Resistance, tolerance, and persistence to antibiotics have mainly been studied at the level of a single microbial isolate. However, in recent years it has become evident that microbial interactions play a role in determining the success of antibiotic treatments, in particular by influencing the occurrence of persistence and tolerance within a population. Additionally, the challenge of resuscitation (the capability of a population to revive after antibiotic exposure) and pathogen clearance are strongly linked to the small size of the surviving population and to the presence of fluctuations in cell counts. Indeed, while large population dynamics can be considered deterministic, small populations are influenced by stochastic processes, making their behaviour less predictable. Our study argues that microbe-microbe interactions within a community affect the mode, tempo, and success of persister resuscitation and that these are further influenced by noise. To this aim, we developed a theoretical model of a three-member microbial community and analysed the role of cell-to-cell interactions on pathogen clearance, using both deterministic and stochastic simulations. Our findings highlight the importance of ecological interactions and population size fluctuations (and hence the underlying cellular mechanisms) in determining the resilience of microbial populations following antibiotic treatment.

Heavy metals represent a significant hazard in textile wastewater, posing a considerable risk to both the ecosystem and human health. The objective of this study was to analyze the removal efficiency of specific heavy metals in a large wastewater treatment plant (WWTP) located in Prato (Tuscany, Italy), where the main Italian textile district is based. To achieve this, the mass balance calculation approach was employed. Therefore, two monitoring campaigns were conducted, collecting wastewater and sludge samples in some specific sections of the WWTP. The concentrations of Pb, Cd, Ni, As, and Sn were consistently below the detection limits. A good removal efficiency was determined for Zn, Cu, Ba, Crtot, and Sb, in the range of 37–79%. These metals are predominantly present in particulate form, facilitating their removal through sedimentation. Conversely, boron is largely present in the dissolved phase, resulting in its complete release through the treated effluent. Subsequently, an excellent linear correlation was identified between the input load and the contaminant load removed. This demonstrated that the plant’s efficiency remains unaffected by an increase in the input load at the observed contaminant concentrations. Finally, a probability law was identified that demonstrates an excellent degree of approximation in representing inlet metal concentrations. The findings of this study indicate that the treatment systems employed by the WWTP are capable of effectively removing heavy metals.

Phytodepuration occurs in the plant-mediated remediation processes exploited to remove pollutants from wastewater, and Phragmites australis is one of the most used plants. This goal is achieved using constructed wetlands (CW), which are engineered systems designed to mimic the natural processes of pollutants removal. The aim of this work was to characterize the bacterial communities associated to P. australis, soils, and permeates of the CW of Calice (Prato, Italy), to evaluate the possible effect of wastewaters on the CW bacterial communities, through a next-generation sequencing-based approach. A total of 122 samples were collected from different tissues of P. australis (i.e., roots, aerial parts, and stem), soil (i.e., rhizospheric and bulk soil), and permeates, and analyzed. All samples were collected during five sampling campaigns, with the first one performed before the activation of the plant. Obtained results highlighted a specific microbiota of P. australis, conserved among the different plant tissues and during time, showing a lower alpha diversity than the other samples and not influenced by the more complex and variable environmental (soils and permeates) bacterial communities. These data suggest that P. australis is able to select and maintain a defined microbiota, a capacity that could allow the plant to survive in hostile environments, such as that of CW.

Lay abstract In the present work, 21 bacteria associated to Phragmites australis roots were tested for their inhibitory activity against 36 human pathogens isolated from food, clinical patients and/or hospitals, which have the ability to escape several commonly used antibiotics. Seven out of 21 bacteria associated to P. australis were able to inhibit the growth of most of target pathogens. This preliminary work could pave the way to the discovery of new antimicrobial compounds active against bacterial pathogens that cannot be killed using several antibiotics. Background: Rising of multidrug-resistant human pathogens demands novel antibiotics: to this aim, unexplored natural sources are investigated to find new compounds. In this context, bacteria associated to medicinal plants, including Phragmites australis, might represent an important source of antimicrobial compounds. Materials & methods: In the present work, 21 bacterial endophytes isolated from P. australis roots were tested, through cross-streaking, for their inhibitory activity against 36 multidrug-resistant pathogens isolated from food, clinical patients and hospitals. Results & conclusion: Seven endophytes, belonging to Pseudomonas and Stenotrophomonas, were able to inhibit the growth of most of target strains. In conclusion, this preliminary work could pave the way to the discovery of new antibiotics active against superbugs.

