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Polyhydroxyalkanoates (PHAs) are well-known biodegradable plastics produced by various bacterial strains, whose major drawback is constituted by the high cost of their synthesis. Producing PHAs from mixed microbial cultures and employing organic wastes as a carbon source allows us to both reduce cost and valorize available renewable resources, such as food waste and sewage sludge. However, different types of pollutants, originally contained in organic matrices, could persist into the final product, thus compromising their safety. In this work, the exploitation of municipal wastes for PHA production is evaluated from the environmental and health safety aspect by determining the presence of polycyclic aromatic hydrocarbons (PAHs) in both commercial and waste-based PHA samples. Quantification of PAHs by gas chromatography-mass spectrometry on 24 PHA samples obtained in different conditions showed very low contamination levels, in the range of ppb to a few ppm. Moreover, the contaminant content seems to be dependent on the type of PHA stabilization and extraction, but independent from the type of feedstock. Commercial PHA derived from crops, selected for comparison, showed PAH content comparable to that detected in PHAs derived from organic fraction of municipal solid waste. Although there is no specific regulation on PAH maximum levels in PHAs, detected concentrations were consistently lower than threshold limit values set by regulation and guidelines for similar materials and/or applications. This suggests that the use of organic waste as substrate for PHA production is safe for both the human health and the environment.

Polyhydroxyalkanoates (PHAs) production at pilot scale has been recently investigated and carried out exploiting different process configurations and organic wastes. More in detail, three pilot platforms, in Treviso (North-East of Italy), Carbonera (North-East of Italy) and Lisbon, produced PHAs by open mixed microbial cultures (MMCs) and different organic waste streams: organic fraction of municipal solid waste and sewage sludge (OFMSW-WAS), cellulosic primary sludge (CPS), and fruit waste (FW), respectively. In this context, two stabilization methods have been applied, and compared, for preserving the amount of PHA inside the cells: thermal drying and wet acidification of the biomass at the end of PHA accumulation process. Afterward, polymer has been extracted following an optimized method based on aqueous-phase inorganic reagents. Several PHA samples were then characterized to determine PHA purity, chemical composition, molecular weight, and thermal properties. The polymer contained two types of monomers, namely 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) at a relative percentage of 92.6–79.8 and 7.4–20.2 w/w, respectively, for Treviso and Lisbon plants. On the other hand, an opposite range was found for 3HB and 3HV monomers of PHA from Carbonera, which is 44.0–13.0 and 56.0–87.0 w/w, respectively. PHA extracted from wet-acidified biomass had generally higher viscosity average molecular weights ( M v ) (on average 424.8 ± 20.6 and 224.9 ± 21.9 KDa, respectively, for Treviso and Lisbon) while PHA recovered from thermally stabilized dried biomass had a three-fold lower M v .

Volatile fatty acid (VFA) rich streams from fermentation of organic residuals and wastewater are suitable feedstocks for mixed microbial culture (MMC) Polyhydroxyalkanoate (PHA) production. However, many such streams have low total VFA concentration (1–10 gCOD/L). PHA accumulation requires a flow-through bioprocess if the VFAs are not concentrated. A flow through bioprocess must balance goals of productivity (highest possible influent flow rates) with goals of substrate utilization efficiency (lowest possible effluent VFA concentration). Towards these goals, dynamics of upshift and downshift respiration kinetics for laboratory and pilot scale MMCs were evaluated. Monod kinetics described a hysteresis between the upshift and downshift responses. Substrate concentrations necessary to stimulate a given substrate uptake rate were significantly higher than the concentrations necessary to sustain the attained substrate uptake rate. A benefit of this hysteresis was explored in Monte Carlo based PHA accumulation bioprocess numerical simulations. Simulations illustrated for a potential to establish continuous flow-through PHA production bioprocesses even at a low (1 gCOD/L) influent total VFA concentration. Process biomass recirculation into an engineered higher substrate concentration mixing zone, due to the constant influent substrate flow, enabled to drive the process to maximal possible PHA production rates without sacrificing substrate utilization efficiency.

