Assessing nitrous oxide emissions from algal-bacterial photobioreactors devoted to biogas upgrading and digestate treatment.

Nitrous oxide (N2O) emissions in High Rate Algal Ponds (HRAP) can negatively affect the sustainability of algal-bacterial processes. N2O emissions from a pilot HRAP devoted to biogas upgrading and digestate treatment were herein monitored for 73 days. The influence of the pH (7.5, 8.5, and 9.5), nitrogen sources (100 mg L-1 of N-NO2-, N-NO3-, and N-NH4+) and illumination on N2O emissions from the algal-bacterial biomass of the HRAP was also assessed in batch tests. Significantly higher N2O gas concentrations of 311.8 ± 101.1 ppmv were recorded in the dark compared to the illuminated period (236.9 ± 82.6 ppmv) in the HRAP. The batch tests revealed that the highest N2O emission rates (49.4 mmol g-1 TSS·h-1) occurred at pH 8.5 in the presence of 100 mg N-NO2-/L under dark conditions. This study revealed significant N2O emissions in HRAPs during darkness.

Exploring resistomes and microbiomes in pilot-scale microalgae-bacteria wastewater treatment systems for use in low-resource settings.

Antibiotic resistance genes (ARGs) released into the environment are an emerging human and environmental health concern, including ARGs spread in wastewater treatment effluents. In low-to-middle income countries (LMICs), an alternate wastewater treatment option instead of conventional systems are low-energy, high-rate algal ponds (HRAP) that use microalgae-bacteria aggregates (MABA) for waste degradation. Here we studied the robustness of ARG removal in MABA-based pilot-scale outdoor systems for 140 days of continuous operation. The HRAP system successfully removed 73 to 88 % chemical oxygen demand and up to 97.4 % ammonia, with aggregate size increasing over operating time. Fourteen ARG classes were identified in the HRAP influent, MABA, and effluent using metagenomics, with the HRAP process reducing total ARG abundances by up to 5-fold from influent to effluent. Parallel qPCR analyses showed the HRAP system significantly reduced exemplar ARGs (p < 0.05), with 1.2 to 4.9, 2.7 to 6.3, 0 to 1.5, and 1.2 to 4.8 log-removals for sul1, tetQ, blaKPC, and intl1 genes, respectively. Sequencing of influent, effluent and MABAs samples showed associated microbial communities differed significantly, with influent communities by Enterobacteriales (clinically relevant ARGs carrying bacteria), which were less evident in MABA and effluent. In this sense, such bacteria might be excluded from MABA due to their good settling properties and the presence of antimicrobial peptides. Microalgae-bacteria treatment systems steadily reduced ARGs from wastewater during operation time, using sunlight as the energetic driver, making them ideal for use in LMIC wastewater treatment applications.

A pilot-scale study of the integrated phycoremediation-photolytic ozonation based municipal solid waste leachate treatment process.

Municipal solid waste (MSW) leachate is a highly polluted liquid that accumulates in the landfill and contains a high concentration of toxic pollutants which can pollute the surrounding surface water and groundwater as well, if not treated properly. In this study, an integrated approach of phycoremediation with photolytic ozonation was employed for the leachate collected from the MSW dumpsite which has high Chemical Oxygen Demand (COD) and ammonium (NH4+) levels. Photolytic ozonation treatment was employed as a pre-treatment step under operating parameters of pH: 9.0; Ozone dosage: 5 g/h; UV-C: λ = 254 nm; and contact time: 60 min, in which the COD and NH4+ in the leachate was reduced up to 81% and 95%, respectively. The selected algae Chlorella vulgaris (C.vulgaris) was employed in a lab-scale study to optimize the inoculum conditions in the photolytic ozonated leachate (POL). The specific growth rate of C.vulgaris was observed as 0.14/d in the POL at the optimized condition (inoculum size of 25% (T25)) during the study period of 11 days. High-rate algal pond (HRAP) was employed for the pilot-scale study in controlled environmental conditions as in the T25 experimental run for the assessment of POL treatment and biomass production. C.vulgaris reduced the concentration of pollutants COD, NH4+, and heavy metals (Cu, Fe) in the POL up to 93%, 94%, and 71%, respectively, with the dry biomass productivity of 0.727 g/L/d which is 3 times higher than the biomass productivity of C.vulgaris in freshwater conditions. The biochemical composition (carbohydrates, proteins, and lipids) of the harvested biomass has higher lipid production with lipid productivity of 120 mg/L/d which can be used as a feedstock for the production of value-a dded products.

Shelterin Components Modulate Nucleic Acids Condensation and Phase Separation in the Context of Telomeric DNA.

