Key factors influencing Hg levels and trends in unperturbed oligotrophic temperate and boreal lakes.

Mercury (Hg) is a toxic metal that presents a major risk to ecosystems, biota, human health, and remains a priority concern. In temperate and boreal lakes Hg and methylmercury (MMHg) are expected to vary as a function of atmospheric Hg deposition, lake water chemistry, catchment characteristics and climate variables. The aim of this study was to quantify Hg and MMHg in unperturbed oligotrophic lakes and to identify the factors controlling their distribution. We first hypothesized that lake Hg (and MMHg to lesser extent) spatial variations are linked to atmospheric deposition, catchment characteristics, and terrestrial exportation of dissolved organic carbon (DOC). We secondly examined if lake Hg concentrations have followed the decrease in atmospheric Hg emission observed between the mid-1990s to the end-2010s. We found that overall, atmospheric Hg has little impact on lake Hg and MMHg concentrations, which are both primarily influenced by DOC input originating from the forest catchment. The relationship between DOC and Hg differed between the spring and the fall, with a Hg-to-DOC ratio twice as high in spring. This seems related to snowmelt input of Hg (with a relatively reduced input of DOC) or the internal lake build-up of Hg during the ice-covered period. Of the 10 lakes intensively visited over a 20-year period, only 3 showed significant lake Hg decreases despite significant negative trends in atmospheric Hg concentrations, suggesting a lag between atmospheric and surface water temporal trends. Overall, terrestrial catchments retain around 80% of atmospheric Hg implying that large Hg pools have been built up in soils in the last decades. As such, the reduction of atmospheric Hg alone will not necessarily result in Hg decreases in lakes, since the Hg concentrations may be modulated by DOC export trends and catchment characteristics. This stresses the need to improve our understanding of the processes governing Hg transfers from catchments into lakes.

Single-step production of autologous bovine platelet concentrate for clinical applications in cattle.

Platelet concentrate (PC) is an alternative therapy to treat mastitis in dairy cattle and is an alternative treatment for reproduction problems such as endometritis. Unfortunately, double-centrifugation processing methods described are time-consuming, require specialized laboratory equipment, and are usually done in a heterologous way, which risks herd health. To overcome this limitation, we evaluated single-step bovine PC processing methods readily applicable to a farm setting using an autologous conditioned plasma (ACP) production system. We investigated the hematologic findings, cytokines, and growth factors of the obtained PC samples. Autologous conditioned plasma was prepared using whole blood (WB) from 4 cows (group 1) using single-step centrifugation and 16 different processing methods. The 2 protocols that yielded the highest ratio of platelet to white blood cell (WBC) concentration were ACP-1 [720 × g (2,200 rpm), 5 min] and ACP-2 [929 × g (2,500 rpm), 3 min]. They were subsequently reproduced and compared using WB from 8 cows (group 2). Hematologic findings were quantified, IL-1ß (cytokine) and growth factors [platelet-derived growth factor (PDGF), transforming growth factor (TGF)-ß, bovine fibroblast growth factor (b-FGF)] were measured, and enrichment factors were compared between samples and processing methods. Hematological characteristics and platelet enrichment varied markedly among tested protocols and all were statistically different from WB. Protocol ACP-2 resulted in significantly greater platelet enrichment (mean 169% of WB) than ACP-1 (125% of WB). We found no significant difference between the 2 ACP preparation protocols with regard to leukocyte reduction (7.53-9.75% WBC compared with WB) or growth factor enrichment (124-125% PDGF, 95-100% TGF-ß, 102-104% b-FGF, and 56-74% IL-1ß compared with WB). In conclusion, both ACP protocols yielded a platelet concentration shown to promote healing for clinical applications in cattle, and the ACP-2 protocol resulted in a greater degree of platelet enrichment. Therefore, this protocol could be used for ACP production for clinical applications in cattle.

Ecotoxicological impacts of oil sand mining activity to endemic caged mussels Pyganodon grandis.