The Sasso Fratino Integral Nature Reserve (Italy) aims to protect nature and territory. Since no anthropic activities are allowed, it represents a good model to study the bacterial community of a wild environment. The aim of this work was to characterise the cultivable and the total bacterial community of soil samples from the reserve in terms of taxonomy, composition, and structure. Seven soil samples were collected at different altitudes, and the chemical composition, the total and the cultivable microbiota, and the antibiotic resistance profiles of isolates were investigated. Total bacterial communities, studied through Next Generation Sequences analysis, included 390 genera. Samples differed in terms of microbial composition basing on the different altitude/vegetation of collection points. Random Amplified Polymorphic DNA Analysis (RAPD) allowed to identify 82 haplotypes out of 158 bacterial isolates. The taxonomic identification through 16S rDNA sequencing revealed that the strains were affiliated to 21 genera. Antibiotic resistance profiles of bacteria were also investigated, highlighting a high resistance against streptomycin and kanamycin. This work represents the first description of the soil bacterial community from the Natural Reserve of Sasso Fratino, and it is the first study considering the soil microbiota of an Italian integral nature reserve.

Improvement of industrial productions through more environment-friendly processes is a hot topic. In particular, land and marine environment pollution is a main concern, considering that recalcitrant compounds can be spread and persist for a long time. In this context, an efficient and cost-effective treatment of wastewater derived from industrial applications is crucial. Phytodepuration has been considered as a possible solution and it is based on the use of plants and their associated microorganisms to remove and/or transform pollutants. In this work we investigated the culturable microbiota of Phragmites australis roots, sampled from the constructed wetlands (CWs) pilot plant in the G.I.D.A. SpA wastewater treatment plant (WWTP) of Calice (Prato, Tuscany, Italy) before and after the CW activation in order to check how the influx of wastewater might affect the resident bacterial community. P. australis specimens were sampled and a panel of 294 culturable bacteria were isolated and characterized. This allowed to identify the dynamics of the microbiota composition triggered by the presence of wastewater. 27 out of 37 bacterial genera detected were exclusively associated to wastewater, and Pseudomonas was constantly the most represented genus. Moreover, isolates were assayed for their resistance against eight different antibiotics and synthetic wastewater (SWW). Data obtained revealed the presence of resistant phenotypes, including multi-drug resistant bacteria, and a general trend regarding the temporal evolution of resistance patterns: indeed, a direct correlation linking the appearance of antibiotic- and SWW-resistance with the time of exposure to wastewater was observed. In particular, nine isolates showed an interesting behavior since their growth was positively affected by the highest concentrations of SWW. Noteworthy, this study is among the few investigating the P. australis microbiota prior to the plant activation.

Among the numerous clean-up techniques for water treatment, sorption methods are widely used for the removal of trace metals. Phragmites australis is a macrophyte commonly used in constructed wetlands for water purification, and in the last decades, its use as biosorbent has attracted increasing attention. In view of a circularly economy approach, this study investigated improvement of trace metal removal by recycling the biomass of P. australis colonizing a constructed wetland, which operates as post-treatment of effluent wastewater from an activated sludge plant serving the textile industrial district of Prato (Italy). After the annual mowing of the reed plants, the biomass was dried and blended to derive a sustainable and eco-friendly biosorbent and its sorption capacity for Fe, Cu, and Zn was investigated comparing the batch system with the easier-to-handle column technique. The possibility of regeneration and reuse of the biosorbent was also evaluated. The biomaterial showed an interesting sorption capacity for Cu, Fe, and Zn, both in batch and in column experiments, especially for Fe ions. The immobilization of the biosorbent in column filters induced some improvement in the removal efficiency, and, in addition, this operation mode has the advantage of being much more suitable for practical applications than the batch process.