Telomeres are nucleoprotein complexes that protect the ends of chromosomes and are essential for chromosome stability in Eukaryotes. In cells, individual telomeres form distinct globules of finite size that appear to be smaller than expected for bare DNA. Moreover, telomeres can cluster together, form telomere-induced-foci or co-localize with promyelocytic leukemia (PML) nuclear bodies. The physical basis for collapse of individual telomeres and coalescence of multiple ones remains unclear, as does the relationship between these two phenomena. By combining single-molecule force spectroscopy measurements, optical microscopy, turbidity assays, and simulations, we show that the telomere scaffolding protein TRF2 can condense individual DNA chains and drives coalescence of multiple DNA molecules, leading to phase separation and the formation of liquid-like droplets. Addition of the TRF2 binding protein hRap1 modulates phase boundaries and tunes the specificity of solution demixing while simultaneously altering the degree of DNA compaction. Our results suggest that the condensation of single telomeres and formation of biomolecular condensates containing multiple telomeres are two different outcomes driven by the same set of molecular interactions. Moreover, binding partners, such as other telomere components, can alter those interactions to promote single-chain DNA compaction over multiple-chain phase separation.

Advanced biokinetic and hydrodynamic modelling to support and optimize the design of full-scale high rate algal ponds.

High rate algal ponds (HRAP) are known for their suitability to treat wastewater and to produce microalgal biomass, which can be converted into bioproducts. However, full-scale application of HRAP is still limited to few cases, and design procedures are not consolidated or standardized. In this study, a demonstrative-scale HRAP system for secondary wastewater treatment to be implemented in India (treatment capacity of 50 m3·d-1) has been designed combining conventional dimensioning techniques and advanced modelling tools. The objective of the study was to assist, verify and optimize the conventional dimensioning of the secondary HRAP by means of simulations predicting the behaviour of the system in the specific local conditions under different configurations and operational strategies. Biokinetic modelling and hydrodynamic analysis using Computational Fluid Dynamics (CFD) were carried out. The simulations performed with the biokinetic model showed that the optimal hydraulic retention time to enhance nutrient removal and biomass production is 4 days. For the hydrodynamic modelling, a 3D model of the HRAP was built to simulate the hydrodynamic behaviour of 36 different designs. Simulations allowed quantifying the presence of low velocity zones as well as the land use efficiency of the different designs in terms of the useful area vs. the total occupied area. Two baffles and tear-shapes with a diameter equal to » of the channel width was the most efficient configuration. Moreover, a technical-economic assessment of the system was carried out, resulting in an investment cost of 483 € per population equivalent and an operational cost of 0.19 € per m3 of treated wastewater.

Graphitic bio-char and bio-oil synthesis via hydrothermal carbonization-co-liquefaction of microalgae biomass (oiled/de-oiled) and multiple heavy metals remediations.

Thermochemical transformation of microalgae biomass into graphitic bio-chars entices as proficient bio-adsorbents for heavy metal contaminants. This study explores the synergistic impact of Chlorella sorokiniana on biomass generation and wastewater remediation in high rate algae pond (HRAP). Biomass produced was applied for hydrothermal carbonization-co-liquefaction (HTCL). The structural and morphological characteristics of HTCL products (i.e. bio-chars and bio-oils) have been systematically studied by XRD, Raman, FTIR, elemental analyzer, SEM, BET, and 1H NMR spectroscopy. The crystallite size of the graphite 2H indexing planes was to be 4.65 nm and 14.07 nm in the bio-chars of oiled biomass (MB-OB) and de-oiled biomass (MB-DOB), respectively. The increase in the ID/IG ratio of MB-DOB indicated the highly disordered graphitic structure due to the appearance of carbonyl, hydroxyl, and epoxy functionalities in the line of high C/N and low C/H ratio. Also, the multiple heavy metals remediation of MB-DOB revealed better efficiency as ~100% in 720 min. The kinetics analysis shows the correlation coefficient of pseudo-second-order is well fitted compared to the pseudo-first-order. The Langmuir adsorption model signifies the adsorption of heavy metal ions in a monolayer adsorption manner. The study proposes the microalgae bio-char potential for multiple heavy metals remediation alongside bio-oils.

Performance of a membrane-coupled high-rate algal pond for urban wastewater treatment at demonstration scale.

