Recent Advances in Microalgal Biorefineries

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Microalgal cultures for the remediation of wastewaters with different nitrogen to phosphorus ratios: Process modelling using artificial neural networks

Microalgae have remarkable potential for wastewater bioremediation since they can efficiently uptake nitrogen and phosphorus in a sustainable and environmentally friendly treatment system. However, wastewater composition greatly depends on its source and has a significant seasonal variability. This study aimed to evaluate the impact of different N:P molar ratios on the growth of Chlorella vulgaris and nutrient removal from synthetic wastewater. Furthermore, artificial neural network (ANN) threshold models, optimised by genetic algorithms (GAs), were used to model biomass productivity (BP) and nitrogen/phosphorus removal rates (RRN/RRP). The impact of various inputs culture variables on these parameters was evaluated. Microalgal growth was not nutrient limited since the average biomass productivities and specific growth rates were similar between the experiments. Nutrient removal efficiencies/rates reached 92.0 +/- 0.6%/6.15 +/- 0.01 mgN L-1 d-1 for nitrogen and 98.2 +/- 0.2%/0.92 +/- 0.03 mgP L-1 d-1 for phosphorus. Low nitrogen concentration limited phosphorus uptake for low N:P ratios (e.g., 2 and 3, yielding 36 +/- 2 mgDW mgP-1 and 39 +/- 3 mgDW mgP-1, respectively), while low phosphorus concentration limited nitrogen uptake with high ratios (e.g., 66 and 67, yielding 9.0 +/- 0.4 mgDW mgN-1 and 8.8 +/- 0.3 mgDW mgN-1, respectively). ANN models showed a high fitting performance, with coefficients of determination of 0.951, 0.800, and 0.793 for BP, RRN, and RRP, respectively. In summary, this study demonstrated that microalgae could successfully grow and adapt to N:P molar ratios between 2 and 67, but the nutrient uptake was impacted by these variations, especially for the lowest and highest N:P molar ratios. Furthermore, GA-ANN models demonstrated to be relevant tools for microalgal growth modelling and control. Their high fitting performance in characterising this biological system can contribute to reducing the experi-mental effort for culture monitoring (human resources and consumables), thus decreasing the costs of microalgae production

HIF-1α-independent hypoxia-induced rapid PTK6 stabilization is associated with increased motility and invasion

This article has been made available through the Brunel Open Access Publishing Fund.PTK6/Brk is a non-receptor tyrosine kinase overexpressed in cancer. Here we demonstrate that cytosolic PTK6 is rapidly and robustly induced in response to hypoxic conditions in a HIF-1-independent manner. Furthermore, a proportion of hypoxic PTK6 subsequently re-localized to the cell membrane. We observed that the rapid stabilization of PTK6 is associated with a decrease in PTK6 ubiquitylation and we have identified c-Cbl as a putative PTK6 E3 ligase in normoxia. The consequences of hypoxia-induced PTK6 stabilization and subcellular re-localization to the plasma membrane include increased cell motility and invasion, suggesting PTK6 targeting as a therapeutic approach to reduce hypoxia-regulated metastatic potential. This could have particular significance for breast cancer patients with triple negative disease.A Breast Cancer Campaign pilot grant awarded to E.M.H. supported this study. I.M.P. is supported by University of Hull HEFCE funding, and Royal Society and Breast Cancer Campaign pilot grants. S.A.E. is supported by ICR HEFCE funding and Cancer Research UK program grant C309/A11566. A.H. is supported by Brunel University HEFCE funding. E.M.H. is supported by a Cancer Research UK grant

Microalgae systems- environmental agents for wastewater treatment and further potential biomass valorisation

