Piggery wastewater treatment with algae-bacteria consortia: Pilot-scale validation and techno-economic evaluation at farm level.

The efficiency of an outdoor pilot-scale raceway pond treating the wastewaters generated by a large-scale piggery farm in Northern Italy was evaluated. The biomass productivity over 208 days of experimentation was 10.7 ± 6.5 g TSS·m-2·d-1, and ammoniacal nitrogen, orthophosphate, and COD average removal efficiencies were 90%, 90%, and 59%, respectively. Results were used to perform a comprehensive techno-economic analysis for integrating algae-based processes in farms of different sizes (100-10000 pigs). The amount of N disposed of on agricultural land could be reduced from 91% to 21%, increasing the fraction returned to the atmosphere from 2.4% to 63%, and the fraction in the biomass from 6.2% to 16%. For intensive farming, the release of 110 t N·ha-1·y-1 contained in the digestate could be avoided by including algae-bacteria processes. The biomass production cost was as low as 1.9 €·kg-1, while the cost for nitrogen removal was 4.3 €·kg N-1.

Metal-based flocculation to harvest microalgae: a look beyond separation efficiency.

Metal-based flocculants are commonly used for biomass harvesting in microalgae-based bio-refineries. Besides the high separation efficiency, additional aspects should be considered, related to the toxicity of metals for the algal biomass. Partitioning tests for commonly used flocculants (i.e., FeCl3 and Al2(SO4)3) showed that metals were mostly transferred to the solid phase with more than 95% of dosed metal ending up into the biomass, and low metal concentrations in the liquid effluent (lower than 0.4 mg L-1 for both metals), thus allowing for water reuse. Photosynthesis inhibition was tested on microalgae and microalgae-bacteria cultures, using a standardized photo-respirometry protocol in which typical concentrations used during coagulation-flocculation were assessed. Modelling dose-response curves, concentrations corresponding to 50% inhibition (IC50) were obtained, describing short-term effects. The obtained IC50 ranged from 13.7 to 28.3 mg Al L-1 for Al, and from 127.9 to 195.8 mg Fe L-1 for Fe, showing a higher toxicity for the Al-based flocculant. The recovery of photosynthesis inhibition was also quantified, to evaluate the possibility of reusing/recycling the harvested biomass. The results highlighted that the residual photosynthetic activities, evaluated after 1 h and 24 h of exposure to metals were partially recovered, especially for Al, passing from 67.3% to 94.6% activity, respectively, while long-term Fe effects were stronger (passing from 64.9% to 77.6% activity). A non-toxic flocculant (cationic starch) was finally tested, excluding potential effects due to biomass aggregation, as the reduction of photosynthetic activity only reached 3.4%, compared to control. Relevant modifications to the light availability and the optical properties of algal suspensions were assessed, identifying a strong effect of iron which caused an increase of the light absorbance up to approximately 40% at high Fe concentrations. Possible implications of dosing metallic flocculants in MBWWT processes are discussed, and suggestions are given to perform inhibition tests on flocculating chemicals.

Selection of photosynthesis and respiration models to assess the effect of environmental conditions on mixed microalgae consortia grown on wastewater.

This study aimed at evaluating the effects of different environmental conditions (irradiance, temperature, pH and dissolved oxygen) on a microalgae-bacteria consortium cultivated in a pilot-scale open pond and fed on the liquid fraction of anaerobic digestate. A standardized photo-respirometry protocol was followed to evaluate the activity of microalgae under different conditions. Two datasets (specific photosynthetic oxygen production rates and respiratory oxygen consumption rates) were obtained for each environmental parameter, throughout the entire range of conditions found in the outdoor cultivation system. Different kinetic models available in literature were fitted to experimental data and the resulting outputs were compared through model selection estimators, in order to select the most appropriate equations. The proposed set of equations constitute a modelling tool for the prediction of algal growth rates in algae-bacteria systems, as a function of environmental conditions.

Lessons learnt from a pilot study on residual dye removal by an aerated treatment wetland.