The objective of this study was to evaluate the performance of an outdoor membrane-coupled high-rate algal pond equipped with industrial-scale membranes for treating urban wastewater. Decoupling biomass retention time (BRT) and hydraulic retention time (HRT) by membrane filtration resulted in improved process efficiencies, with higher biomass productivities and nutrient removal rates when operating at low HRTs. At 6 days of BRT, biomass productivity increased from 30 to 66 and to 95 g·m-3·d-1 when operating at HRTs of 6, 4 and 2.5 days, respectively. The corresponding nitrogen removal rates were 4, 8 and 11 g N·m-3·d-1 and the phosphorous removal rates were 0.5, 1.3 and 1.6 g P·m-3·d-1. The system was operated keeping moderate specific air demands (0.25 m3·m-2·h-1), resulting in reasonable operating and maintenance costs (€0.04 per m3) and energy requirements (0.29 kWh per m3). The produced water was free of pathogens and could be directly used for reusing purposes.

Small-scale phyco-mitigation of raw urban wastewater integrated with biodiesel production and its utilization for aquaculture.

A low-cost small-scale high-rate algal pond (HRAP) was constructed to investigate the synergistic potential of a novel oleaginous microalga, Chlorella sorokiniana for phyco-mitigation, and biodiesel production using raw urban wastewater. An enhanced nutrient removal (97%), total organic carbon (74%), alkalinity (70%) and hardness (75%) from the wastewater was obtained. The microalga dominated in the HRAP as ~90% increase in the dissolved oxygen with high biomass (1.13 g/L) was noted. The microalga biomass showed sufficient lipid content (~31% of dry cell weight) as compared to control (Bold's Basal media). The total lipid profiling of the microalga cultivated in wastewater showed augmentation in the levels of both storage and neutral lipids with good quality fatty acids composition. Moreover, the sucker fishes grew healthy in the treated wastewater with an increase in body weight.

A conceptual scenario for the use of microalgae biomass for microgeneration in wastewater treatment plants.

Microalgae are a potential source of biomass for the production of energy, which is why the amount of research on this topic has increased in recent years. This work describes the state of the art of microalgae production from wastewater treatment plants (WWTP), its potential to generate electricity and the scale in which it is possible. The methodology used was a systematic review of the gasification of microalgae from 49 articles selected. Based on the review, a conceptual scenario for microgeneration in WWTP using as feedstock microalgae for thermal gasification was developed. The most consistent assumptions for a real scale microgeneration are microalgae production in open ponds using domestic sewage as a nutritional medium; the use of the flocculation process in process of harvesting; microalgae to energy through thermal gasification process using a downdraft gasifier. Considering a WWTP with a 3000 m3/d flux capacity, 860 kg/d of dry microalgae biomass might be produced. For which, gasification has a production potential of 0.167 kWh/m3 of treated sewage, but the energy balance is compromised by the drying process. However, when the biogas produced in anaerobic treatment enter in the model, it is possible to add a surplus of electricity of 0.14 kWh/m3 of treated sewage. Finally, a cost estimate is made for the acquisition of drying and gasification-electricity generation systems. For this scenario, the results suggest that the investments may be financially returned after five years, with additional potential for further optimization.

Technology validation of photosynthetic biogas upgrading in a semi-industrial scale algal-bacterial photobioreactor.

The performance of photosynthetic biogas upgrading coupled to wastewater treatment was evaluated in an outdoors high rate algal pond (HRAP) interconnected to an absorption column at semi-industrial scale. The influence of biogas flowrate (274, 370 and 459 L h-1), liquid to biogas ratio (L/G = 1.2, 2.1 and 3.5), type of wastewater (domestic versus centrate) and hydraulic retention time in the HRAP (HRT) on the quality of the biomethane produced was assessed. The highest CO2 and H2S removal efficiencies (REs) were recorded at the largest L/G due to the higher biogas-liquid mass transfer at increasing liquid flowrates. No significant influence of the biogas flowrate on process performance was observed, while the type of wastewater was identified as a key operational parameter. CO2 and H2S-REs of 99% and 100% at a L/Gmax = 3.5 were recorded using centrate. The maximum CH4 content in the biomethane (90%) was limited by N2 and O2 desorption.

Combining sun-based technologies (microalgae and solar disinfection) for urban wastewater regeneration.

