Abundance, composition, and distribution of microplastics in intertidal sediment and soft tissues of four species of Bivalvia from Southeast Brazilian urban beaches.

In coastal areas, microplastics (MPs) can deposit in sediment, allowing it to be ingested by benthic organisms, like mussels, thus creating a possible transfer to humans. The aim of this study is to evaluate MPs pollution in sediment as a function of shoreline elevation in two urbanized beaches and to evaluate the abundance/frequency of MPs in 4 different species of bivalves commonly used in the human diet, such as the oyster Crassostrea brasiliana, the mussels Mytella strigata and Perna perna and the clam Tivela mactroides, and identify the polymers via µ-FTIR technique. A total of 3337 MPs were found in this study, of which 1488 were found in the sediment at the five sites analyzed, and 1849 in the bivalve tissues at the two sampling sites. MPs contamination was observed in all sediment samples and species of the pool and in each of the 10 specimens of the four species. Thus, the frequency of contamination by MPs reached 100 % for the analyzed samples. The number of filaments is higher than fragments in sediment samples and in each bivalve species. Regarding types and colors, the blue were greater than fragment-type in sediments and samples. In an effort to classify the polymers via µ-FTIR, our study was able to identify polypropylene, polyethylene and polyethylene terephthalate, besides a great number of cellulose fibers.

Under ice spills of conventional crude oil and diluted bitumen: Physiological resilience of the blue mussel and transgenerational effects.

Spillages at sea of diluted bitumen (dilbit) from oil sands have received little attention until now. To our best knowledge, there are no reports on the impact of a severe exposure to dilbit on the Blue mussel (Mytilus edulis). In this study, adult Blue mussels were exposed to one conventional crude oil (Heidrun) and two dilbits (Cold Lake Blend and Access Western Blend) for a period of 7 days in an ice-covered environment and then maintained for three months until the spawning season. The exposed mussels were monitored for aromatic hydrocarbon bioaccumulation, physiological energetic budget, cellular stress, byssus production and gametogenesis. In spring, spawning was induced to characterize breeding success. Bioaccumulation of polycyclic aromatic hydrocarbons (PAHs) was detected after three days of exposure, with higher concentrations of PAHs associated to the conventional oil (5.49 ± 0.12 µg·g-1 d.w.) compared to both dilbits (0.91 ± 0.02 µg·g-1; 0.51 ± 0.03 µg·g-1 d.w.). Despite a fast depuration rate and a good resilience of the exposed mussels, significant negative effects were observed at the cellular, physiological and fitness levels, especially in offspring. Our results suggest a higher toxicity of the diluted bitumen compared to the conventional crude despite the lower bioaccumulation of total PAHs. Dilbit treatments caused evident negative transgenerational effects on unexposed F1 generation.

Toxicological interactions of silver nanoparticles and organochlorine pesticides in mouse peritoneal macrophages.

Nanotechnology occupies a prominent space in economy and science due to the beneficial properties of nanomaterials. However, nanoparticles may pose risks to living organisms due to their adsorption and pro-oxidative properties. The aim of the current study was to investigate the effects of polymer-coated silver nanoparticles (AgNPs) and organochlorine pesticides (OCPs), as well as their combined effects on mouse peritoneal macrophages. Macrophages were isolated and exposed to three concentrations of AgNPs (groups: N1 = 30, N2 = 300 and N3 = 3000 ng.ml(-1)), two concentrations of OCPs (groups: P1 = 30 and P2 = 300 ng.ml(-1)) and the six possible combinations of these two contaminants for 24 h. AgNPs had irregular shape, Feret diameter of 8.7 ± 7.5 nm and zeta potential of -28.7 ± 3.9 mV in water and -10.7 ± 1.04 mV in culture medium. OCP mixtures and the lower concentrations of AgNPs had no detectable effects on cell parameters, but the highest AgNPs concentration showed high toxicity (trypan blue and MTT assays) resulting in morphological changes, increase of nitric oxide levels and phagocytic index. Foremost, the association of N3 and P2 led to distinct effects from those observed under single exposure.

