Growth of Rhodococcus sp. strain BCP1 on gaseous n-alkanes: New metabolic insights and transcriptional analysis of two soluble di-iron monooxygenase genes

Rhodococcus sp. strain BCP1 was initially isolated for its ability to grow on gaseous n-alkanes, which act as inducers for the co-metabolic degradation of low-chlorinated compounds. Here, both molecular and metabolic features of BCP1 cells grown on gaseous and short-chain n-alkanes (up to n-heptane) were examined in detail. We show that propane metabolism generated terminal and sub-terminal oxidation products such as 1- and 2-propanol, whereas 1-butanol was the only terminal oxidation product detected from n-butane metabolism. Two gene clusters, prmABCD and smoABCD-coding for Soluble Di-Iron Monooxgenases (SDIMOs) involved in gaseous n-alkanes oxidation-were detected in the BCP1 genome. By means of Reverse Transcriptase-quantitative PCR (RT-qPCR) analysis, a set of substrates inducing the expression of the sdimo genes in BCP1 were assessed as well as their transcriptional repression in the presence of sugars, organic acids, or during the cell growth on rich medium (Luria-Bertani broth). The transcriptional start sites of both the sdimo gene clusters were identified by means of primer extension experiments. Finally, proteomic studies revealed changes in the protein pattern induced by growth on gaseous- (n-butane) and/or liquid (n-hexane) short-chain n-alkanes as compared to growth on succinate. Among the differently expressed protein spots, two chaperonins and an isocytrate lyase were identified along with oxidoreductases involved in oxidation reactions downstream of the initial monooxygenase reaction step

Continuous flow adsorption of phenolic compounds from olive mill wastewater with resin XAD16N: life cycle assessment, cost\u2013benefit analysis and process optimization

BACKGROUND: Olive mill wastewaters (OMWs) represent a major environmental concern due to their high organic load and phytotoxic activity. The selective recovery of phenolic compounds (PCs) from OMW is promising, thanks to the antioxidant and antimicrobial properties of PCs. The goal of this work was to perform a life cycle assessment (LCA) and cost\u2013benefit analysis (CBA) of a full-scale process of PC adsorption/desorption on resin Amberlite XAD16N. The industrial process was designed on the basis of laboratory tests aimed at performing a preliminary process optimization. RESULTS: Adsorption tests were conducted at different velocities in a 1.8-m column packed with XAD16N. The optimal superficial velocity and retention time (2.78 m h \u20131 and 0.56 h) allowed the attainment of satisfactory performances in terms of resin operating capacity (0.46), PC adsorption yield (0.92), PC mass fraction in the sorbed product (0.50 g PC /g VS ) and specific antioxidant activity (3\u20136 g ascorbic acid /g PC ). Six consecutive adsorption/desorption cycles, operated with the same resin load, resulted in stable process performances. The LCA indicated that the environmental impact of the process could be decreased markedly through the addition of an anaerobic digestion step for the production of irrigation-quality water and fertilizers from the dephenolized OMW. The PC market price required for the generation of a positive business case resulted relatively low (\u20ac1.7\u201313.5 kg PC\u20131 ). CONCLUSION: The results indicate that the proposed PC adsorption/desorption technology, if integrated with an anaerobic digestion step, represents a promising solution for the treatment and valorization of OMW, a major agro-industrial waste in Mediterranean countries

A new Rhodococcus aetherivorans strain isolated from lubricant-contaminated soil as a prospective phenol biodegrading agent

