Search

Supplementary data to this article can be found online at https://doi. org/10.1016/j.scitotenv.2021.151783, Environmental DNA (eDNA) metabarcoding (parallel sequencing of DNA/RNA for identification of whole communities within a targeted group) is revolutionizing the field of aquatic biomonitoring. To date, most metabarcoding studies aiming to assess the ecological status of aquatic ecosystems have focused on water eDNA and macroinvertebrate bulk samples. However, the eDNA metabarcoding has also been applied to soft sediment samples, mainly for assessing microbial or meiofaunal biota. Compared to classical methodologies based on manual sorting and morphological identification of benthic taxa, eDNA metabarcoding offers potentially important advantages for assessing the environmental quality of sediments. The methods and protocols utilized for sediment eDNA metabarcoding can vary considerably among studies, and standardization efforts are needed to improve their robustness, comparability and use within regulatory frameworks. Here, we review the available information on eDNA metabarcoding applied to sediment samples, with a focus on sampling, preservation, and DNA extraction steps. We discuss challenges specific to sediment eDNA analysis, including the variety of different sources and states of eDNA and its persistence in the sediment. This paper aims to identify good-practice strategies and facilitate method harmonization for routine use of sediment eDNA in future benthic monitoring., The work is the result of the DNAqua-Net workshop held in Rome, on June 10–12, 2019, organised by IRSA-CNR and supported by European Cooperation in Science and Technology (COST) grant CA15219 (DNAqua-Net). It largely benefited from the discussions with DNAqua-Net members during this event as well as other DNAqua-Net conferences.This study was carried out within the UNIVPM VIRIDE Project (n. 2017EKFA98) funded by the Ministry of University and Research of Italy (MUR) and the Italian National Project on the use of environmental DNA for the identification of marine non-indigenous species funded by the Istituto Superiore per la Protezione e la Ricerca Ambientale (ISPRA). JP, TC, and LAPG were supported by the Swiss National Science Foundation (grant 31003A_179125) as well as FL with an Early Postdoc.Mobility grant (P2GEP3_171829). MS-M was supported by the Ministry of Education, Youth and Sports of the Czech Republic, European Regional Development Fund-Project CZ.02.1.01/0.0/0.0/16_019/0000845 and LTC 20063. KP was supported by the European Regional Development Fund within National Programme for Addressing Socio-Economic Challenges through R&D (L190003VLVI RITA1/02-60-05) and base financed project P190250PKKH of Estonian University of Life Sciences, IM was supported by the Slovenian Research Agency (ARRS) programme P4-0085. SD was supported by the Portuguese Foundation for Science and Technology (FCT) (CEECIND/00667/2017). LC was supported by the PON AIM 1854833 – PON Ricerca e Innovazione 2014–2020 – Azione I.2 – D.D. n. 407, 27/02/2018 – Attraction and International Mobility.VK was supported by grants PUT1389 (ETAg) and Institute of Technology, University of Tartu basic funding grant.

BACKGROUND: Microbial electrochemical technologies (METs) represent a novel platform to harvest the energy trapped in municipal wastewater. At the anode of METs, electro‐active bacteria (EAB) anaerobically oxidize wastewater constituents using the electrode as the terminal electron acceptor and, by so doing, generate an electric current. To convert complex wastewater constituents into electricity, EAB must not only establish syntrophic relationships with other members of the microbial community, but also compete with methanogens for consumption of hydrogen and acetate. Here, we examined the addition of magnetite nanoparticles (NPs) (250 mg Fe L⁻¹) as a novel strategy to manipulate such metabolic interactions and in turn maximize the efficiency of wastewater treatment and the yield of electric current generation. RESULTS: Batch experiments carried out either in the presence of a mixture of volatile fatty acids or of a synthetic sewage demonstrated that magnetite addition accelerate the rate of electrogenic oxidation of specific compounds, particularly propionate (up to 120%), an intermediate which frequently accumulates during anaerobic treatment processes, while correspondingly enhancing electric current generation (up to 90%), and diminishing the rate of competing methane generation (up to 50%). Notably, the composition of the microbial community was not substantially affected by the presence of magnetite nanoparticles, possibly suggesting that these latter facilitated extracellular electron transfer mechanisms (among microbes and with the electrode), rather than enriching conditions for specific microorganisms. CONCLUSION: The addition of magnetite NPs may represent a practical strategy to kick‐start a bioelectrochemical system designed for wastewater treatment and improve the effectiveness of electrogenic substrate oxidation processes. © 2019 Society of Chemical Industry

