Low nitrogen (N) content and retention in bauxite-processing residue sand (BRS) disposal areas pose a great challenge to the establishment of sustainable vegetation cover in this highly alkaline environment. The budget and fate of applied N in BRS and its potential environmental impacts are largely unknown. We investigated the effect of combined application of biochars [aged acidic (AC) vs alkaline pine (PC)] and di-ammonium phosphate (DAP) fertiliser on ammonia (NH₃) volatilisation, nitrous oxide (N₂O) emission and N retention in a 116-day glasshouse study. The application of AC to BRS decreased pH (≈0.5 units) in BRS, while PC biochar increased pH (≈0.3 units). The application of AC reduced NH₃ volatilisation by ca. 80%, while PC by ca. 25%. On the other hand, the AC treatment increased N₂O emission by 5 folds. However, the N loss via N₂O emission in the AC treatment only accounted for ca. 0.4% of applied N. The reduction in BRS pH and increased retention of mineral N due to the presence of oxygen-containing (phenolic and carboxylic) functional groups in AC may be responsible for reduced NH₃ volatilisation and increased N₂O emission. This study has highlighted the potential of biochar (particularly aged biochar) in improving N retention and minimising environmental impacts in highly alkaline environments.

Low nitrogen (N) content and retention in bauxite-processing residue sand (BRS) disposal areas pose a great challenge to the establishment of sustainable vegetation cover in this highly alkaline environment. The budget and fate of applied N in BRS and its potential environmental impacts are largely unknown. We investigated the effect of combined application of biochars [aged acidic (AC) vs alkaline pine (PC)] and di-ammonium phosphate (DAP) fertiliser on ammonia (NH3) volatilisation, nitrous oxide (N2O) emission and N retention in a 116-day glasshouse study. The application of AC to BRS decreased pH (≈0.5 units) in BRS, while PC biochar increased pH (≈0.3 units). The application of AC reduced NH3 volatilisation by ca. 80%, while PC by ca. 25%. On the other hand, the AC treatment increased N2O emission by 5 folds. However, the N loss via N2O emission in the AC treatment only accounted for ca. 0.4% of applied N. The reduction in BRS pH and increased retention of mineral N due to the presence of oxygen-containing (phenolic and carboxylic) functional groups in AC may be responsible for reduced NH3 volatilisation and increased N2O emission. This study has highlighted the potential of biochar (particularly aged biochar) in improving N retention and minimising environmental impacts in highly alkaline environments.

The response of sediment bacterial communities in subtropical freshwater benthic microcosms to sediment-associated triclosan (TCS; 28 d exposure) was analysed using Illumina high-throughput sequencing. This study highlights the interactive effects of TCS and the presence of benthic macroinvertebrates (Limnodrilus hoffmeisteri and Viviparidae bellamya) on sediment bacterial communities. Our results show that TCS alone significantly altered the taxonomic composition and decreased alpha diversity of sediment bacterial communities at concentrations ≥80 μg TCS/g dry weight (dw) sediment (sed). Regarding dominant phyla, TCS significantly reduced the relative abundance of Bacteroidetes and Firmicutes at these concentrations, whereas the relative abundance of Chloroflexi and Cyanobacteria increased. In the presence of benthic macroinvertebrates, the sediment bacterial community was affected by 8 μg TCS/g dw sed as well. However, the presence of benthic macroinvertebrates did not cause measurable changes to bacterial community in unspiked (i.e., control) sediment. These results indicate that TCS alone would not alter the sediment bacterial community at environmentally relevant concentrations (up till 8 μg/g dw sed), but may have an effect in combination with the presence of benthic macroinvertebrates. Therefore, we recommend to include benthic macroinvertebrates when assessing the response of sediment bacterial communities during exposure to environmental stress such as organic contaminants.

