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The use of phototrophic organisms in wastewater treatment is gaining increasing interest, due to the possibility of exploiting photosynthetic oxygenation, thus lowering treatment costs and environmental impacts. Mixed consortia of phototrophic organisms and aerobic bacteria rapidly develop in photobioreactors receiving wastewaters; in this case, the complex mutualistic interactions among microorganisms cannot be easily described, making difficult to forecast removal efficiencies and biomass productivities. Respirometric methods are applied for evaluating bacterial activity in activated sludge (conventional respirometry) or photosynthetic activity in phototrophic organisms (photorespirometry). Photo-respirometric methods can be also exploited for monitoring algae-bacteria systems and for calibrating mathematical models, allowing for a better comprehension of the interactions involved and a more accurate prediction of system efficiencies. A general protocol and a series of guidelines for conducting photorespirometric tests on algal-bacterial suspensions are here proposed, based on the review of experimental protocols applied to evaluate the effects of several parameters on these systems.

Tetraselmis suecica was cultivated in a semi-continuously operated tubular photobioreactor fed on aquaculture wastewater (AW) testing two hydraulic retention times (HRT): 10 and 7 days (RUN_1 and RUN_2, respectively). The integrated mechanistic model BIO_ALGAE was validated with experimental data in order to simulate the biomass production and nutrient uptake of T. suecica. Moreover, AW was used as substitute synthetic cultivation medium to test the production of lipids, proteins, and carbohydrates in the microalgal biomass. Preliminary photo-respirometric tests were carried out on the AW suspension containing microalgae and bacteria. Dissolved Inorganic Nitrogen (DIN) and Dissolved Inorganic Phosphorus (DIP) were analyzed for the two RUNs, and no significant difference was highlighted (p > 0.05). On the contrary, the productivity of the Total suspended solids (TSS) was significantly higher (p < 0.05) for RUN_1 (900 mg TSS/L) than for RUN_2 (550 mg TSS/L). The analysis of the biochemical composition of biomass has demonstrated a higher content of proteins than of lipids and carbohydrates for the two RUNs. BIO_ALGAE model was validated by comparing simulated results to experimental data. The model was able to reproduce the pattern of these experimental data quite well, for both nutrient uptake and biomass production. The simulated curve follows the same pattern as the experimental data for both RUNs. The wavelike trend indicates the good accuracy of the simulated curves to reproduce the microalgae growth and nutrient uptake that occurring during daytime and at night. With this study, BIO_ALGAE Model was demonstrated to be useful to simulate bioremediation and microalgae production in aquaculture wastewater in a semi-continuous system with different environmental factors. The photo-respirometric outputs were compared with the process rates affecting dissolved oxygen dynamics computed by the mathematical model. The authors would like to thank the Italian Ministry of University and Research (MIUR) and Fondazione Cariplo, for funding part of this research (“Vademecum” and “Il polo delle microalghe” projects) and Dr. Nadia Margariti for her valuable help in laboratory.

Milk whey processing wastewaters (MWPWs) are characterized by high COD and organic nitrogen content, the concentrations of phosphorus are also relevant. A microalgal-based process was tested at lab scale in order to assess the feasibility of treating MWPW without any dilution or pre-treatment. Different microalgal strains and populations were tested. Based on the obtained results, Scenedesmus acuminatus (SA) and a mixed population (PM) chiefly made of Chlorella, Scenedesmus, and Chlamydomonas spp. were grown in duplicate for 70 days in Plexiglas column photobioreactors (PBRs), fed continuously (2.5 L culture volume, 7 days hydraulic retention time). Nutrient removal, microalgae growth, photosynthetic efficiency, and the composition of microalgal populations in the columns were monitored. At steady state, the microalgal growth was similar for SA and PM. The average removal efficiencies for the main pollutants were: 93% (SA), 94% (PM) for COD, 88% (SA) and 90% (PM) for total N, and 69% (SA) and 73% (PM) for total P. The residual pollution levels in the effluent from the PBRs were low enough to allow their discharge into surface waters, such good results were achieved thanks to the synergy between the microalgae and bacteria in the CO2 and oxygen production/consumption and in the nitrogen mineralization.

