Wastewater from textile digital printing as a substrate for microalgal growth and valorization.

This study aims at evaluating an innovative biotechnological process for the concomitant bioremediation and valorization of wastewater from textile digital printing technology based on a microalgae/bacteria consortium. Nutrient and colour removal were assessed in lab-scale batch and continuous experiments and the produced algae/bacteria biomass was characterized for pigment content and biomethane potential. Microbial community analysis provided insight of the complex community structure responsible for the bioremediation action. Specifically, a community dominated by Scenedesmus spp. and xenobiotic and dye degrading bacteria was naturally selected in continuous photobioreactors. Data confirm the ability of the microalgae/bacteria consortium to grow in textile wastewater while reducing the nutrient content and colour. Improvement strategies were eventually identified to foster biomass growth and process performances. The experimental findings pose the basis of the integration of a microalgal-based process into the textile sector in a circular economy perspective.

Microbial community and performance of a partial nitritation/anammox sequencing batch reactor treating textile wastewater.

Implementation of onsite bioremediation technologies is essential for textile industries due to rising concerns in terms of water resources and quality. Partial nitritation-anaerobic ammonium oxidation (PN/A) processes emerged as a valid, but unexplored, solution. In this study, the performance of a PN/A pilot-scale (9 m3) sequencing batch reactor treating digital textile printing wastewater (10-40 m3 d-1) was monitored by computing nitrogen (N) removal rate and efficiencies. Moreover, the structure of the bacterial community was assessed by next generation sequencing and quantitative polymerase chain reaction (qPCR) analyses of several genes, which are involved in the N cycle. Although anaerobic ammonium oxidation activity was inhibited and denitrification occurred, N removal rate increased from 16 to 61 mg N g VSS-1 d-1 reaching satisfactory removal efficiency (up to 70%). Ammonium (18-70 mg L-1) and nitrite (16-82 mg L-1) were detected in the effluent demonstrating an unbalance between the aerobic and anaerobic ammonia oxidation activity, while constant organic N was attributed to recalcitrant azo dyes. Ratio between nitrification and anammox genes remained stable reflecting a constant ammonia oxidation activity. A prevalence of ammonium oxidizing bacteria and denitrifiers suggested the presence of alternative pathways. PN/A resulted a promising cost-effective alternative for textile wastewater N treatment as shown by the technical-economic assessment. However, operational conditions and design need further tailoring to promote the activity of the anammox bacteria.

Assessment of anammox, microalgae and white-rot fungi-based processes for the treatment of textile wastewater.

The treatability of seven wastewater samples generated by a textile digital printing industry was evaluated by employing 1) anammox-based processes for nitrogen removal 2) microalgae (Chlorella vulgaris) for nutrient uptake and biomass production 3) white-rot fungi (Pleurotus ostreatus and Phanerochaete chrysosporium) for decolorization and laccase activity. The biodegradative potential of each type of organism was determined in batch tests and correlated with the main characteristics of the textile wastewaters through statistical analyses. The maximum specific anammox activity ranged between 0.1 and 0.2 g N g VSS-1 d-1 depending on the sample of wastewater; the photosynthetic efficiency of the microalgae decreased up to 50% during the first 24 hours of contact with the textile wastewaters, but it improved from then on; Pleurotus ostreatus synthetized laccases and removed between 20-62% of the colour after 14 days, while the enzymatic activity of Phanerochaete chrysosporium was inhibited. Overall, the findings suggest that all microbes have great potential for the treatment and valorisation of textile wastewater after tailored adaptation phases. Yet, the depurative efficiency can be probably enhanced by combining the different processes in sequence.

Performance Analysis and Microbial Community Evolution of In Situ Biological Biogas Upgrading with Increasing H2/CO2 Ratio.

The effect of the amount of hydrogen supplied for the in situ biological biogas upgrading was investigated by monitoring the process and evolution of the microbial community. Two parallel reactors, operated at 37°C for 211 days, were continuously fed with sewage sludge at a constant organic loading rate of 1.5 gCOD∙(L∙d)-1 and hydrogen (H2). The molar ratio of H2/CO2 was progressively increased from 0.5 : 1 to 7 : 1 to convert carbon dioxide (CO2) into biomethane via hydrogenotrophic methanogenesis. Changes in the biogas composition become statistically different above the stoichiometric H2/CO2 ratio (4 : 1). At a H2/CO2 ratio of 7 : 1, the methane content in the biogas reached 90%, without adversely affecting degradation of the organic matter. The possibility of selecting, adapting, and enriching the original biomass with target-oriented microorganisms able to biologically convert CO2 into methane was verified: high throughput sequencing of 16S rRNA gene revealed that hydrogenotrophic methanogens, belonging to Methanolinea and Methanobacterium genera, were dominant. Based on the outcomes of this study, further optimization and engineering of this process is feasible and needed as a means to boost energy recovery from sludge treatment.