Quorum sensing signals regulate various functions within activated sludge processes such as formation of microbial aggregates. Disturbance of this signaling system, known as quorum quenching (QQ), provides opportunities for eliminating some problems related to biological wastewater treatment (e.g., biofouling and excess sludge production). However, it is poorly understood how and to what extent QQ systems can affect the microbial aggregation processes and the following floc formation. In particular, an in-depth structural characterization at the scale of microbial aggregate while considering nutrient conditions in the reactor is still largely disregarded. Here, we evaluated the QQ effects at the short-term time scale (i.e., after 4 h for the exogenous period and 19 h for exogenous/endogenous period), by combining advanced techniques for microbial characterization (flow cytometry, CARD-FISH, and confocal laser scanning microscopy) and conventional physical-chemical assessments. The results indicated that by implementing QQ agents (immobilized Acylase I enzyme in porous alginate beads) the abundance of single cells and suspended microbial aggregates in the supernatant did not show significant changes during the exogenous period. Conversely, at the end of the exogenous/endogenous period a significant increase of single prokaryotic cells, small and large microbial aggregates favored the growth of grazers, including free-living nanoflagellates and ciliates. Flocs became looser and thinner than those in the control reactor, thus affecting the sludge settling behavior. Inability of microbial community in degradation of soluble protein during the endogenous period confirmed that the QQ agents are likely to inhibit the secretion of protease enzyme within microbial communities of activated sludge.

Direct and indirect effects of extremely high geogenic CO2 levels, commonly occurring in volcanic and hydrothermal environments, on biogeochemical processes in soil are poorly understood. This study investigated a sinkhole in Italy where long-term emissions of thermometamorphic-derived CO2 are associated with accumulation of carbon in the topsoil and removal of inorganic carbon in low pH environments at the bottom of the sinkhole. The comparison between interstitial soil gasses and those collected in an adjacent bubbling pool and the analysis of the carbon isotopic composition of CO2 and CH4 clearly indicated the occurrence of CH4 oxidation and negligible methanogenesis in soils at the bottom of the sinkhole. Extremely high CO2 concentrations resulted in higher microbial abundance (up to 4 × 109 cell g-1 DW) and a lower microbial diversity by favoring bacteria already reported to be involved in acetogenesis in mofette soils (i.e., Firmicutes, Chloroflexi, and Acidobacteria). Laboratory incubations to test the acetogenic and methanogenic potential clearly showed that all the mofette soil supplied with hydrogen gas displayed a remarkable CO2 fixation potential, primarily due to the activity of acetogenic microorganisms. By contrast, negligible production of acetate occurred in control tests incubated with the same soils, under identical conditions, without the addition of hydrogen. In this study, we report how changes in diversity and functions of the soil microbial community - induced by high CO2 concentration - create peculiar biogeochemical profile. CO2 emission affects carbon cycling through: (i) inhibition of the decomposition of the organic carbon and (ii) promotion of CO2-fixation via the acetyl-CoA pathway. Sites naturally exposed to extremely high CO2 levels could potentially represent an untapped source of microorganisms with unique capabilities to catalytically convert CO2 into valuable organic chemicals and fuels.

