Thermochemical valorization of greenhouse cucumber, tomato and pepper as biofuel.

Agricultural residues adequate managing contributes to reduce CO2 emissions and easy the circular economy. In this paper, cucumber, tomato and pepper greenhouse crop residues were characterized both chemically and energetically to potential of waste as a biofuel. When compared to other herbaceous and woody biomass, the data show higher moisture, ash and chlorine contents. The gross calorific values of these residues are in the same range as the herbaceous biomass (14.92 MJ/kg), with net values of 13.71, 14.58 and 15.73 MJ/kg for cucumber, tomato and pepper, respectively. The study also included other not common elements, finding the highest values for calcium (13-25 mg/kg) and potassium (23-30 mg/kg). To date, empirical correlations have been defined to predict the CV as a function of total C content (expressed as % in dry weight) for these residues individually. The empirical correlations obtained had maximum absolute errors (MAE) of 0.11, 0.32 and 0.58 % for cucumber, tomato and pepper whereas the marginal mean bias errors (MBE) were 0.016, 0.002 and 0.004 %, respectively, which confirms the value of using this method to determinate the gross calorific value of these residues. The correlations shown provide a useful tool for developing energy production processes based on crop residues, that would appear to be an interesting source of renewable energy to produce heat.

Identification of Marine Biotechnology Value Chains with High Potential in the Northern Mediterranean Region.

Marine (blue) biotechnology is an emerging field enabling the valorization of new products and processes with massive potential for innovation and economic growth. In the Mediterranean region, this innovation potential is not exploited as well as in other European regions due to a lack of a clear identification of the different value chains and the high fragmentation of business innovation initiatives. As a result, several opportunities to create an innovative society are being missed. To address this problem, eight Northern Mediterranean countries (Croatia, France, Greece, Italy, Montenegro, Portugal, Slovenia and Spain) established five national blue biotechnology hubs to identify and address the bottlenecks that prevent the development of marine biotechnology in the region. Following a three-step approach (1. Analysis: setting the scene; 2. Transfer: identification of promising value chains; 3. Capitalization: community creation), we identified the three value chains that are most promising for the Northern Mediterranean region: algae production for added-value compounds, integrated multi-trophic aquaculture (IMTA) and valorization aquaculture/fisheries/processing by-products, unavoidable/unwanted catches and discards. The potential for the development and the technical and non-technical skills that are necessary to advance in this exciting field were identified through several stakeholder events which provided valuable insight and feedback that should be addressed for marine biotechnology in the Northern Mediterranean region to reach its full potential.

Photosynthesis Monitoring in Microalgae Cultures Grown on Municipal Wastewater as a Nutrient Source in Large-Scale Outdoor Bioreactors.

Microalgae cultures were used for a WW treatment to remediate nutrients while producing biomass and recycling water. In these trials, raceway ponds (RWPs; 1 and 0.5 ha) were located next to a municipal (WW) treatment plant in Mérida, Spain. The ponds were used for continuous, all-year-round microalgae production using WW as a source of nutrients. Neither CO2 nor air was supplied to cultures. The objective was to validate photosynthesis monitoring techniques in large-scale bioreactors. Various in-situ/ex-situ methods based on chlorophyll fluorescence and oxygen evolution measurements were used to follow culture performance. Photosynthesis variables gathered with these techniques were compared to the physiological behavior and growth of cultures. Good photosynthetic activity was indicated by the build-up of dissolved oxygen concentration up to 380% saturation, high photochemical yield (Fv/Fm = 0.62-0.71), and relative electron transport rate rETR between 200 and 450 µmol e- m-2 s-1 at midday, which resulted in biomass productivity of about 15-25 g DW m-2 day-1. The variables represent reliable markers reflecting the physiological status of microalgae cultures. Using waste nutrients, the biomass production cost can be significantly decreased for abundant biomass production in large-scale bioreactors, which can be exploited for agricultural purposes.

Centrate as a sustainable growth medium: Impact on microalgal inocula and bacterial communities in tubular photobioreactor cultivation systems.

