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Deep-sea mining may lead to the release of high concentrations of metals into the surrounding seabed, which can disturb important ecosystem functions provided by microbial communities. Among these, the production of N2O and its reduction to N2 is of great relevance since N2O is an important greenhouse gas. Metal impacts on net N2O production by deep-sea bacteria are, however, currently unexplored. Here, we evaluated the effects of cadmium (Cd) on net N2O production by a deep-sea isolate, Shewanella loihica PV-4. We performed a series of Cd exposure incubations in oxic conditions and determined N2O fluxes during induced anoxic conditions, as well as the relative expression of the nitrite reductase gene (nirK), preceding N2O production, and N2O reductase gene (nosZ), responsible for N2O reduction. Net N2O production by S. loihica PV-4 exposed to Cd was strongly inhibited when compared to the control treatment (no metal). Both nirK and nosZ gene expression were inhibited in reactors with Cd, but nirK inhibition was stronger, supporting the lower net N2O production observed with Cd. The Cd inhibition of net N2O production observed in this study poses the question whether other deep-sea bacteria would undergo the same effects. Future studies should address this question as well as its applicability to complex communities and other physicochemical conditions, which remain to be evaluated. [ABSTRACT FROM AUTHOR]

Following a fuel leakage inside a Portuguese maritime port, we conducted parallel 30-day experiments using contaminated seawater and fuel, sampled five days after the incident. This study aimed to (i)survey the native microbial community response to the spilled fuel and (ii)evaluate the efficacy of bioremediation, both biostimulation and bioaugmentation with a lyophilized bacterial consortium (Rhodococcus erythropolis , Pseudomonas sp.), in accelerating hydrocarbon degradation. Metabarcoding analysis revealed a shift in microbial communities, with increased abundance of hydrocarbon-degraders (e.g. Alcanivorax , Thalassospira). Ninety-five hydrocarbonoclastic bacteria were isolated, including key groups from the enriched communities. The lyophilized bacteria added in bioaugmentation, enhanced the abundance of hydrocarbon-degraders over time and were recovered throughout time. Bioremediation treatments favoured biodegradation, achieving over 60 % removal of total petroleum hydrocarbons after 15 days, contrasting with natural attenuation where almost no TPH was removed. This work highlights the potential of bioremediation technologies to accelerate hydrocarbon-degrading activity, for oil spills inside ports. [Display omitted] • Spilled fuel and seawater from a port were used for enrichment experiments. • Enriched communities were dominated by Proteobacteria and Bacteroidota members. • Lyophilized bacterial consortium enhanced the abundance of HC-degraders early on. • Bioremediation treatments favoured the biodegradation process (>60 % TPH removal). [ABSTRACT FROM AUTHOR]

This study aimed to evaluate the potential of a salt marsh plant and its rhizosphere microorganisms for the removal of two pharmaceutical compounds, bezafibrate and paroxetine, from estuarine environment. Plants were exposed for 7 days to a simplified estuarine medium, elutriate solution with or without sediment, doped with bezafibrate or paroxetine. Tests were done in absence and presence of nutrients or copper. Phragmites australis (Cav.) Trin. Ex Steud, alone or with the sediment microbial communities, contributed for pharmaceuticals removal. In the presence of P. australis, for paroxetine a 65% removal was observed. Removal increased up to 90% when sediment was present. For bezafibrate, removals reached ca. 47% in P. australis presence, increasing to ca. 70% when nutrients were added to the medium, indicating a good nutritional state can contribute for a higher compound removal. When Cu was added, 75% removal for bezafibrate and 95% removal for paroxetine were observed indicating the metal might influence the removal of the pharmaceuticals. Overall, the plant and its rhizosediments and associated microorganisms showed potential for pharmaceuticals removal from estuaries, eventually degrading the selected compounds, a feature requiring more research. Results indicate that phytoremediation could be a viable option for eliminating/diminishing the environmental impact of pharmaceutical compounds in estuarine areas. [ABSTRACT FROM AUTHOR]

