Thermal response analysis and compilation of cardinal temperatures for 424 strains of microalgae, cyanobacteria, diatoms and other species.

Microalgae and other phototrophic microorganisms can be cultivated to produce food and valuable bioproducts, also allowing to remove nutrients from wastewater and CO2 from biogas or polluted gas streams. Among other environmental and physico-chemical parameters, microalgal productivity is strongly influenced by the cultivation temperature. In this review, cardinal temperatures identifying the thermal response, i.e., the optimal growth condition (TOPT), and the lower and upper limits for microalgae cultivation (TMIN and TMAX), have been included in a structured and harmonized database. Literature data for 424 strains belonging to 148 genera of green algae, cyanobacteria, diatoms, and other phototrophs were tabulated and analysed, with a focus on the most relevant genera that are currently cultivated at the industrial scale in Europe. The dataset creation aimed at facilitating the comparison of different strain performances for different operational temperatures and assisting in the process of thermal and biological modelling, to reduce energy consumption and biomass production costs. A case study was presented, to illustrate the effect of temperature control on the energetic expenditure for cultivating different Chorella sp. strains under a greenhouse located in different European sites.

Respirometric assessment of bacterial kinetics in algae-bacteria and activated sludge processes.

Algae-bacteria (AB) consortia can be exploited for effective wastewater treatment, based on photosynthetic oxygenation to reduce energy requirements for aeration. While algal kinetics have been extensively evaluated, bacterial kinetics in AB systems are still based on parameters taken from the activated sludge models, lacking an experimental validation for AB consortia. A respirometric procedure was therefore proposed, to estimate bacterial kinetics in both activated sludge and AB, under different conditions of temperature, pH, dissolved oxygen, and substrate availability. Bacterial activities were differently influenced by operational/environmental conditions, suggesting that the adoption of typical activated sludge parameters could be inadequate for AB modelling. Indeed, respirometric results show that bacteria in AB consortia were adapted to a wider range of conditions, compared to activated sludge, confirming that a dedicated calibration of bacterial kinetics is essential for effectively modelling AB systems, and respirometry was proven to be a powerful and reliable tool to this purpose.

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.

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.

Biogas from mono- and co-digestion of microalgal biomass grown on piggery wastewater.

Biogas production has been suggested as a valid valorization solution for microalgal/bacteria biomass (MAB) grown on wastewater. This research is aimed at assessing the feasibility to use MAB grown in an outdoor raceway fed on piggery wastewater for biogas production. Batch and continuous anaerobic tests were conducted on the sole MAB and on a blend of MAB and carbonaceous substrates (deproteinated cheese whey and cellulose) to improve the carbon/nitrogen ratio. Results of batch biochemical methane potential tests confirmed that the sole microalgal/bacteria biomass was poorly degradable (119 NmLCH4·gCOD -1), while blending it with deproteinated cheese whey or cellulose (80% of carbonaceous material and 20% of MAB, as chemical oxygen demand (COD)) had no synergistic effects on the methane yield, although slight improvements in the degradation kinetics were observed. Continuous anaerobic degradation tests (at an organic loading rate of 1.5 gCOD·L-1·d-1, 35 °C and 30 days of hydraulic retention time) increased the overall methane yield from 81 NmLCH4·gCOD -1 (sole MAB) to 216 NmLCH4·gCOD -1 (MAB and deproteinated cheese whey) and 122 NmLCH4·gCOD -1 (MAB and cheese whey). However, data confirm that no evident synergistic effects were obtained.

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.

Inhibition on anammox bacteria upon exposure to digestates from biogas plants treating the organic fraction of municipal solid waste and the role of conductivity.

The aim of this research was to evaluate the applicability of the anammox process for removing nitrogen from the supernatant originating from the anaerobic digestion of the organic fraction of municipal solid waste (OFMSW). The short term inhibitory potential of this concentrated wastewater was evaluated by means of batch tests in terms of maximum specific anammox activity reduction. A total of 20 real wastewater samples were tested originating from 4 different full scale anaerobic digestion plants treating OFMSW. Activity reduction between 73% and 89% was observed in the presence of undiluted real wastewaters. The specific activity remained stable for 6-7days after the initial reduction, thus suggesting its treatability even without dilution. The inhibitory effect of both the real and synthetic saline media tested could be modelled as a function of conductivity. IC50 of 6.1mS/cm was obtained for exposure to the tested liquid fraction of biowaste digestate.

Autotrophic nitrogen removal by a two-step SBR process applied to mixed agro-digestate.