Endocrine status of a migratory bird potentially exposed to the Deepwater Horizon oil spill: a case study of northern gannets breeding on Bonaventure Island, Eastern Canada.

The Deepwater Horizon oil spill caused the death of a large number of seabirds in the Gulf of Mexico in 2010. However, the long term consequences of oil exposure on migratory birds overwintering in this area have received limited attention. The present study aimed to investigate the impact of oil contamination (e.g., polycyclic aromatic hydrocarbons (PAHs)) on the circulating status of prolactin and corticosterone, two hormones that influence reproductive success in birds, in Northern gannets (Morus bassanus) breeding on Bonaventure Island, Eastern Canada. Using light-based geolocators, it was found that 23.5% of Northern gannets from Bonaventure Island overwintered in the Gulf of Mexico in 2010-2011; the remainder of this population overwintered along the Atlantic Coast of the United States. PAH concentrations (eight compounds) in gannet blood cells were all found to be under the method limits of quantification, which could be the result of the ability of seabirds to metabolize these compounds and the time elapsed between oil exposure and blood sampling. Corticosterone and prolactin levels as well as body mass did not differ between the two major birds' wintering sites. Moreover, levels of both these hormones did not vary from early to late incubation period. Present results suggest that if Bonaventure Island-breeding Northern gannets had been exposed to oil in the Gulf of Mexico in the aftermath of this historical spill, this exposure could not be associated with changes in hormonal status and body mass in breeding individuals.

Biosorption of thorium on the external shell surface of bivalve mollusks: the role of shell surface microtopography.

External shell surface (ESS) of bivalve mollusks is known to adsorb various metals dissolved in ambient water in high concentration. It is hypothesized here that the surface microtopography of the thin organic coating layer, periostracum, or calcareous shell (if periostracum was destroyed) plays a major role in the adsorption of actinides on ESS. Thorium (natural alpha-emitter) was used in short-term biosorption experiment with shell fragments of five bivalve mollusks. After a 72 h exposure to Th (~6 kBq L(-1)), thorium concentration was measured on ESS using laser ablation inductively coupled plasma mass spectrometry; the distribution and density of alpha tracks were subsequently visualized by α-track autoradiography. A trend in reduced Th concentrations on the ESS was observed depending upon the species tested: (group 1 ~4000 µg g(-1)) Chlamys islandica (M.), Mercenaria mercenaria (L.), Dreissena polymorpha (P.)>(group 2 ~1200 µg g(-1)) Crassostrea virginica (G.)≫(group 3 ~150 µg g(-1)) Mytilus edulis L. The microtopography of ESS was characterized by scanning electron microscopy revealing the high porosity of the calcareous surface of C. islandica and M. mercenaria, lamellate surface of periostracum in D. polymorpha, uneven but a weakly porous surface of periostracum of C. virginica, and a nearly smooth surface of the periostracum of M. edulis. This work has demonstrated, for the first time, the presence of a strong correlation between concentration of adsorbed Th and ESS microtopography, and the role of the periostracum in this process is discussed.

Effects of pesticide compounds (chlorothalonil and mancozeb) and benzo[a]pyrene mixture on aryl hydrocarbon receptor, p53 and ubiquitin gene expression levels in haemocytes of soft-shell clams (Mya arenaria).