Treatment wetlands (TWs) have shown good capacity in dye removal from textile wastewater. However, the high hydraulic retention times (HRTs) required by these solutions and the connected high area requirements, remain a big drawback towards the application of TWs for dye treatment at full scale. Aerated TWs are interesting intensified solutions that attempt to reduce the TW required area. Therefore, an aerated CW pilot plant, composed of a 20 m2 horizontal subsurface flow TW (HF) and a 21 m2 Free Water System (FWS), equipped with aeration pipelines, was built and monitored to investigate the potential reduction of required area for dye removal from the effluent wastewater of a centralized wastewater treatment plant (WWTP). During a 8 months long study, experimenting with different hydraulic retention times (HRTs - 1.2, 2.6 and 3.5 days) and aeration modes (intermittent and continuous), the pilot plant has shown a normal biological degradation for organic matter and nutrients, while the residual dye removal has been very low, as demonstrated by the absorbance measure at three wavelengths: at 426 nm (blue) the removal varies from -55% at influent absorbance of 0.010 to 41% at 0.060; at 558 nm (yellow) the removal is negative at 0.005 (-58%) and high at higher influent concentrations (72% at 0.035 of absorbance for the inlet); at 660 nm (red) -82% of removal efficiency was obtained at influent absorbance of 0.002 and 74% at 0.010. These results are a consequence of the biological oxidation processes taking place in the WWTP, so that the residual dye seems to be resistant to further aerobic degradation. Therefore, TWs enhanced by aeration can provide only a buffer effect on peak dye concentrations.

Digestate treatment with algae-bacteria consortia: A field pilot-scale experimentation in a sub-optimal climate area.

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 ±â€¯33.1 mg TSS·L-1·d-1 (8.2 ±â€¯8.5 g TSS·m-2·d-1) while in the PBR it was 25.6 ±â€¯26.8 mg TSS·L-1·d-1; the average nitrogen removal was 20 ±â€¯29% (maximum 78%) and 22 ±â€¯29% (maximum 71%) in the raceway and in the column, respectively. Nevertheless, nitrification had a key role as 61 ±â€¯24% and 52 ±â€¯32% of the nitrogen load was oxidized in the raceway and in the column, respectively.

Activity assessment of microalgal-bacterial consortia based on respirometric tests.

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.

Antibiotic resistant bacteria in urban sewage: Role of full-scale wastewater treatment plants on environmental spreading.

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.

Toxicity of Quaternary Ammonium Compounds (QACs) as single compounds and mixtures to aquatic non-target microorganisms: Experimental data and predictive models.

The toxic effects of five Quaternary Ammonium Compounds (QACs) that are widely used as active ingredients in personal care products were assessed using the bioluminescent bacterium Aliivibrio fischeri (formerly Vibrio fischeri) (Microtox® test system). The experimental results showed a relevant toxicity for almost all of the single QACs, with IC50 values lower than 1mgL-1. Analysis of the mode of action through the application of the Quantitative Structure-Activity Relationship (QSAR) models indicated an a-specific reactivity for most of the QACs toward A. fischeri. Only hexadecyl trimethyl ammonium chloride (ATMAC-16) behaved as a polar-narcotic, with a low reactivity toward the bacterial cell membrane. The concentration response curves of the different binary and multicomponent mixtures of QACs were also evaluated with respect to the predictions from the Concentration Addition (CA) and Independent Action (IA) models. For almost all of the binary and multicomponent mixtures (7 out of 11 mixtures tested), an agreement between the experimental and predicted ICx was observed and confirmed via application of the Model Deviation Ratio (MDR). In four cases, some deviations from the expected behaviour were observed (potential antagonistic and synergistic interactions) at concentrations on the order of hundreds of µgL-1, which could be of environmental concern, especially in the case of synergistic effects. The analysis of aquatic ecotoxicity data and the few available values of the measured environmental concentrations (MECs) from the literature for wastewaters and receiving waterbodies suggest that a potential risk toward aquatic life cannot be excluded.

Growth of microalgal biomass on supernatant from biosolid dewatering.