This study represents the first example in which benthic heterotrophic and photoautotrophic microbial communities (prokaryotes and microphytobenthos-MPB, respectively) were simultaneously investigated. To explore how they synergistically respond to multiple natural and anthropogenic stressors, in the framework of the Project RITMARE four lagoons of the Po River Delta were sampled in May 2016: two with more marine features, i.e. Scardovari (SC) and Caleri (CL), and two more directly affected by the Po River flow, i.e. Canarin (CN) and Vallona-Marinetta (MV). The abundance and structure of benthic communities were related to physical-chemical parameters i.e. grain-size, total N, total organic C, stable C and N isotopes, and synthetic organic contaminants. Stations were gathered into three distinct groups (R-ansosim = 0.540 p < 0.001) according to their different physical-chemical features: outer, inner-freshwater and inner-marine sites. Contamination levels did not seem to severely affect the microbial abundances that were overall stimulated by the combined effect of high organic and nutrient loads: prokaryotes up to + 42% and MPB up to +93%. Bacteria and Archaea displayed high densities at sites directly influenced by the freshwater input and anthropogenically derived nitrogen. Delta- (30.8%), Alpha- (12.8%), Gamma- (11.8%) and Beta-Proteobacteria (7.1%) were the dominant classes at all sites. For both communities, a significant inter-lagoonal (among different lagoons) and infra-lagoonal (among stations within the same lagoon) pattern was highlighted by PERMANOVA. nMDS and SIMPER analyses revealed distinct assemblages in the inner and outer parts of the lagoons. We applied a novel functional approach based on diatom life modes. At inner and outer sites, different diatom living forms developed: epipsammic (on sand) nearby the lagoonal mouths, epipelic (on mud) at the innermost sites and non-benthic forms (planktonic, tychopelagic and epiphytic) at sites directly influenced by freshwater. Distance-based Linear Models further indicated that salinity, clay and temperature were the significant drivers of the prokaryotic spatial distribution whereas clay, PAHs, PCBs and salinity best explained the MPB structure. The dominance of non-benthic diatom life modes in the more polluted lagoon (CN) suggests a negative influence of contamination on the MPB structure. Two diatom keystone species likely contribute to restore the oxic gradient in sediments frequently exposed to anoxia, allowing the subsequent microbial aerobic degradation and the recolonization of higher trophic organisms. The capacity of re-oxygenation after anoxia has important ecological and economic implications in lagoons exploited for aquaculture.

An integrated geochemical and microbiological investigation of soils from the Solfatara Crater (Campi Flegrei, southern Italy) demonstrated that interstitial soil gases dominated by CO2 and other typical hydrothermal gaseous species (e.g. H2S, CH4, ethane, benzene, alkenes and S-bearing organic compounds) influenced the composition of microbial communities. The relatively high concentrations of hydrothermal fluids permeating the soil produced acidic conditions and whitish deposits that characterize the Solfatara Crater floor. Archaea and Bacteria showed almost equal cell abundance (up to 3.2 x 10(7) and 4.2 x 10(7) cell/g, respectively) with relatively low levels of biodiversity and equitability in sites characterized by elevated temperatures (up to 70 degrees C), very low pH values (up to 2.2) and reducing conditions. In these sites, high-throughput sequencing showed the marked selection of microorganisms, mainly affiliated with the genera Thermoplasma, Ferroplasma and Acidithiobacillus. A relatively high biodiversity and concomitant distinctive structure of the microbial community were observed in soils poorly affected by fumarolic emissions that were oxic and rich in organic matter.

Arsenic (As) contamination in drinking water represents a worldwide threat to human health. During last decades, the exploitation of microbial As-transformations has been proposed for bioremediation applications. Among biological methods for As-contaminated water treatment, microbial As(III)-oxidation is one of the most promising approaches since it can be coupled to commonly used adsorption removal technologies, without requiring the addition of chemicals and producing toxic by-products. Despite the As(III) oxidation capability has been described in several bacterial pure or enrichment cultures, very little is known about the real potentialities of this process when mixed microbial communities, naturally occurring in As contaminated waters, are used. This study highlighted the contribution of native groundwater bacteria to As(III)-oxidation in biofilters, under conditions suitable for a household-scale treatment system. This work elucidated the influence of a variety of experimental conditions (i.e., various filling materials, flow rates, As(III) inflow concentration, As(III):As(V) ratio, filter volumes) on the microbially-mediated As(III)-oxidation process in terms of oxidation efficiency and rate. The highest oxidation efficiencies (up to 90% in 3 h) were found on coarse sand biofilters treating total initial As concentration of 100 μg L−1. The detailed microbial characterization of the As(III) oxidizing biofilms revealed the occurrence of several OTUs affiliated with families known to oxidize As(III) (e.g., Burkholderiaceae, Comamonadaceae, Rhodobacteraceae, Xanthomonadaceae). Furthermore, As-related functional genes increased in biofilter systems in line with the observed oxidative performances.

