Martinique Islands. Due to its physical and chemical properties, this molecule is particularly persistent in the natural environment. Volcanic soil of Guadeloupe and Martinique contain allophanes (amorphous clays), which favor chlordecone trapping due to their structure and physical properties. Thus, with this trapping ability, allophanes serve as a vector allowing chlordecone to contaminate runoff waters and, finally, the sea. In the present publication, several studies recently conducted in the Lesser Antilles have been compiled in order to evaluate the desorption of chlordecone from allophanes when arriving in the estuarine environment and to determine the transfer of chlordecone along marine trophic food webs. The experiments showed that 20% of the initial quantity of chlordecone was released from allophanes in estuarine conditions and 10% in the marine environment. These results could explain the high level of contamination found in the suspended organic matter and zooplankton in the coastal areas located downstream of the contaminated watersheds. The contamination of the marine food webs of mangroves, seagrass beds, and coral reefs is dominated by a contamination "by bath" in littoral waters containing chlordecone and by bioamplification seawards.

Contamination of marine fauna by chlordecone in Guadeloupe: evidence of a seaward decreasing gradient.

Chlordecone is an organochlorine pesticide, used in the Lesser Antilles from 1972 to 1993 to fight against a banana weevil. That molecule is very persistent in the natural environment and ends up in the sea with runoff waters. The objective of the present study is to evaluate the level of contamination in several trophic groups of marine animals according to their distance from the source of pollution. Samples of suspended matter, macroalgae, herbivorous fishes, detrivorous crustaceans, zooplanktivorous fishes, first- and second-order of carnivorous fishes, and piscivorous fishes have been collected in two sites, located downstream the contaminated sites (Goyave and Petit-Bourg), in three marine habitats (coastal mangroves, seagrass beds located 1.5 km from the shoreline, and coral reefs at 3 km offshore). Animals collected in mangroves were the most contaminated (mean concentrations 193 µg kg-1 in Goyave and 213 µg kg-1 in Petit-Bourg). Samples from seagrass beds presented intermediate concentrations of chlordecone (85 µg kg-1 in Goyave and 107 µg kg-1 in Petit-Bourg). Finally, samples from coral reefs were the less contaminated (71 µg kg-1 in Goyave and 74 µg kg-1 in Petit-Bourg). Reef samples, collected 3 km offshore, were two to three times less contaminated than those collected in mangroves.

Bioaccumulation, distribution and elimination of chlordecone in the giant freshwater prawn Macrobrachium rosenbergii: Field and laboratory studies.

Chlordecone is a persistent organochlorine pesticide that has been widely used in Guadeloupe (French West Indies) to control the banana weevil Cosmopolites sordidus from 1972 to 1993. A few years after its introduction, widespread contamination of soils, rivers, wild animals and aquatic organisms was reported. Although high chlordecone concentrations have been reported in several crustacean species, its uptake, internal distribution, and elimination in aquatic species have never been described. This study aimed at investigating the accumulation and tissue distribution of chlordecone in the giant freshwater prawn Macrobrachium rosenbergii, using both laboratory (30 days exposure) and field (8 months exposure) approaches. In addition, depuration in chlordecone-free water was studied. Results showed that chlordecone bioconcentration in prawns was dose-dependent and time-dependent. Moreover, females appeared to be less contaminated than males after 5 and 7 months of exposure, probably due to successive spawning leading in the elimination of chlordecone through the eggs. Chlordecone distribution in tissues of exposed prawns showed that cephalothorax organs, mainly represented by the hepatopancreas, was the most contaminated. Results also showed that chlordecone was accumulated in cuticle, up to levels of 40% of the chlordecone body burden, which could be considered as a depuration mechanism since chlordecone is eliminated with the exuviae during successive moults. Finally, this study underlined the similarity of results obtained in laboratory and field approaches, which highlights their complementarities in the chlordecone behaviour understanding in M. rosenbergii.

Level of contamination by metallic trace elements and organic molecules in the seagrass beds of Guadeloupe Island.

Seagrass bed ecosystems occupy the most important part of coastal shelf in the French West Indies. They also constitute nurseries for many invertebrates and fishes harvested by local fisheries. In Guadeloupe, coastal fish stocks are declining meanwhile several agroecosystems revealed to be heavily contaminated by pollutants (agricultural lands, rivers, mangroves, seagrass beds, and coral reefs). Considering these facts, a study of the contamination of seagrass beds (8000 ha) of the Grand Cul-de-Sac Marin (GCSM) bay was conducted on their sediments and marine phanerogams. The analyses concerned six metals (Cd, Cu, Hg, Pb, V, Zn), tributyltin, 18 polycyclic aromatic hydrocarbons (PAHs), eight polybrominated diphenyl ethers (PBDEs), 38 polychlorobiphenyls (PCBs), dithiocarbamates (CS2 residues), and 225 pesticide molecules.Overall, the level of contamination of the seagrass beds was low for both sediments and phanerogams. Metallic trace elements were the main pollutants but with higher concentrations recorded in coastal sites, and their distribution can be explained by the proximity of river mouths and current patterns. The level of contamination was lower in plants than in sediments. However, the level of contamination between these two compartments was significantly correlated. The conclusion of this study is that, unlike other coastal ecosystems of Guadeloupe such as mangroves, the seagrass beds in the GCSM present a low degree of pollution. The observed level of contaminants does not seem to threaten the role of nursery played by the seagrass beds and does not likely present a risk for the reintroduction of manatees.

Assessment of the contamination of marine fauna by chlordecone in Guadeloupe and Martinique (Lesser Antilles).

