Chemical-physical parameters and microbial community changes induced by electrodes polarization inhibit PCB dechlorination in a marine sediment.

Microbial reductive dechlorination of organohalogenated pollutants is often limited by the scarcity of electron donors, that can be overcome with microbial electrochemical technologies (METs). In this study, polarized electrodes buried in marine sediment microcosms were investigated to stimulate PCB reductive dechlorination under potentiostatic (-0.7 V vs Ag/AgCl) and galvanostatic conditions (0.025 mA·cm-2-0.05 mA·cm-2), using graphite rod as cathode and iron plate as sacrificial anode. A single circuit and a novel two antiparallel circuits configuration (2AP) were investigated. Single circuit polarization impacted the sediment pH and redox potential (ORP) proportionally to the intensity of the electrical input and inhibited PCB reductive dechlorination. The effects on the sediment's pH and ORP, along with the inhibition of PCB reductive dechlorination, were mitigated in the 2AP system. Electrodes polarization stimulated sulfate-reduction and promoted the enrichment of bacterial clades potentially involved in sulfate-reduction as well as in sulfur oxidation. This suggested the electrons provided were consumed by competitors of organohalide respiring bacteria and specifically sequestered by sulfur cycling, which may represent the main factor limiting the applicability of METs for stimulating PCB reductive dechlorination in marine sediments.

Site-specific response of sediment microbial community to supplementation of polyhydroxyalkanoates as biostimulants for PCB reductive dechlorination.

The use of biodegradable plastics is constantly raising, increasing the likeliness for these polymers to end up in the environment. Environmental applications foreseeing the intentional release of biodegradable plastics have been also recently proposed, e.g., for polyhydroxyalkanoates (PHAs) acting as slow hydrogen releasing compounds to stimulate microbial reductive dehalogenation processes. However, the effects of their release into the environment on the ecosystems still need to be thoroughly explored. In this work, the use of PHAs to enhance the microbial reductive dechlorination of polychlorobiphenyls (PCBs) and their impact on the metabolic and compositional features of the resident microbial community have been investigated in laboratory microcosms of a polluted marine sediment from Mar Piccolo (Taranto, Italy), and compared with recent findings on a different contaminated marine sediment from Pialassa della Baiona (Ravenna, Italy). A decreased biostimulation efficiency of PHAs on PCBs reductive dechlorination was observed in the sediment from Mar Piccolo, with respect to the sediment from Pialassa della Baiona, suggesting that the sediments' physical-chemical characteristics and/or the biodiversity and composition of its microbial community might play a key role in determining the outcome of this biostimulation strategy. Regardless of the sediment origin, PHAs were found to have a specific and pervasive effect on the sediment microbial community, reducing its biodiversity, defining a newly arranged microbial core of primary degraders and consequently affecting, in a site-specific way, the abundance of subdominant bacteria, possibly cross-feeders. Such potential to dramatically change the structure of autochthonous microbial communities should be carefully considered, since it might have secondary effects, e.g., on the natural biogeochemical cycles.

Triggering of Polymer-Degrading Enzymes from Layered Double Hydroxides for Recycling Strategies.

The use of degrading enzymes in polymer formulation is a very attractive strategy to manage the end-of-life of plastics. However, high temperatures cause the denaturation of enzymes and the loss of their catalytic activity; therefore, protection strategies are necessary. Once protected, the enzyme needs to be released in appropriate media to exert its catalytic activity. A successful protection strategy involves the use of layered double hydroxides: cutinase, selected as a highly degrading polyester hydrolytic enzyme, is thermally protected by immobilization in Mg/Al layered double hydroxide structures. Different triggering media are here evaluated in order to find the best releasing conditions of cutinase from LDH. In detail, phosphate and citrate-phosphate buffers, potassium carbonate, sodium chloride, and sodium sulfate solutions are studied. After the comparison of all media in terms of protein release and activity retained, phosphate buffer is selected as the best candidate for the release of cutinase from LDH, and the effect of pH and concentration is also evaluated. The amount of the enzyme released is determined with the Lowry method. Activity tests are performed via spectrophotometry.

Effect of polyhydroxyalkanoates on the microbial reductive dechlorination of polychlorinated biphenyls and competing anaerobic respirations in a marine microbial culture.