The intense mining extraction of oil sand (OS) has increased over the last few decades, raising concerns about the release of OS contaminants and toxicity in resident aquatic organisms in the Athabasca River (Alberta, Canada). To address this, endemic Pyganodon grandis mussels were caged for 6 weeks at various upstream and downstream sites of industrial OS mining activities. Post-exposure mussels were then analyzed for light/medium/heavy polyaromatic hydrocarbons (PAHs) in tissues, general health (weight to length ratio, growth rate, air survival time), biotransformation (cytochrome P4501A and 3A and glutathione S-transferase activities), oxidative stress/inflammation (lipid peroxidation-LPO and arachidonate cyclooxygenase-COX), genotoxicity (DNA strand breaks), and gonad status (triglycerides, GSI and vitellogenin-like proteins). The following effects significantly differed between OS mining area and natural/background sites: health condition, growth rate, air survival time, COX (immune/inflammation) activity, P4501A/GST activity, LPO and DNA breaks in the digestive gland and vitellogenin-like proteins in the gonad. Correlation analysis revealed that the biochemical responses were scaled to at least one of the following impacts at the individual level: air survival time, weight to length ratio, growth rate and vitellogenin-like proteins. These indices were therefore identified as key adverse outcome pathways of mussels impacted by OS mining activities. Based on the relative levels of light/medium/heavy PAHs in tissues, the observed effects appears to be associated rather to the disturbance of OS in this area than contamination from OS tailing ponds leaching into the aquatic environment.

Modeling one-handed grip strangulation: Intentionality of the gesture and age influence.

Strangulation is a violent act which can be lethal and is often studied in forensic context. The neck includes several anatomical elements that can evolve with aging. We therefore created a numerical human neck model including the main anatomical elements and simulated one-handed grip strangulation cases. In addition, we created 3 models each representing age groups: 20-30 years old, 30-50 years old and over 50 years old. The main changes between the different age groups are the ossification of the cartilages and the muscles mechanical properties. Several initial and boundary conditions have been tested to perform a realistic simulation of one-handed grip strangulation. Stress analysis and fracture observation were compared with the grip strength of an average man, 552 N, to look at the intentionality of the gesture. In each age group, the results show no model fracture for a force of 552 N. It is necessary to reach a minimum of 1406 N before observing a first fracture on the hyoid bone. However, it is possible to get stresses on the hyoid bone and on the thyroid cartilage way before 552 N. It thus appears that the force created by one-handed grip strangulation is not sufficient to cause fractures of the bony elements of the neck, but it remains sufficient to compress the larynx and at least reduce airflow.

The Rare Knowledge Mining Methodological Framework for the Development of Practice Guidelines and Knowledge Translation Tools for Rare Diseases.

Rare diseases bring on a heavy health, social and economic burden that impacts patients' lives and puts pressure on the healthcare system. Furthermore, they are often associated with limited published studies to inform multidisciplinary clinical practice thus limiting evidence-based practice. Moreover, the development of knowledge translation products including clinical care guidelines are often very challenging based on the current available methodological frameworks relying mostly on critical appraisal of the published research evidence where randomized clinical trial design is considered as the gold standard. To overcome this barrier, we proposed the Rare Knowledge Mining Methodological Framework (RKMMF). The RKMMF is one possible answer to improve the development of knowledge translation products for rare diseases. This framework includes other sources of evidence including registry information and qualitative studies and the involvement of expert patients. This article documents the RKMMF structure and its application is exemplified through knowledge translation products developed for a neuromuscular population.

Prevalence of Coxiella burnetii seropositivity and shedding in farm, pet and feral cats and associated risk factors in farm cats in Quebec, Canada.