Chlordecone is an organochlorine pesticide, used in the Lesser Antilles from 1972 to 1993 to fight against a banana weevil. That molecule is very persistent in the natural environment and ends up in the sea with runoff waters. From 2003 to 2013, seven campaigns of samplings have been conducted to evaluate the level of contamination of fish, crustaceans, and mollusks. The present study is the first assessment and the first comparison of the concentrations of chlordecone between marine areas, taxonomic groups, and ecological factors like trophic groups or preferential habitat of fish species. The four most contaminated marine areas are located downstream the contaminated rivers and banana plantations. Crustaceans seemed to be more sensitive to the contamination than fish or mollusks. Finally, when comparing contamination of fish according to their ecology, we found that fish usually living at the border of mangrove and presenting detritivores-omnivores diets were the most contaminated by chlordecone. These results are particularly useful to protect the health of the local population by controlling the fishing and the commercialization of seafood products, potentially contaminated by chlordecone.

Effects of chlordecone on 20-hydroxyecdysone concentration and chitobiase activity in a decapod crustacean, Macrobrachium rosenbergii.

Chlordecone (CLD) is an organochlorine insecticide abundant in aquatic environment of the French West Indies. However, few studies have investigated its impact on freshwater invertebrates. Whereas CLD is suspected of inducing endocrine disruption, this work aimed to study the effects of environmentally relevant concentrations of CLD on the 20-hydroxyecdysone (20-HE) hormone concentration and on the chitobiase activity, both having key roles in the molting process of crustaceans. In addition, the bioaccumulation of CLD was measured in the muscle tissue of Macrobrachium rosenbergii to underline potential dose-response relationship. The results have shown that CLD was bioaccumulated in exposed organisms according to a trend to a dose-response relationship. Moreover, it was observed that CLD decreased the 20-HE concentration in exposed prawns when compared to control, whatever the duration of exposure, as well as it inhibited the chitobiase activity after 30days of exposure. The present study indicates that CLD could interfere with molting process of M. rosenbergii by disturbing the 20-HE concentration and the activity of chitobiase, suggesting consequences at the long term on the shrimp development. This study also confirmed that CLD could be an endocrine disruptor in decapod crustaceans, as it was already observed in vertebrates.

Expression of biotransformation and oxidative stress genes in the giant freshwater prawn Macrobrachium rosenbergii exposed to chlordecone.

Chlordecone is a persistent organochlorine pesticide widely used between 1972 and 1993 in the French West Indies to control the root borer in banana fields. Chlordecone use resulted in long-term pollution of soils, contamination of waters, of aquatic organisms, and of fields. Chlordecone is known to be neurotoxic, to increase prostate cancer, and to have negative effects on cognitive and motor development during infancy. In Guadeloupe, most of the freshwater species living in contaminated rivers exceed the French legal limit of 20 µg·kg(-1) wet weight. In the present study, we chose a transcriptomic approach to study the cellular effects of chlordecone in the giant freshwater prawn Macrobrachium rosenbergii, an important economical species in Guadeloupe. Quantitative PCR revealed an induction of genes involved in defense mechanism against oxidative stress (catalase and selenium-dependent glutathione peroxidase) in prawns exposed to low environmental concentrations of chlordecone after 12 and 24 h of exposure. In prawns reared in a contaminated farm, transcription of genes involved in the biotransformation process (cytochrome P450 and glutathione-S-transferase (GST)) were induced after 8 days of exposure. Our results provide information on the mechanims of defense induced by chlordecone in aquatic crustacean species. This gene expression study of selected genes should be further strengthened by proteomic analyses and enzymatic activity assays to confirm the response of these biomarkers of stress in crustaceans and to give new insights into the mechanism of toxicity by chlordecone.

Kinetics of the inhibition of neutrophil proteinases by recombinant elafin and pre-elafin (trappin-2) expressed in Pichia pastoris.

Elafin and its precursor, trappin-2 or pre-elafin, are specific endogenous inhibitors of human neutrophil elastase and proteinase 3 but not of cathepsin G. Both inhibitors belong, together with secretory leukocyte protease inhibitor, to the chelonianin family of canonical protease inhibitors of serine proteases. A cDNA coding either elafin or its precursor, trappin-2, was fused in frame with yeast alpha-factor cDNA and expressed in the Pichia pastoris yeast expression system. Full-length elafin or full-length trappin-2 were secreted into the culture medium with high yield, indicating correct processing of the fusion proteins by the yeast KEX2 signal peptidase. Both recombinant inhibitors were purified to homogeneity from concentrated culture medium by one-step cationic exchange chromatography and characterized by N-terminal amino acid sequencing, Western blot and kinetic studies. Both recombinant elafin and trappin-2 were found to be fast-acting inhibitors of pancreatic elastase, neutrophil elastase and proteinase 3 with k(ass) values of 2-4 x 10(6) m(-1).s(-1), while dissociation rate constants k(diss) were found to be in the 10(-4) s(-1) range, indicating low reversibility of the complexes. The equilibrium dissociation constant K(i) for the interaction of both recombinant inhibitors with their target enzymes was either directly measured for pancreatic elastase or calculated from k(ass) and k(diss) values for neutrophil elastase and proteinase 3. K(i) values were found to be in the 10(-10) molar range and virtually identical for both inhibitors. Based on the kinetic parameters determined here, it may be concluded that both recombinant elafin and trappin-2 may act as potent anti-inflammatory molecules and may be of therapeutic potential in the treatment of various inflammatory lung diseases.