The effect of polyhydroxyalkanoates (PHAs) with different composition on the reductive dechlorination activity of a polychlorinated biphenyls (PCBs) dechlorinating marine microbial community and on the activity of sulfate-reducing (SRB) and methanogenic bacteria (MB), were investigated in marine sediment microcosms and compared with the main monomer, 3-hydroxybutyric acid (3HB). Despite PHAs were fermented more slowly than 3HB, all electron donors stimulated constantly sulfate-reduction, methanogenesis and, only transiently, PCB reductive dechlorination. No relevant differences were observed with different compositions of PHAs. According to electron balances, the majority of the supplied electrons (50 %) were consumed by SRB and to less extent by MB (9-31 %), while a small percentage (0.01 %) was delivered to OHRB. In the studied conditions PHAs were confirmed as potential slow­hydrogen releasing compounds in marine environment but their fermentation rate was sufficiently high to mainly stimulate the competitors of organohalide respring bacteria for electron donors.

Bacterial colonization dynamics of different microplastic types in an anoxic salt marsh sediment and impact of adsorbed polychlorinated biphenyls on the plastisphere.

Plastic debris dispersed into the environment provide a substrate for microbial colonization, constituting a new human-made ecosystem called "plastisphere", and altering the microbial species distribution in aquatic, coastal and benthic ecosystems. The study aims at exploring the interaction among microplastics (MPs) made of different polymers, a persistent organic contaminant (polychlorinated biphenyls, PCBs), and the environmental microbial communities, in an anoxic marine sediment. Plastic pellets were incubated in the field in a salt marsh anoxic sediment, to observe the stages of plastisphere formation, by quantitative PCR and 16S rRNA gene sequencing, and PCB dechlorination activity on the MPs surface. Microbes from the sediment rapidly colonized the different microplastics types, with PVC recruiting a peculiar community enriched in sulfate-reducing bacteria. The composition of the plastisphere varied along the 1-year incubation possibly in response either to warmer temperatures in spring-summer or to microhabitat's changes due to the progressive plastic surface weathering. Even if PCB contaminated MPs were able to recruit potentially dehalogenating taxa, actual dechlorination was not detectable after 1 year. This suggests that the concentration of potentially dehalorespiring bacteria in the natural environment could be too low for the onset of the dechlorination process on MP-sorbed contaminants. Our study, which is among very few available longitudinally exploring the plastisphere composition in an anoxic sediment context, is the first exploring the fate and possible biodegradation of persistent organic pollutants sorbed on MPs reaching the seafloor.

Enzymatic Degradation of the Most Common Aliphatic Bio-Polyesters and Evaluation of the Mechanisms Involved: An Extended Study.

Commercial hydrolytic enzymes belonging to different subclasses (several lipases, proteinase k, cutinase) were investigated for their ability to degrade different aliphatic polyesters, i.e., poly(butylene succinate) (PBS), poly(butylene succinate-co-adipate) (PBSA), two poly(caprolactone), having two different molecular weights, poly(lactic acid) (PLA) and poly(propylene carbonate) (PPC). The enzyme screening was first carried out by investigating the capacity of fully degrading the target polymers in 24 h, then weight loss measurements of selected polyesters and target enzymes were performed. Solid residues after enzyme degradation were characterized by proton nuclear magnetic resonance (1H NMR), gel permeation chromatography (GPC), infrared spectroscopy (FT-IR), differential scanning calorimetry (DSC) and thermogravimetry (TGA). Liquid fractions were studied via GPC, 1H NMR and high-performance liquid chromatography (HPLC). PCL and PBSA were found to be the most biodegradable polyesters, under the conditions used in this study. PBS was fully degraded only by cutinase, whereas none of the tested enzymes were able to completely degrade PLA and PPC, in the conditions assessed here. Cutinase exhibited the highest hydrolytic activity on PBSA, while lipase from Candida sp. (CALB) on low molecular weight PCL. Chemical analyses on residual solids showed that the enzymatic degradation occurred homogeneously from the surface through an erosion mechanism and did not significantly affect the macromolecular structure and thermal stability. Cleaving action mode for each enzyme (endo- and/or exo-type) on the different polyesters were also proposed based on the evaluation of the degradation products in the liquid fraction.

Hexabromocyclododecanes Are Dehalogenated by CYP168A1 from Pseudomonas aeruginosa Strain HS9.