Cats represent a potential source of Coxiella burnetii, the aetiological agent of Q fever in humans. The prevalence and risk factors of C. burnetii infection in farm, pet and feral cats were studied in Quebec, Canada, using a cross-sectional study. Serum samples were tested using a specific enzyme-linked immunosorbent assay (ELISA) for the presence of antibodies against C. burnetii, whereas rectal swabs were assayed using real-time quantitative polymerase chain reaction (qPCR) for the molecular detection of the bacteria. Potential risk factors for farm cats were investigated using clinical examinations, questionnaires and results from a concurrent study on C. burnetii farm status. A total of 184 cats were tested: 59 from ruminant farms, 73 pets and 52 feral cats. Among farm cats, 2/59 (3.4%) were ELISA-positive, 3/59 (5.1%) were ELISA-doubtful and 1/59 (1.7%) was qPCR-positive. All pets and feral cats were negative to C. burnetii ELISA and qPCR. Farm cat positivity was associated with a positive C. burnetii status on the ruminant farm (prevalence ratio = 7.6, P = 0.03). Our results suggest that although pet and feral cats do not seem to pose a great C. burnetii risk to public health, more active care should be taken when in contact with cats from ruminant farms.

Changes in spinal bone density, back muscle size, and visceral adipose tissue and their interaction following a multi-component exercise program in older men: secondary analysis of an 18-month randomized controlled trial.

In middle-aged and older men, an 18-month multi-component exercise program improved spinal trabecular BMD, paraspinal, and psoas muscle cross-sectional area (CSA) but not visceral adipose tissue (VAT). However, changes in both muscle and VAT CSA were associated with changes in spinal BMD, independent of the exercise intervention. INTRODUCTION: In older men, we previously reported that a multi-component exercise program improved lumbar spine (LS) trabecular volumetric BMD (Tb.vBMD) compared with no exercise. This study aimed to investigate the following: (1) the effect of the exercise program on paraspinal and psoas (back) muscle CSA and VAT, and 2) if any exercise-related changes in muscle CSA and/or VAT were associated with changes in spinal BMD. METHODS: Men (n = 180) aged 50-79 years were randomized to an exercise or no-exercise group. Exercise involved high-intensity progressive resistance training (60-85% max) with weight-bearing impact exercise (3 days/week) for 18 months. Quantitative computed tomography was used to assess L1-L3 Tb.vBMD, paraspinal, and psoas muscle CSA and VAT. RESULTS: Exercise resulted in a 2.6% ((95% CI, 1.1, 4.1), P < 0.01) net gain in back muscle CSA, but no effect on VAT (-1.6% (95% CI, -7.3, 4.2)) relative to no exercise. Robust regression indicated that percentage changes in Tb.vBMD were positively associated with changes (expressed as z-scores) in back muscle CSA in both the exercise (beta (ß)-coefficient = 1.9, 95% CI 0.5, 3.2, P = 0.007) and no-exercise (ß = 2.6, 95% CI, 1.1, 4.1, P = 0.001) group, and negatively with the changes in VAT (ß = -2.0, 95% CI -3.3, -0.7, P = 0.003) in the exercise only group. There were no group differences in the slopes for the muscle-bone or VAT-bone relationships. Regression analysis (pooled data) revealed that back muscle CSA and VAT were independent predictors of the change in Tb.vBMD, explaining 14% of the variance. CONCLUSION: A multi-component exercise program in middle-aged and older men improved spinal BMD and back muscle size but not visceral fat. However, changes in back muscle size and VAT were associated with the changes in spinal BMD, independent of exercise. TRIAL REGISTRATION: ACTRN 12617001224314, 22/08/2017 retrospectively registered.

The influence of surface coatings on the toxicity of silver nanoparticle in rainbow trout.

Silver nanoparticles (nAg) are often produced with different coatings that could influence bioavailability and toxicity in aquatic organisms. The purpose of this study was to examine the influence of 4 surface coatings of nAg of the same core size towards bioavailability and toxicity in juvenile rainbow trout (Oncorhynchus mykiss). Juveniles were exposed to 50 µg/L of 50 nm diameter nAg for 96 h at 15 °C with the following coatings: branched polyethylenimine (bPEI), citrate, polyvinylpyrrolidone (PVP) and silicate (Si). The data revealed that the coatings influenced hepatic Ag loadings in the following trend PVP > citrate > bPEI and Si with estimated bioavailability factors of 28, 18, 6 and 2 L/kg respectively. Hepatic Ag levels were significantly associated with DNA damage and inflammation as determined by arachidonate cyclooxygenase activity. The bPEI and citrate-coated nAg consistently produced the observed effects above in addition to increased mitochondrial electron transport activity and glutathione S-transferase activity. The absence of metallothionein and lipid peroxidation suggests that mechanisms other than the liberation of Ag+ were at play. In conclusion, surface coatings were shown to significantly influence bioavailability and toxic properties of nAg to rainbow trout juveniles.