Centrate is a low-cost alternative to synthetic fertilizers for microalgal cultivation, reducing environmental burdens and remediation costs. Adapted microalgae need to be selected and characterised to maximise biomass production and depuration efficiency. Here, the performance and composition of six microalgal communities cultivated both on synthetic media and centrate within semi-open tubular photobioreactors were investigated through Illumina sequencing. Biomass grown on centrate, exposed to a high concentration of ammonium, showed a higher quantity of nitrogen (5.6% dry weight) than the biomass grown on the synthetic media nitrate (3.9% dry weight). Eukaryotic inocula were replaced by other microalgae while cyanobacterial inocula were maintained. Communities were generally similar for the same inoculum between media, however, inoculation with cyanobacteria led to variability within the eukaryotic community. Where communities differed, centrate resulted in a higher richness and diversity. The higher nitrogen of centrate possibly led to higher abundance of genes coding for N metabolism enzymes.

Simulation and Techno-Economical Evaluation of a Microalgal Biofertilizer Production Process.

Due to population growth in the coming years, an increase in agricultural production will soon be mandatory, thus requiring fertilizers that are more environmentally sustainable than the currently most-consumed fertilizers since these are important contributors to climate change and water pollution. The objective of this work is the techno-economic evaluation of the production of biofertilizer concentrated in free amino acids from microalgal biomass produced in a wastewater treatment plant, to determine its economic viability. A process proposal has been made in six stages that have been modelled and simulated with the ASPEN Plus simulator. A profitability analysis has been carried out using a Box-Behnken-type response surface statistical design with three factors-the cost of the biomass sludge, the cost of the enzymes, and the sale price of the biofertilizer. It was found that the most influential factor in profitability is the sale price of the biofertilizer. According to a proposed representative base case, in which the cost of the biomass sludge is set to 0.5 EUR/kg, the cost of the enzymes to 20.0 EUR/kg, and the sale price of the biofertilizer to 3.5 EUR/kg, which are reasonable costs, it is concluded that the production of the biofertilizer would be economically viable.

Profiling microalgal cultures growing on municipal wastewater and fertilizer media in raceway photobioreactors.

Microalgae cultivation is proposed as an effective system for pathogens reduction and wastewater depuration, however, a full characterisation of the risks is still needed. Two raceways were inoculated with Scenedesmus, one using wastewater and the other using a fertilizer medium. Microbial community and pathogen presence were explored by next generation sequencing (NGS), commercial qPCR array and plate counts. These methods proved to be complementary for a full characterization of community structure and potential risks. Media and sampling locations contributed to shape communities and pathogenic loads. The main pathogenic genera detected were Arcobacter and Elizabethkingia (mainly in wastewater) with an important presence of Aeromonas (all samples). A lower presence of pathogens was detected in fertilizer samples, while wastewater showed a reduction from inlet to outlet. Raceways showed potential as an effective biotreatment, with most of the retained pathogens released in the outlet and only a minor part settled in the biomass.

Pathogens and predators impacting commercial production of microalgae and cyanobacteria.

Production of phytoplankton (microalgae and cyanobacteria) in commercial raceway ponds and other systems is adversely impacted by phytoplankton pathogens, including bacteria, fungi and viruses. In addition, cultures are susceptible to productivity loss, or crash, through grazing by contaminating zooplankton such as protozoa, rotifers and copepods. Productivity loss and product contamination are also caused by otherwise innocuous invading phytoplankton that consume resources in competition with the species being cultured. This review is focused on phytoplankton competitors, pathogens and grazers of significance in commercial culture of microalgae and cyanobacteria. Detection and identification of these biological contaminants are discussed. Operational protocols for minimizing contamination, and methods of managing it, are discussed.

Microalgae based wastewater treatment coupled to the production of high value agricultural products: Current needs and challenges.

One of the main social and economic challenges of the 21st century will be to overcome the worlds' water deficit expected by the end of this decade. Microalgae based wastewater treatment has been suggested as a strategy to recover nutrients from wastewater while simultaneously producing clean water. Consortia of microalgae and bacteria are responsible for recovering nutrients from wastewater. A better understanding of how environmental and operational conditions affect the composition of the microalgae-bacteria consortia would allow to maximise nutrient recoveries and biomass productivities. Most of the studies reported to date showed promising results, although up-scaling of these processes to reactors larger than 100 m2 is needed to better predict their industrial relevance. The main advantage of microalgae based wastewater treatment is that valuable biomass with unlimited applications is produced as a co-product. The aim of the current paper was to review microalgae based wastewater treatment processes focusing on strategies that allow increasing both biomass productivities and nutrient recoveries. Moreover, the benefits of microalgae based agricultural products were also discussed.

Effect of operational parameters, environmental conditions, and biotic interactions on bacterial communities present in urban wastewater treatment photobioreactors.