The response of sediment bacterial communities in subtropical freshwater benthic microcosms to sediment-associated triclosan (TCS; 28 d exposure) was analysed using Illumina high-throughput sequencing. This study highlights the interactive effects of TCS and the presence of benthic macroinvertebrates (Limnodrilus hoffmeisteri and Viviparidae bellamya) on sediment bacterial communities. Our results show that TCS alone significantly altered the taxonomic composition and decreased alpha diversity of sediment bacterial communities at concentrations ≥80 μg TCS/g dry weight (dw) sediment (sed). Regarding dominant phyla, TCS significantly reduced the relative abundance of Bacteroidetes and Firmicutes at these concentrations, whereas the relative abundance of Chloroflexi and Cyanobacteria increased. In the presence of benthic macroinvertebrates, the sediment bacterial community was affected by 8 μg TCS/g dw sed as well. However, the presence of benthic macroinvertebrates did not cause measurable changes to bacterial community in unspiked (i.e., control) sediment. These results indicate that TCS alone would not alter the sediment bacterial community at environmentally relevant concentrations (up till 8 μg/g dw sed), but may have an effect in combination with the presence of benthic macroinvertebrates. Therefore, we recommend to include benthic macroinvertebrates when assessing the response of sediment bacterial communities during exposure to environmental stress such as organic contaminants.

Microplastics debris (<5 mm) are increasingly abundant in the marine environment, therefore, potentially becoming a growing threat for different marine organisms. Through aquatic animals, these can enter in the human food chain, and can be perceived as a risk for consumers’ health.Different studies report the presence of particles in marketable shellfish including the world wide commercially grown Pacific oyster Magallana gigas (Thunberg, 1793). The aim of this study is to examine the potential risk of microplastics entering in the human food chain through this shellfish species, investigating the dynamics of the uptake, egestion (faeces) and rejection (pseudofaeces) of microplastics in Pacific oysters under controlled conditions.M. gigas collected from a farm in the San Teodoro lagoon (Italy), were exposed to 60 fluorescent orange polystyrene particles L⁻¹ of known sizes (100, 250 and 500 μm). The uptake of each particle size was 19.4 ± 1.1%, 19.4 ± 2% and 12.9 ± 2% respectively. After exposure M. gigas were left to depurate for 72 h, during which 84.6 ± 2% of the particles taken up were released whilst 15.4 ± 2% were retained inside the shell cavity. No microplastic particles were found in the animals’ soft tissues.The results of this study, suggest that depuration is an effective method to reduce presence of large microplastic particles, in the size range 100–500 μm, in M. gigas. Importantly, the data suggests that the burden that could theoretically be up taken by consumers from these shellfish is negligible when compared to other routes.

The Italian territory is covered of approximately 384,000 ha of coastal lagoons and these could represent potential location for Pacific oyster, Crassostrea gigas (Thunberg, 1793) farming. Approximately 10,000 ha of the coastal Italian lagoons are located in the Sardinian region, these are currently not utilized for shellfish farming, and present suitable environmental conditions that allow for good growth rates and optimal market quality for Pacific oysters farming. Therefore, these lagoons can become potential farming sites to increase this industry in the Italian territory, where the demand for this shellfish species cannot be met by domestic production alone. The aim of this PhD study was to enhance Pacific oyster farming in Mediterranean coastal lagoons taking into account local aquaculture industry’s ambitions and challenges, therefore different Pacific oyster culture aspects were investigated. To achieve the goals of this PhD study, novel farming tools and their benefits to Pacific oyster farming were investigated. Moreover, the validation of ShellSIM® growth model in a new environment was carried out, providing a new validated tool to farmers and stakeholders to monitor oysters’ performances and estimate productivity in local waters. Finally, a site selection methodology was developed to identify suitable sites for Pacific oyster farming, and potential risks to consumers linked to microplastics pollution in coastal areas and uptake in Pacific oyster were investigated. Taken together, the results of this PhD study provide new insight on ways to improve C. gigas sustainable aquaculture industry in Mediterranean coastal lagoons, with particular focus on the Italian territory.