Biomechanical study of the thyroid cartilage: A model of bi-digital strangulation.

The presence of fracture on neck elements is an indication of violence. Both the hyoid bone and the larynx can be damaged by a strangulation mechanism. Thyroid cartilage, more specifically, may present lesions in response to this mechanical stress. These lesions result in fractures at the bases of the horns of the thyroid cartilage. This study focuses on the thyroid cartilage behavior in cases of bi-digital strangulation, using an anthropometric and biomechanical approach. To develop a biomechanical model, we performed an anthropometric study taking into account 14 distances measurements as well as 3 measurements of angles. These measures allowed us to determine a significant sexual dimorphism between individuals. Then, we define 6 morphologies models, composed of 3 females and 3 males individuals. In order to visualize the ossification of the cartilage, each model has been tested with bone properties. Strangulation cases were simulated by applying an imposed velocity of 0.4m/s then 1m/s. We observed different behaviors of the thyroid cartilage according to the sex and the morphology.

A major release of urban untreated wastewaters in the St. Lawrence River (Quebec, Canada) altered growth, reproduction, and redox status in experimentally exposed Daphnia magna.

In 2015, five billion liters of untreated urban wastewater (UWW) were released into the St. Lawrence River (Quebec, Canada) over the course of four days in order to repair the Montreal's sewer interceptor network related to the city's primary wastewater treatment plant. The UWW discharge originated mainly from household, industrial, and hospital sources. The objective of this study was to investigate the toxicological effects of this unprecedented punctual UWW release on aquatic invertebrates to gather information that could help understand the potential impacts to the receiving environment of overflow episodes occurring during heavy rain events. Water samples were collected at four impacted and non-impacted sites during and four weeks after the release. The freshwater crustacean Daphnia magna were experimentally exposed to surface water collected from UWW-impacted sites for 13 days and analyzed for life-history endpoints and suitable biomarkers related to oxidative stress (i.e., catalase, superoxide dismutase, lipid peroxidation, and glutathione-s-transferase) and reproduction (chitinase). Results indicated that D. magna growth and reproduction were significantly increased by exposure to UWWs. These effects were correlated with an increase in chitinase activity, which is primarily controlled by reproductive hormones and involved in growth, suggesting potential impacts on these processes. Results also indicated that some UWW samples might have caused oxidative stress during the release but that it was overcome by antioxidant defenses and did not lead to cellular damage. Overall, current results contribute to a better understanding of the biological impacts of UWW to aquatic invertebrates for a better stormwater management.

Gene expression changes and toxicity of selected rare earth elements in rainbow trout juveniles.

Rare earth elements (REEs) are increasingly used in electronics industry and other areas of our economy and questions were raised about their impacts to the environment. The purpose of this study was to examine the lethal and sublethal toxicity of REEs in juvenile rainbow (Oncorhynchus mykiss) trout. The fish were exposed to increasing concentrations (0.064, 0.32, 1.6, 8 and 40 mg/L) of the following 7 REEs for 96 h at 15 °C: cerium (CeCl3), erbium (ErCl3), gadolinium (GdCl3), lanthanum (LaCl3), neodymium (NdCl3), samarium (SmCl3) and yttrium (YCl3). The mortality were determined and in the surviving fish, 10 target gene transcripts were measured in the liver to track changes in oxidative stress, DNA repair, tissue growth/proliferation, protein chaperoning, xenobiotic biotransformation and ammonia metabolism. The data revealed that Y, Sm, Er and Gd formed a distinct group based on toxicity (mortality) and gene expression changes. Electronegativity was significantly correlated (r = -0.8, p < 0.01) with the lethal concentration (LC50). Gene expression changes occurred at concentration circa 120 times lower than the LC50 and the following transcripts in protein chaperoning (heat shock proteins), DNA repair (growth arrest DNA Damage) and CYP1A1 gene expression involved in the metabolism of coplanar aromatic hydrocarbons were involved. In conclusion, the study revealed that the more electronegative REEs were the most toxic to trout juveniles and produced sublethal effects at concentrations 2 orders of magnitude lower than the lethal concentrations. The toxicity of REEs depends on the elements were toxicity involves specific pathways at the gene expression level.