Autotrophic nitrogen removal for decentralized treatment of ammonia-rich industrial textile wastewater: process assessment, stabilization and modelling.

Digital textile printing (DTP) is a game-changer technology that is rapidly expanding worldwide. On the other hand, process wastewater is rich in ammoniacal and organic nitrogen, resulting in relevant issues for discharge into sewer system and treatment in centralized plants. The present research is focused on the assessment of the partial nitritation/anammox process in a single-stage granular sequencing batch reactor for on-site decentralized treatment. The technical feasibility of the process was assessed by treating wastewater from five DTP industries in a laboratory-scale reactor, in one case investigating long-term process stabilization. While experimental results indicated nitrogen removal efficiencies up to about 70%, complying with regulations on discharge in sewer system, these data were used as input for process modelling, whose successful parameter calibration was carried out. The model was applied to the simulation of two scenarios: (i) the current situation of a DTP company, in which wastewater is discharged into the sewer system and treated in a centralized plant, (ii) the modified situation in which on-site decentralized treatment for DTP wastewater is implemented. The second scenario resulted in significant improvements, including reduced energy consumption (- 15%), reduced greenhouse gases emission, elimination of external carbon source for completing denitrification at centralized WWTP and reduced sludge production (- 25%).

Comparison of the Performance and Microbial Community Structure of Two Outdoor Pilot-Scale Photobioreactors Treating Digestate.

This study aimed at examining and comparing the nutrient removal efficiency, biomass productivity and microbial community structure of two outdoor pilot-scale photobioreactors, namely a bubble column and a raceway pond, treating the liquid fraction of an agricultural digestate. Bacterial and eukaryotic communities were characterized using a metabarcoding approach and quantitative PCR. The abundance, composition, diversity, and dynamics of the main microbes were then correlated to the environmental conditions and operational parameters of the reactors. Both photobioreactors were dominated either by Chlorella sp. or Scenedesmus sp. in function of temperature, irradiance and the nitrogen compounds derived by nitrification. Other species, such as Chlamydomonas and Planktochlorella, were sporadically present, demonstrating that they have more specific niche requirement. Pseudomonas sp. always dominated the bacterial community in both reactors, except in summertime, when a bloom of Calothrix occurred in the raceway pond. In autumn, the worsening of the climate conditions decreased the microalgal growth, promoting predation by Vorticella sp. The study highlights the factors influencing the structure and dynamics of the microbial consortia and which ecological mechanisms are driving the microbial shifts and the consequent reactor performance. On these bases, control strategies could be defined to optimize the management of the microalgal-based technologies.

Outdoor pilot-scale raceway as a microalgae-bacteria sidestream treatment in a WWTP.

This study aims at demonstrating the feasibility of using microalgae-bacteria consortia for the treatment of the sidestream flow of the supernatant from blackwater dewatering (centrate) in an urban wastewater treatment plant in Northern Italy. A 1200 L raceway reactor was used for the outdoor cultivation of a diverse community of Chlorella spp., Scenedesmus spp. and Chlamydomonas spp. in continuous operation mode with 10 days hydraulic retention time. During the trial, an average daily areal productivity of 5.5 ± 7.4 g TSS m-2 day-1 was achieved while average nutrient removal efficiencies were 86% ± 7% and 71% ± 10% for NH4-N and PO4-P, respectively. The microalgal nitrogen assimilation accounted for 10% of the nitrogen in the centrate while 34% was oxidized to nitrite and nitrate. The oxygen produced by microalgae fully covert the oxygen demand for nitrification. This suggests that the proposed process would reduce the aeration demand for nitrification in the water line of the plant, while producing algal biomass to be further valorized for energy or material recovery.

Metataxonomy and functionality of wood-tar degrading microbial consortia.