The objective of this paper was to define a new and reliable approach to the Ammonia Oxidizing Bacteria kinetic parameters estimation process. The proposed approach is based on the use of the information provided by a continuous on-line differential titrimeter developed for the estimation of the Actual Maximum Ammonia Oxidation Rate. The instrument was tested in a 200000 p.e. WRRF (Water Resources Recovery Facility) treating urban and industrial (textile) sewage wastewater and truck-transported liquid wastes in the Tuscan textile district (Italy). Maximum Ammonia Oxidation Rate was used as an innovative data series for maximum specific growth rate (mu(maxAOB)) and decay coefficient (b(AOB)) estimation. Titrimetric tests were conducted under non-limiting condition for oxygen and substrate and half-saturation constant of ammonia (K-NH) was calibrated referring to the effluent quality of the WRRF. The proposed procedure allows to define a new kinetic parameters data set for AOB biomass (mu(maxAOB) = 0.72 d(-1), b(AOB) = 0.25 d(-1), K-NH = 0.4 mg N-NH4+ L-1). The new data set was used to produce a new model that allows to improve the accuracy of the description of the process efficiency of the WRRF when compared with a model using kinetic parameters estimated through conventional methods.

In this work the occurrence and fate of polycyclic aromatic hydrocarbons (PAHs), phthalic acid esters (PAEs), mono and diethoxylate alkylphenols (AP1-2EOs) and alkylphenols (APs) have been investigated during a two-weeks period in a facility treating mixed textile-domestic wastewater (Prato, Italy). The wastewater treatment plant (WWTP) consists of primary sedimentation, activated sludge biological oxidation, secondary sedimentation, clariflocculation and ozonation. The sludge is treated within the facility by thickening, dewatering and final incineration, thus providing the almost quantitative removal of the adsorbed micropollutants. Naphthalene (50%), di(2-ethylhexyl) phthalate (74%) and branched 4-nonylphenols (59%) were the individual main representative compounds of each class in the influent wastewater, which showed concentration ranges of 5.6-66, 85-290 and 21-133μg/L for PAHs, PAEs and APs+AP1-2EOs, respectively. The WWTP efficiently removed PAHs, PAEs and APs+AP1-2EOs, providing effluent concentrations of 0.075-0.16ng/L 0.38-9.9μg/L and 0.53-1.4μg/L. All targeted priority and priority-hazardous micropollutants showed effluent concentrations in line with the European environmental quality standards (EQS), even though for di(2-ethylhexyl) phthalate and benzo(a)pyrene after correction for the dilution factor of the recipient. The WWTP performance was evaluated by mass balance, verifying its accuracy by monitoring Pb and Cd as conservative species. The biological treatment sections provided mass losses of 85.5%, 74.5% and 56.8% for APs+AP1-2EOs, PAEs and PAHs, highlighting efficient biotransformation performances of the activated sludge process. However, for the more volatile PAHs (e.g. naphthalene), a significant contribution of stripping cannot be excluded. A remarkable mass loss was also determined in the ozonation stage for PAEs (72.9%) and especially PAHs (97.0%), whereas a lower efficiency was observed for APs+AP1-2EOs (41.3%). The whole plant allowed for obtaining an almost quantitative removal (96.7-98.4%) for all targeted compounds.

The objective of this paper is to find the optimum solid retention time (SRT) of a wastewater treatment plant (WWTP), which minimizes operating costs, using a modeling approach with WEST software by MIKE DHI®. For the determination of the kinetic and stoichiometric parameters (used for the correct calibration of the model implemented), respirometric and kinetic batch tests were carried out. Each Oxidation ditch was modeled by a sequence of four aerated activated sludge units (ASUs) and four anoxic ASUs with recirculation. The model is able to simulate the separation efficiency of the secondary settler, which is generally quite low: in fact, the industrial origin of the wastewater induces the formation of small flocs, the dimensions of which can be further reduced by the presence of surface aerators. The nitrification/denitrification process is also accurately predicted. Using data obtained from the model, mass balances at the steady state for COD and N were made and compared to the ones obtained using measured data. After calibration and validation of the model, steady-state simulations were carried out by increasing and decreasing the SRT of the system under two different operational conditions used by the managing company and by evaluating the costs related to the water treatment line and the sludge treatment line for each scenario. It is interesting to note how the total costs are lower in summer than in winter (7.2 €cent/m3 in summer and 8.7 €cent/m3 in winter, in scenario 0). In general, the increase in the SRT led to a decrease in the total management costs. In fact, differences between scenario 0 and the scenario with the lowest total treatment costs (corresponding to an SRT of 11.4 d in winter and 10.0 d in summer) could give rise to total savings of about 44·000€/year in summer and 93·000€/year in winter.