The influence of surface waters on the bioavailability and toxicity of zinc oxide nanoparticles in freshwater mussels.

The release of engineered nanoparticles in the aquatic environment could pose a threat to the biota. The purpose of the study was to examine the influence of surface water characteristics on zinc oxide nanoparticles (nZnO) and ZnS04 toxicity to the freshwater mussel Dreissena polymorpha. Mussels were exposed to an equivalent concentration of 25 µg/L Zn as either nZnO or ZnSO4 for 96 h at 15 °C in 4 types of surface waters: green water (high conductivity and pH with low natural organic matter content), brown water (low conductivity and pH with high natural organic matter content), diluted municipal effluent (high conductivity and pH with high urban organic matter content) and aquarium water (treated green water with organic matter removed). After the exposure period, mussels were analyzed for air-time survival, total and labile Zn levels in tissues, lipid metabolism (phospholipase A2, triglycerides levels) and oxidative stress (glutathione S-transferase, arachidonate cyclooxygenase, lipid peroxidation). The data revealed that mussels exposed to ZnSO4 in controlled aquarium water accumulated more total and labile Zn tissues, decreased oxidative stress and triglycerides and increased air time survival. While nZnO had few effects in aquarium water, oxidative stress was enhanced and total Zn in tissues were decreased in brown water and diluted municipal effluent and triglycerides were higher in nZn-exposed mussels in brown water. Air-time survival was decreased in mussels kept in green water and nZnO. It was also decreased in mussels exposed to ZnSO4 in green water and diluted municipal effluent. In conclusion, the fate and toxic effects of Zn could be influenced by both the chemical form (nanoparticles or ionic Zn) and surface water properties in freshwater mussels.

Effects of Ionic Strength, Salt, and pH on Aggregation of Boehmite Nanocrystals: Tumbler Small-Angle Neutron and X-ray Scattering and Imaging Analysis.

The US government currently spends significant resources managing the legacies of the Cold War, including 300 million liters of highly radioactive wastes stored in hundreds of tanks at the Hanford (WA) and Savannah River (SC) sites. The materials in these tanks consist of highly radioactive slurries and sludges at very high pH and salt concentrations. The solid particles primarily consist of aluminum hydroxides and oxyhydroxides (gibbsite and boehmite), although many other materials are present. These form complex aggregates that dramatically affect the rheology of the solutions and, therefore, efforts to recover and treat these wastes. In this paper, we have used a combination of transmission and cryo-transmission electron microscopy, dynamic light scattering, and X-ray and neutron small and ultrasmall-angle scattering to study the aggregation of synthetic nanoboehmite particles at pH 9 (approximately the point of zero charge) and 12, and sodium nitrate and calcium nitrate concentrations up to 1 m. Although the initial particles form individual rhombohedral platelets, once placed in solution they quickly form well-bonded stacks, primary aggregates, up to ∼1500 Å long. These are more prevalent at pH = 12. Addition of calcium nitrate or sodium nitrate has a similar effect as lowering pH, but approximately 100 times less calcium than sodium is needed to observe this effect. These aggregates have fractal dimension between 2.5 and 2.6 that are relatively unaffected by salt concentration for calcium nitrate at high pH. Larger aggregates (>∼4000 Å) are also formed, but their size distributions are discrete rather than continuous. The fractal dimensions of these aggregates are strongly pH-dependent, but only become dependent on solute at high concentrations.

