Cell and tissue level responses in mussels Mytilus galloprovincialis dietarily exposed to PVP/PEI coated Ag nanoparticles at two seasons

Silver nanoparticles (Ag NPs) are present in numerous consumer products due to their antimicrobial and other unique properties, thus concerns about their potential input into aquatic ecosystems are increasing. Toxicity of Ag NPs in waterborne exposed aquatic organisms has been widely investigated, but studies assessing the potential toxic effects caused after ingestion through the food web, especially at low realistic concentrations, remain scarce. Moreover, it is not well known whether season may influence toxic effects of Ag NPs. The main objective of this study was to determine cell and tissue level responses in mussels Mytilus galloprovincialis dietarily exposed to poly-N-vinyl-2-pirrolidone/polyethyleneimine (PVP/PEI) coated 5 nm Ag NPs for 1, 7 and 21 days both in autumn and spring. Mussels were fed every day with microalgae Isochrysis galbana exposed for 24 h to a low dose (1 mu g Ag/L Ag NPs) in spring and to a higher dose (10 mu g Ag/L Ag NPs) in spring and autumn. Mussels fed with microalgae exposed to the high dose accumulated Ag significantly after 21 days in both seasons, higher levels being measured in autumn compared to spring. Intralysosomal metal accumulation measured in mussel digestive gland and time- and dose-dependent reduction of mussels health status was similar in both seasons. DNA strand breaks increased significantly in hemocytes at both exposure doses along the 21 days in spring and micronuclei frequency showed an increasing trend after 1 and 7 days of exposure to 1 mu g Ag/L Ag NPs in spring and to 10 mu g Ag/L in both seasons. Values decreased after 21 days of exposure in all the cases. In conclusion, PVP/PEI coated 5 nm Ag NPs ingested through the food web were significantly accumulated in mussel tissues and caused adverse cell and tissue level effects both in autumn and in spring.This work has been funded by the Spanish Ministry of Economy and Competitiveness (Nano Silver Omics project MAT2012-39372), Basque Government (SAIOTEK project S-PE13UN142 and Consolidated Research Group GIC IT810-13 and IT1302-19), the University of the Basque Country UPV/EHU (UFI 11/37 and PhD fellowship to N.D.) and French Ministry of Higher Education and Research (PhD fellowship to M.M.)

Changes in protein expression in mussels Mytilus galloprovincialis dietarily exposed to PVP/PEI coated silver nanoparticles at different seasons

Potential toxic effects of Ag NPs ingested through the food web and depending on the season have not been addressed in marine bivalves. This work aimed to assess differences in protein expression in the digestive gland of female mussels after dietary exposure to Ag NPs in autumn and spring. Mussels were fed daily with microalgae previously exposed for 24 hours to 10 µg/L of PVP/PEI coated 5 nm Ag NPs. After 21 days, mussels significantly accumulated Ag in both seasons and Ag NPs were found within digestive gland cells and gills. Two-dimensional electrophoresis distinguished 104 differentially expressed protein spots in autumn and 142 in spring. Among them, chitinase like protein-3, partial and glyceraldehyde-3-phosphate dehydrogenase, that are involved in amino sugar and nucleotide sugar metabolism, carbon metabolism, glycolysis/gluconeogenesis and the biosynthesis of amino acids KEGG pathways, were overexpressed in autumn but underexpressed in spring. In autumn, pyruvate metabolism, citrate cycle, cysteine and methionine metabolism and glyoxylate and dicarboxylate metabolism were altered, while in spring, proteins related to the formation of phagosomes and hydrogen peroxide metabolism were differentially expressed. Overall, protein expression signatures depended on season and Ag NPs exposure, suggesting that season significantly influences responses of mussels to NP exposure.This work has been funded by the Spanish Ministry of Economy and Competitiveness (NanoSilverOmics project MAT2012-39372), Basque Government (SAIOTEK project S-PE13UN142 and Consolidated Research Group GIC IT810-13) and the University of the Basque Country UPV/EHU (UFI 11/37 and PhD fellowship to N.D.). This study had also the support of Fundação para a Ciência e Tecnologia (FCT) from Portugal through the Strategic Project UID/MAH00350/2013 granted to CIMA. The contribution of K. Mehennaoui was possible within the project NanoGAM (AFR-PhD-9229040) and M. Mikolaczyk was supported by a PhD fellowship from the French Ministry of Higher Education and Research.info:eu-repo/semantics/acceptedVersio