Wood-tar is a liquid material obtained by wood gasification process, and comprises several polycyclic aromatic hydrocarbons (PAH). Tar biodegradation is a very challenging task, due to its toxicity and to its complex chemistry. The 'microbial resource management' concerns the use of environmental microbial communities potentially able to provide us services. We applied this concept in tar biodegradation. Tar composed by several PAH (including phenanthrene, acenaphthylene and fluorene) was subjected to a biodegradation process in triplicate microcosms spiked with a microbial community collected from PAH-rich soils. In 20 days, 98.9% of tar was mineralized or adsorbed to floccules, while negative controls showed poor PAH reduction. The dynamics of fungal and bacterial communities was assessed through Automated Ribosomal Intergenic Spacer Analysis (ARISA), 454 pyrosequencing of the fungal ITS and of the bacterial 16S rRNA. Quantification of the degrading bacterial communities was performed via quantitative Real Time PCR of the 16S rRNA genes and of the cathecol 2,3-dioxygenase genes. Results showed the importance of fungal tar-degrading populations in the first period of incubation, followed by a complex bacterial dynamical growth ruled by co-feeding behaviors.

Impact of the reusing of food manufacturing wastewater for irrigation in a closed system on the microbiological quality of the food crops.

In order to evaluate if the reuse of food industry treated wastewater is compatible for irrigation of food crops, without increased health risk, in the present study a cropping system, in which ground water and treated wastewater were used for irrigation of tomato and broccoli, during consecutive crop seasons was monitored. Water, crop environment and final products were monitored for microbial indicators and pathogenic bacteria, by conventional and molecular methods. The microbial quality of the irrigation waters influenced sporadically the presence of microbial indicators in soil. No water sample was found positive for pathogenic bacteria, independently from the source. Salmonella spp. and Listeria monocytogenes were detected in soil samples, independently from the irrigation water source. No pathogen was found to contaminate tomato plants, while Listeria monocytogenes and E. coli O157:H7 were detected on broccoli plant, but when final produce were harvested, no pathogen was detected on edible part. The level of microbial indicators and detection of pathogenic bacteria in field and plant was not dependent upon wastewater used. Our results, suggest that reuse of food industry wastewater for irrigation of agricultural crop can be applied without significant increase of potential health risk related to microbial quality.

Inoculum pre-treatment affects the fermentative activity of hydrogen-producing communities in the presence of 5-hydroxymethylfurfural.

To enhance the productivity of mixed microbial cultures for fermentative bio-hydrogen production, chemical-physical pre-treatments of the original seed are needed to suppress the activity of hydrogen (H2)-consuming microbes. This approach might influence negatively the composition and diversity of the hydrogen-producing community with consequences on the functional stability of the H2-producing systems in case of perturbations. In this study, we aimed at investigating the effect of different types of pre-treatment on the performance of hydrogen production systems in the presence of an inhibitor, such as 5-hydroxymethylfurfural (HMF). The efficiency and the microbial community structure of batch reactors amended with HMF and inoculated with non-pretreated and pretreated (acid, heat shock, and aeration) anaerobic sludge were evaluated and compared with control systems. The type of pre-treatments influenced the microbial community assembly and activity in inhibited systems, with significant effect on the performance. Cumulative H2 production tests showed that the pre-aerated systems (control and HMF inhibited) were the most efficient, while the difference of the lag phase of the pre-acidified control and HMF-added test was negligible. Analyses of the structure of the enriched microbial community in the systems through PCR-denaturing gradient gel electrophoresis (DGGE) followed by band sequencing revealed that the differences in performance were mostly related to shifts in the metabolic pathways rather than in the predominant species. In conclusion, the findings suggest that the use of specific inoculum pre-treatment could contribute to regulate the metabolic activity of the fermentative H2-producing bacteria in order to enhance the bio-energy production.

Invasibility of resident biofilms by allochthonous communities in bioreactors.

Invasion of non-native species can drastically affect the community composition and diversity of engineered and natural ecosystems, biofilms included. In this study, a molecular community fingerprinting method was used to monitor the putative establishment and colonization of allochthonous consortia in resident multi-species biofilms. To do this, biofilms inoculated with tap water or activated sludge were grown for 10 days in bubble column reactors W1 and W2, and S, respectively, before being exposed to non-native microbial consortia. These consortia consisted of fresh activated sludge suspensions for the biofilms inoculated with tap water (reactors W1 and W2) and of transplanted mature tap water biofilm for the activated sludge biofilm (reactor S). The introduction of virgin, unoccupied coupons into W1 and W2 enabled us to additionally investigate the competition for new resources (space) among the resident biofilm and the allochthonous consortia. CE-SSCP revealed that after the invasion event changes were mostly observed in the abundance of the dominant species in the native biofilms rather than their composition. This suggests that the resident communities within a bioreactor immediately outcompete the allochthonous microbes and shape the microbial community assemblage on both new coupons and already colonized surfaces for the short term. However, with time, latent members of the allochthonous community might grow up affecting the diversity and composition of the original biofilms.