Metal bioaccumulation and biomarkers of effects in caged mussels exposed in the Athabasca oil sands area.

The Athabasca oil sands deposit is the world's largest known reservoir of crude bitumen and the third-largest proven crude oil reserve. Mining activity is known to release contaminants, including metals, and to potentially impact the aquatic environment. The purpose of this study was to determine the impacts of oil sands mining on water quality and metal bioaccumulation in mussels from the Fort McMurray area in northern Alberta, Canada. The study presents two consecutive years of contrasting mussel exposure conditions (low and high flows). Native freshwater mussels (Pyganodon grandis) were placed in cages and exposed in situ in the Athabasca River for four weeks. Metals and inorganic elements were then analyzed in water and in mussel gills and digestive glands to evaluate bioaccumulation, estimate the bioconcentration factor (BCF), and determine the effects of exposure by measuring stress biomarkers. This study shows a potential environmental risk to aquatic life from metal exposure associated with oil sands development along with the release of wastewater from a municipal treatment plant nearby. Increased bioaccumulation of Be, V, Ni and Pb was observed in mussel digestive glands in the Steepbank River, which flows directly through the oil sands mining area. Increased bioaccumulation of Al, V, Cr, Co, Ni, Mo and Ni was also observed in mussel gills from the Steepbank River. These metals are naturally present in oil sands and generally concentrate and increase with the extraction process. The results also showed different pathways of exposure (particulate or dissolved forms) for V and Ni resulting from different river water flows, distribution coefficient (Kd) and BCF. Increasing metal exposure downstream of the oil sands mining area had an impact on metallothionein and lipid peroxidation in mussels, posing a potential environmental risk to aquatic life. These results confirm the bioavailability of some metals in mussel tissues associated with detoxification of metals (metallothionein levels), and oxidative stress in mussels located downstream of the oil sands mining area. These results highlight a potential ecotoxicological risk to biota and to the aquatic environment downstream of the oil sands mining area, even at low metal exposure levels.

Determination of the bioavailability of selected pharmaceutical residues in fish plasma using liquid chromatography coupled to tandem mass spectrometry.

Aquatic systems near major urban centers are constantly contaminated with effluent from wastewater treatment plants. Pharmaceuticals are part of the contamination and several classes of drugs have been detected in surface waters in the last decade. To better understand the impact of those pharmaceuticals in ecosystems, the exposure to aquatic species needs to be investigated. This study presents a new simple and rugged quantitative method for the determination of several classes of drugs using 100µL of plasma from fish environmentally exposed to a major but highly diluted urban effluent. Six common drugs (i.e., diclofenac, ibuprofen, naproxen, salbutamol, sulfamethoxazole and trimethoprim) and one major metabolite (2-hydroxy-ibuprofen), present in significant amount in impacted waterways have been selected for the development and validation of the method. First, all drugs were extracted using cation exchange solid phase extraction (SPE) and eluted with two solvent mixtures. Then, the extracts were analyzed using a reverse-phase analytical column Waters® CORTECS C18+ (150×2.1mm, 2.7µm) within 14min. MS/MS was performed with an electrospray (ESI) interface in positive ion mode, with multiple reaction monitoring (MRM) experiment acquiring two product ions per drugs. Quantification has been made with standard curves for each analyte using isotopically labeled internal standards. This method has high sensitivity with limits of quantification of 1ngmL-1 for each drug, except for ibuprofen and its metabolite 2-hydroxy-ibuprofen at 2ngmL-1. The precision of the method was below 11%, the accuracy between 94 and 105% and overall recovery between 94 and 111% for all drugs, with high selectivity. Application of the method to plasma samples from wild northern pike inhabiting the St. Lawrence River collected over a three-year period showed the presence of naproxen, diclofenac, trimethoprim and salbutamol at very low concentrations (around 1ngmL-1).