Hexabromocyclododecanes (HBCDs) are widely used brominated flame retardants that cause antidiuretic hormone syndrome and even induce cancer. However, little information is available about the degradation mechanisms of HBCDs. In this study, genomic and proteomic analyses, reverse transcription-quantitative PCR, and gene knockout assays reveal that a cytochrome P450-encoding gene is responsible for HBCD catabolism in Pseudomonas aeruginosa HS9. The CO difference spectrum of the enzyme CYP168A1 was matched to P450 characteristics via UV visibility. We demonstrate that the reactions of debromination and hydrogenation are carried out one after another based on detection of the metabolites pentabromocyclododecanols (PBCDOHs), tetrabromocyclododecadiols (TBCDDOHs), and bromide ion. In the 18O isotope experiments, PBCD18OHs were only detected in the H218O group, proving that the added oxygen is derived from H2O, not from O2. This study elucidates the degradation mechanism of HBCDs by Pseudomonas. IMPORTANCE Hexabromocyclododecanes (HBCDs) are environmental pollutants that are widely used in industry. In this study, we identified and characterized a novel key dehalogenase, CYP168A1, that is responsible for HBCD degradation from Pseudomonas aeruginosa strain HS9. This study provides new insights into understanding biodegradation of HBCDs.

Unraveling the Metabolic Potential of Asgardarchaeota in a Sediment from the Mediterranean Hydrocarbon-Contaminated Water Basin Mar Piccolo (Taranto, Italy).

Increasing number of metagenome sequencing studies have proposed a central metabolic role of still understudied Archaeal members in natural and artificial ecosystems. However, their role in hydrocarbon cycling, particularly in the anaerobic biodegradation of aliphatic and aromatic hydrocarbons, is still mostly unknown in both marine and terrestrial environments. In this work, we focused our study on the metagenomic characterization of the archaeal community inhabiting the Mar Piccolo (Taranto, Italy, central Mediterranean) sediments heavily contaminated by petroleum hydrocarbons and polychlorinated biphenyls (PCB). Among metagenomic bins reconstructed from Mar Piccolo microbial community, we have identified members of the Asgardarchaeota superphylum that has been recently proposed to play a central role in hydrocarbon cycling in natural ecosystems under anoxic conditions. In particular, we found members affiliated with Thorarchaeota, Heimdallarchaeota, and Lokiarchaeota phyla and analyzed their genomic potential involved in central metabolism and hydrocarbon biodegradation. Metabolic prediction based on metagenomic analysis identified the malonyl-CoA and benzoyl-CoA routes as the pathways involved in aliphatic and aromatic biodegradation in these Asgardarchaeota members. This is the first study to give insight into the archaeal community functionality and connection to hydrocarbon degradation in marine sediment historically contaminated by hydrocarbons.

A Pseudomonas sp. strain uniquely degrades PAHs and heterocyclic derivatives via lateral dioxygenation pathways.

Polycyclic aromatic hydrocarbons (PAHs) and heterocyclic derivatives are organic pollutants that pose a serious health risk to human beings. In this study, a newly isolated Pseudomonas brassicacearum strain MPDS could effectively degrade PAHs and heterocyclic derivatives, including naphthalene, fluorene, dibenzofuran (DBF) and dibenzothiophene (DBT). Notably, strain MPDS is able to degrade fluorene, DBF and DBT uniquely via a lateral dioxygenation pathway, while most reported strains degrade fluorene, DBF and DBT via an angular dioxygenation pathway or co-metabolize them via a lateral dioxygenation pathway. Strain MPDS completely degraded 50 mg naphthalene (in 50 mL medium) in 84 h, and OD600 reached 1.0-1.1; while, it stabilized at OD600 0.5-0.6 with 5 mg fluorene or DBF or DBT. Meanwhile, 65.7% DBF and 32.1% DBT were degraded in 96 h, and 40.3% fluorene was degraded in 72 h, respectively. Through genomic and transcriptomic analyses, and comparative genomic analysis with another DBF degradation strain, relevant gene clusters were predicted, and a naphthalene-degrading gene cluster was identified. This study provides understanding of degradation of PAHs and their heterocyclic derivatives, as well as new insights into the lateral dioxygenation pathway of relevant contaminants.

Comparative Preliminary Evaluation of 2 In-stream Water Treatment Technologies for the Agricultural Reuse of Drainage Water in the Nile Delta.