Uncertainties exist regarding the magnitude of in situ dissolved organic matter (DOM) processing in lotic systems. In addition, little is known about the effects of extreme hydrological events on in-stream DOM retention or release during downriver transport. This study quantified the net in-stream retention/release efficiencies (η) of dissolved organic carbon (DOC) and its humic and protein-like fluorescent fractions along a Mediterranean river during drought, baseflow and flood conditions. High performance size exclusion chromatography was used to describe the apparent size distributions of the humic and protein-like DOM moieties. A snapshot mass balance allowed estimating the η values of DOC and humic and protein-like fractions. Significant DOM net retention (η < 0) was detected during the drought condition and the protein-like fraction was more retained than the humic-like fraction and bulk DOC. In addition, small substances were more efficiently retained than larger substances. DOC retention decreased under baseflow conditions, but it remained significant. The humic and protein-like net efficiencies exhibited high variability, but the net retention were not significant. From a longitudinal perspective, the entire fluvial corridor contributed net retention of DOC and humic and protein-like moieties net retention during drought condition. In contrast, net retention/release efficiencies exhibited spatial variability during baseflow condition. The flood preferentially mobilized large size DOM molecules and the fluvial corridor behaved as a homogeneous passive DOM (η = 0) conduit. This research highlights the relevance of hydrological extreme events on the magnitude of DOM retention/release mass balance and emphasizes the need to perform measurements during these conditions to quantify the impact of fluvial corridors on DOM fate and transport This research is funded by the Spanish Ministry of Education and Science (MEC) (CGL2011-30151-C02 and CGL2014-5876-C3-R) and European Community 7th Framework Programme (No. 603629- ENV-2013-6.2.1-Globaqua)

The presence of arsenic (As) in aquatic environments is often attributable to geogenic processes occurring within aquifers. Arsenic exists in two prevalent oxidation states, with the trivalent arsenite [As(III)] exerting stronger toxicity effects on the aquatic biota than the pentavalent arsenate [As(V)]. The review evaluates the literature available on the main arsenic removal technologies and the application of combined chemical and biological treatments. We provide a synthetic outlook on the potential strategies of biological As(III) oxidation to As(V) by means of cell-detoxifying mechanisms or metabolic processes with the aim to enhance As removal efficiency. Furthermore, the role of microorganisms in the mobility of arsenic in natural systems as well as the distribution of As-resistant bacteria, potentially suitable for arsenic removal, is discussed in the context of a case study carried out in Latium region (Italy), which is known for arsenic contamination of waters.

Arsenic contamination of soil and groundwater is a serious problem worldwide. Here we show that anaerobic oxidation of As(III) to As(V), a form which is more extensively and stably adsorbed onto metal-oxides, can be achieved by using a polarized (+497 mV vs. SHE) graphite anode serving as terminal electron acceptor in the microbial metabolism. The characterization of the microbial populations at the electrode, by using in situ detection methods, revealed the predominance of gammaproteobacteria. In principle, the proposed bioelectrochemical oxidation process would make it possible to provide As(III)-oxidizing microorganisms with a virtually unlimited, low-cost and low-maintenance electron acceptor as well as with a physical support for microbial attachment This work was supported by MIUR PRIN 2010-2011 project (grant agreement No 2010JBNLJ7-002). The authors thank the Advanced Centre for Microscopy "P. Albertano" of University of Rome "Tor Vergata" for CLSM analyses. Narcis Pous and Sebastia Puig were supported by a project grant from the Catalan Government (2012 FI-B 00941) and the Spanish Government (CTQ2011-23632). Narcis Pous was also supported by a COST Action ES1202 in the context of Water 2020 (COST-STSM-ECOST-STSM-ES1202-011013-035423)