The aim of this study is to investigate the effects of the pesticides/polycyclic aromatic hydrocarbon mixture on aryl hydrocarbon receptor (AhR), p53 and ubiquitin mRNA level in haemocytes of Mya arenaria exposed to a mixture of chlorothalonil, mancozeb and benzo[a]pyrene (BaP) for 48 and 72 h. AhR, p53 and ubiquitin gene expression levels were quantified using quantitative Real-time PCR. For robust and accurate quantification of transcripts, suitable housekeeping genes were selected from four sets of ribosomal and elongation factors transcripts previously sequenced from Mya arenaria using geNorm open source software. Quantitative Real-time PCR data exhibited a significantly high expression of AhR after 72 h of exposure (P ≤ 0.05). p53 gene expression seems to be up-regulated by the mixture after 48 h, however not significantly; but the level of p53 mRNA is down-regulated by the xenobiotics between 48 and 72 h after exposure. This study postulates that AhR mRNA levels could be used as an indicator of the exposure of clams' haemocytes to a mixture of xenobiotics such as chlorothalonil, mancozeb and BaP. However, further studies have to be pursued in order to unravel the molecular mechanisms involved in the p53 signaling pathway.

Effects of temperature and fertilization on total vs. active bacterial communities exposed to crude and diesel oil pollution in NW Mediterranean Sea.

The dynamics of total and active microbial communities were studied in seawater microcosms amended with crude or diesel oil at different temperatures (25, 10 and 4 degrees C) in the presence/absence of organic fertilization (Inipol EAP 22). Total and hydrocarbon-degrading microbes were enumerated by fluorescence microscopy and Most Probable Number (MPN) method, respectively. Total (16S rDNA-based) vs. active (16S rRNA) bacterial community structure was monitored by Capillary-Electrophoresis Single Strand Conformation Polymorphism (CE-SSCP) fingerprinting. Hydrocarbons were analyzed after 12 weeks of incubation by gas chromatography-mass spectrometry. Total and hydrocarbon-degrading microbial counts were highly influenced by fertilization while no important differences were observed between temperatures. Higher biodegradation levels were observed in fertilized microcosms. Temperature and fertilization induced changes in structure of total bacterial communities. However, fertilization showed a more important effect on active bacterial structure. The calculation of Simpson's diversity index showed similar trends among temperatures whereas fertilization reduced diversity index of both total and active bacterial communities.

Potential link between exposure to fungicides chlorothalonil and mancozeb and haemic neoplasia development in the soft-shell clam Mya arenaria: a laboratory experiment.

The aetiology of haemic neoplasia (HN) is unknown, so far but many causative factors are suggested such as viral, pollution and genetics. The aim of this study was to determine if, under chronic exposure, two major pesticides (chlorothalonil and mancozeb) which are used in potato production could induce HN in soft-shell clams (Mya arenaria). Short-term experiments with acute exposure were also performed. Clams were collected from an epizootic site (North River, PEI) and from a site free of the disease (Magdalen Islands, Quebec). The tetraploid level of haemocytes was assessed by flow cytometry for each clam to determine the HN status. The bioaccumulation of pesticides in tissues was quantified by gas chromatography/mass spectrometry (GC/MS) for chlorothalonil while mancozeb and manganese were quantified by inductively coupled plasma-mass spectrometer (ICP/MS). Long term exposure to fungicide Bravo 500((R)) did not induce high tetraploid levels on negative calm from North River and the analysis of the digestive gland and the mantle did not reveal any detectable level of chlorothalonil. In the Manzate 200 DF((R)), some clams revealed high level of tetraploid cells but no difference were observed between the treatments and the control. The analysis of the digestive gland and the mantle for manganese did not highlight any significant difference in tissue concentration (p=0.05). For the acute exposure, chlorothalonil analysis showed that the active ingredient is distributed between four chlorinated compounds: 99.5% for chlorothalonil isomers, 0.4% for pentachlorothalonil and 0.1% for trichlorothalonil isomers. For a 72 h experiment, the accumulation was within 4h; the higher tissue concentration of chlorothalonil was 59.2 microg g(-1) in the mantle after 48 h, following by a decrease to an undetectable level at the end. For the manganese, the accumulation was detected after 4h; the higher tissue concentration was 48.8 microg g(-1) in the mantle after 24h and, over the following 48 h, the accumulation decreased until the end of the trial. Based on the data, the accumulation of these fungicides seems to be transitory. Chlorothalonil and mancozeb are both oxidative-stress promoters and could have induced cell dysfunction while in the tissue. Study on the effect of these fungicides on the p53 protein system is an example of strategy that would provide information on cellular events promoting neoplasia.