A preliminary and qualitative study of resource ratio theory to nitrifying lab-scale bioreactors.

The incorporation of microbial diversity in design would ideally require predictive theory that would relate operational parameters to the numbers and distribution of taxa. Resource ratio-theory (RRT) might be one such theory. Based on Monod kinetics, it explains diversity in function of resource-ratio and richness. However, to be usable in biological engineered system, the growth parameters of all the bacteria under consideration and the resource supply and diffusion parameters for all the relevant nutrients should be determined. This is challenging, but plausible, at least for low diversity groups with simple resource requirements like the ammonia oxidizing bacteria (AOB). One of the major successes of RRT was its ability to explain the 'paradox of enrichment' which states that diversity first increases and then decreases with resource richness. Here, we demonstrate that this pattern can be seen in lab-scale-activated sludge reactors and parallel simulations that incorporate the principles of RRT in a floc-based system. High and low ammonia and oxygen were supplied to continuous flow bioreactors with resource conditions correlating with the composition and diversity of resident AOB communities based on AOB 16S rDNA clone libraries. Neither the experimental work nor the simulations are definitive proof for the application of RRT in this context. However, it is sufficient evidence that such approach might work and justify a more rigorous investigation.

Multi-scale modelling of bioreactor-separator system for wastewater treatment with two-dimensional activated sludge floc dynamics.

A simple "first generation" multi-scale computational model of the formation of activated sludge flocs at micro-scale and reactor performance at macro-scale is proposed. The model couples mass balances for substrates and biomass at reactor scale with an individual-based approach for the floc morphology, shape and micro-colony development. Among the novel model processes included are the group attachment/detachment of micro-flocs to the core structure and the clustering of nitrifiers. Simulation results qualitatively describe the formation of micro-colonies of ammonia and nitrite oxidizers and the extracellular polymeric substance produced by heterotrophic microorganisms, as typically observed in fluorescence in situ hybridization images. These results are the first step towards realistic multi-scale multispecies models of the activated sludge wastewater treatment systems and a generic modelling strategy that could be extended to other engineered biological systems.

Low-dissolved-oxygen nitrifying systems exploit ammonia-oxidizing bacteria with unusually high yields.

In wastewater treatment plants, nitrifying systems are usually operated with elevated levels of aeration to avoid nitrification failures. This approach contributes significantly to operational costs and the carbon footprint of nitrifying wastewater treatment processes. In this study, we tested the effect of aeration rate on nitrification by correlating ammonia oxidation rates with the structure of the ammonia-oxidizing bacterial (AOB) community and AOB abundance in four parallel continuous-flow reactors operated for 43 days. Two of the reactors were supplied with a constant airflow rate of 0.1 liter/min, while in the other two units the airflow rate was fixed at 4 liters/min. Complete nitrification was achieved in all configurations, though the dissolved oxygen (DO) concentration was only 0.5 ± 0.3 mg/liter in the low-aeration units. The data suggest that efficient performance in the low-DO units resulted from elevated AOB levels in the reactors and/or putative development of a mixotrophic AOB community. Denaturing gel electrophoresis and cloning of AOB 16S rRNA gene fragments followed by sequencing revealed that the AOB community in the low-DO systems was a subset of the community in the high-DO systems. However, in both configurations the dominant species belonged to the Nitrosomonas oligotropha lineage. Overall, the results demonstrated that complete nitrification can be achieved at low aeration in lab-scale reactors. If these findings could be extended to full-scale plants, it would be possible to minimize the operational costs and greenhouse gas emissions without risk of nitrification failure.

Ammonia-oxidizing bacteria in wastewater.

Ammonia-oxidizing bacteria (AOB) have a key role in the conversion of ammonia to nitrite in wastewater treatment plants (WWTPs). The characterization of AOB communities in such systems requires the use of genomic methods as AOB are difficult to isolate from environmental samples. Fluorescence in situ hybridization (FISH) using fluorescently labeled probes targeting 16S rRNA molecules provides a robust tool for the detection and quantification of AOB populations in biofilms and activated sludge flocs. The abundance of AOB may be also determined by real-time quantitative polymerase chain reaction (qPCR) using primers that amplify either the 16S rRNA or amoA genes. The evaluation of changes in the AOB community in time and space can be undertaken by PCR amplification of these gene fragments followed by denaturing gradient gel electrophoresis (PCR-DGGE). In this chapter, we summarize the most commonly applied procedures for the analysis of the AOB in wastewater, emphasizing their advantages and limitations.