Microbial electrochemical systems (MES) represent an innovative reagent-free technology for in situ remediation of groundwater contaminated by petroleum hydrocarbons. Here we describe the long-term (>160 days) anaerobic treatment of synthetic groundwater containing toluene (25 mg L−1) in a novel laboratory-scale, continuous-flow bioelectrochemical reactor, termed the “bioelectric well”. Under optimal operating conditions (i.e., anode potential potentiostatically controlled at +0.2 V vs. SHE and recycle flow-rate set at 75 mL min−1), the observed electrogenic toluene oxidation rate was 67.2 ± 5.7 mg L d−1, a value which is among the highest reported in the literature for laboratory-scale anaerobic treatment systems. Correspondingly, electric current was 5.1 ± 0.1 mA and the Coulombic efficiency (i.e., the yield of toluene conversion into electric current) was 79 ± 7%. Electrogenic toluene oxidation was most likely catalyzed by Geobacter species, which were found to dominate the surface of the graphite anode. The combined application of GC-MS (for detection and identification of metabolites) and qPCR (for quantification of functional genes) revealed that toluene degradation was initiated by fumarate addition, an activation pathway that is widely distributed among anaerobic hydrocarbon-degrading microorganisms. Collectively, the results of this study indicate that the bioelectric well holds remarkable potential for in situ treatment of groundwater contaminated by petroleum hydrocarbons.

The interaction of anaerobic dechlorinating cultures with soil and aquifer geochemical components is largely unknown, although this has potentially a major impact on the bioremediation of chlorinated solvent-contaminated sites. In this study, we found that addition of magnetite (Fe 3 O 4 ) – the end-product of Fe(III)-reduction by dissimilatory iron reducing bacteria – to anaerobic dechlorinating cultures enhances the kinetics of trichloroethene dechlorination up to 1.5-times, compared to unamended controls. Specifically, a low concentration (approx. 10 mg/L as total Fe) of small size particles (200 nm-filtered) resulted in a greater stimulatory effect compared to the addition of a higher concentration (approx. 300 mg/L as total Fe) of unfiltered particles. Notably, Desulforomonas spp. were substantially enriched in microcosms supplemented with magnetite, whereas Dehalococcoides mccartyi spp. was found to be markedly inhibited or outcompeted. Multiple lines of evidence, including the direct visualization of microbial cells and magnetite particles via Confocal Laser Scanning Microscopy (CLSM), suggest that electrically conductive particles promoted the establishment of a cooperative metabolism, based on direct interspecies electron transfer, between dechlorinating and non-dechlorinating microorganisms.

Antibiotics are emerging contaminants, which wing to their bioactivity, may lead to short-term and long-term alterations of natural microbial communities in aquatic environment. We investigated the effects of antibiotics on biofilm bacterial communities in the Llobregat River (Northeast Spain). Three sampling sites were selected: two less polluted sites and one hotspot. River water was collected from each site and used both as inoculum and medium for growing biofilms in independent mesocosms. After 25 d of biofilm colonization, we exposed the colonized biofilms to river waters from the downstream sites (progressively contaminated by antibiotics). A control from each site was maintained where the growing biofilm was always exposed to water from the same site. The bacterial community composition, bacterial live/dead ratio and extracellular enzyme activities of the biofilms were measured before and 9 d after exposing the biofilms to increasing contaminated waters. Sixteen antibiotic compounds were detected in the water from the three sampling sites. At each site, the antibiotics present in the highest concentrations were sulfonamides, followed by quinolones and macrolides. Bacterial communities of biofilms grown with the three river waters differed markedly in their structure, but less so in terms of functional descriptors. After switching the medium water to increasing pollution, biofilms exhibited increased levels of actinobacteria (HGC), a trend that was associated to the higher antibiotic concentrations in the water. These biofilms also showed increased bacterial mortality, and decreased extracellular leucine-aminopeptidase and alkaline phosphatase. There was a significant correlation between antibiotic concentrations and biofilm responses. Our results indicate that the continuous entrance of antibiotics in running waters cause significant structural and functional changes in microbial attached communities.