Actions for improving water quality are critical and include the remediation of polluted groundwater. The effectiveness of the remediation strategy to remove contamination by chlorinated solvents may be increased by combining physicochemical treatments (i.e., adsorption) and biological degradation (i.e., biological reductive dechlorination (BRD)). Recent studies have shown the potentialities of bio-based materials for bioremediation purposes, including polyhydroxybutyrate (PHB), a biodegradable microbial polyester tested as a fermentable source of slow-release electron donors. Further, a low-cost biochar derived from the pyrolysis of pinewood waste (PWB), used as sorbent material, has recently been proposed to accelerate reductive microbial dehalogenation. Here, we propose a coupled adsorption and biodegradation (CAB) process for trichloroethylene (TCE) removal in a mini pilot-scale reactor composed of two reactive zones, the first one filled with PHB and the second one with PWB. This work aimed to evaluate the performance of the CAB process with particular regard to the effectiveness of the PWB in sustaining the biofilm, mostly enriched by Dehalococcoides mccartyi. The main results showed the CAB system treated around 1300 L of contaminated water, removing 102 mg TCE per day. Combining PHB and PWB had a positive effect on the growth of the dechlorinating community with a high abundance of Dhc cells.

Groundwater remediation is one of the main objectives to minimize environmental impacts and health risks. Chlorinated aliphatic hydrocarbons contamination is prevalent and presents particularly challenging scenarios to manage with a single strategy. Different technologies can manage contamination sources and plumes, although they are usually energy-intensive processes. Interesting alternatives involve in-situ bioremediation strategies, which allow the chlorinated contaminant to be converted into non-toxic compounds by indigenous microbial activity. Despite several advantages offered by the bioremediation approaches, some limitations, like the relatively low reaction rates and the difficulty in the management and control of the microbial activity, can affect the effectiveness of a bioremediation approach. However, those issues can be addressed through coupling different strategies to increase the efficiency of the bioremediation strategy. This mini review describes different strategies to induce the reduction dechlorination reaction by the utilization of innovative strategies, which include the increase or the reduction of contaminant mobility as well as the use of innovative strategies of the reductive power supply. Subsequently, three future approaches for a greener and more sustainable intervention are proposed. In particular, two bio-based materials from renewable resources are intended as alternative, long-lasting electron-donor sources (e.g., polyhydroxyalkanoates from mixed microbial cultures) and a low-cost adsorbent (e.g., biochar from bio-waste). Finally, attention is drawn to novel bio-electrochemical systems that use electric current to stimulate biological reactions.

In this work, polyhydroxy butyrate (PHB) and biochar from pine wood (PWB) are used in a mini-pilot scale biological reactor (11.3 L of geometric volume) for trichloroethylene (TCE) removal (80 mgTCE/day and 6 L/day of flow rate). The PHB-biochar reactor was realized with two sequential reactive areas to simulate a multi-reactive permeable barrier. The PHB acts as an electron donor source in the first “fermentative” area. First, the thermogravimetric (TGA) and differential scanning calorimetry (DSC) analyses were performed. The PHB-powder and pellets have different purity (96% and 93% w/w) and thermal properties. These characteristics may affect the biodegradability of the biopolymer. In the second reactive zone, the PWB works as a Dehalococcoides support and adsorption material since its affinity for chlorinated compounds and the positive effect of the “coupled adsorption and biodegradation” process has been already verified. A specific dechlorinating enriched culture has been inoculated in the PWB zone to realize a coupled adsorption and biodegradation process. Organic acids were revealed since the beginning of the test, and during the monitoring period the reductive dichlorination anaerobic pathway was observed in the first zone; no chlorinated compounds were detected in the effluent thanks to the PWB adsorption capacity.

This study uses a membrane‐less reactor to explore the bioelectrochemical remediation of real contaminated groundwater from chlorinated aliphatic hydrocarbons (CAHs) and nitrates. The research focuses on testing a column‐type bioelectrochemical reactor to stimulate in situ degradation of contaminants through the supply of electrons by a graphite granules biocathode. After a preliminary laboratory characterization and operation with a synthetic feeding solution, a field test is conducted in a real contaminated site, where the reactor demonstrates effective degradation of CAHs and inorganic anions. Notably, the cathodic potential promotes the reductive dechlorination of chlorinated species. Simultaneously, nitrate reduction, sulfate reduction, and methanogenesis occurr, influencing the overall coulombic efficiency of the process. The use of real groundwater, compared to the synthetic medium, significantly decreases the coulombic efficiency of reductive dechlorination, dropping from 2.43% to 0.01%. Concentration profiles along the bioelectrochemical reactor allow for a deeper description of the reductive dechlorination rate at different flow rates, as well as increase the knowledge about reduction and oxidation mechanisms. Scaling up the technology presents several challenges, including the optimization of coulombic efficiency and the management of competing microbial metabolisms. The study provides a valuable contribution toward advancing bioelectrochemical technologies for the bioremediation of complex contaminated sites.