Toxic effects of tributyltin and its metabolites on harbour seal (Phoca vitulina) immune cells in vitro.

The widespread environmental contamination, bioaccumulation and endocrine disruptor effects of butyltins (BTs) to wildlife are well documented. Although suspected, potential effects of BTs exposure on the immune system of marine mammals have been little investigated. In this study, we assessed the effects of tributyltin (TBT) and its dealkylated metabolites dibutyltin (DBT) and monobutyltin (MBT) on the immune responses of harbour seals. Peripheral blood mononuclear cells isolated from pup and adult harbour seals were exposed in vitro to varying concentrations of BTs. DBT resulted in a significant decrease at 100 and 200 nM of phagocytotic activity and reduced significantly phagocytic efficiency at 200 nM in adult seals. There was no effect in phagocytosis with TBT and MBT. In pups, the highest concentration (200 nM) of DBT inhibited phagocytic efficiency. A reduction of tumor-killing capacity of adult natural killer (NK) cells occurred when leukocytes were incubated in vitro with 50 nM DBT and 200 nM TBT for 24h. In adult seals, T-lymphocyte proliferation was significantly suppressed when the cells were exposed to 200 nM TBT and 100 nM DBT. In pups, the proliferative response increased after an exposure to 100 nM TBT and 50 nM DBT, but decreased with 200 nM TBT and 100 nM DBT. The immune functions were more affected by BTs exposure in adults than in pups, suggesting that other unsuspected mechanisms could trigger immune parameters in pups. The toxic potential of BTs followed the order of DBT>TBT>MBT. BT concentrations of harbour seal pups from the St. Lawrence Estuary (Bic National Park) ranged between 0.1-0.4 ng Sn/g wet weight (ww) and 1.2-13.4 ng Sn/g ww in blood and blubber, respectively. For these animals, DBT concentrations were consistently below the quantification limit of 0.04 ng Sn/g ww in blood and 0.2 ng Sn/g ww in blubber. Results suggest that concentrations measured in pups are considered too low to induce toxic effects to their immune system during first days of life. However, based on our in vitro results, we hypothesize that BTs, and DBT in particular, could pose a serious threat to the immune functions in free-ranging harbour seal adults.

Sorption characteristics of polycyclic aromatic hydrocarbons in aluminum smelter residues.

High temperature carbon oxidation in primary aluminum smelters results in the release of polycyclic aromatic hydrocarbons (PAH) into the environment. The main source of PAH are the anodes, which are composed of petroleum coke (black carbon, BC) and coal tar pitch. To elucidate the dominant carbonaceous phase controlling the environmental fate of PAH in aluminum smelter residues (coke BC and/or coal tar), the sorptive behavior of PAHs has been determined, using passive samplers and infinitesink desorption methods. Samples directly from the wet scrubber were studied as well as ones from an adjacent 20-year old storage lagoon and roof dust from the smelter. Carbon-normalized distribution coefficients of native PAHs were 2 orders of magnitude higher than expected based on amorphous organic carbon (AOC)/water partitioning, which is in the same order of magnitude as reported literature values for soots and charcoals. Sorption isotherms of laboratory-spiked deuterated phenanthrene showed strong (-100 times stronger than AOC) but nonetheless linear sorption in both fresh and aged aluminum smelter residues. The absence of nonlinear behavior typical for adsorption to BC indicates that PAH sorption in aluminum smelter residues is dominated by absorption into the semi-solid coal tar pitch matrix. Desorption experiments using Tenax showed that fresh smelter residues had a relatively large rapidly desorbing fraction of PAH (35-50%), whereas this fraction was strongly reduced (11-16%) in the lagoon and roof dust material. Weathering of the coal tar residue and/or redistribution of PAH between coal tar and BC phases could explain the reduced availability in aged samples.

TBT toxicity on a natural planktonic assemblage exposed to enhanced ultraviolet-B radiation.