The role of P content on the treatment and valorization of the liquid fraction of digestate, namely centrate, through microalgae-based technologies was evaluated in this study. The performance of four column photobioreactors, which were fed on diluted centrate with corrected (10 mg N/ mg P) and not modified (129 mg N/ mg P) N:P ratio, were monitored and compared. The results demonstrated that P shortage in the centrate affected neither the total nitrogen and COD removal rate nor the volumetric biomass productivity, suggesting that expensive addition of P salts is not necessary to maximize the efficiency of the process. On the contrary, the addition of P to the centrate promoted the ammonia oxidation process as higher nitrite production was observed in the photobioreactors with adjusted N:P ratio than in the ones fed with the non-adjusted N:P ratio. These findings were confirmed by fluorescence in-situ hybridization and quantitative PCR assays, which revealed a higher number of ammonia-oxidizing bacteria in the microalgal suspensions cultivated on centrate with P addition. In conclusion, the N:P ratio in the centrate seems to have a role in controlling the nitrification process rather than in the overall nutrient removal rate and biomass productivity of the microalgae-based system.

The definition of relationships among the attributes of macroinvertebrate communities and environmental gradients is fundamental to understand ecological dynamics. Testing the response of macroinvertebrate communities and the related metrics to the action of multiple drivers is crucial to provide reliable tools for the management and conservation of freshwater ecosystems. In this study habitat conditions, seasons and physical–chemical water characteristics were evaluated as potential limiting factors for macroinvertebrate communities. 42 sites, along 11 pre-Alpine rivers and streams, were surveyed for macroinvertebrate communities and water chemistry, collecting a total of 387 samples. Hydromorphological characteristics and habitat availability in each sampling site were assessed to describe different features of channel, banks and vegetation that can be relevant for management purposes. Macroinvertebrate community characteristics were summarized by both structural metrics (i.e. density, diversity and taxonomy-based) and ecosystem attributes derived from functional trait analysis. Functional traits are based on morphological and behavioural attributes associated with feeding, modes of attachment, concealment and locomotion, together with voltinism and drift propensity. In this framework, quantitative models have been developed to predict the potential characteristics of macroinvertebrate communities as a function of selected environmental variables. These relationships can be used to predict the effects of river restoration programs on macroinvertebrate diversity and ecosystem processes. The various metrics considered in this study may help to raise a better awareness in the natural functioning of river ecosystems, the selection of effective strategies for the conservation of biological communities, and the evaluation of river restoration success, thus supporting decision-making processes.

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.

Respirometric techniques are useful tools to evaluate bacterial activities in activated sludge processes due to their fast execution and the possibility to obtain several kinetic parameters from a single test. Using such techniques in microalgae-bacteria consortia treating wastewater could allow a better understanding of mutual interactions between the microbial populations as a function of environmental parameters. This work aims at developing and testing a novel experimental respirometric protocol to determine oxygen uptake rates and oxygen production rates by a microalgae-bacteria consortium. The defined protocol is characterized by alternating light/dark regimes and by dosing substrates/inhibitors to selectively activate/inactivate microalgal and bacterial metabolisms. The protocol was then applied on microalgal and bacterial consortia, which were grown on the liquid fraction of black water from biogas plants fed on agricultural and municipal waste sludge. Results elucidate the presence and activity of microalgae and nitrifying bacteria in the tested systems, suggesting that the respirometric tests could be included into monitoring procedures of photobioreactors/algal ponds.

The presence of antibiotic resistant bacteria (ARB) in wastewater was investigated and the role of wastewater treatment plants (WWTPs) in promoting or limiting antibiotic resistance was assessed. Escherichia coli (E. coli) and total heterotrophic bacteria (THB) resistance to ampicillin, chloramphenicol and tetracycline was monitored in three WWTPs located in Milan urban area (Italy), differing among them for the operating parameters of biological process, for the disinfection processes (based on sodium hypochlorite, UV radiation, peracetic acid) and for the discharge limits to be met. Wastewater was collected from three sampling points along the treatment sequence (WWTP influent, effluent from sand filtration, WWTP effluent). Antibiotic resistance to ampicillin was observed both for E. coli and for THB. Ampicillin resistant bacteria in the WWTP influents were 20–47% of E. coli and 16–25% of THB counts. A limited resistance to chloramphenicol was observed only for E. coli, while neither for E. coli nor for THB tetracycline resistance was observed. The biological treatment and sand filtration led to a decrease in the maximum percentage of ampicillin-resistant bacteria (20–29% for E. coli, 11–21% for THB). However, the conventionally adopted parameters did not seem adequate to support an interpretation of WWTP role in ARB spread. Peracetic acid was effective in selectively acting on antibiotic resistant THB, unlike UV radiation and sodium hypochlorite. The low counts of E. coli in WWTP final effluents in case of agricultural reuse did not allow to compare the effect of the different disinfection processes on antibiotic resistance.