An investigation of the immunotoxicity of oil sands processed water and leachates in trout leukocytes.

Increased oil sands (OS) mining activity has raised concerns about impacts on aquatic organisms. This study sought to examine the effects of single representative compounds from OS (benzo(a)pyrene, naphthalene), a mixture of naphthenic acids (NAs), OS-processed water (OSPW) and OS leachate (OSL) extracts on rainbow trout leukocytes. Primary cultures of trout leukocytes were exposed to increasing concentrations of benzo(a)pyrene, naphthalene, NAs, OSPW and OSL for 48h at 18°C. Immunocompetence was followed by measuring changes in lymphocyte and macrophage viability and phagocytosis. Changes in the expression of 10 transcripts were also followed: interleukin 1, 2 and 6 (Il-1, Il-2 and Il-6), calreticulin (CRT), caspase 9 (Cas9), aryl hydrocarbon receptor (AhR), cyclooxygenase-2 (COX2), glutathione S-transferase (GST), catalase (CAT) and p53 tumor suppressor. The results revealed that exposure to OSPW extracts decreased the capacity of macrophages to engulf three beads or more, while the other compounds generally increased phagocytosis activity. Lymphocyte apoptosis was increased by all compounds and mixtures except naphthalene. Both OSPW and OSL induced apoptosis in macrophages. At the gene expression level, Cas9, CRT, Il-1 (inhibition) and Il-2 were specifically influenced by OSPW, while CAT, p53, COX2 and Il-1 (induction) transcripts were specifically expressed by OSL. Leukocyte exposure to OSPW produced characteristic changes in immunocompetence and genes involved in proinflammatory, apoptosis and protein damage (CRT) pathways which could not be explained by OSL, benzo(a)pyrene, naphthalene and NA mixture.

Elemental profiles of freshwater mussels treated with silver nanoparticles: A metallomic approach.

Nanoparticles released into the environment could pose a risk to resident organisms that feed on suspended particles in aquatic ecosystems. The purpose of this study was to examine the effects of silver nanoparticles (nanoAg) of different sizes in freshwater mussels using a multi-elemental (metallomic) approach in order to determine signature effects of nanoparticulate and ionic Ag. Mussels were exposed to three concentrations (0.8, 4 and 20µg/L) of 20-nm and 80-nm nanoAg and AgNO3 for 48h at 15°C. After the exposure period, mussels were placed in clean, aerated water for a depuration step and analyzed for the following total elements in gill, digestive gland and gonad tissues: Al, Ag, As, Ba, Be, Ca, Cd, Co, Cr, Cu, Fe, K, Mg, Mn, Mo, Pb, Na, Ni, Se, Sr, Th, U, V and Zn. Metallothioneins (MT; digestive gland only) and lipid peroxidation (LPO) were also determined in gills, digestive glands and gonads. The 20-nm-diameter nanoAg was detected in all three tissues at 20µg/L, while the 80-nm nanoAg was detected more strongly in the digestive gland. Ionic Ag was found at higher levels in gills than in other tissues. Correlation analysis revealed that gonad Ag levels were significantly correlated with Al (r=0.28), V (r=0.28), Cr (r=0.31), Co (r=0.32), Se (r=0.34) and MT levels (r=0.28). Indeed, the MT levels in the digestive gland were significantly increased by 20-nm nanoAg (20µg/L) and 80-nm nanoAg (4µg/L) and AgNO3 (<0.8µg/L). LPO was observed in gills, digestive glands and even gonads for all Ag forms. Discriminant function analysis revealed that all forms of Ag differed from each other and from unexposed mussels, where ionic Ag was more closely related to the 80-nm-diameter nanoAg. Factorial analysis revealed that Ba, Ca, Co, Mn, Sr, U and Zn had consistently high factorial weights in all tissues; that explained 80% of the total variance. Moreover, the following elements showed strong correlations (r>0.7) with each other: Sr, Ba, Zn, Ca, Mg Cr, Mn and U. Comparisons of these elements with other elements showing low or no correlations (e.g., transition elements) revealed that these elements had significantly lower standard reduction potential and electronegativity, suggesting that stronger reducing elements were most influenced by the oxidizing effects of nanoAg and ionic Ag in tissues. Indeed, tissues with oxidative stress (LPO) had decreased levels for most of these reducing elements. We conclude that exposure to Ag nanoparticles produces a characteristic change in the elemental composition of gills, digestive gland and gonad tissues in freshwater mussels. Elements most responsive to oxidative stress were more influenced by both nanoAg and ionic Ag. Sr and Ba were readily decreased by Ag and appeared to respond more sensitively to nanoAg than to ionic Ag. The metallomic approach could contribute in the understanding of fundamental mode of action of nanoparticles in mussels.