A microcosm approach was designed to study the combined effects of tributyltin (TBT) from antifouling paints and ultraviolet-B radiation (UVBR: 280-320 nm), on a natural planktonic assemblage (<150 microm) isolated from the St. Lawrence Estuary at the end of the springtime. Microcosms (9l, cylindrical Teflon bags, 75 cm heightx25 cm width) were immersed in the water column of mesocosms (1800 l, polyethylene bags, 2.3 m depth) and exposed to two different UVBR regimes: natural ambient UVBR (NUVBR), and enhanced level of UVBR (HUVBR). During consecutive 5 days, effects of TBT (120 ng l -1) and enhanced UVBR (giving a biologically weighted UVBR 2.15-fold higher than natural light condition) were monitored in the samples coming from following treatments: (i) NUVBR light condition without TBT (NUVBR), (ii) NUVBR light condition with TBT-added (NUVBR+TBT), (iii) HUVBR light condition without TBT (HUVBR) and (iv) HUVBR light condition with TBT-added (HUVBR+TBT). Each treatment was conducted in triplicate microcosms. Different parameters were then measured during 5 days, including TBT analysis, bacterial abundance and productivity, phytoplankton abundance, cellular characteristics and growth rates, as well as in vivo chlorophyll a (Chl a) fluorescence. Following TBT addition (NUVBR+TBT treatment), Chl a concentrations never exceeded 1 microg l-1 whereas final values as high as 54 microg l-1 were observed in TBT-free treatments (NUVBR and HUVBR). TBT addition resulted also in the lost of fluorescence signal of the maximum efficiency of the photosystem II in phytoplankton assemblage. TBT toxicity caused on phytoplankton <20 microm an increase of mean cell size and changes in shape reflected a drastic disturbance of the cell cycle leading to an inhibition of the apparent growth rate. These negative effects of TBT resulted in a final abundance of phytoplankton <20 microm of 591+/-35 cells ml-1 in NUVBR+TBT relative to NUVBR treatment (i.e., 31,846+/-312 cells ml-1). Moreover, when cells were submitted to TBT under enhanced UVBR (HUVBR+TBT treatment), final abundance of phytoplankton <20 microm was only 182+/-90 cells ml-1, with a significant interaction between TBT and UVBR during the last 2 days of the experiment. The same type of interaction was also observed for bacterial abundance in NUVBR+TBT and HUVBR+TBT with stimulation of 226 and of 403%, respectively due to TBT addition relative to NUVBR treatment. When considering bacterial productivity, TBT addition resulted in an inhibition of 32%, and this inhibition was significantly more pronounced under dual stresses (i.e., 77% in HUVBR+TBT). These results clearly demonstrate that the combination of TBT and UVBR stresses have synergistic effects affecting the first trophic levels of the marine food web.

Degradation of petroleum hydrocarbons in two sub-antarctic soils: influence of an oleophilic fertilizer.

In order to determine the long-term effects of fertilizer on the degradation rate and the toxicity of hydrocarbons in sub-Antarctic soils contaminated by petroleum hydrocarbons, a field study was initiated in December 2000 on two different soils of the Kerguelen Islands (69 degrees 42'E, 49 degrees 19'S). The number of hydrocarbon-degrading bacteria (HDB) increased greatly after crude-oil and diesel-fuel contamination, and the fertilizer addition had a favorable effect on HDB growth and activity. Hydrocarbon-degrading bacteria counts remained high until the end of the experiment although the total hydrocarbon content in all contaminated soils was reduced to 80 to 90% of their initial value after 330 d. Degradation of n-alkanes was enhanced significantly in the presence of the fertilizer, while the degradation of polycyclic aromatic hydrocarbons (PAHs) was only barely enhanced. Toxicity results showed a noticeable reduction with time, although toxicity remained present and important in both soils at the end of the experiment. In addition, fertilized plots showed a toxic signal greater than unfertilized ones. Overall results clearly show that fertilizer addition improves the rate of degradation of both oil contaminants. However, remaining toxic residues may constitute a drawback of the fertilizer-assisted biodegradation process at low temperatures.