The paper reports the results of an experiment to assess the feasibility of including a photobioreactor within the design of a wastewater treatment plant, growing microalgae on the centrate from anaerobic sludge dewatering. The growth of algal biomass would take advantage of the available nitrogen and provide a substrate for biogas production by anaerobic digestion. Tests were carried out by semi-continuously feeding a photobioreactor with a centrate-effluent blend and by increasing the fraction of centrate. The experimental results show that the centrate does not induce any toxicity and, on the contrary, can be well utilized by microalgae, whose average specific growth rate (µ), on centrate as such, was between 0.04 and 0.06 d(-1). The maximum biomass concentration in the photobioreactor effluent was 1.6 gSS/L at 10 days HRT (hydraulic retention time). Methane production tests led to biochemical methane production values of 335 ± 39, and 284 ± 68 mL 0°C, 1 atm CH4/g VS for the two tested samples, in agreement with literature values. Settling tests show that the settling capacity of microalgae, although satisfactory, could be effectively improved after mixing with activated sludge, confirming the potential to use the existing primary settler for microalgae thickening in order to feed microalgae for anaerobic digestion with primary/secondary sludge.

Peracetic acid for secondary effluent disinfection: a comprehensive performance assessment.

The paper is a review of previous research on secondary effluent disinfection by peracetic acid (PAA) integrated with new data about the effect of a preliminary flash-mixing step. The process was studied at bench and pilot scale to assess its performance for discharge in surface water and agricultural reuse (target microorganisms: Escherichia coli and faecal coliform bacteria). The purposes of the research were: (1) determining PAA decay and disinfection kinetics as a function of operating parameters, (2) evaluating PAA suitability as a disinfectant, (3) assessing long-term disinfection efficiency, (4) investigating disinfected effluent biological toxicity on some aquatic indicator organisms (Vibrio fischeri, Daphnia magna and Selenastrum capricornutum), (5) comparing PAA with conventional disinfectants (sodium hypochlorite, UV irradiation). PAA disinfection was capable of complying with Italian regulations on reuse (10 CFU/100 mL for E. coli) and was competitive with benchmarks. No regrowth phenomena were observed, as long as needed for agricultural reuse (29 h after disinfection), even at negligible concentrations of residual disinfectant. The toxic effect of PAA on the aquatic environment was due to the residual disinfectant in the water, rather than to chemical modification of the effluent.

Searching for a compromise between ecological quality targets, and social and ecosystem costs for heavily modified water bodies (HMWBs): the Lambro-Seveso-Olona system case study.

The Lambro-Seveso-Olona (L-S-O) system derives from the human regulation of the natural hydrology of the territory around Milan city area. The average population density in the L-S-O area is among the highest in Italy and Europe. Industry is also highly developed in this basin: chemical, textile, paper, pulp and food industries being the most important ones. Although, at present, the L-S-O system no longer receives the untreated wastewaters of the Milan urban area, treated wastewaters constitute about half of the streamflow. Biotic communities in this river have a long history of poor quality status, having suffered great damage due to domestic and industrial discharges. Recently, new chemical quality standards for macropollutants have been set by the Italian legislation as support for the good ecological status according to the Water Framework Directive (WFD). This new index is very restrictive, and it makes it extremely challenging to achieve the water quality objectives for the L-S-O system. The aim of this study is to analyse through a modelling exercise the restoration possibilities of the L-S-O system, investigating both the source apportionment of the macropollutants, the discharge limits that should be set to achieve the good quality status and their corresponding cost.

A multivariate approach to assess habitat integrity in urban streams using benthic macroinvertebrate metrics.

Benthic macroinvertebrates are widely used as indicators of the health of freshwater ecosystems, responding both to water quality and to the hydromorphological integrity. In urban streams, evaluations can be tricky for the synergistic effects of multiple stressors and confounding factors. In these situations, the most broadly used multimetric indices can be used to assess the overall damage to the invertebrate community and, thus, the overall anthropogenic pressure, but they do not allow to understand the specific causal effects. Particularly, habitat loss due to morphological alterations can be difficult to evaluate, especially due to the often concurrent disturbance caused by water pollution. We used a multivariate approach to focus on the characteristics of the streams and rivers in an urban district and to define which macroinvertebrate metrics should be used to assess the influence of the different kinds of alteration in a severely damaged environment. Some metrics enabling the assessment of habitat loss (ratio of oligochaeta, ratio of filterers) were identified. These metrics may help to raise a better awareness in the evaluation of river restoration success and, thus, in the support of decision-making processes.