Oil spills are among the most catastrophic events to marine ecosystems and current remediation techniques are not suitable for ecological restoration. Bioremediation approaches can take advantage of the activity of microorganisms with biodegradation capacity thus helping to accelerate the recovery of contaminated environments. The use of native microorganisms can increase the bioremediation efficiency since they have higher potential to survive in the natural environment while preventing unpredictable ecological impacts associated with the introduction of non-native organisms. In order to know the geographical scale to which a native bioremediation consortium can be applied, we need to understand the spatial heterogeneity of the natural microbial communities with potential for hydrocarbon degradation. In the present study, we aim to describe the genetic diversity and the potential of native microbial communities to degrade petroleum hydrocarbons, at an early stage of bioremediation, along the NW Iberian Peninsula coast, an area particularly susceptible to oil spills. Seawater samples collected in 47 sites were exposed to crude oil for 2 weeks, in enrichment experiments. Seawater samples collected in situ , and samples collected after the enrichment with crude oil, were characterized for prokaryotic communities by using 16S rRNA gene amplicon sequencing and predictive functional profiling. Results showed a drastic decrease in richness and diversity of microbial communities after the enrichment with crude oil. Enriched microbial communities were mainly dominated by genera known to degrade hydrocarbons, namely Alcanivorax , Pseudomonas , Acinetobacter , Rhodococcus , Flavobacterium , Oleibacter , Marinobacter , and Thalassospira , without significant differences between geographic areas and locations. Predictive functional profiling of the enriched microbial consortia showed a high potential to degrade the aromatic compounds aminobenzoate, benzoate, chlorocyclohexane, chlorobenzene, ethylbenzene, naphthalene, polycyclic aromatic compounds, styrene, toluene, and xylene. Only a few genera contributed for more than 50% of this genetic potential for aromatic compounds degradation in the enriched communities, namely Alcanivorax , Thalassospira , and Pseudomonas spp. This work is a starting point for the future development of prototype consortia of hydrocarbon-degrading bacteria to mitigate oil spills in the Iberian NW coast. [ABSTRACT FROM AUTHOR]

The importance of planktonic microbial communities is well acknowledged, since they are fundamental for several natural processes of aquatic ecosystems. Microorganisms naturally control the flux of nutrients, and also degrade and recycle anthropogenic organic and inorganic contaminants. Nevertheless, climate change effects and/or the runoff of nutrients/pollutants can affect the equilibrium of natural microbial communities influencing the occurrence of microbial pathogens and/or microbial toxin producers, which can compromise ecosystem environmental status. Therefore, improved microbial plankton monitoring is essential to better understand how these communities respond to environmental shifts. The study of marine microbial communities typically involves highly cost and time-consuming sampling procedures, which can limit the frequency of sampling and data availability. In this context, we developed and validated an in situ autonomous biosampler (IS-ABS) able to collect/concentrate in situ planktonic communities of different size fractions (targeting prokaryotes and unicellular eukaryotes) for posterior genomic, metagenomic, and/or transcriptomic analysis at a home laboratory. The IS-ABS field prototype is a small size and compact system able to operate up to 150 m depth. Water is pumped by a micropump (TCS MG2000) through a hydraulic circuit that allows in situ filtration of environmental water in one or more Sterivex filters placed in a filter cartridge. The IS-ABS also includes an application to program sampling definitions, allowing pre-setting configuration of the sampling. The efficiency of the IS-ABS was tested against traditional laboratory filtration standardized protocols. Results showed a good performance in terms of DNA recovery, as well as prokaryotic (16S rDNA) and eukaryotic (18S rDNA) community diversity analysis, using either methodologies. The IS-ABS automates the process of collecting environmental DNA, and is suitable for integration in water observation systems, what will contribute to substantially increase biological surveillances. Also, the use of highly sensitive genomic approaches allows a further study of the diversity and functions of whole or specific microbial communities. [ABSTRACT FROM AUTHOR]

Hydrocarbon degradation (HD) potential by autochthonous microorganisms in the coastal sediments of the NW area of Portugal (coastal sandy beaches and estuaries of the rivers Minho and Douro) was evaluated, investigating if water and sediment contamination/characteristics influence it. Sediments were characterized for microbial abundance (by DAPI), HD microorganisms' abundance (by MPN), microbial community structure (by ARISA), hydrocarbons (by FTIR and SPME-GC–MS), hazardous and noxious substances (SPME-GC–MS) and metals (by AAS). To our knowledge, this is the first time all these pollutants, including the selected HNS, were measured simultaneously in sediments of the selected coastal area. Higher contaminants concentrations were, generally, registered in Douro samples. A clear differentiation of the microbial community structure between beaches and estuaries was observed, as well as, between Douro and Minho river estuaries. BIO-ENV analysis indicated both sediment characteristics (e.g. OM content) and contaminants presence/concentrations (e.g. tetrachloroethylene presence) affected the structure of the microbial community along the studied areas. In all the selected sites, the characterized autochthonous microbial communities showed potential for hydrocarbons degradation, with HD microorganisms being found in all collected sediments. These microorganisms can be a valuable asset to recover contaminated areas, but sediment characteristics and contaminants presence/levels need to be taken into account as they can affect their bioremediation potential and the success of their application as biotechnological tool. [ABSTRACT FROM AUTHOR]