The effects of water depth, operational and environmental conditions on bacterial communities were analyzed in microalgal-bacterial outdoor photobioreactors treating urban wastewaters from March to August 2014. Three raceway photobioreactors inoculated with Scenedesmus sp. and with different water depths (20, 12, and 5 cm) were used at different dilution rates (0.15, 0.3, 0.4, and 0.5 d-1). A thin-layer reactor with 2 cm water depth and operated at 0.3 d-1 was used as a control. The results showed that biomass productivity increased as water depth decreased. The highest biomass productivity was 0.196 gL-1d-1, 0.245 gL-1d-1, and 0.457 gL-1d-1 for 20, 12, and 5 cm depth raceway photobioreactors, respectively. These values were lower than the maximum productivity registered in the control reactor (1.59 gL-1d-1). Bacterial communities, analyzed by high-throughput 16S rRNA sequencing, were not affected by water depth. A decrease in community evenness was related to a decrease in nutrient removal. Hetetrotrophs and phototrophs, mainly from the family Rhodobacteraceae, dominated bacterial diversity. The community changed due to increasing temperatures, irradiance, and organic carbon, ammonia, and phosphate contents in the photobioreactor-influent as well as, microalgae inhibition and higher organic carbon in the effluent. The photobioreactors shared a core-biome that contained five clusters of co-occurring microorganisms. The bacteria from the different clusters were taxonomically and ecologically different but functionally redundant. Overall, the drivers of the community changes could be related to abiotic variables and complex biological interactions, likely mediated by microalgae excretion of organic substances and the microorganisms' competence for substrates.

OPTIMIZATION OF A NEW CULTURE MEDIUM FOR THE LARGE-SCALE PRODUCTION OF PROTEIN-RICH ARTHROSPIRA PLATENSIS (OSCILLATORIALES, CYANOPHYCEAE).

Our aim was to develop a novel medium for the large-scale production of protein-rich Arthrospira with potential applications as a biofertilizer. The novel culture medium, termed as FM-II, was formulated using low-cost commercial chemicals and specifically designed to improve protein production. Both Arthrospira platensis and Arthrospira maxima were produced using FM-II and Arnon medium, which was used as a control. Photosynthetic status of the cells, which was checked by measuring chlorophyll fluorescence, biomass dry weight and protein content, was assessed daily. Arthrospira platensis had higher biomass and protein productivities than A. maxima when cultured in both control and FM-II media. Incorporation of varied micronutrients into FM-II formulation did not improve biomass productivity. Maximum biomass dry weight in FM-II and control medium was 2.9 and 2.5 g · L-1 , respectively. Total protein content of the biomass ranged between 55% and 65%, suggesting potential for being used in the development of high-value agricultural products. As some nutrients were discarded unused, the initial content of phosphates and bicarbonates was reduced by 75% and 50%, respectively, without affecting the process productivity. Results reported herein could promote the production and utilization of Arthrospira platensis by significantly reducing productions costs and therefore increasing the feasibility of the process.

Spirulina for the food and functional food industries.

Humans are no strangers to the consumption of microalgae as already in the sixteenth century Spirulina was harvested from Lake Texcoco and consumed in markets in Tenochtitlan (today Mexico City). Nowadays, microalgae are being incorporated into many food formulations. Most of these use microalgae as a marketing strategy or as a colouring agent. However, Spirulina (and compounds derived thereof) show potential for being used as ingredients in the development of novel functional foods, which are one of the top trends in the food industry. Several human intervention studies demonstrated the potential of Spirulina for being used in the prevention or treatment of disorders related to metabolic syndrome. The aim of the current paper was to review current and potential applications of this microalga in the food and functional food industries. Health benefits associated with consuming Spirulina and/or some of the most important compounds derived from Spirulina were also discussed.

Utilization of centrate for the outdoor production of marine microalgae at the pilot-scale in raceway photobioreactors.