BACKGROUND: Secondary effluent wastewaters still contain resources including water and nutrients that must be safely reused and recovered. In this study, the combined role of microalgae as disinfectant and nutrient removal agent was evaluated for the potential application of a microalgae-based process as a cost-effective tertiary treatment. Nutrient removal, biomass productivity and disinfection performances were monitored in laboratory-scale photobioreactors (batch and continuous) fed on a secondary effluent mixed with a 10% on influent collected at a large municipal wastewater treatment plant where tertiary disinfection is performed by UV treatment. RESULTS: In microalgae-based batch disinfection tests, Escherichia coli counts (0.5 ± 0.7 log CFU 100 mL−1) were comparable to those after traditional UV process (0.7 ± 0.84 log CFU 100 mL−1) and lower than in tests where light was applied without microalgae. In the following continuous test, E. coli counts were reduced by one order of magnitude and the pathogenic strain of E. coli O157:H7/H−, Salmonella spp. and indicators such as Bacteroides spp. and Enterococcus spp. were never detectable in the effluents by molecular tools. Total nitrogen and phosphorus removals reached 93 and 100%, respectively, while the algal biomass productivity of the system averaged 50 ± 30 mg TSS L−1 day−1. CONCLUSIONS: The effluents of the photobioreactors reached quality standards appropriate for water reuse. Moreover, nutrients could be recovered through the generation of algal biomass suitable for further valorization. © 2019 The Authors. Journal of Chemical Technology & Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

BACKGROUND: Microalgae–bacteria‐based processes are among the most promising low‐cost technologies to treat livestock wastewaters. The current literature reports the need for pretreatment or dilution of piggery wastewater for adequate microalgal growth. The aim of this study is to optimize the potential of microalgal–bacterial communities to treat undiluted and untreated piggery wastewater by investigating the influence of some operational parameters such as phosphorus and CO₂ availability and hydraulic retention time on the nitrogen removal efficiency and biomass productivity. RESULTS: The microalgal community (dominated by Chlorella spp.) developed quickly and remained quite stable. The rates of biomass production and NH₄‐N removal were 55 ± 30 mg TSS L⁻¹ day⁻¹ and 13 ± 3 mg NH₄‐N L⁻¹ day⁻¹ respectively. CO₂ adjustment had a positive effect on microalgal growth and NH₄‐N removal. CONCLUSION: Data confirm the ability of the microalgal–bacterial consortium to grow on undiluted and untreated piggery wastewater under semi‐continuous conditions. Synergy between algae and bacteria seems positive since photosynthesis produces the oxygen needed for ammonia oxidation. © 2019 Society of Chemical Industry

This study was carried out to assess the efficiency of a pilot-scale bubble-column reactor to remove nitrogen in centrate from the biosolid dewatering of a municipal wastewater treatment plant whilst producing biomass for agricultural purposes. The column was inoculated with a mixed community of Scenedesmus and Chlorella spp. and operated outdoor in batch for 55 days and in continuous for further 130 days. In continuous, the average daily biomass productivity was 40 ± 62 mg TSS L −1 d−1 and the average NH4+-N removal was 20 ± 10 mg L −1 d−1. Nitrification was fostered by photo‑oxygenation leading to the oxidation of 34 ± 27% of the incoming ammonia nitrogen. Microalgal and bacterial activity inside the column was analyzed by the Generalized Linear Models in order to understand the main factors affecting the process performances. Microalgal growth was affected positively by the NH4+-N content in the influent and negatively by the amount of TSS entering the system, probably due to the competition between microalgae and bacteria for phosphorus and other nutrients. The removal rate of NH4+-N was positively affected by NH4+-Nin (influent concentration) and by pH, whose increase fosters stripping, and decreased for increasing NH3-N concentrations, responsible for inhibiting nitrifying bacteria. NH4+-N oxidation was the result of complex interactions between algae and bacteria and was also affected by flow and solar radiation. No other specific limiting factors have been highlighted. The possibility of improving the process performance by controlling pH, by supplying off-gas as CO2 additional source, appears as an interesting option. In view of a scale-up, the most relevant expected result would be the energy saving due to the decrease in the oxygen demand for nitrification in the water line. The microalgal biomass grown on centrate was suitable for agricultural use due to its low contamination by heavy metals.