The authors wish to make the following corrections to this paper [[1]]: After the publication of the manuscript, the authors recognized that one reference in the discussion session (in the main text) was wrong and should be replaced with a new one. Reference 1 Tomasino M.P., Aparício M., Ribeiro I., Santos F., Caetano M., Almeida C.M.R., de Fátima Carvalho M., Mucha A.P. Diversity and hydrocarbon-degrading potential of deep-sea microbial community from the Mid-Atlantic Ridge, south of the Azores (North Atlantic Ocean). Changes in Discussion Furthermore, Potts et al. [58] in a recent work reported differences in hydrocarbons removal between shallow and deep sediment consortia (50-58% and 33-42%, respectively), highlighting the role of consortium origin. [Extracted from the article]

Deep-sea sediments (DSS) are one of the largest biotopes on Earth and host a surprisingly diverse microbial community. The harsh conditions of this cold environment lower the rate of natural attenuation, allowing the petroleum pollutants to persist for a long time in deep marine sediments raising problematic environmental concerns. The present work aims to contribute to the study of DSS microbial resources as biotechnological tools for bioremediation of petroleum hydrocarbon polluted environments. Four deep-sea sediment samples were collected in the Mid-Atlantic Ridge, south of the Azores (North Atlantic Ocean). Their autochthonous microbial diversity was investigated by 16S rRNA metabarcoding analysis. In addition, a total of 26 deep-sea bacteria strains with the ability to utilize crude oil as their sole carbon and energy source were isolated from the DSS samples. Eight of them were selected for a novel hydrocarbonoclastic-bacterial consortium and their potential to degrade petroleum hydrocarbons was tested in a bioremediation experiment. Bioaugmentation treatments (with inoculum pre-grown either in sodium acetate or petroleum) showed an increase in degradation of the hydrocarbons comparatively to natural attenuation. Our results provide new insights into deep-ocean oil spill bioremediation by applying DSS hydrocarbon-degrading consortium in lab-scale microcosm to simulate an oil spill in natural seawater. [ABSTRACT FROM AUTHOR]

Floating wetland islands (FWI) are considered nature-based solutions with great potential to promote several ecosystem services, such as biodiversity and water quality enhancement through phytoremediation processes. To our knowledge, the present work is the first to scientifically document the in-situ establishment of an FWI in a seawater port marina. The establishment and performance of a cork floating platform with a polyculture (Sarcocornia perennis, Juncus maritimus, Phragmites australis, Halimione portulacoides, Spartina maritima, Limonium vulgare) was evaluated. The diversity of organisms present in the FWI was undertaken based on the macrofauna assessment, taking into consideration marine water characterization, with a focus on hydrocarbons. Microbial communities were assessed based on metabarcoding approach to study 16S rRNA gene from environmental DNA retrieved from biofilm (from the planting media), marine biofouling (from the submerged platform) and surface marina water. S. perennis was the species with the highest survival rate and growth. The structure of the microbial community showed clear differences between those established in the FWI and those in the surrounding water, showing the presence of some bacterial groups that can be relevant for bioremediation processes (e.g., Saprospiraceae family). Concerning the macrofauna analysis, Mytilus sp. was the predominant taxa. To be of relevance, total petroleum hydrocarbons were detected at the marina up to ca. 6 mg/L. This study gives new insights into broadening FWI application to the saline environments of port marinas and to supporting a management strategy to promote several ecosystem services such biodiversity, species habitat, water quality enhancement and added aesthetic value to the marina landscape. [ABSTRACT FROM AUTHOR]