In this paper, the design and start-up of a constructed wetland (CW) for the tertiary treatment of landfill leachates is presented. The flux is characterized by high salinity, high concentration of nitrogen (almost completely in the form of nitrate) and a biochemical oxygen demand (BOD)/chemical oxygen demand (COD) ratio close to zero. The CW pilot plant suffered from mechanical and hydraulic malfunctions which led to an uneven growth of plants inside the tanks. Despite this, COD has been reduced in the range of 0-30%, reduced forms of N (ammonia and nitrite) are also oxidized and removed by 50-80% and 20-26% on average. Considering the low number of plants and the loading rate, CW pilot plant allowed to remove more than 16 kg of COD, leading to a specific removal of 10 gCOD/d · m2. Moreover, bacterial communities associated to plants were isolated and analyzed in order to evaluate the influence of such communities on phytoremediation. Bulk soil registered the lowest bacterial titers, while plant compartments and rhizospheric soil showed to be more suitable for bacterial colonization. Twenty-five different bacterial genera were observed among the analyzed isolates, with the predominance of Pseudomonas genus.

Acquarossa river (Viterbo, Italy) was the site of a prospering Etruscan civilization thanks to metallurgical activity around 625–550 B.C. This caused the spread of heavy metals throughout the area. Rocks along the river probably act as a filter for these elements and they are covered by two different biofilms (epilithons). They differ for both color and bacterial composition. One is red and is enriched with Pseudomonas strains, while the other one is black and Acinetobacter is the most represented genus. Along the river lay the Infernaccio waterfalls, whose surrounding rocks are covered only by the red epilithon. The bacterial composition of this biofilm was analyzed through high throughput sequencing and compared to those ones of red and black epilithons of Acquarossa river. Moreover, cultivable bacteria were isolated and their phenotype (i.e., resistance against antibiotics and heavy metals) was studied. As previously observed in the case of Acquarossa river, characterization of bacterial composition of the Infernaccio red epilithon revealed that the two most represented genera were Acinetobacter and Pseudomonas. Nonetheless, these strains differed from those isolated from Acquarossa, as revealed by RAPD analysis. This work, besides increasing knowledge about the ecological properties of this site, allowed to isolate new bacterial strains, which could potentially be exploited for biotechnological applications, because of their resistance against environmental pollutants.

This research is intended to study the possible transfer of the residual chemical contamination from treated wastewaters reused for irrigation purposes of Fragaria x ananassa strawberry (cv. Camarosa). Different sewages from urban, and mixed urban-textile origins treated according to different treatment trainswere used for the irrigation of strawberry in pots. Organic and inorganic chemical contamination indicators, i.e.: PCBs, including dioxin-like congeners, PAHs, and Cr(VI), were monitored along the whole agricultural production chain (wastewater treatment effluents, soil and crop). Robust analytical procedures were specifically developed for the determination of contaminants in the above-mentioned matrices with quantitation limits (MQLs) ranging from 1.3 (Phe) to 11.0 ng/L (PCB169) for wastewaters; from 3 (PCB180) to 10 mu g/kg (BbFl) for soils; from 1.0 (Phe) to 10.9 mu g/Kg (PCB169) for strawberries. For Cr(VI), limits were 0.15 mu g/L (waters) and 0.018mg/kg (soils). These performances fully satisfy limits fixed by Italian or European regulations on maximum admitted concentration of pollutants in treated water intended for irrigation, in soils and crops. Even if selected PAHs and PCBs were detected in wastewaters (highest concentrations observed for phenanthrene, 429 mu g/L, and PCB52, 110 mu g/L) their presence was not observed in soils and in strawberries above the MQLs. On the contrary, chromium content in strawberries and soils irrigated with TWs suggested a possible transfer of the metal during irrigation, which however does not represent a hazardous situation for consumers since calculated daily intake does not exceed the Tolerable Daily Intake of 300 mu g/kg b.w.