Accumulation de l’argent et du cuivre chez l’huître japonaise Crassostrea gigas : outils géochimiques pour une optimisation du message environnemental

The data-base analyses of the French biomonitoring program (RNO/ROCCH, IFREMER, 2003-2014) in terms of silver (Ag) and copper (Cu) concentrations accumulation in oysters from 13 sites along the French Atlantic Coast showed spatial and time-dependent variations reflecting local pressures. The high Ag and Cu relations seems to be partly due to physiological processes involved in their bioaccumulation by oysters. The Cu/Ag elementary ratios showed an interesting potential for Ag and Cu sources detection, which were invisible through the use of only absolute concentrations characterized by a high individual variability. Chemical measurements and histological observations performed in wild oysters from Gironde Estuary showed a preferential Ag and Cu accumulation in the digestive gland with important cellular damages. The innovative isotope spiking method is highly sensitive and precise to observe rapid in vivo Ag and Cu accumulation kinetics through direct pathway using close-to-real exposure concentrations. The isotopic signals allowed to quantify the rapid MTE accumulation in the different organs and to observe the Ag and Cu co-regulation. Furthermore, it represents a real step forward in ecotoxicological experiments allowing to obtain the initial and final contamination state inside the same organism. The study of the trophic pathway suggested a higher Ag accumulation in algae than for Cu. The Ag and Cu potentially bioavailable fraction in estuarine particles was estimated respectively at 60% and 82%. The results showed that this estimation depends on the exposure time and sediment concentration, which can lead to important under-estimations.L’analyse des données de biosurveillance (RNO/ROCCH, IFREMER, 2003-2014) des concentrations en argent (Ag) et cuivre (Cu) dans les huîtres sauvages de 13 sites de la côte atlantique française a montré des variations spatio-temporelles reflétant les pressions locales. De fortes relations entre Ag et Cu semblent en partie dues à des facteurs physiologiques impliqués dans leur bioaccumulation chez les huîtres. Le rapport élémentaire Cu/Ag a montré un potentiel intéressant pour détecter des sources de Ag et Cu, invisibles à travers les concentrations absolues qui sont caractérisées par une très forte variabilité inter-individuelle. Les mesures chimiques et les observations histologiques dans les huîtres sauvages de la Gironde, ont montré une accumulation préférentielle de Ag et Cu dans la glande digestive, accompagnée d’importantes dégradations cellulaires. La méthode innovante du dopage isotopique est très sensible et précise pour observer les cinétiques rapides d’accumulation in vivo de Ag et Cu par voie directe à des niveaux d’exposition réalistes. Les signaux isotopiques ont permis de quantifier l’accumulation rapide des ETM dans les différents organes et d’observer des phénomènes de co-régulation entre Ag et Cu. De plus, elle représente une réelle avancée pour des expérimentations écotoxicologiques permettant d’analyser l’état de contamination initial et après exposition dans le même individu. L’étude de la voie trophique suggère une accumulation de Ag supérieure à celle de Cu chez les algues. La fraction de Ag et Cu particulaire potentiellement biodisponible des particules estuariennes a été estimée à 60% et 82%, respectivement. Les résultats ont montré que cette estimation dépend du temps d’exposition et de la concentration en sédiment, pouvant entraîner d’importantes sous-estimations

Silver and copper accumulation in oyster Crassostrea gigas : geochemical tools for environmental message optimization