Fluoroquinolones and cephalosporins are two classes of veterinary antibiotics arising as pollutants of emerging concern. In this work, the microbial degradation of two representative antibiotics of both these classes, enrofloxacin (ENR) and ceftiofur (CEF), is reported. Biodegradation of the target antibiotics was investigated by supplementing the culture medium with ENR and CEF, individually and in mixture. Microbial inocula were obtained from rhizosphere sediments of plants derived from experimental constructed wetlands designed for the treatment of livestock wastewaters contaminated with trace amounts of these antibiotics. Selected microbial inocula were acclimated during a period of 5 months, where the antibiotics were supplemented every three weeks at the concentration of 1 mg L − 1 , using acetate as a co-substrate. After this period, the acclimated consortia were investigated for their capacity to biodegrade 2 and 3 mg L − 1 of ENR and CEF. Complete removal of CEF from the inoculated culture medium was always observed within 21 days, independently of its concentration or the concomitant presence of ENR. Biodegradation of ENR decreased with the increase in its concentration in the culture medium, with defluorination percentages decreasing from ca. 65 to 4%. Ciprofloxacin and norfloxacin were detected as biodegradation intermediates of ENR in the microbial cultures supplemented with this antibiotic, indicating that defluorination of at least part of ENR in these cultures is not an immediate catabolic step. Abiotic mechanisms showed high influence in the removal of CEF, affecting less ENR degradation. The acclimation process with the target antibiotics led to significant shifts in the structure and diversity of the microbial communities, predominantly selecting microorganisms belonging to the phyla Proteobacteria (e.g. Achromobacter , Variovorax and Stenotrophomonas genera) and Bacteroidetes (e.g. Dysgonomonas, Flavobacterium and Chryseobacterium genera). The results presented in this study indicate that biodegradation can be an important mechanism for the environmental removal of the tested compounds. [ABSTRACT FROM AUTHOR]

This study investigates the potential of an indigenous estuarine microbial consortium to degrade two polycyclic aromatic hydrocarbons (PAHs), naphthalene and fluoranthene, under nitrate-reducing conditions. Two physicochemically diverse sediment samples from the Lima Estuary (Portugal) were spiked individually with 25 mg L −1 of each PAH in laboratory designed microcosms. Sediments without PAHs and autoclaved sediments spiked with PAHs were run in parallel. Destructive sampling at the beginning and after 3, 6, 12, 30 and 63 weeks incubation was performed. Naphthalene and fluoranthene levels decreased over time with distinct degradation dynamics varying with sediment type. Next-generation sequencing (NGS) of 16 S rRNA gene amplicons revealed that the sediment type and incubation time were the main drivers influencing the microbial community structure rather than the impact of PAH amendments. Predicted microbial functional analyses revealed clear shifts and interrelationships between genes involved in anaerobic and aerobic degradation of PAHs and in the dissimilatory nitrate-reducing pathways (denitrification and dissimilatory nitrate reduction to ammonium - DNRA). These findings reinforced by clear biogeochemical denitrification signals (NO 3 − consumption, and NH 4 + increased during the incubation period), suggest that naphthalene and fluoranthene degradation may be coupled with denitrification and DNRA metabolism. The results of this study contribute to the understanding of the dissimilatory nitrate-reducing pathways and help uncover their involvement in degradation of PAHs, which will be crucial for directing remediation strategies of PAH-contaminated anoxic sediments. [ABSTRACT FROM AUTHOR]

Abstract: Salt marshes are important sinks for contaminants, namely metals that tend to accumulate around plant roots and could eventually be taken up in a process known as phytoremediation. On the other hand, microbial communities display important roles in the salt marsh ecosystems, such as recycling of nutrients and/or degradation of organic contaminants. Thus, plants can benefit from the microbial activity in the phytoremediation process. Nevertheless, above certain levels, metals are known to be toxic to microorganisms, fact that can eventually compromise their ecological functions. In this vein, the aim of present study was to investigate, in the laboratory, the effect of selected metals (Cd, Cu and Pb) on the microbial communities associated to the roots of two salt marsh plants. Sediments colonized by Juncus maritimus and Phragmites australis were collected in the River Lima estuary (NW Portugal), and spiked with each of the metals at three different Effects Range-Median (ERM) concentrations (1, 10×, 50×), being ERM the sediment quality guideline that indicates the concentration above which adverse biological effects may frequently occur. Spiked sediments were incubated with a nutritive saline solution, being left in the dark under constant agitation for 7 days. The results showed that, despite the initial sediments colonized by J. maritimus and P. australis displayed significant (p < 0.05) differences in terms of microbial community structure (evaluated by ARISA), they presented similar microbial abundances (estimated by DAPI). Also, in terms of microbial abundance, both sediments showed a similar response to metal addition, with a decrease in number of cells only observed for the higher addition of Cu. Nevertheless, both Cu and Pb, at intermediate metals levels promote a shift in the microbial community structure, with possibly effect on the ecological function of these microbial communities in salt marshes. These changes may affect plants phytoremediation potential and further work on this subject is in need. [Copyright &y& Elsevier]