The aim of this research was to evaluate the applicability of partial-nitritation/anammox processes for biological N removal from a centrifuge supernatant coming from a full scale anaerobic digester fed on a mixture of piggery manure, poultry manure, and agro-wastes. Stable partial nitritation was achieved at pilot-scale (650L SBR), obtaining a suitable influent for the anammox lab-scale SBR reactor (3L). The anammox lab scale reactor was fed with increasing fractions of the partial nitritation effluent, blended with synthetic wastewater. In the last 100days no dilution was used. The nitrogen loading rate applied to the anammox reactor was 0.5-0.6gNL(-)(1)d(-)(1) and the average nitrogen removal was 91±10%. During the first days of operation with undiluted supernatant, the maximum anammox activity in the SBR decreased, but recovered afterwards, suggesting the ability of the anammox biomass to acclimate to the wastewater. N2O emissions in both reactors were also measured.

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.

Enzymatic and metabolic activities of four anaerobic sludges and their impact on methane production from ensiled sorghum forage.

Biochemical methane potential (BMP) tests were run on ensiled sorghum forage using four inocula (urban, agricultural, mixture of agricultural and urban, granular) and differences on their metabolic and enzymatic activities were also discussed. Results indicate that no significant differences were observed in terms of BMP values (258±14NmLCH4g(-1)VS) with a slightly higher value when agricultural sludge was used as inoculum. Significant differences can be observed among different inocula, in terms of methane production rate. In particular the fastest biomethanization occurred when using the urban sludge (hydrolytic kinetic constant kh=0.146d(-1)) while the slowest one was obtained from the agricultural sludge (kh=0.049d(-1)). Interestingly, positive correlations between the overall enzymatic activities and methane production rates were observed for all sludges, showing that a high enzymatic activity may favour the hydrolysis of complex substrate and accelerate the methanization process of sorghum.

Benefit of sodium hydroxide pretreatment of ensiled sorghum forage on the anaerobic reactor stability and methane production.

The assessment of the pretreatment effect on the anaerobic digestion process is generally based on the results of batch tests, which may fail in truly predicting full-scale anaerobic reactors performance. Therefore, in this study, the effect of alkaline pretreatment on the anaerobic digestion of ensiled sorghum forage was evaluated by comparing the results of two semi-continuous CSTR (Continuously Stirred Tank Reactor) anaerobic reactors. Results showed that an alkaline pretreatment step, prior to the anaerobic digestion of ensiled sorghum forage, can have a beneficial effect both in enhancing methane production (an increase of 25% on methane production was observed, if compared to that of untreated sorghum) and in giving more stability to the anaerobic digestion process.

Sodium hydroxide pretreatment of ensiled sorghum forage and wheat straw to increase methane production.

The aim of this study was to determine the effect of sodium hydroxide pretreatment on the chemical composition and the methane production of ensiled sorghum forage and wheat straw. NaOH pretreatment was conducted in closed bottles, at 40 °C for 24 h. Samples were soaked in a NaOH solution at different dosages (expressed in terms of total solids (TS) content) of 1 and 10% gNaOH/gTS, with a TS concentration of 160 gTS/L. At the highest NaOH dosage the reduction of cellulose, hemicelluloses and lignin was 31, 66 and 44%, and 13, 45 and 3% for sorghum and wheat straw, respectively. The concentration of soluble chemical oxygen demand (CODs) in the liquid phase after the pretreatment was also improved both for wheat straw and sorghum (up to 24 and 33%, respectively). Total sugars content increased up to five times at 10% gNaOH/gTS with respect to control samples, suggesting that NaOH pretreatment improves the hydrolysis of cellulose and hemicelluloses. The Biochemical Methane Potential (BMP) tests showed that the NaOH pretreatment favoured the anaerobic degradability of both substrates. At 1 and 10% NaOH dosages, the methane production increased from 14 to 31% for ensiled sorghum forage and from 17 to 47% for wheat straw. The first order kinetic constant increased up to 65% for sorghum and up to 163% for wheat straw.

Long-term effects of the ozonation of the sludge recycling stream on excess sludge reduction and biomass activity at full-scale.