Microplastics debris (< 5 mm) are increasingly abundant in the marine environment, therefore, potentially becoming a growing threat for different marine organisms. Through aquatic animals, these can enter in the human food chain, and can be perceived as a risk for consumers’ health. Different studies report the presence of particles in marketable shellfish including the world wide commercially grown Pacific oyster Magallana gigas (Thunberg, 1793). The aim of this study is to examine the potential risk of microplastics entering in the human food chain through this shellfish species, investigating the dynamics of the uptake, egestion (faeces) and rejection (pseudofaeces) of microplastics in Pacific oysters under controlled conditions. M. gigas collected from a farm in the San Teodoro lagoon (Italy), were exposed to 60 fluorescent orange polystyrene particles L−1 of known sizes (100, 250 and 500 μm). The uptake of each particle size was 19.4 ± 1.1%, 19.4 ± 2% and 12.9 ± 2% respectively. After exposure M. gigas were left to depurate for 72 h, during which 84.6 ± 2% of the particles taken up were released whilst 15.4 ± 2% were retained inside the shell cavity. No microplastic particles were found in the animals’ soft tissues. The results of this study, suggest that depuration is an effective method to reduce presence of large microplastic particles, in the size range 100–500 μm, in M. gigas. Importantly, the data suggests that the burden that could theoretically be up taken by consumers from these shellfish is negligible when compared to other routes.

Various industrial activities lead to environmental pollution by heavy metals. Toxic heavy metals enter the food chain of dairy cows through feed and water, then transferred into milk. This study investigated the correlations of heavy metal contents between individual cows’ milk, water, silage and soil. The relationships between heavy metal contents in individual cows’ milk with milk protein, fat, lactose, solid nonfat (SNF), and total solids (TS) were analysed. Concentrations of Pb, As, Cr, and Cd in milk, silage and water were measured by Inductively Coupled Plasma Mass Spectrometry (ICP-MS). Lead, Cr, and Cd in soil were measured by Atomic Absorption Spectrometry (AAS), and As was detected by Atomic Fluorescence Spectrometry (AFS). One-way non-parametric tests and Spearman correlation analyses were performed using SAS 9.4 software. Levels of Pb and Cd in milk from the unpolluted area were significantly lower (P < 0.01) than those from industrial area. Significantly higher (P < 0.01) As residue was recorded in milk from unpolluted area. Positive correlation of Pb was observed between milk and silage, and As in milk was positively correlated with As in water. Content of As in milk was slightly (r = 0.09) correlated with As in silage, even though strong positive correlation (r = 0.78) was observed between silage and water. Positive correlations were observed for Cr and Cd between milk and silage, as well as milk and soil. Positive correlations were observed in Pb-protein, Cr-protein, and Cd-lactose; other positive correlation coefficients were nearly equal to zero. The results suggest that industrial activities lead to possible Pb and Cd contamination in milk. Drinking water could be the main source of As contamination in cows. No clear relationship was found between milk composition and heavy metals contents in milk. Water and soil on the farm had a partial contribution to heavy metal contamination in milk.