Colour removal and carbonyl by-production in high dose ozonation for effluent polishing.

Experimental tests have been conducted to investigate the efficiency and the by-product generation of high dose ozonation (10-60 mg O3 L(-1)) for complete colour removal from a treated effluent with an important component of textile dyeing wastewater. The effluent is discharged into an effluent-dominated stream where no dilution takes place, and, thus, the quality requirement for the effluents is particularly strict. 30, 60 and 90 min contact times were adopted. Colour was measured as absorbance at 426, 558 and 660 nm wavelengths. pH was monitored throughout the experiments. The experimental work showed that at 50 mg L(-1) colour removal was complete and at 60 mg O3 L(-1) the final aldehyde concentration ranged between 0.72 and 1.02 mg L(-1). Glyoxal and methylglyoxal concentrations were directly related to colour removal, whereas formaldehyde, acetaldehyde, acetone and acrolein were not. Thus, the extent of colour removal can be used to predict the increase in glyoxal and methylglyoxal concentrations. As colour removal can be assessed by a simple absorbance measurement, in contrast to the analysis of specific carbonyl compounds, which is much longer and complex, the possibility of using colour removal as an indicator for predicting the toxic potential of ozone by-products for textile effluents is of great value.

Assessment of peracetic acid disinfected effluents by microbiotests.

Bioassays were performed by commercially available kits on peracetic acid (PAA) solutions, at different concentrations, and on secondary effluents (from two different wastewater treatment plants) after disinfection at bench-scale, considering both samples containing residual active PAA and the same samples where residual PAA was quenched. Four indicator organisms were used: Vibrio fischeri, Thamnocephalus platyurus, Daphnia magna, and Selenastrum capricornutum. The experiments lead to conclude that Thamnocephalus platyurus is a very sensitive organism, probably not adequate to perform a reliable toxicity assessment of effluents for monitoring purposes. The presence of specific organic compounds deriving from human metabolism and urban pollution, even at very low concentrations, can affect the results of bioassays, especially those performed on Vibrio fischeri. PAA is toxic for bacteria and crustaceans even at concentrations lower than the ones commonly used in wastewater disinfection (2-5 mg/L), while its effect on algae is smaller. The toxic effect on bacteria was expected, as PAA is used for disinfection, but its possible influence on biological processes in the receiving aquatic environment should be considered. Toxicity on crustaceans would confirm the fact that discharging disinfected effluents could raise some environmental problems.

Wastewater disinfection alternatives: chlorine, ozone, peracetic acid, and UV light.

Disinfection tests were carried out at pilot scale to compare the disinfection efficiency of ozone, sodium hypochlorite (NaOCl), peracetic acid (PAA), and UV irradiation. Total coliforms, fecal coliforms, and Escherichia coli were monitored as reference microorganisms. Total heterotrophic bacteria (THB) were also enumerated by cytometry. At similar doses, NaOCl was more effective than PAA, and its action was less affected by contact time. The results obtained by ozonation were comparable for total coliforms, fecal coliforms, and E. coli. On the contrary, some differences among the three indicators were observed for NaOCl, PAA, and UV. Differences increased with increasing values of the disinfectant concentration times contact time (C x t) and were probably the result of different initial counts, as total coliforms include fecal coliforms, which include E. coli. The UV irradiation lead to complete E. coli removals, even at low doses (10 to 20 mJ/cm2). Total heterotrophic bacteria appeared to be too wide a group to be a good disinfection indicator; no correlation was found among THB inactivation, dose, and contact time.

Peracetic acid disinfection: a feasible alternative to wastewater chlorination.