The final publication is available at Springer via http://dx.doi.org/10.1007/s11270-019-4122-0 This study investigates and compares the uptake of nutrients (nitrogen, phosphorus) and the growth of Tetraselmis suecica and Dunaliella tertiolecta in aquaculture wastewater. The obtained data were used to implement and calibrate the microalgae-bacteria model BIO_ALGAE to simulate the bioremediation and the biomass production of these species. The microalgae were cultivated in batch conditions for 7 days using 120-L vertical column photobioreactors. In the first 4 days, after which the algal density reached a steady state, the average biomass production was 83.7 ± 4.4 mg/L/day for T. suecica and 56.4 ± 5.1 mg/L/day for D. tertiolecta. The two species were able to remove more than 96% of dissolved inorganic nitrogen (DIN) and dissolved inorganic phosphorus (DIP). The total lipid content was analyzed at the end of the 7 days; T. suecica and D. tertiolecta had different lipid content: 75.8 ± 1.6% and 23.2 ± 2.0%, respectively. The BIO_ALGAE model fits very well the experimental data of both species in terms of biomass and nutrient uptake and could be an effective tool to predict the production of microalgae using aquaculture wastewater as growth media, obtaining at the same time the removal of nutrients from wastewater and the production of biomass to be used as feed. In particular, this mathematical model can be applied to forecast the performance under different operating conditions, for the design, optimization, and control of the process in aquaculture systems.

Dreissena polymorpha is a widespread filter-feeder species, resistant to a broad range of environmental conditions and different types of pollutants,which has recently colonized Italian freshwaters. Although widely used to monitor pollution in freshwater environments, this species is also an important food source for some fish and water birds. It can also be used to concentrate or remove particulate organic matter to interrupt avian-to-human transmission of pollutants and control health risks for animals and humans. In this study, the accumulation/inactivation in D. polymorpha of human health-related spiked enteric viruses was described. The removal of endogenous Escherichia coli, the classical indicator of fecal contamination,was tested as well.Our preliminary lab-scale results demonstrate that zebra mussels can reduce significantly poliovirus titer after 24 h and rotavirus titer after 8 h. E. coli counts were also reduced in the presence of zebra mussels by about 1.5 log after 4 h and nearly completely after 24 h. The fate of the two enteric viruses after concentration by zebra mussels was also investigated after mechanical disruption of the tissues. To our knowledge, the accumulation from water and inactivation of human health-related enteric viruses by zebra mussels has never been reported.

This paper addresses the efficiency of a microalgae-based agricultural digestate treatment at pilot-scale in an outdoor raceway pond (880 L, pH-dependent CO2 dosage) and in a bubble column (74.5 L, air-bubbling). Specifically, nitrogen removal, evolution of the algae-bacteria consortium, and the actual process applicability in the Po Valley climate are discussed. The performance of the two reactors varied seasonally. The average algal productivity in the raceway was 32.4 ± 33.1 mg TSS·L−1·d−1 (8.2 ± 8.5 g TSS·m−2·d−1) while in the PBR it was 25.6 ± 26.8 mg TSS·L−1·d−1; the average nitrogen removal was 20 ± 29% (maximum 78%) and 22 ± 29% (maximum 71%) in the raceway and in the column, respectively. Nevertheless, nitrification had a key role as 61 ± 24% and 52 ± 32% of the nitrogen load was oxidized in the raceway and in the column, respectively.

A constructed wetland system composed of a subsurface flow wetland, a surface flow wetland and a facultative pond was studied from July 2008 until May 2012. It was created to treat the domestic sewage produced by a hamlet of 150 inhabitants. Monthly physicochemical and microbiological analyses were carried out in order to evaluate the removal efficiency of each stage of the process and of the total treatment system. Pair-wise Student's t-tests showed that the mean removal of each considered parameter was significantly different (α = 0.05) between the various treatment phases. Two-way ANOVA and Tukey's HSD tests were used to find significant differences between wetland types and seasons in the removal efficiency of the considered water quality parameters. Significant differences in percent removal efficiency between the treatment phases were observed for total phosphorus, total nitrogen, ammonia nitrogen and organic load (expressed as Chemical Oxygen Demand). In general, the wastewater treatment was carried by the sub-superficial flow phase mainly, both in growing season and in quiescence season. Escherichia coli removal ranged from 98% in quiescence season to >99% in growing season (approximately 2–3 orders of magnitude). The inactivation of fecal bacteria was not influenced by the season, but only by the treatment phase.

This paper discusses the possibility of including the culturing of microalgae within a conventional wastewater treatment sequence by growing them on the blackwater (BW) from biosolid dewatering to produce biomass to feed the anaerobic digester. Two photobioreactors were used: a 12 L plexiglas column for indoor, lab-scale tests and a 85 L plexiglas column for outdoor culturing. Microalgae (Chlorella sp. and Scenedesmus sp.) could easily grow on the tested blackwater. The average specific growth rate in indoor and outdoor batch tests was satisfactory, ranging between 0.14 and 0.16 day−1. During a continuous test performed under outdoor conditions from May to November, in which the off-gas from the combined heat and power unit was used as the CO2 source, an average biomass production of 50 mgTSS L−1 day−1 was obtained. However, statistical analyses confirmed that microalgal growth was affected by environmental conditions (temperature and season) and that it was negatively correlated with the occurrence of nitrification. Finally, the biochemical methane potential of the algal biomass was slightly higher than that from waste sludge (208 mLCH4 gVS−1 vs. 190 mLCH4 gVS−1).