In the Nile Delta, a complex network of canals collects drainage water from surface-irrigated fields but also from municipal wastewater. The goal of this work was to assess the technical, environmental, and financial feasibility of the upgrade of a drainage canal (DC) into either an in-stream constructed wetland (ICW) or a canalized facultative lagoon (CFL), in order to produce a water reusable in agriculture according to Egyptian law. The model-based design of the proposed technologies was derived from field experimental data for the ICW and laboratory data for the CFL. Both technologies, integrated by a sedimentation pond and a disinfection canal, led to the attainment of the water quality standards imposed by Egyptian Law 92/2013 for the reuse of drainage water. The life cycle assessment indicated that the upgrade of an existing DC to either an ICW or a CFL results in an extremely small environmental burden, ≤0.3% of that of a traditional activated sludge process. The cost-benefit analysis (CBA) was based on the assumptions that 1) farmers currently irrigate a nonfood crop (cotton) with the low-quality drainage water present in the DC, and 2) thanks to the upgrade to a ICW or CFL, farmers will irrigate a food crop characterized by a higher market price (rice). The CBA indicated that the DC upgrade to an ICW represents an attractive investment because it leads to a financial rate of return >10% over a wide range of cotton market prices. Conversely, the upgrade to a CFL is less attractive due to high investment costs. In conclusion, the upgrade of DCs to ICWs appears a promising option for the treatment of drainage canal water in the Nile Delta, thanks to the high pollutant removal performances, low cost, and negligible environmental burden. Integr Environ Assess Manag 2020;16:920-933. © 2020 SETAC.

Microbial colonization of different microplastic types and biotransformation of sorbed PCBs by a marine anaerobic bacterial community.

We investigated the colonization dynamics of different microplastic (MP) pellets, namely, polyethylene (PE), polyethylene terephthalate (PET), polystyrene (PS), polypropylene (PP) and polyvinyl chloride (PVC), either pristine or contaminated with polychlorinated biphenyls (PCBs), by an organohalide respiring marine microbial community and its biotransformation activity towards PCBs sorbed on MPs, in anaerobic laboratory microcosms of a marine sediment. All MPs were rapidly colonized by the microbial community within 2 weeks of incubation, when approximately 1010 16S rRNA gene copies cm-2 were detected on PVC, 109 copies cm-2 on PE, and 108 copies cm-2 on PET, PP and PS. A greater biofilm growth on PVC pellets than other MPs was confirmed by quantification of the reducing sugars of the EPS and biofilm staining with crystal violet. Illumina sequencing of the 16S rRNA genes and Principal Coordinate Analysis (PCoA) revealed that the biofilm community on MPs significantly differed from the sediment community, being enriched of chemoorganotrophic fermenting species, and was significantly affected by the type of polymer. The presence of sorbed PCBs did not significantly affect the overall community composition, and mainly resulted in the enrichment of Dehalococcoidia, i.e., of the organohalide respiring members of the community. Reductive dechlorination of PCBs sorbed to MPs was observed after 2 weeks of incubation, when the average number of chlorines per biphenyl molecule was reduced from 5.2 to 4.8-4.3, and was faster (35.2 ± 1.9 to 61.2 ± 5.8 µmol of Cl removed kgMP-1 week-1) than that of sediment-sorbed ones (33.9 ± 9.1 µmol of Cl removed kgsediment-1 week-1), which started only after 10 weeks of incubation. These data suggest that microbial colonization of contaminated MPs might change the composition of sorbed PCB mixtures and therefore the toxicity associated to PCB-polluted MPs.

Containment of a genetically modified microorganism by an activated sludge system.

The effectiveness of physical, chemical and biological barriers to the diffusion of genetically modified microorganisms (GMMs) to prevent their release into the environment is currently under scrutiny worldwide because of the associated potential ecological impacts. An industrial discharge of a non-sterilized fermentation broth containing GMM biomass into a conventional municipal wastewater treatment plant would deliver the GMMs into the activated sludge system process (ASSP). The present work aimed to model and evaluate the containment capability of a small ASSP (part of a 20,000 people equivalent municipal plant) in the event of receiving GMM biomass from a medium-small biotechnological plant dedicated to the production of polyhydroxyalkanoates (3000 t/year of biopolymer). An actual GMM (Pseudomonas putida KTOY06) was injected into a bench-scale ASSP (ASSPLab) in a quantity proportional to the relative dimensions of the plants mentioned. The experimental and model results indicated that the ASSP of the target municipal treatment plant would not be capable of holding back such a sudden input of GMM; 6 h after the discharge, 11-15 % of injected GMM cells were released through the clarified stream of the ASSPLab, with the rest being gradually released over time. Since the GMM employed did not exhibit any growth in the ASSPLab, its concentration in the clarified water stream would not represent a substantial risk of release into the environment if appropriate tertiary treatments were integrated. This study confirmed the necessity of a thorough risk assessment of biotechnological processes prior to their implementation.