This study explores the use of pine wood biochar (BC) waste gasified at 950 °C as fillers in polymer matrices to create BC@biopolymer composites with perspectives in groundwater remediation. Four biochar samples underwent different sieving and grinding processes and were extensively characterized via UV–Vis, FTIR, and FESEM–EDS, highlighting the fact that that BCs are essentially graphitic in nature with a sponge-like morphology. The grinding process influences the particle size, reducing the specific surface area by about 30% (evaluated by BET). The adsorption performances of raw BC were validated via an adsorption isotherm using trichloroethylene (TCE) as a model contaminant. A selected BC sample was used to produce hydrophilic, stable polymer composites with chitosan (CS), alginate (ALG), potato starch (PST), and sodium carboxymethylcellulose (CMC) via a simple blending approach. Pilot sedimentation tests over 7 days in water identified BC@PST and BC@CMC as the most stable suspensions due to a combination of both hydrogen bonds and physical entrapment, as studied by FTIR. BC@CMC showed optimal distribution and retention properties without clogging in breakthrough tests. The study concludes that biopolymer-based biochar composites with improved stability in aqueous environments hold significant promise for addressing various groundwater pollution challenges.

In the context of the surfactant-enhanced remediation of polluted sites, this work focuses on the development of non-ionic alkylpolyglucosidic (APG) surfactant formulations with different additives for the solubilization and mobilization of strongly adsorbed organic pollutants. The study involved three consecutive phases to evaluate the effect of the additives on surface behavior and the potential improvement in alkylpolyglycoside surfactant’s capability to solubilize toluene and perchloroethylene (PCE), selected as reference contaminants. After a chemical–physical characterization phase, the APG-based formulations were first used in a batch configuration test, in which the formulations’ solubilization ability was indirectly assessed by observing the effect on pollutants’ adsorption. Lastly, a continuous configuration column experiment was performed to simulate the flushing process of a synthetic matrix previously contaminated with strongly adsorbed toluene or PCE. The results showed that the presence of additives firstly reduced the ability of the surfactant to form micelles, increasing the CMC, but at the same time improved the ability to reduce surface tension. Moreover, the addition of the additives overall resulted in a significant improvement in adsorbed pollutant removal in a minimal volume of fed solution, reaching 96% and 99% efficiencies for toluene and PCE, respectively, compared with 76% and 92%, for toluene and PCE, respectively, in the presence of free-additive APG surfactant.

Volatile fatty acids obtained from the fermentation of the organic fraction of municipal solid waste can be used as raw materials for non-toxic ethyl ester (EE) synthesis as well as feedstock for the production of polyhydroxyalkanoates (PHAs). Taking advantage of the concept of an integrated process of a bio-refinery, in the present paper, a systematic investigation on the extraction of intracellular poly(3-hydroxybutyrate-co-3-hydroxyvalerate), produced by mixed microbial culture by using EEs was reported. Among the tested EEs, ethyl acetate (EA) was the best solvent, dissolving the copolymer at the lowest temperature. Then, extraction experiments were carried out by EA at different temperatures on two biomass samples containing PHAs with different average molecular weights. The parallel characterization of the extracted and non-extracted PHAs evidenced that at the lower temperature (100 °C) EA solubilizes preferentially the polymer fractions richer in 3HV comonomers and with the lower molecular weight. By increasing the extraction temperature from 100 °C to 125 °C, an increase of recovery from about 50 to 80 wt% and a molecular weight reduction from 48% to 65% was observed. The results highlighted that the extracted polymer purity is always above 90 wt% and that it is possible to choose the proper extraction condition to maximize the recovery yield at the expense of polymer fractionation and degradation at high temperatures or use milder conditions to maintain the original properties of a polymer.