This paper presents a full-scale experience of sludge minimization by means of short contact time ozonation in a wastewater treatment plant (WWTP) mainly fed on textile wastewater. The WWTP performance over a 3-year operational data series was analysed and compared with a two-year operation with sludge ozonation. Lab-scale respirometric tests were also performed to characterize biomass activity upstream and downstream of the ozone contact reactor. Results suggest that sludge ozonation: (1) is capable of decreasing excess sludge production by 17%; (2) partially decreases both N removal, by lowering the denitrification capacity, and P removal, by reducing biomass synthesis; (3) increases the decay rate from the typical value of 0.62 d(-1) to 1.3 d(-1); (4) decreases the heterotrophic growth yield from the typical value of 0.67 to 0.58 gCOD/gCOD.

Microcalorimetric and manometric tests to assess anammox activity.

The present study compares two experimental methods to evaluate Anammox activity based on the assessment of (1) the N(2) production rate by a manometric device, as previously proposed, and (2) the heat production rate by a microcalorimeter. Two samples of Anammox suspended biomass were taken from a pilot-plant, and their specific Anammox activity measured by both techniques. Both methods were successfully applied. As for calorimetric tests, they were performed for the first time on Anammox enriched sludge samples. Comparisons between the specific Anammox activities estimated by manometry and calorimetry and between expected (from the reaction enthalpy) and measured heat productions were performed. Promising results were obtained.

A simple method to evaluate the short-term biogas yield in anaerobic codigestion of WAS and organic wastes.

The present study was aimed at setting and applying a procedure to measure the anaerobic degradability of different organic substrates by short-term tests (2-7 days) carried out at lab-scale with a low food to biomass (F/M) ratio. All tests were carried out using an acclimated sludge taken from a pilot-plant anaerobic digester (200 L). Trials were performed with a manometric system. The experimental reliability of the device in measuring the anaerobic degradability was assessed by several preliminary tests carried out using acetate and glucose as reference substrates. The average conversion to methane was 99% for acetate and of 83% for glucose. The results of tests in triplicate showed the high repeatability of the method with an average coefficient of variation lower than 2%. Then, the lab-scale procedure was applied to study the short-term anaerobic degradability of complex organic substrates: thickened waste activated sludge, two kinds of organic fraction of municipal solid waste (a kitchen waste and a fruit and vegetable waste collected at the wholesale market of Florence), olive mill wastewater and freshly harvested grass. Results indicated that organic fraction of municipal solid waste, olive mill wastewater and grass were characterized by a much higher anaerobic degradability if compared to the thickened activated sludge, well in agreement with literature data.

Automatic set-point titration for monitoring nitrification in SBRs.

Nitrification is usually the bottleneck of biological nitrogen removal processes. In SBRs systems, it is not often enough to monitor dissolved oxygen, pH and ORP to spot problems which may occur in nitrification processes. Therefore, automated supervision systems should be designed to include the possibility of monitoring the activity of nitrifying populations. Though the applicability of set-point titration for monitoring biological processes has been widely demonstrated in the literature, the possibility of an automated procedure is still at its early stage of industrial development. In this work, the use of an at-line automated titrator named TITAAN (TITrimetric Automated ANalyser) is presented. The completely automated sensor enables us to track nitrification rate trend with time in an SBR, detecting the causes leading to slower specific nitrification rates. It was also possible to perform early detection of toxic compounds in the influent by assessing their effect on the nitrifying biomass. Nitrifications rates were determined with average errors+/-10% (on 26 tests), never exceeding 20% as compared with UV-spectrophotometric determinations.

SBR on-line monitoring by set-point titration.

The applicability of set-point titration for monitoring biological processes has been widely demonstrated in the literature. Based on published and on-going experiences, some operating procedures have been specifically developed to be applied to SBRs, so that real-time information about the process and/or the influent can be obtained. This, in turn, would allow plant operators to select the most appropriate actions properly and timely. Five operating modes are described for the monitoring of (1) influent toxicity, (2) influent N-content, (3) nitrification capacity, (4) end of the nitrification reaction, and (5) nitrate effluent concentration, and are currently tested on the on-line titrator TITAAN (TITrimetric Automated ANalyser) which is in operation on a pilot scale SBR.

Coupling pH-stat and DO-stat titration to monitor degradation of organic substrates.

A combined titration methodology at constant pH and dissolved oxygen concentration to monitor the degradation of organics as carbon dioxide evolution rate (CER) and oxygen uptake rate (OUR) is presented. Equations necessary to assess CER from alkali titration rates and the instrument used to test the technique are described. Experiments were performed on samples of activated sludge from a domestic wastewater treatment plant using glucose and ethanol as substrates. OUR and CER were calculated for both substrates, as well as the respiratory quotient RQ = CER/OUR and biomass to substrate yields coefficients. RQ data for ethanol compared better with corresponding stoichiometric values than those for glucose.