Phenol biodegradation by Acinetobacter radioresistens APH1 and its application in soil bioremediation.

Phenol accounts for a large proportion of the contamination in industrial wastewater discharged from chemical plants due to its wide use as a raw chemical. Residual phenol waste in water and soil significantly endangers human health and the natural environment. In this study, an Acinetobacter radioresistens strain, APH1, was isolated and identified for its efficient capability of utilizing phenol as sole carbon source for growth. A draft genome sequence containing 3,290,330 bases with 45 contigs was obtained, and 22 genes were found to be involved in phenol metabolism and 51 putative drug-resistance genes were annotated by genomic analysis. The optimal conditions for cell culture and phenol removal were determined to be 30 °C, pH 6.0, and a phenol concentration of 500 mg/L; the upper limit of phenol tolerance was 950 mg/L. Based on GC-MS analysis, the key metabolites including cis,cis-muconic acid, catechol, and succinic acid were detected. During bioremediation experiment using 450 mg/kg (dry weight) of phenol-contaminated soil, the strain APH1 removed 99% of the phenol within 3 days. According to microbial diversity analysis, the microbial abundance of Chungangia, Bacillus, Nitrospira, Lysinibacillus, and Planomicrobium increased after the addition of phenol. Furthermore, at day 23, the abundance of strain APH1 was greatly reduced, and the microbial diversity and structure of the whole microbial community were gradually recovered, indicating that strain APH1 would not affect this microbial ecosystem. These findings provide insights into the bioremediation of soil contaminated with phenol.

Advances in combined enzymatic extraction of ferulic acid from wheat bran.

Wheat bran could be utilised as feedstock for innovative and sustainable biorefinery processes. Here, an enzymatic hydrolysis process for ferulic acid (FA) extraction was optimised step by step for total wheat bran (Tritello) and then also applied to the outer bran layer (Bran 1). Proteins, reducing sugars, total phenols and FA were quantified. The highest FA yields (0.82-1.05 g/kg bran) were obtained either by rehydrating the bran by autoclaving (Tritello) or by steam explosion (Bran 1) using a bran/water ratio of 1:20, followed by enzymatic pre-treatment with Alcalase and Termamyl, to remove protein and sugars, and a final enzymatic hydrolysis with Pentopan and feruloyl esterase to solubilise phenol. FA was recovered from the final digestate via solid phase extraction. A 40-fold scale-up was also performed and the release of compounds along all the process steps and at increasing incubation times was monitored. Results showed that FA was initially present at a minimum level while it was specifically released during the enzymatic treatment. In the final optimized process, the FA extraction yield was higher than that obtained with NaOH control hydrolysis while, in comparison with other FA enzymatic extraction methods, fewer process steps were required and no buffers, strong acid/alkali nor toxic compounds were used. Furthermore, the proposed process may be easily scaled-up, confirming the feasibility of wheat bran valorisation by biorefinery processes to obtain valuable compounds having several areas of potential industrial exploitation.

Enhancing Bioremediation Potential of Pseudomonas putida by Developing Its Acid Stress Tolerance With Glutamate Decarboxylase Dependent System and Global Regulator of Extreme Radiation Resistance.

The extensive use of acids in a variety of manufacturing industries results in the increase of discharged acidic waste stream into the environment. Such co-pollution of acids and other organic pollutants limits the biodegradation capability of neutrophilic degraders. With high-throughput genetic techniques, we aim to improve the acid tolerance of a pollutant-degrading bacterium, Pseudomonas putida S16 by genetically engineering it with the glutamate decarboxylase (GAD)-dependent system and the global regulator (IrrE) of extreme radiation resistance. The engineered strains holding either GAD system or irrE regulator could grow under pH 4.5, compared to the wild type. They could also degrade over 90% of a selected pollutant (benzoate or nicotine) under pH 5.0 in 48 h, while no biodegradation was detected with the wild type under the same conditions. We conclude that acid stress tolerance by the possession of the GAD system or IrrE regulator in pollutant-degrading bacteria would be a promising approach to enhance their viability and biodegrading activities in bioremediation of acidic wastes.

Decoding the ocean's microbiological secrets for marine enzyme biodiscovery.