Solar disinfection (SODIS) of urban wastewater can be a suitable technology for improving the microbiological quality of reclaimed water as a complement to other extensive and environmentally friendly technologies such as microalgae biotreatment. The objective of this work is to evaluate the feasibility of incorporating the SODIS technology at the end of a pilot scale urban wastewater treatment plant (WWTP) where the processes are based on microalgae biotechnology and comprising three Upflow Anaerobic Sludge Blanket (UASB, 20m3 each one) reactor, six High Rate Algal Ponds (HRAP, 32m2 each one), and a Dissolved Air Flotation (DAF, 1m3) unit. E. coli concentration was monitored at the effluent of the different units (UASB, HRAP, DAF) of the pilot WWTP. The efficiency of the SODIS process was studied for the inactivation of three of the commonly employed indicator microorganisms (Escherichia coli, Enterococcus spp. and Clostridium perfringens) using a compound parabolic collector (CPC) for five months under various conditions of irradiance and temperature. E. coli and Enterococcus spp. were more effectively disinfected by the SODIS unit (2.9 and 2.5 logarithms of reduction on average, respectively) than by the HRAP (2 and 1.1) or the DAF (0.9 and 0.1). On the contrary, the DAF technology achieved better reduction rates of C. perfringens (1.7) than the SODIS (0.9) and the HRAP (0.1). No regrowth of any microorganisms was detected during dark storage after the SODIS treatment. Incorporating a SODIS unit after the non-conventional WWTP processes substantially increases the possibilities for reuse of the treated water after receiving a cumulative UV radiation dose of 25W·h/m2 (50min of normalized time of solar illumination). The surface requirement of the SODIS equipment would be 3.5 times smaller than the HRAP's surface.

Influence of Water Depth on Microalgal Production, Biomass Harvest, and Energy Consumption in High Rate Algal Pond Using Municipal Wastewater.

The high rate algal ponds (HRAP) powered and mixed by a paddlewheel have been widely used for over 50 years to culture microalgae for the production of various products. Since light incidence is limited to the surface, water depth can affect microalgal growth in HRAP. To investigate the effect of water depth on microalgal growth, a mixed microalgal culture constituting three major strains of microalgae including Chlorella sp., Scenedesmus sp., and Stigeoclonium sp. (CSS), was grown at different water depths (20, 30, and 40 cm) in the HRAP, respectively. The HRAP with 20cm of water depth had about 38% higher biomass productivity per unit area (6.16 ± 0.33 g·m⁻²·d⁻¹) and required lower nutrients and energy consumption than the other water depths. Specifically, the algal biomass of HRAP under 20c m of water depth had higher settleability through larger floc size (83.6% settleability within 5 min). These results indicate that water depth can affect the harvesting process as well as cultivation of microalgae. Therefore, we conclude that water depth is an important parameter in HRAP design for mass cultivation of microalgae.

Bioinformatics analysis to assess potential risks of allergenicity and toxicity of HRAP and PFLP proteins in genetically modified bananas resistant to Xanthomonas wilt disease.

Banana Xanthomonas wilt (BXW) disease threatens banana production and food security throughout East Africa. Natural resistance is lacking among common cultivars. Genetically modified (GM) bananas resistant to BXW disease were developed by inserting the hypersensitive response-assisting protein (Hrap) or/and the plant ferredoxin-like protein (Pflp) gene(s) from sweet pepper (Capsicum annuum). Several of these GM banana events showed 100% resistance to BXW disease under field conditions in Uganda. The current study evaluated the potential allergenicity and toxicity of the expressed proteins HRAP and PFLP based on evaluation of published information on the history of safe use of the natural source of the proteins as well as established bioinformatics sequence comparison methods to known allergens (www.AllergenOnline.org and NCBI Protein) and toxins (NCBI Protein). The results did not identify potential risks of allergy and toxicity to either HRAP or PFLP proteins expressed in the GM bananas that might suggest potential health risks to humans. We recognize that additional tests including stability of these proteins in pepsin assay, nutrient analysis and possibly an acute rodent toxicity assay may be required by national regulatory authorities.

Emerging contaminant degradation and removal in algal wastewater treatment ponds: Identifying the research gaps.

Whereas the fate of emerging contaminants (ECs) during 'conventional' and 'advanced' wastewater treatment (WWT) has been intensively studied, little research has been conducted on the algal WWT ponds commonly used in provincial areas. The long retention times and large surface areas exposed to light potentially allow more opportunities for EC removal to occur, but experimental evidence is lacking to enable definite predictions about EC fate across different algal WWT systems. This study reviews the mechanisms of EC hydrolysis, sorption, biodegradation, and photodegradation, applying available knowledge to the case of algal WWT. From this basis the review identifies three main areas that need more research due to the unique environmental and ecological conditions occurring in algal WWT ponds: i) the effect of diurnally fluctuating pH and dissolved oxygen upon removal mechanisms; ii) the influence of algae and algal biomass on biodegradation and sorption under relevant conditions; and iii) the significance of EC photodegradation in the presence of dissolved and suspended materials. Because of the high concentration of dissolved organics typically found in algal WWT ponds, most EC photodegradation likely occurs via indirect mechanisms rather than direct photolysis in these systems.