Microbe-based decontamination of phenol-polluted environments has significant advantages over physical and chemical approaches by being relatively cheaper and ensuring complete phenol degradation. There is a need to search for commercially prospective bacterial strains that are resistant to phenol and other co-pollutants, e.g. oil hydrocarbons, in contaminated environments, and able to carry out efficient phenol biodegradation at a variable range of concentrations. This research characterizes the phenol-biodegrading ability of a new actinobacteria strain isolated from a lubricant-contaminated soil environment. Phenotypic and phylogenetic analyses showed that the novel strain UCM Ac-603 belonged to the species Rhodococcus aetherivorans, and phenol degrading ability was quantitatively characterized for the first time. R. aetherivorans UCM Ac-603 tolerated and assimilated phenol (100% of supplied concentration) and various hydrocarbons (56.2–94.4%) as sole carbon sources. Additional nutrient supplementation was not required for degradation and this organism could grow at a phenol concentration of 500 mg L −1 without inhibition. Complete phenol assimilation occurred after 4 days at an initial concentration of 1750 mg L −1 for freely-suspended cells and at 2000 mg L −1 for vermiculite-immobilized cells: 99.9% assimilation of phenol was possible from a total concentration of 3000 mg L −1 supplied at daily fractional phenol additions of 750 mg L −1 over 4 days. In terms of phenol degradation rates, R. aetherivorans UCM Ac-602 showed efficient phenol degradation over a wide range of initial concentrations with the rates (e.g. 35.7 mg L −1 h −1 at 500 mg L −1 phenol, and 18.2 mg L −1 h −1 at 1750 mg L −1 phenol) significantly exceeding (1.2–5 times) reported data for almost all other phenol-assimilating bacteria. Such efficient phenol degradation ability compared to currently known strains and other beneficial characteristics of R. aetherivorans UCM Ac-602 suggest it is a promising candidate for bioremediation of phenol-contaminated environments. </p

Development of a continuous-flow anaerobic co-digestion process of olive mill wastewater and municipal sewage sludge

BACKGROUND: Olive mill wastewater (OMW) represents an environmental problem due to its high organic load and relevant concentration of phenolic compounds (PCs). OMW treatment and disposal represents a relevant challenge and cost for olive mills and multi-utilities in charge of waste management in Mediterranean countries. The goal of this study was to develop an anaerobic co-digestion (co-AD) process of OMW and sewage sludge (SwS) from municipal wastewater treatment. RESULTS: Different volumetric OMW:SwS ratios up to 100% OMW were fed in continuous 1.7-L bioreactors. The reactors fed with raw OMW (rOMW) performed better than those fed with OMW dephenolized by adsorption (dOMW). At a 23-day hydraulic retention time, the best performances were obtained in the reactor fed with 25% rOMW, with a 105% increase in methane yield in comparison to the 100% SwS test. At a 40-day hydraulic retention time, the reactor fed with 40% rOMW attained a 268 NLCH4/kgvolatile solids methane yield. The conversion of phenolic compoundsreached 70% when the hydraulic retention time was increased from 23 to 40 days. A cost\u2013benefit analysis indicated that both rOMW co-AD in existing digesters and phenolic compounds recovery from OMW followed by co-AD of dOMW can lead to relevant additional revenues for the multi-utilities in charge of wastewater management. CONCLUSION: This work proves that, using the existing network of SwS anaerobic digesters, it is feasible to co-digest the entire OMW production in regions characterized by intense olive oil production, thus attaining a relevant increase in methane production yield (a 144% increase in comparison to 100% SwS)

Unravelling the role of the group 6 soluble di-iron monooxygenase (SDIMO) SmoABCD in alkane metabolism and chlorinated alkane degradation