In this study, the outdoor production of marine microalgae in raceway photobioreactors was investigated, modifying the centrate percentage in the culture medium (20, 30, 40 and 50%) and using two different dilution rates (0.2 day-1 and 0.3 day-1). The data obtained showed that microalgae are capable of producing biomass in addition to recovering the nutrients contained in the centrate. The best results for biomass productivity and light efficiency were obtained when the centrate was set at 20% with a dilution rate of 0.3 day-1. The biomass productivity was 32.42 g m-2·day-1 while the photosynthetic efficiency was 0.74 gbiomass·E-1 (3.66%). Regarding the nutrients, nitrogen (the majority being in the form of ammonium [NH4+]) and phosphorus were only fixed into biomass when optimal conditions were set; if this was not the case, they were lost to stripping or precipitation. The maximal nutrient removal capacities under the optimal conditions were 28.72 mgN·l-1·day-1 and 3.99 mgP·l-1·day-1. Population changes were determined by the dilution rate set whilst the centrate percentage had little effect. Four strains were present in the culture, Nannochloropsis g. being the main one. Biochemical changes did not vary greatly between the conditions set for the culture, with a composition rich in proteins and carbohydrates being observed. One can conclude that to produce marine microalgal biomass for a range of potential commodities such as feed, biofertilizers and biofuels, it is possible to use centrate from anaerobic digestion as the sole nutrient source, as a way of reducing costs.

A new approach for detection and quantification of microalgae in industrial-scale microalgal cultures.

In industrial-scale microalgal cultures, non-target microalgae compete with the desired species for nutrients and CO2, thus reducing the growth rate of the target species and the quality of the produced biomass. Microalgae identification is generally considered a complicated issue; although, in the last few years, new molecular methods have helped to rectify this problem. Among the different techniques available, DNA barcoding has proven very useful in providing rapid, accurate, and automatable species identification; in this work, it is used to assess the genomic identity of the microalga species Scenedesmus sp. 'almeriensis', a common strain in industrial-scale cultures. Barcode markers rbcL and ITS1-5.8S-ITS2 were sequenced and the obtained genomic information was used to design a quantitative PCR assay to precisely quantify the S. almeriensis concentration in microalgal cultures of industrial interest. TaqMan chemistry was used to quantify down to 1 µg/L dry weight of S. almeriensis cells, including in the presence of concentrated mixed cultures of other microalgae. A simple direct qPCR approach was also investigated to avoid classic DNA extraction and to reduce total assay time to approximately 2 h. The objective was to design strain-specific tools able to confirm and quantify the presence of different strains in whatever microalgae culture so as to achieve maximal productivity and quality of the produced biomass.

A whole biodiesel conversion process combining isolation, cultivation and in situ supercritical methanol transesterification of native microalgae.

A coupled process combining microalgae production with direct supercritical biodiesel conversion using a reduced number of operating steps is proposed in this work. Two newly isolated native microalgae strains, identified as Chlorella sp. and Nannochloris sp., were cultivated in both batch and continuous modes. Maximum productivities were achieved during continuous cultures with 318mg/lday and 256mg/lday for Chlorella sp. and Nannochloris sp., respectively. Microalgae were further characterized by determining their photosynthetic performance and nutrient removal efficiency. Biodiesel was produced by catalyst-free in situ supercritical methanol transesterification of wet unwashed algal biomass (75wt.% of moisture). Maximum biodiesel yields of 45.62wt.% and 21.79wt.% were reached for Chlorella sp. and Nannochloris sp., respectively. The analysis of polyunsaturated fatty acids of Chlorella sp. showed a decrease in their proportion when comparing conventional and supercritical transesterification processes (from 37.4% to 13.9%, respectively), thus improving the quality of the biodiesel.

New approach to solar photo-Fenton operation. Raceway ponds as tertiary treatment technology.

The photo-Fenton process has proven its efficiency in the removal of micropollutants. However, the high costs usually associated with it prevent a spread of this technology. An important factor affecting costs is the kind of photoreactor used, usually tubular with a reflecting surface. Tubular reactors like compound parabolic collectors, CPCs, involve high capital costs. In comparison, the application of less costly reactors such as the extensive raceway ponds (RPRs) would help to spread the use of the photo-Fenton process as tertiary treatment at commercial scale. As far as the authors know, RPRs have never been used in advanced oxidation processes (AOPs) applications. This work is aimed at studying the applicability of RPRs to remove micropollutants with solar photo-Fenton. For this purpose, a pesticide mixture of commercial acetamiprid (ACTM) and thiabendazole (TBZ) (100µg/L each) was used in simulated secondary effluent. Iron concentration (1, 5.5 and 10mg/L) and liquid depth (5, 10 and 15cm) were studied as process variables. TBZ was removed at the beginning of the treatment (less than 5min), although ACTM removal times were longer (20-40min for the highest iron concentrations). High treatment capacity per surface area was obtained (48mg/hm(2) with 5.5mg Fe/L and 15cm liquid depth), proving the feasibility of using RPRs for micropollutant removal.