Arsenic (As) is a toxic element released in aquatic environments by geogenic processes or anthropic activities. To counteract its toxicity, several microorganisms have developed mechanisms to tolerate and utilize it for respiratory metabolism. However, still little is known about identity and physiological properties of microorganisms exposed to natural high levels of As and the role they play in As transformation and mobilization processes. This work aims to explore the phylogenetic composition and functional properties of aquatic microbial communities in As-rich freshwater environments of geothermal origin and to elucidate the key microbial functional groups that directly or indirectly may influence As-transformations across a natural range of geogenic arsenic contamination. Distinct bacterial communities in terms of composition and metabolisms were found. Members of Proteobacteria, affiliated to Alpha- and Betaproteobacteria were mainly retrieved in groundwaters and surface waters, whereas Gammaproteobacteria were the main component in thermal waters. Most of the OTUs from thermal waters were only distantly related to 16S rRNA gene sequences of known taxa, indicating the occurrence of bacterial biodiversity so far unexplored. Nitrate and sulphate reduction and heterotrophic As(III)-oxidization were found as main metabolic traits of the microbial cultivable fraction in such environments. No growth of autotrophic As(III)-oxidizers, autotrophic and heterotrophic As(V)-reducers, Fe-reducers and oxidizers, Mn-reducers and sulfide oxidizers was observed. The ars genes, involved in As(V) detoxifying reduction, were found in all samples whereas aioA (As(III) oxidase) and arrA genes (As(V) respiratory reductase) were not found. Overall, we found that As detoxification processes prevailed over As metabolic processes, concomitantly with the intriguing occurrence of novel thermophiles able to tolerate high levels of As.

Understanding DOM transport and reactivity in rivers is essential to having a complete picture of the global carbon cycle. In this study, we explore the effects of hydrological variability and downstream transport on dissolved organic matter (DOM) dynamics in a Mediterranean river. We sampled the main stem of the river Tordera from the source to the sea, over a range of fifteen hydrological conditions including extreme events (flood and drought). By exploring spatial and temporal gradients of DOM fluorescence properties, river hydrology was found to be a significant predictor of DOM spatial heterogeneity. An additional space-resolved mass balance analysis performed on four contrasting hydrological conditions revealed that this was due to a shift in the biogeochemical function of the river. Flood conditions caused a conservative transport of DOM, generating a homogeneous, humic-like spatial profile of DOM quality. Lower flows induced a non-conservative, reactive transport of DOM, which enhanced the spatial heterogeneity of DOM properties. Moreover, the downstream evolution of DOM chemostatic behaviour revealed that the role of hydrology in regulating DOM properties increased gradually downstream, indicating an organised inter-dependency between the spatial and the temporal dimensions. Overall, our findings reveal that riverine DOM dynamics is in constant change owing to varying hydrological conditions, and emphasize that in order to fully understand the role of rivers in the global carbon cycle, it is necessary to take into account the full range of hydrological variability, from floods to droughts.

Sediment resuspension represents a key process in all natural aquatic systems, owing to its role in nutrient cycling and transport of potential contaminants. Although suspended solids are generally accepted as an important quality parameter, current monitoring programs cover quantitative aspects only. Established methodologies do not provide information on origin, fate, and risks associated with uncontrolled inputs of solids in waters. Here we discuss the analytical approaches to assess the occurrence and ecological relevance of resuspended particulate matter in freshwaters, with a focus on the dynamics of associated contaminants and microorganisms. Triggered by the identification of specific physical–chemical traits and community structure of particle-associated microorganisms, recent findings suggest that a quantitative determination of microorganisms can be reasonably used to trace the origin of particulate matter by means of nucleic acid-based assays in different aquatic systems.