Soluble di-iron monooxygenases (SDIMOs) are multi-component enzymes catalysing the oxidation of various substrates. These enzymes are characterized by high sequence and functional diversity that is still not well understood despite their key role in biotechnological processes including contaminant biodegradation. In this study, we analysed a mutant of Rhodoccocus aetherivorans BCP1 (BCP1-2.10) characterized by a transposon insertion in the gene smoA encoding the alpha subunit of the plasmid-located SDIMO SmoABCD. The mutant BCP1-2.10 showed a reduced capacity to grow on propane, lost the ability to grow on butane, pentane and n-hexane and was heavily impaired in the capacity to degrade chloroform and trichloroethane. The expression of the additional SDIMO prmABCD in BCP1-2.10 probably allowed the mutant to partially grow on propane and to degrade it, to some extent, together with the other short-chain n-alkanes. The complementation of the mutant, conducted by introducing smoABCD in the genome as a single copy under a constitutive promoter or within a plasmid under a thiostreptone-inducible promoter, allowed the recovery of the alkanotrophic phenotype as well as the capacity to degrade chlorinated n-alkanes. The heterologous expression of smoABCD allowed a non-alkanotrophic Rhodococcus strain to grow on pentane and n-hexane when the gene cluster was introduced together with the downstream genes encoding alcohol and aldehyde dehydrogenases and a GroEL chaperon. BCP1 smoA gene was shown to belong to the group 6 SDIMOs, which is a rare group of monooxygenases mostly present in Mycobacterium genus and in a few Rhodococcus strains. SmoABCD originally evolved in Mycobacterium and was then acquired by Rhodococcus through horizontal gene transfer events. This work extends the knowledge of the biotechnologically relevant SDIMOs by providing functional and evolutionary insights into a group 6 SDIMO in Rhodococcus and demonstrating its key role in the metabolism of short-chain alkanes and degradation of chlorinated n-alkanes

Development and validation of an adsorption process for phosphate removal and recovery from municipal wastewater based on hydrotalcite-related materials

In the current international context characterized by the tendency to stricter limits for P concentration in treated wastewater and a strong drive towards phosphate recovery, it is crucial to develop cost-effective technologies to remove and recover phosphate from municipal wastewater (MWW). In this study, an initial screening of the phosphate adsorption performances of 9 sorbents including several hydrotalcites led to the selection of calcined pyroaurite - an innovative material composed of mixed Mg/Fe oxides - as the best-performing one. The assessment of calcined pyroaurite by means of isotherms and continuous-flow adsorption/desorption tests conducted with actual MWW resulted in a high P sorption capacity (12 mgP g- 1 at the typical phosphate concentration in MWW), the capacity to treat 730 BVs at the 1 mgP L- 1 breakpoint imposed by the current EU legislation, and a 93 % phosphate recovery. Calcined pyroaurite resulted in satisfactory performances also in a test conducted with a saline MWW deriving from a hotspot of seawater intrusion, a rapidly increasing phenomenon as a result of climate change. Five consecutive adsorption/desorption cycles conducted in a 20-cm column at a 5-min empty bed contact time resulted stable in terms of P adsorption/recovery performances, specific surface area and chemical structure of calcined pyroaurite. In the perspective to apply phosphate recovery with calcined pyroaurite at full scale, the process scale-up to a 60-cm packed bed - close to the column heights of industrial applications- resulted in stable performances. Calcium phosphate, widely used to produce phosphate-based fertilizers, can be obtained from the desorbed product by precipitation with Ca(OH)2. 2 . These results point to calcined pyroaurite as a very promising material for phosphate removal and recovery from MWW and from other P-rich effluents in a circular economy perspective

Development of an ion exchange process for ammonium removal and recovery from municipal wastewater using a metakaolin K-based geopolymer

Ion exchange represents a promising process for ammonium removal from municipal wastewater (MWW), in order to recover it for fertilizer production. Previous studies on ammonium ion exchange neglected the assessment of process robustness and the optimization the desorption/recovery step. This study aimed at developing a continuous-flow process of ammonium removal/recovery based on a metakaolin K-based geopolymer, named G13. Process robustness was assessed by operating 7 adsorption/desorption cycles with two types of MWW. These tests resulted in satisfactory and constant performances: operating capacity at 40 mgN L-1 in the inlet&nbsp;=&nbsp;12 mgN gdry sorbent-1, bed volumes of treated MWW at the selected breakpoint&nbsp;=&nbsp;199-226, ammonium adsorption yield&nbsp;=&nbsp;88-91%. Empty bed contact time (EBCT) was decreased from 10 to 5&nbsp;min without any reduction in performances. The NH4+ adsorption process was effectively simulated by the Thomas model, allowing a model-based assessment of the effect of EBCT reductions on process performances. An innovative desorption procedure led to high ammonium recovery yields (86-100%) and to a desorbed product composed primarily of KNO3 (54%w) and NH4NO3 (39%w), two salts largely used in commercial fertilizers. The energy consumption of ammonium removal/recovery with G13 resulted 0.027 kWh m-3treated WW, with a relevant reduction in comparison to traditional nitrification/denitrification, whereas the operational cost resulted equal to 60-110% of the cost of the benchmark process. These results show that G13 is a promising material to recover ammonium in a circular economy approach