Degradation of the pharmaceuticals diclofenac and sulfamethoxazole and their transformation products under controlled environmental conditions.

Contamination of the aquatic environment by pharmaceuticals via urban effluents is well known. Several classes of drugs have been identified in waterways surrounding these effluents in the last 15years. To better understand the fate of pharmaceuticals in ecosystems, degradation processes need to be investigated and transformation products must be identified. Thus, this study presents the first comparative study between three different natural environmental conditions: photolysis and biodegradation in aerobic and anaerobic conditions both in the dark of diclofenac and sulfamethoxazole, two common drugs present in significant amounts in impacted surface waters. Results indicated that degradation kinetics differed depending on the process and the type of drug and the observed transformation products also differed among these exposure conditions. Diclofenac was nearly degraded by photolysis after 4days, while its concentration only decreased by 42% after 57days of exposure to bacteria in aerobic media and barely 1% in anaerobic media. For sulfamethoxazole, 84% of the initial concentration was still present after 11days of exposure to light, while biodegradation decreased its concentration by 33% after 58days of exposure under aerobic conditions and 5% after 70days of anaerobic exposure. In addition, several transformation products were observed and persisted over time while others degraded in turn. For diclofenac, chlorine atoms were lost primarily in the photolysis, while a redox reaction was promoted by biodegradation under aerobic conditions. For sulfamethoxazole, isomerization was favored by photolysis while a redox reaction was also favored by the biodegradation under aerobic conditions. To summarize this study points out the occurrence of different transformation products under variable degradation conditions and demonstrates that specific functional groups are involved in the tested natural attenuation processes. Given the complexity of environmental samples more analytical effort is needed to fully identify new products of potential toxicity.

Fate of silver nanoparticles in wastewater and immunotoxic effects on rainbow trout.

Silver nanoparticles (AgNPs) are currently used in technology, medicine and consumer products, even though the fate and the ecotoxicological risks on aquatic organisms of these new materials are not well known. The purpose of this study was to investigate the fate, bioavailability of AgNPs and their effects on fish in presence of municipal effluents. Juvenile rainbow trout were exposed for 96h to 40µg/L of AgNPs or 4µg/L of dissolved silver (AgNO3) in diluted (10%) municipal wastewater. Silver (Ag) concentrations were measured both on water samples and fish tissues (liver and gills). Toxicity was investigated by following immunological parameters in the pronephros (viability, phagocytosis) and biomarkers in liver and gills (cyclooxygenase activity, lipid peroxidation, glutathione-S-transferase, metallothioneins, DNA strand breaks and labile zinc). Results indicated that AgNPs appeared as small non-charged aggregates in wastewaters (11.7±1.4nm). In gills, the exposure to AgNPs induced morphological modifications without visible nanoparticle bioaccumulation. Dissolved Ag(+) was bioavailable in diluted effluent and induced oxidative stress (lipid peroxidation), labile zinc and a marginal decrease in superoxide dismutase in fish gills. Ag(+) also increased significantly metallothionein levels and inhibited the DNA repair activity in the liver. Finally, the two silver forms were found in liver and induced immunosuppression and inflammation (increase in cyclooxygenase activity). This study demonstrated that both forms of Ag produced harmful effects and AgNPs in wastewater were bioavailable to fish despite of their formation of aggregates.