In Mediterranean regions, drought is one of the main factors shaping fluvial ecosystems. Droughts cause a shift from lotic to lentic conditions, triggering a gradual fragmentation of the longitudinal hydrological continuum, and a severe alteration of water chemical properties. However, within a biogeochemical perspective, little is known about how and to which extend droughts modify the chemical properties of dissolved organic matter (DOM). In this study, the variability of DOM properties along a fragmented fluvial system is explored, during a summer severe drought, by means of (a) the ratio between dissolved organic carbon and nitrogen concentrations (DOC:DON); (b) DOC bioavailability (BDOC) and (c) DOM optical properties (SUVA index, fluorescence index, and excitation–emission fluorescence matrices). DOM and water measurements were collected from isolated water parcels that became disconnected from the fluvial continuum at different times, and were compared with data obtained in the following autumn, when the fluvial continuum was re-established. Analysis of DOM chemical properties evidenced that these properties during drought clearly differed from those observed in autumn, but changes did not follow an arbitrary pattern. Thus, the sampling sites with lotic water bodies showed DOM properties similar to those observed in autumn reflecting the dominance of terrestrial inputs. But, once hydrological fragmentation occurred, there was a gradual increase in the contribution of autochthonous DOM as the time elapsed since the pools were established, and the geochemical conditions shifted from oxidized to reduced conditions. In consequence, the fragmentation of fluvial continuum generates a set of distinct biochemical hot spots (i.e., each water parcel), revealing that extreme drought greatly amplifies the qualitative heterogeneity of organic matter in a fluvial system.

Temporary rivers are characterized by dry-wet phases and represent an important water resource in semi-arid regions worldwide. The fate and effect of contaminants have not been firmly established in temporary rivers such as in other aquatic environments. In this study, we assessed the effects of sediment amendment with Polycyclic Aromatic Hydrocarbons (PAHs) on benthic microbial communities. Experimental microcosms containing natural (Control) and amended sediments (2 and 20 mg PAHs kg(-1) were incubated for 28 days. The PAH concentrations in sediments were monitored weekly together with microbial community structural (biomass and phylogenetic composition by TGGE and CARD-FISH) and functional parameters (ATP concentration, community respiration rate, bacterial carbon production rate, extracellular enzyme activities). The concentration of the PAH isomers did not change significantly with the exception of phenanthrene. No changes were observed in the TGGE profiles, whereas the occurrence of Alpha- and Beta-Proteobacteria was significantly affected by the treatments. In the amended sediments, the rates of carbon production were stimulated together with aminopeptidase enzyme activity. The community respiration rates showed values significantly lower than the Control after 1 day from the amendment then recovering the Control values during the incubation. A negative trend between the respiration rates and ATP concentration was observed only in the amended sediments. This result indicates a potential toxic effect on the oxidative phosphorylation processes. The impoverishment of the energetic resources that follows the PAH impact may act as a domino on the flux of energy from prokaryotes to the upper level of the trophic chain, with the potential to alter the temporary river functioning.

We studied the efficiency of two hybridization techniques for the analysis of benthic bacterial community composition under varying sediment water content. Microcosms were set up with sediments from four European temporary rivers. Wet sediments were dried, and dry sediments were artificially rewetted. The percentage of bacterial cells detected by fluorescence in situ hybridization with fluorescently monolabeled probes (FISH) significantly increased from dry to wet sediments, showing a positive correlation with the community activity measured via incorporation of 3H leucine. FISH and signal amplification by catalyzed reporter deposition (CARD-FISH) could significantly better detect cells with low activity in dried sediments. Through the application of an optimized cell permeabilization protocol, the percentage of hybridized cells by CARD-FISH showed comparable values in dry and wet conditions. This approach was unrelated to 3H leucine incorporation rates. Moreover, the optimized protocol allowed a significantly better visualization of Gram-positive Actinobacteria in the studied samples. CARD-FISH is, therefore, proposed as an effective technique to compare bacterial communities residing in sediments with contrasting water content, irrespective of differences in the activity state of target cells. Considering the increasing frequencies of flood and drought cycles in European temporary rivers, our approach may help to better understand the dynamics of microbial communities in such systems.