Genetic damage induced by trophic doses of lead in the neotropical fish Hoplias malabaricus (Characiformes, Erythrinidae) as revealed by the comet assay and chromosomal aberrations

The effects of clastogenic or mutagenic agents have rarely been studied in neotropical fish species exposed to contaminated water. In this study, the genetic damage caused by lead in the widely distributed South American fish, Hoplias malabaricus, was assessed using the comet (SCGE) assay and by testing for chromosomal aberrations. Eighteen specimens were acclimatized to laboratory conditions and then chronically exposed to contaminated food by feeding prey (Cyprinus sp.) injected intraperitoneally with doses of inorganic lead adjusted to give a contamination level of 21 mg of Pb2+.g-1 net weight of H. malabaricus. Three fish were sampled for chromosomal analysis after four doses (18 days) and another three after eight doses (41 days) of lead and the results then compared with three untreated controls kept under lead-free conditions. An additional six treated fish and three controls were sampled for the comet assay after 13 doses (64 days). Exposure to lead significantly increased the frequency of chromosomal aberrations and the frequency of tailed cell nuclei, the latter indicating DNA damage. These results show that H. malabaricus is a useful biological model for screening the clastogenic effects of lead and possibly other xenobiotics. The genetic damage seen here illustrates the need to investigate the potential effects of heavy metals on fish species in South America.

Metabolic interactions between low doses of benzo[a]pyrene and tributyltin in arctic charr (Salvelinus alpinus): a long-term in vivo study.

We have previously reported that short-term, single exposure to a high dose of tributyltin (TBT), a widely used antifouling biocide, inhibited both the in vivo metabolism and metabolic activation of the carcinogenic polycyclic aromatic hydrocarbon benzo[a]pyrene (BaP) in fish; (BaP), in turn, stimulated TBT metabolism. Here, we provide further mechanistic evidence of mutual metabolic interactions between BaP and TBT in response to long-term, repeated exposures to low doses. Juvenile Arctic charr (Salvelinus alpinus) received 10 separate i.p. injections (a single injection every 6 days) of BaP (3 mg/kg), TBT (0.3 mg/kg), or both in combination; control fish received corn oil vehicle only. Two days after the 2nd (Day 8), 6th (Day 32), and 10th dose (Day 56), blood, bile, and liver samples were collected and analyzed for a suite of biomarkers. HPLC/fluorescence analysis indicated that TBT cotreatment inhibited the formation of (+)-anti-BaP diol-epoxide adducts with plasma albumin (53%, Day 8), hepatic DNA (27%, Day 32), or both albumin and globin (50 and 58%, Day 56) compared to BaP alone. This antagonistic interaction was associated with a time-dependent modulation (inhibition at Day 8, enhancement at Day 32) of both cytochrome P450 (P450)1A-mediated ethoxyresorufin O-deethylase (EROD) activity and biliary BaP metabolite formation. TBT cotreatment also inhibited (39%) the BaP-mediated induction of hepatic glutathione S-transferase (GST) activity observed at Day 8. Treatment with TBT alone increased EROD activity (60%) at Day 32, but decreased both GST activity (70 and 37%) and glutathione content (24% and 16%) at Days 32 and 56, respectively. GC/MS analysis revealed that, at Day 56, BaP cotreatment increased (200%) the levels of biliary butyltin compounds, including mono- and dibutyltin metabolites. This potentiative interaction was associated with a protective effect of BaP cotreatment against the TBT-mediated decreases in GST activity and glutathione content. The current results demonstrate that, whereas TBT inhibited BaP bioactivation via a time-dependent modulation of P4501A induction, BaP stimulated the Phase II metabolism of TBT and/or its biliary excretion. The mutual metabolic interactions between these two widespread aquatic pollutants reinforce the need for long-term in vivo interactive studies at low doses.