L’analyse des données de biosurveillance (RNO/ROCCH, IFREMER, 2003-2014) des concentrations en argent (Ag) et cuivre (Cu) dans les huîtres sauvages de 13 sites de la côte atlantique française a montré des variations spatio-temporelles reflétant les pressions locales. De fortes relations entre Ag et Cu semblent en partie dues à des facteurs physiologiques impliqués dans leur bioaccumulation chez les huîtres. Le rapport élémentaire Cu/Ag a montré un potentiel intéressant pour détecter des sources de Ag et Cu, invisibles à travers les concentrations absolues qui sont caractérisées par une très forte variabilité inter-individuelle. Les mesures chimiques et les observations histologiques dans les huîtres sauvages de la Gironde, ont montré une accumulation préférentielle de Ag et Cu dans la glande digestive, accompagnée d’importantes dégradations cellulaires. La méthode innovante du dopage isotopique est très sensible et précise pour observer les cinétiques rapides d’accumulation in vivo de Ag et Cu par voie directe à des niveaux d’exposition réalistes. Les signaux isotopiques ont permis de quantifier l’accumulation rapide des ETM dans les différents organes et d’observer des phénomènes de co-régulation entre Ag et Cu. De plus, elle représente une réelle avancée pour des expérimentations écotoxicologiques permettant d’analyser l’état de contamination initial et après exposition dans le même individu. L’étude de la voie trophique suggère une accumulation de Ag supérieure à celle de Cu chez les algues. La fraction de Ag et Cu particulaire potentiellement biodisponible des particules estuariennes a été estimée à 60% et 82%, respectivement. Les résultats ont montré que cette estimation dépend du temps d’exposition et de la concentration en sédiment, pouvant entraîner d’importantes sous-estimations.The data-base analyses of the French biomonitoring program (RNO/ROCCH, IFREMER, 2003-2014) in terms of silver (Ag) and copper (Cu) concentrations accumulation in oysters from 13 sites along the French Atlantic Coast showed spatial and time-dependent variations reflecting local pressures. The high Ag and Cu relations seems to be partly due to physiological processes involved in their bioaccumulation by oysters. The Cu/Ag elementary ratios showed an interesting potential for Ag and Cu sources detection, which were invisible through the use of only absolute concentrations characterized by a high individual variability. Chemical measurements and histological observations performed in wild oysters from Gironde Estuary showed a preferential Ag and Cu accumulation in the digestive gland with important cellular damages. The innovative isotope spiking method is highly sensitive and precise to observe rapid in vivo Ag and Cu accumulation kinetics through direct pathway using close-to-real exposure concentrations. The isotopic signals allowed to quantify the rapid MTE accumulation in the different organs and to observe the Ag and Cu co-regulation. Furthermore, it represents a real step forward in ecotoxicological experiments allowing to obtain the initial and final contamination state inside the same organism. The study of the trophic pathway suggested a higher Ag accumulation in algae than for Cu. The Ag and Cu potentially bioavailable fraction in estuarine particles was estimated respectively at 60% and 82%. The results showed that this estimation depends on the exposure time and sediment concentration, which can lead to important under-estimations

The estuarine geochemical reactivity of Zn isotopes and its relevance for the biomonitoring of anthropogenic Zn and Cd contaminations from metallurgical activities: Example of the Gironde fluvial-estuarine system, France

Zinc stable isotopes measurements by MC-ICP-MS, validated by laboratory intercalibrations, were performed on wild oysters, suspended particles and filtered river/estuarine water samples to provide new constraints for the use of Zn isotopes as environmental tracers. The samples selected were representative of the long range (400 km) transport of metal (Zn, Cd, etc.) contamination from former Zn-refining activities at Decazeville (i.e. δ66Zn > 1 ‰) and its phasing out, recorded during 30 years in wild oysters from the Gironde Estuary mouth (RNO/ROCCH sample bank). The study also addresses additional anthropogenic sources (urban and viticulture) and focuses on geochemical reactivity of Zn in the turbidity gradient and the maximum turbidity zone (MTZ) of the fluvial Gironde Estuary. In this area, dissolved Zn showed a strong removal onto suspended particulate matter (SPM) and progressive enrichment in heavy isotopes with increasing SPM concentrations varying from δ66Zn = -0.02 ‰ at 2 mg/L to +0.90 ‰ at 1310 mg/L. These signatures were attributed to kinetically driven adsorption due to strongly increasing sorption sites in the turbidity gradient and MTZ of the estuary. Oysters from the estuary mouth, contaminated sediments from the Lot River and SPM entering the estuary showed parallel historical evolutions (1979-2010) for Zn/Cd ratios but not for δ66Zn values. Oysters had signatures varying from δ66Zn = 1.43 ‰ in 1983 to 1.18 ‰ in 2010 and were offset by δ66Zn = 0.6 - 0.7 ‰ compared to past (1988) and present SPM from the salinity gradient. Isotopic signatures in river-borne particles entering the Gironde Estuary under contrasting freshwater discharge regimes during 2003-2011 showed similar values (δ66Zn ≈ 0.35 ± 0.03 ‰; 1SD, n=15), i.e. they were neither related to former metal refining activities at least for the past decade nor clearly affected by other anthropogenic sources. Therefore, the Zn isotopic signatures in Gironde oysters reflect the geochemical reactivity of Zn in the estuary rather than signatures of past metallurgical contaminations in the watershed as recorded in contaminated river sediments. The study also shows that the isotopic composition of Zn is strongly fractionated by its geochemical reactivity in the Gironde Estuary, representative of meso-macrotidal estuarine systems