peer reviewed | This works estimated the relationships between heavy metals in milk from individual cows, drinking water, silage and soil as well as the links between those elements and the milk composition. © 2019Various industrial activities lead to environmental pollution by heavy metals. Toxic heavy metals enter the food chain of dairy cows through feed and water, then transferred into milk. This study investigated the correlations of heavy metal contents between individual cows’ milk, water, silage and soil. The relationships between heavy metal contents in individual cows’ milk with milk protein, fat, lactose, solid nonfat (SNF), and total solids (TS) were analysed. Concentrations of Pb, As, Cr, and Cd in milk, silage and water were measured by Inductively Coupled Plasma Mass Spectrometry (ICP-MS). Lead, Cr, and Cd in soil were measured by Atomic Absorption Spectrometry (AAS), and As was detected by Atomic Fluorescence Spectrometry (AFS). One-way non-parametric tests and Spearman correlation analyses were performed using SAS 9.4 software. Levels of Pb and Cd in milk from the unpolluted area were significantly lower (P < 0.01) than those from industrial area. Significantly higher (P < 0.01) As residue was recorded in milk from unpolluted area. Positive correlation of Pb was observed between milk and silage, and As in milk was positively correlated with As in water. Content of As in milk was slightly (r = 0.09) correlated with As in silage, even though strong positive correlation (r = 0.78) was observed between silage and water. Positive correlations were observed for Cr and Cd between milk and silage, as well as milk and soil. Positive correlations were observed in Pb-protein, Cr-protein, and Cd-lactose; other positive correlation coefficients were nearly equal to zero. The results suggest that industrial activities lead to possible Pb and Cd contamination in milk. Drinking water could be the main source of As contamination in cows. No clear relationship was found between milk composition and heavy metals contents in milk. Water and soil on the farm had a partial contribution to heavy metal contamination in milk. © 2019 | Project of Risk Assessment on Raw Milk (GJFP2019008)

The increasing consumption of anticancer drugs through single and/or combinatory chemotherapy worldwide raised concern regarding their toxicity burden in coastal zones. The toxicity of a mixture of three compounds involving the drugs cisplatin (CisPt), cyclophosphamide (CP) and tamoxifen (TAM) was determined on the marine polychaete Nereis diversicolor exposed to an increasing range of their concentrations, respectively: Mix A: 0.1 + 10 + 0.1 ng L−1; Mix B: 10 + 100 + 10 ng L−1; Mix C: 100 + 500 + 25 ng L−1; Mix D: 100 + 1000 + 100 ng L−1. Different endpoints were assessed, including disturbance in the burrowing behaviour, neurotoxicity (acetylcholinesterase – AChE activity), antioxidant enzymes (superoxide dismutase – SOD; catalase – CAT; selenium-dependent glutathione peroxidase – Se-GPx and total glutathione peroxidases T-GPx activities), biotransformation metabolism (glutathione-S-transferases - GST), lipid peroxidation (LPO) and genotoxicity (DNA damage). Biological effects of the mixtures of anticancer compounds on N. diversicolor were compared with previous studies about effects on the same biological model under single-drug exposure conducted with the same molecules. Regarding SOD activity, TAM showed an antagonist effect over CisPt and CP in mixtures C and D. In Mix D, there was a synergistic effect of TAM and CisPt that inhibited CAT activity and an additive interaction of CisPt and CP on the Phase II biotransformation enzyme. Drugs in Mix A also suppressed polychaetes' GST activity, although different from the respective single-drug responses, besides able to induce T-GPx activity, that was not sufficient to avoid oxidative damage and mid-grade DNA damage. Due to the absence of burrowing impairment in Mix A, mechanisms involved in neurotoxicity were other than the one driven by AChE alterations. At the intermediary concentrations (Mix B and C), only LPO occurred. Data from drugs individually may not predict the risks provided by mixtures.