The liquid fraction of digestate produced by agricultural biogas plants is rich in macro and micronutrients that are valuable for the culturing of microalgae. Nonetheless, the high ammonium concentration may cause toxicity and the high optical density may reduce light penetration, negatively affecting the biomass production rate. Dilution with fresh water has been frequently suggested as a mean for improving the digestate characteristics in view of microalgal culturing. In this paper, the feasibility of culturing microalgae on undiluted raw digestate or on digestate after pretreatment by stripping and adsorption was investigated. First, adsorption tests were performed using commercial activated carbon from wood in order to identify appropriate conditions for optical density (OD) reduction. Up to 88% reduction was obtained by dosing 40gL−1 after 24h of contact time. Then, culturing tests were performed on a microalgal inoculum including mainly Chlorella spp. and Scenedesmus spp. under controlled temperature and light conditions during 6–14weeks. Raw, stripped, and stripped and adsorbed digestate samples were tested. The biomass production rate increased from 27±13mgTSSL−1d−1 on raw digestate, to 82±18mgTSSL−1d−1 by using stripped digestate, and to 220±78mgTSSL−1d−1 by using the stripped and adsorbed digestate. Moreover, nitrification was constantly suppressed when using the stripped and adsorbed digestate, while relevant nitrite built-up was observed when using raw and stripped digestate. These results suggest that microalgae are able to grow on the raw digestate, provided that long hydraulic retention times are applied. A much faster growth (up to 10 times) can be obtained by pretreating the liquid fraction of digestate by stripping and adsorption, which may be an effective means of improving the areal productivity of microalgal culturing on digestates.

PAH (Polycyclic Aromatic Hydrocarbons) analyses were carried out on samples from two Wastewater Treatment Plants (WWTPs) in Lombardy, similar for treatment sequences but fed on different influents: industrial component accounts for 70% at Alto Seveso plant while it is absent in Nosedo plant. Sampling concerned the influent and the effluent from activated sludge reactor and the final effluent after disinfection (ozonation for Alto Seveso and peracetic acid treatment for Nosedo). The concentrations of total PAHs were 5.3 ± 4.0 μg L−1 and 2.4 ± 1.3 μg L−1 in Alto Seveso and Nosedo influent, respectively. The lowest molecular weight PAHs had the highest concentrations in both plants; acenaphthene and naphthalene were the most important components in the influent to Alto Seveso and Nosedo WWTPs, respectively. The higher molecular weight compounds had the lowest concentrations and benzo(g,h,i)perylene and dibenzo(a,h)anthracene were never detected. Most of the PAH load entered biological treatment in dissolved form. For both plants PAHs were mostly removed in the biological section (96.5% and 89.5% for Alto Seveso and Nosedo, respectively), while disinfection had a minor role. Peracetic acid (Nosedo) seemed more efficient than ozone (Alto Seveso) in the removal of PAHs (4.18% and 0.89%, respectively). It is now necessary to confirm this result by using the same effluent for the two disinfection treatments.

The emergence and diffusion of antibiotic-resistant bacteria has been a major public health problem for many years now. In this study, antibiotic-resistance of coliforms and Escherichia coli were investigated after their isolation from samples collected in a municipal wastewater treatment plant in the Milan area (Italy) along different points of the treatment sequence: inflow to biological treatment; outflow from biological treatment following rapid sand filtration; and outflow from peracetic acid disinfection. The presence of E. coli that showed resistance to ampicillin (AMP) and chloramphenicol (CAF), used as representative antibiotics for the efficacy against Gram-positive and Gram-negative bacteria, was evaluated. After determining E. coli survival using increasing AMP and CAF concentrations, specific single-resistant (AMPR or CAFR) and double-resistant (AMPR/CAFR) strains were identified among E. coli colonies, through amplification of the β-lactamase Tem-1 (bla) and acetyl-transferase catA1 (cat) gene sequences. While a limited number of CAFR bacteria was observed, most AMPR colonies showed the specific resistance genes to both antibiotics, which was mainly due to the presence of the bla gene sequence. The peracetic acid, used as disinfection agent, showed to be very effective in reducing bacteria at the negligible levels of less than 10 CFU/100 mL, compatible with those admitted for the irrigation use of treated waters.