A global census of marine microbial life has been underway over the past several decades. During this period, there have been scientific breakthroughs in estimating microbial diversity and understanding microbial functioning and ecology. It is estimated that the ocean, covering 71% of the earth's surface with its estimated volume of about 2 × 1018 m3 and an average depth of 3800 m, hosts the largest population of microbes on Earth. More than 2 million eukaryotic and prokaryotic species are thought to thrive both in the ocean and on its surface. Prokaryotic cell abundances can reach densities of up to 1012 cells per millilitre, exceeding eukaryotic densities of around 106 cells per millilitre of seawater. Besides their large numbers and abundance, marine microbial assemblages and their organic catalysts (enzymes) have a largely underestimated value for their use in the development of industrial products and processes. In this perspective article, we identified critical gaps in knowledge and technology to fast-track this development. We provided a general overview of the presumptive microbial assemblages in oceans, and an estimation of what is known and the enzymes that have been currently retrieved. We also discussed recent advances made in this area by the collaborative European Horizon 2020 project 'INMARE'.

Effect of oxygen mass transfer rate on the production of 2,3-butanediol from glucose and agro-industrial byproducts by Bacillus licheniformis ATCC9789.

BACKGROUND: 2,3-Butanediol (BD) is a largely used fossil-based platform chemical. The yield and productivity of bio-based BD fermentative production must be increased and cheaper substrates need to be identified, to make bio-based BD production more competitive. As BD bioproduction occurs under microaerobic conditions, a fine tuning and control of the oxygen transfer rate (OTR) is crucial to maximize BD yield and productivity. Very few studies on BD bioproduction focused on the use of non-pathogenic microorganisms and of byproducts as substrate. The goal of this work was to optimize BD bioproduction by the non-pathogenic strain Bacillus licheniformis ATCC9789 by (i) identifying the ranges of volumetric and biomass-specific OTR that maximize BD yield and productivity using standard sugar and protein sources, and (ii) performing a preliminary evaluation of the variation in process performances and cost resulting from the replacement of glucose with molasses, and beef extract/peptone with chicken meat and bone meal, a byproduct of the meat production industry. RESULTS: OTR optimization with an expensive, standard medium containing glucose, beef extract and peptone revealed that OTRs in the 7-15 mmol/L/h range lead to an optimal BD yield (0.43 ± 0.03 g/g) and productivity (0.91 ± 0.05 g/L/h). The corresponding optimal range of biomass-specific OTR was equal to 1.4-7.9 [Formula: see text], whereas the respiratory quotient ranged from 1.8 to 2.5. The switch to an agro-industrial byproduct-based medium containing chicken meat and bone meal and molasses led to a 50% decrease in both BD yield and productivity. A preliminary economic analysis indicated that the use of the byproduct-based medium can reduce by about 45% the BD production cost. CONCLUSIONS: A procedure for OTR optimization was developed and implemented, leading to the identification of a range of biomass-specific OTR and respiratory quotient to be used for the scale-up and control of BD bioproduction by Bacillus licheniformis. The switch to a byproduct-based medium led to a relevant decrease in BD production cost. Further research is needed to optimize the process of BD bioproduction from the tested byproduct-based medium.

Integrated technological and management solutions for wastewater treatment and efficient agricultural reuse in Egypt, Morocco, and Tunisia.

Mediterranean-African countries (MACs) face a major water crisis. The annual renewable water resources are close to the 500 m3 /capita threshold of absolute water scarcity, and water withdrawals exceed total renewable water resources by 30%. Such a low water availability curbs economic development in agriculture, which accounts for 86% of freshwater consumption. The analysis of the current situation of wastewater treatment, irrigation, and water management in MACs and of the research projects targeted to these countries indicates the need for 1) an enhanced capacity to analyze water stress, 2) the development of water management strategies capable of including wastewater reuse, and 3) development of locally adapted water treatment and irrigation technologies. This analysis shaped the MADFORWATER project (www.madforwater.eu), whose goal is to develop a set of integrated technological and management solutions to enhance wastewater treatment, wastewater reuse for irrigation, and water efficiency in agriculture in Egypt, Morocco, and Tunisia. MADFORWATER develops and adapts technologies for the production of irrigation-quality water from drainage canals and municipal, agro-industrial, and industrial wastewaters and technologies for water efficiency and reuse in agriculture, initially validated at laboratory scale, to 3 hydrological basins in the selected MACs. Selected technologies will be further adapted and validated in 4 demonstration plants of integrated wastewater treatment and reuse. Integrated strategies for wastewater treatment and reuse targeted to the selected basins are developed, and guidelines for the development of integrated water management strategies in other basins of the 3 target MACs will be produced. The social and technical suitability of the developed technologies and nontechnological tools in relation to the local context is evaluated with the participation of MAC stakeholders and partners. Guidelines on economic instruments and policies for the effective implementation of the proposed water management solutions in the target MACs will be developed. Integr Environ Assess Manag 2018;14:447-462. © 2018 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals, Inc. on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Optimization of washing conditions with biogenic mobilizing agents for marine fuel-contaminated beach sands.