The increasing consumption of anticancer drugs through single and/or combinatory chemotherapy worldwide raised concern regarding their toxicity burden in coastal zones. The toxicity of a mixture of three compounds involving the drugs cisplatin (CisPt), cyclophosphamide (CP) and tamoxifen (TAM) was determined on the marine polychaete Nereis diversicolor exposed to an increasing range of their concentrations, respectively: Mix A: 0.1 + 10 + 0.1 ng L-1; Mix B: 10 + 100 + 10 ng L-1; Mix C: 100 + 500 + 25 ng L-1; Mix D: 100 + 1000 + 100 ng L-1. Different endpoints were assessed, including disturbance in the burrowing behaviour, neurotoxicity (acetylcholinesterase - AChE activity), antioxidant enzymes (superoxide dismutase - SOD; catalase - CAT; selenium-dependent glutathione peroxidase - Se-GPx and total glutathione peroxidases T-GPx activities), biotransformation metabolism (glutathione-S-transferases - GST), lipid peroxidation (LPO) and genotoxicity (DNA damage). Biological effects of the mixtures of anticancer compounds on N. diversicolor were compared with previous studies about effects on the same biological model under single-drug exposure conducted with the same molecules. Regarding SOD activity, TAM showed an antagonist effect over CisPt and CP in mixtures C and D. In Mix D, there was a synergistic effect of TAM and CisPt that inhibited CAT activity and an additive interaction of CisPt and CP on the Phase II biotransformation enzyme. Drugs in Mix A also suppressed polychaetes' GST activity, although different from the respective single-drug responses, besides able to induce T-GPx activity, that was not sufficient to avoid oxidative damage and mid-grade DNA damage. Due to the absence of burrowing impairment in Mix A, mechanisms involved in neurotoxicity were other than the one driven by AChE alterations. At the intermediary concentrations (Mix B and C), only LPO occurred. Data from drugs individually may not predict the risks provided by mixtures. | 202360/2014-8; 308533/2018-6; 311609/2014-7 | info:eu-repo/semantics/publishedVersion

MicroRNAs (miRNAs) are a class of small non-coding RNA that control multiple biological processes through negative post-transcriptional regulation of gene expression. Recently a role of miRNAs in the response of aquatic organisms to environmental toxicants emerged. Toxicant-induced changes in miRNA expression might then represent novel biomarkers to evaluate the health status of these organisms. In this study, we aimed to identify the miRNA repertoire in the liver of the European eel Anguilla anguilla and to compare their differential expression between a polluted site located in the Gironde Estuary and a pristine site in Arcachon Bay (France).A total of 299 mature miRNAs were identified. In polluted water, 19 miRNAs were up-regulated and 22 were down-regulated. We predicted that these differentially expressed miRNAs could target 490 genes that were involved in ribosome biogenesis, response to hormones, response to chemical and chromatin modification. Moreover, we observed only few examples (29) of negative correlation between the expression levels of miRNAs and their targets suggesting that, in the system studied, miRNAs might not only regulate gene expression directly by degrading mRNA but also by inhibiting protein translation or by regulating other epigenetic processes.This study is the first example of in situ investigation of the role of miRNAs in the response of a fish species to water quality. Our findings provide new insights into the involvement of epigenetic mechanisms in the response of animals chronically exposed to pollution and pave the way for the utilization of miRNAs in aquatic ecotoxicology.

MicroRNAs (miRNAs) are a class of small non-coding RNA that control multiple biological processes through negative post-transcriptional regulation of gene expression. Recently a role of miRNAs in the response of aquatic organisms to environmental toxicants emerged. Toxicant-induced changes in miRNA expression might then represent novel biomarkers to evaluate the health status of these organisms. In this study, we aimed to identify the miRNA repertoire in the liver of the European eel Anguilla anguilla and to compare their differential expression between a polluted site located in the Gironde Estuary and a pristine site in Arcachon Bay (France). A total of 299 mature miRNAs were identified. In polluted water, 19 miRNAs were up-regulated and 22 were down-regulated. We predicted that these differentially expressed miRNAs could target 490 genes that were involved in ribosome biogenesis, response to hormones, response to chemical and chromatin modification. Moreover, we observed only few examples (29) of negative correlation between the expression levels of miRNAs and their targets suggesting that, in the system studied, miRNAs might not only regulate gene expression directly by degrading mRNA but also by inhibiting protein translation or by regulating other epigenetic processes. This study is the first example of in situ investigation of the role of miRNAs in the response of a fish species to water quality. Our findings provide new insights into the involvement of epigenetic mechanisms in the response of animals chronically exposed to pollution and pave the way for the utilization of miRNAs in aquatic ecotoxicology.