The paper summarizes the results of a bench-scale study to evaluate the feasibility of using peracetic acid (PAA) as a substitute for sodium hypochlorite both for discharge into surface water and for agricultural reuse. Trials were carried out with increasing doses (1, 2, 3, 5, 10, and 15 mg/L) and contact times (6, 12, 18, 36, 42, and 54 minutes) to study disinfectant decay and bacterial removal and regrowth, using fecal coliform and Escherichia coli (E. coli) as process efficiency indicators. Peracetic acid decay kinetics was evaluated in tap water and wastewater; in both cases, PAA decays according to first-order kinetics with respect to time, and a correlation was found between PAA oxidative initial consumption and wastewater characteristics. The PAA disinfection efficiency was correlated with operating parameters (active concentration and contact time), testing different kinetic models. Two data groups displaying a different behavior on the basis of initial active concentration ranges (1 to 2 mg/L and 5 to 15 mg/L, respectively) can be outlined. Both groups had a "tailing-off" inactivation curve with respect to time, but the second one showed a greater inactivation rate. Moreover, the effect of contact time was greater at the lower doses. Hom's model, used separately for the two data groups, was found to best fit experimental data, and the disinfectant active concentration appears to be the main factor affecting log-survival ratios. Moreover, the S-model better explains the initial resistance of E. coli, especially at low active concentrations (< 2 mg/L) and short contact times (< 12 minutes). Microbial counts, performed by both traditional methods and flow cytometry, immediately and 5 hours after sample collection (both with or without residual PAA inactivation), showed that no appreciable regrowth took place after 5 hours, neither for coliform group bacteria, nor for total heterotrophic bacteria.

Secondary effluent disinfection: PAA long term efficiency.

The paper summarizes the results of a bench-scale study to evaluate the long-term disinfection efficiency of peracetic acid (PAA). Bacterial counts were repeated 5, 24, and 29 h after the end of the disinfection test, to simulate real re-growth conditions (no residual quenching) and, for the 5 h interval, the potential re-growth (quenching of residual PAA). Fecal coliforms, Escherichia coli, and total heterotrophic bacteria (THB) were enumerated by traditional plate count technique; THB were also enumerated by cytometry. After disinfection, the residual PAA concentration became negligible in about 5 to 11 h, depending on the tested doses. Microbial counts showed that no appreciable re-growth took place after 29 h for coliform group bacteria. For THB, the previously cited enumeration techniques gave different results in re-growth tests, especially for the lowest PAA doses. Indeed plate count technique evaluates the ability to form colonies, while cytometry enumerates intact membrane cells. No regrowth took place, even when no residual disinfectant was present, suggesting that bacteria are unable, even at the lowest doses, to repair damage caused by the PAA disinfecting action. PAA was found to be an efficient disinfecting agent, not only as a bacteriostatic, but also as a bactericide.

Study on anaerobic and aerobic degradation of different non-ionic surfactants.

Six alcohol ethoxylates (C5E2, C6E4, C7E4, C8E2, C8E4, C10E4) and two fatty acid esters were tested at lab-scale for degradation in anaerobic and aerobic conditions and oxygen uptake rate (OUR). Anaerobic removal of C5E2, C6E4 and C7E4 improved with increasing number of ethoxy groups (E) and decreasing length of the alkyl chain (C). Their aerobic removal was also great but lower than the anaerobic values. C8E2, C8E4 and C10E4 were adsorbed on sludge but not degraded in anaerobic conditions, while they were efficiently removed under aerobiosis. The fatty acid esters were removed to a level between the two alcohol ethoxylates groups in both anaerobiosis and aerobiosis. The measured OUR confirmed the different behaviours of the three groups of compounds.

Anaerobic removal of linear alcohol ethoxylates.

The present paper deals with a laboratory-scale study of anaerobic treatment of two commercial mixtures (LS2, LT7) of alcohol ethoxylates with 8-14 carbon atoms and 2 and 7 ethoxy groups. Tests were carried out in batch, with a 2 g l(-1) single dose, and in semibatch, with daily 0.2 g l(-1) doses. The behaviour of the tested mixtures was different: anaerobic sludge adsorption was the main removal process for LS2, while adsorption was less significant and biodegradation was more important for LT7. These differences appeared to be mainly related to the ethoxy portion length determining the extent of biodegradability and adsorption.