Interspecies electron transfer mechanisms between Bacteria and Archaea play a pivotal role during methanogenic degradation of organic matter in natural and engineered anaerobic ecosystems. Growing evidence suggests that in syntrophic communities electron transfer does not rely exclusively on the exchange of diffusible molecules and energy carriers such as hydrogen or formate, rather microorganisms have the capability to exchange metabolic electrons in a more direct manner. Here, we show that supplementation of micrometer-size magnetite (Fe3O4) particles to a methanogenic sludge enhanced (up to 33%) the methane production rate from propionate, a key intermediate in the anaerobic digestion of organic matter and a model substrate to study energy-limited syntrophic communities. The stimulatory effect most probably resulted from the establishment of a direct interspecies electron transfer (DIET), based on magnetite particles serving as electron conduits between propionate-oxidizing acetogens and carbon dioxide-reducing methanogens. Theoretical calculations revealed that DIET allows electrons to be transferred among syntrophic partners at rates which are substantially higher than those attainable via interspecies H2 transfer. Besides the remarkable potential for improving anaerobic digestion, which is a proven biological strategy for renewable energy production, the herein described conduction-based DIET could also have a role in natural methane emissions from magnetite-rich soils and sediments.

Chlorinated ethenes contamination of soil and groundwater is a widespread problem in most industrialized countries. To date, there is a general consensus in the literature that members of the genus Dehalococcoides are required for complete dechlorination of these compounds. The availability of specific identification tools to track their distribution in the field is therefore a topic of particular relevance in environmental studies. These microorganisms have been successfully visualized by fluorescence in situ hybridization (FISH) in highly active dechlorinating cultures. However, FISH detection of Dehalococcoides under low activity conditions can be strongly hampered by their small cell size and low ribosome content. In this study, catalyzed reporter deposition (CARD)-FISH was employed as an alternative detection method. In a trichloroethene (TCE) dechlorinating enrichment culture, CARD-FISH, using proteinase K as a permeabilization pre-treatment, was found to be significantly superior to conventional FISH in terms of both microscopic visualization and quantification efficiency (about 30%). An application of this method on contaminated aquifer samples is also presented and discussed.

Bacterial degradation of mucilaginous aggregates (creamy layers, stringers and macroflocs) collected during two summer events (2001-2002) was tested. The objective was to describe the temporal trend of the bacterial activity, abundance and composition in the aggregated and dissolved organic matter under different trophic conditions. In the native aggregates proteins and organic phosphorous were actively hydrolyzed as aminopeptidase and alkaline phosphatase activities represented up to 87% and 25% of total activity, respectively; polysaccharides were less hydrolyzed and the highest activities were observed for beta-glucosidase (5% of the total). This hydrolysation pattern tends to a progressive accumulation of long persistent polysaccharides. During short term incubations nutrient addition (P, N and Glucose) differently stimulated bacterial growth in the seawater: P played the main role in stimulating bacterial production from 3 to 6 folds higher than in the control, whereas a secondary C-limitation was observed only for bacteria growing on seawater from macroflocs. This scarce dissolved organic carbon (DOC) bioavailability was confirmed by the lower DOC removal (13% macroflocs, 36% stringers). The total amount of carbon incorporated by bacteria living on aggregates was similar (0.58 mg C L(-1)) both in the control and under P enrichments showing a more balanced condition with respect to the seawater. Hence the well-known P limitation in the Northern Adriatic Sea affects only dissolved organic carbon uptake without influencing the uptake of aggregated organic matter. Organic matter limitation was observed only on stringers--total C incorporated raised to 0.96 mg C L(-1) after PNG addition. Macroflocs release of refractory compounds leads to DOC accumulation (73 microM DOC) contributing to inflate the pool of refractory DOC in the surrounding waters. Several evidences, including different monosaccharide composition of stringers and macroflocs (glucose 15% and 56% on stringers and macroflocs, respectively), bring to the conclusion that stringers are in an older stage in comparison with macroflocs. Community composition described by fluorescence in situ hybridization did not show significant differences between free-living and attached bacteria but it was modified by the different enrichment conditions: Cytophaga-Flavobacteria increased after inorganic nutrients enrichments while organics advantaged gamma-Proteobacteria.