This paper presents a case study describing a full-scale membrane bioreactor (MBR) for the pretreatment of landfill leachates. The treatment train includes an aerated equalization tank, a denitrification tank, an oxidation/nitrification tank, and two ultrafiltration units. The plant has worked continuously since 2008 treating landfill leachates at a flux of 2–11 L·h−1·m−2. The old train of membranes worked in these conditions for more than seven years prior to being damaged and replaced. The permeability (K) of the membrane varied between 30 and 80 L·h−1·m−2·bar−1 during the years of operation. In 2010, after two years of operation, the oxidation/nitrification tank was changed to work in alternate cycles of aerated and anoxic conditions, in order to improve the denitrification process. The MBR, working at a mean sludge retention time of 144 days and with mixed liquor suspended solids of 17 g/L, achieved high removal rates of conventional contaminants, with more than 98% for Biochemical Oxygen Demand, 96% for ammonium, and 75% for Chemical Oxygen Demand (COD). From the COD balance, half the COD entering was determined to be biologically oxidized into carbon dioxide, while another 24% remains in the sludge. In order to obtain these results, the company used 5.2 KWh·m−3, while spending 0.79 €·m−3.

Epilithic river biofilms characterize the rock surfaces along the Acquarossa river (Viterbo, Italy); they are in part red and in part black colored, maintaining a well-defined borderline. This peculiarity has raised questions about the biotic and abiotic phenomena that might avoid the mixing of the two biofilms. In this study, the structuring of bacterial communities in black and red epilithic biofilm in the Acquarossa river has been investigated with both culture dependent and independent approaches. Data obtained highlighted a (very) different taxonomic composition of black and red epilithons bacterial communities, dominated by Acinetobacter sp. and iron-oxidizing bacteria, respectively. The chemical characterization of both river water and biofilms revealed a substantial heavy metals pollution of the environment; heavy metals were also differentially accumulated in red and black epilithons. Overall, our data revealed that the structuring of red and black epilithons might be affected mainly by the antagonistic interactions exhibited by bacterial genera dominating the two biofilms. These findings suggest that biotic factors might be responsible for the structuring of natural bacterial communities, suggesting that there is a selection of populations at very small scale, and that different populations might compete for different niches.

The textile sector plays a critical role in this context, because textile companies are large users of water - typically 0.2-0.5 m3 are needed to produce 1 kg of finished product [1] – and they are historically concentrated in districts (e.g. Prato in Italy) located in Southern Member States, such as Italy, Spain and Portugal, which are facing more and more longer periods of drought and therefore are strongly in the need to preserve the availability of water for potable use. The finishing processes (i.e. pre-treatment, dyeing, printing and special finishing) are especially polluting and resource consuming in terms of water, energy and chemicals. Almost the entire of water used in textile production is discharged and, on average, the effluents volume covers 90-95% of the water used [2]. According to that, a new Ultrafiltration/Sonichemical combined process has been investigated on textile wastewater by evaluating the reduction of the pollution load induced by physical separation and by chemical oxidation promoted by Ultrasound cavitation. The experimental campaign carried out at the Municipal Wastewater treatment Plant in Prato showed that the quality of the treated water reaches the target reported in literature [3] for textile manufacturing sectors. An estimation of the costs of the purified water has been assessed and the combined process seems to be promising for further development. The authors would like to thank the European Commission to funding this project (contract number LIFE07/IT/000439) within the LIFE+ Environmental Policy and Governance 2007 Programme.

The occurrence of pharmaceuticals and other hazardous substances in surface waters and drinking water and the long-term exposure may represent a potential risk for both the environment and human health. Many studies have evidenced that conventional technologies used for wastewater treatment do not completely remove pharmaceuticals and personal care products (PPCPs) residues, which can be detected in receiving waters at concentrations ranging from few ng L-1 until more than mu kg L-1. Nanomaterials are of fundamental importance in the current research efforts to develop more efficient water treatment and remediation systems in place of conventional technologies. This review provides a comprehensive overview of the most relevant works available in literature reporting the use of nanosorbents (nanotubes and zeolites), nanofiltration (NF) and advanced oxidation processes (AOPs) for the removal of PPCPs compounds from water and wastewater. The literature review of laboratory- and pilot-scale studies have shown that nanomaterials are promising tools in environmental cleaning processes and water purification, even if more researches are necessary.