Washing is a rapid and effective treatment to remediate contaminated sands impacted by oil spills, although synthetic additives used to increase extraction efficiency may cause additional pollution issues due to their intrinsic toxicity and very often low biodegradability. In this study, different biogenic mobilizing agents (soybean lecithins, cyclodextrins, cholic acids, plant-derived cleaners, rhamnolipids and sophorolipids) were tested in the washing of beach sands artificially contaminated with the Intermediate Fuel Oil IFO-180. Among these, a de-oiled soybean lecithin (SL-1), hydroxypropyl-ß-cyclodextrins (HPB-CD) and sophorolipids (SR) achieved hydrocarbon removals close to those attained with the synthetic surfactant Triton™ X-100 (TX) in preliminary washing tests carried out at constant mixing rate, water/sand ratio and IFO-180 contamination level using agents concentrations close to their critical micelle concentration (0.1% and 1% w/v for microbial and non-microbial agents, respectively). The effects of agent concentration, water/sand ratio, mixing rate and IFO-180 contamination on hydrocarbons removal were modelled using face-centred central composite design and ANOVA. Optimal washing parameters for sand contamination levels in the range 0.5-20 g/kg were identified with response surface methodology. While HPB-CD and SR performed equally to TX only at low sand contaminations, SL-1 attained hydrocarbon removal higher or equal to that of TX at any IFO-180 contamination and at lower application rates. SL-1 also outperformed TX when minimizing the water/sand ratio, i.e., the volume of water used. Considering its lower toxicity, higher biodegradability and higher hydrocarbon removal efficiencies, SL-1 is an effective and environmentally sustainable alternative to synthetic surfactants in washing treatments for marine fuel-contaminated sands.

Identification of two organohalide-respiring Dehalococcoidia associated to different dechlorination activities in PCB-impacted marine sediments.

BACKGROUND: Microbial reductive dechlorination of polychlorinated biphenyls (PCBs) plays a major role in detoxifying anoxic contaminated freshwater and marine sediments from PCBs. Known members of the phylum Chloroflexi are typically responsible for this activity in freshwater sediments, whereas less is known about the microorganisms responsible for this activity in marine sediments. PCB-respiring activities were detected in PCB-impacted marine sediments of the Venice Lagoon. The aim of this work was to identify the indigenous organohalide-respiring microorganisms in such environments and assess their dechlorination specificity against spiked Aroclor™ 1254 PCBs under laboratory conditions resembling the in situ biogeochemistry. RESULTS: High PCB dechlorination activities (from 150 ± 7 to 380 ± 44 µmol of chlorine removed kg-1 week-1) were detected in three out of six sediments sampled from different locations of the lagoon. An uncultured non-Dehalococcoides phylotype of the class Dehalococcoidia closely related to Dehalobium chlorocoercia DF-1, namely phylotype VLD-1, was detected and enriched up to 109 16S rRNA gene copies per gram of sediment where dechlorination activities were higher and 25-4/24-4 and 25-2/24-2/4-4 chlorobiphenyls (CB) accumulated as the main tri-/dichlorinated products. Conversely, a different phylotype closely related to the SF1/m-1 clade, namely VLD-2, also enriched highly where lower dechlorination activity and the accumulation of 25-3 CB as main tri-chlorinated product occurred, albeit in the simultaneous presence of VLD-1. Both phylotypes showed growth yields higher or comparable to known organohalide respirers and neither phylotypes enriched in sediment cultures not exhibiting dechlorination. CONCLUSIONS: These findings confirm the presence of different PCB-respiring microorganisms in the indigenous microbial communities of Venice Lagoon sediments and relate two non-Dehalococcoides phylotypes of the class Dehalococcoidia to different PCB dechlorination rates and specificities.