Comparative preliminary evaluation of two in-stream water treatment technologies for the agricultural reuse of drainage water in the Nile delta

In the Nile Delta, a complex network of canals collects drainage water from surface-irrigated fields, but also municipal wastewater. The goal of this work was to assess the technical, environmental and financial feasibility of the upgrade of a drainage canal (DC) into either an in-stream constructed wetland (ICW) or a canalized facultative lagoon (CFL), in order to produce a water re-usable in agriculture according to the Egyptian law. The model-based design of the proposed technologies was derived from field experimental data for the ICW and laboratory data for the CFL. Both technologies, integrated by a sedimentation pond and a disinfection canal, led to the attainment of the water quality standards imposed by Egyptian Law 92/2013 for the reuse of drainage water. The life cycle assessment indicated that the upgrade of an existing DC to either an ICW or a CFL results in an extremely small environmental burden, 64 0.3% of that of a traditional activated sludge process. The cost/benefit analysis (CBA) was based on the assumptions that (i) farmers currently irrigate a non-food crop (cotton) with the low-quality drainage water present in the DC, and (ii) thanks to the upgrade to a ICW or CFL, farmers will irrigate a food crop characterized by a higher market price (rice). The CBA indicated that the DC upgrade to an ICW represents an attractive investment, as it leads to a financial rate of return &gt; 10% over a wide range of cotton market prices. Conversely, the upgrade to a CFL is less attractive due to high investment costs. In conclusion, the upgrade of DCs to ICWs appears a promising option for the treatment of drainage canal water in the Nile Delta, thanks to the high pollutant removal performances, low cost and negligible environmental burden. This article is protected by copyright. All rights reserved

Establishing correlations between time series of wastewater parameters under extreme and regular weather conditions

This study investigates the correlations between key wastewater parameters - water level, turbidity, and electrical conductivity - under varying weather conditions, including extreme rainfall events such as the May 2023 flood event in Bologna, Italy. Data collected via IoT-based sensors are analyzed using Detrended Cross-Correlation Analysis and Autoregressive Distributed Lag (ARDL) models. The results highlight significant correlations between water level and other parameters, with distinct patterns emerging during extreme and regular weather periods. Notably, water level correlates negatively with electrical conductivity, particularly during flood events, due to the dilution effect of rainwater. Turbidity shows a complex relationship with water level, influenced by weather conditions and the opposing effects of different factors. ARDL models further demonstrate the potential to predict turbidity and electrical conductivity from water level data, offering valuable insights for wastewater management in urban areas

Negotiated matter: a robotic exploration of craft-driven innovation

This paper introduces a novel approach to craft-driven robotic innovation in architectural research. Here craft is not portrayed as a source of ornamental or historical inspiration, but instead as an open-ended process described by a framework involving material properties, diverging modes of knowledge production and representation, emergent tectonic configurations and embodied interaction with technology. To do so, this paper firstly contrasts a definition of craft (Pye 1968) with practices of robotic architectural production. Additionally, the notion of emergent tectonics resulting from negotiated material and technological processes is addressed by critically situating the theories of architectural tectonics by Kenneth Frampton (2001) and digital tectonics by Leach, Turnbull and Williams (2004) in the context of robotic fabrication in architecture. Finally, the ongoing project “Computing Craft” is presented as a case study illustrating a proposed framework for robotic craft-driven innovation