Water is the most valuable resource on the planet. However, massive anthropogenic activities generate threat-ening levels of biological, organic, and inorganic pollutants that are not efficiently removed in conventional wastewater treatment systems. High levels of conventional pollutants (carbon, nitrogen, and phosphorus), emerging chemical contaminants such as antibiotics, and pathogens (namely antibiotic-resistant ones and related genes) jeopardize ecosystems and human health. Conventional wastewater treatment systems entail several environmental issues: (i) high energy consumption; (ii) high CO2 emissions; and (iii) the use of chemicals or the generation of harmful by-products. Hence, the use of microalgal systems (entailing one or several microalgae species, and in consortium with bacteria) as environmental agents towards wastewater treatment has been seen as an environmentally friendly solution to remove conventional pollutants, antibiotics, coliforms and antibiotic resistance genes. In recent years, several authors have evaluated the use of microalgal systems for the treatment of different types of wastewater, such as agricultural, municipal, and industrial. Generally, microalgal systems can provide high removal efficiencies of: (i) conventional pollutants, up to 99%, 99%, and 90% of total nitrogen, total phosphorus, and/or organic carbon, respectively, through uptake mechanisms, and (ii) antibiotics frequently found in wastewaters, such as sulfamethoxazole, ciprofloxacin, trimethoprim and azithromycin at 86%, 65%, 42% and 93%, respectively, through the most desirable microalgal mechanism, biodegradation. Although pathogens removal by microalgal species is complex and very strain-specific, it is also possible to attain total coliform and Escherichia coli removal of 99.4% and 98.6%, respectively. However, microalgal systems' effectiveness strongly relies on biotic and abiotic conditions, thus the selection of operational conditions is critical. While the combination of selected species (microalgae and bacteria), ratios and inoculum concentration allow the efficient removal of conventional pollutants and generation of high amounts of biomass (that can be further converted into valuable products such as biofuels and biofertilisers), abiotic factors such as pH, hydraulic retention time, light intensity and CO2/O2 supply also have a crucial role in conventional pollutants and anti-biotics removal, and wastewater disinfection. However, some rationale must be considered according to the purpose. While alkaline pH induces the hydrolysis of some antibiotics and the removal of faecal coliforms, it also decreases phosphates solubility and induces the formation of ammonium from ammonia. Also, while CO2 supply increases the removal of E. coli and Pseudomonas aeruginosa, as well as the microalgal growth (and thus the conventional pollutants uptake), it decreases Enterococcus faecalis removal. Therefore, this review aims to pro-vide a critical review of recent studies towards the application of microalgal systems for the efficient removal of conventional pollutants, antibiotics, and pathogens; discussing the feasibility, highlighting the advantages and challenges of the implementation of such process, and presenting current case-studies of different applications of microalgal systems

Sustainable Microalgal Harvesting Process Applying Opuntia cochenillifera: Process Parameters Optimization

Microalgae harvesting by coagulation can use coagulant agents such as alum, synthetic polymers or biocoagulants. Biocoagulants have attracted the attention of researchers because they are natural, biodegradable, and promote high microalgal harvesting efficiencies. This study aims to optimize the harvesting of Chlorella vulgaris based on the dosage of the Opuntia cochenillifera extract and the choice of eluent for biopolymer extraction. The outdoor cultivation of C. vulgaris achieved a specific growth rate of 0.455 d(-1) and a maximum biomass concentration of 1.28 g(DW) L-1. In order to harvest the microalgal biomass, the polymer present in the mucilage of O. cochenillifera was extracted using NaOH and HCl. Coagulation and sedimentation assays were performed with different coagulant dosages: 3.5, 5.9, and 8.2 g L-1. The maximum harvesting efficiencies using the acid and alkaline extract coagulant solutions were 80.8% and 99.5%, respectively, with a dosage of 3.5 g L-1. According to the results, the C. vulgaris biomass can be harvested with the mucilage from O. cochenillifera in acid and alkaline eluents. The application of this biocoagulant constitutes a sustainable solution for microalgal harvesting

Microalgal Systems, a Green Solution for Wastewater Conventional Pollutants Removal, Disinfection, and Reduction of Antibiotic Resistance Genes Prevalence?

The low-efficiency rate of urban wastewater (UWW) treatment generates tons of discharged water with a high concentration of pollutants, pathogens and antibiotic-resistance genes (ARGs). Microalgal systems may be a green alternative to be implemented as a UWW polishing treatment. This study assessed the ability of Chlorella vulgaris and UWW autochthonous microalgal species (AMS) to simultaneously remove PO4-P, and reduce the proliferation of coliforms and ARGs. AMS seems to be more promising due to: (i) the higher specific growth rate, mu(max) (0.687 +/- 0.065 d(-1)); (ii) efficient PO4-P removal (92.62 +/- 0.10%); (iii) faster reduction of coliforms proliferation achieving concentrations below the limits of quantification (6 d); (iv) the reduction of intl1 and the ARGs sul1 and blaTEM abundance in ca. of 70.4%, 69.2%, and 75.7%, respectively (9 d); and (v) the additional reduction of these genes in ca. of 97.1%, 94.2%, and 99.9%, respectively, after 5 d storage in the dark and at room temperature. Results also revealed that the high pH values in both microalgal systems (due to microalgal growth) were highly correlated with a reduction in the proliferation of coliforms, including Escherichia coli. In conclusion, using AMS as a final polishing treatment of UWW seems to be very promising

Microalgal Growth in Aquaculture Effluent: Coupling Biomass Valorisation with Nutrients Removal

Natural resources are becoming increasingly scarce, and the need to control their consumption and recycle their use is growing. Water is one of the essential resources for human survival. Therefore, there has been an increasing interest in ways to save, recycle and treat water supplies. Aquaculture is one of the most polluting activities as it produces a significant wastewater volume, which needs proper treatment before being discharged into the environment or recycled. Microalgae are a potential solution for wastewater treatment. Due to their numerous advantages, the use of microalgal biomass is being studied, and, at present, there is already a market and room for profit in the sale of microalgal components in various forms, such as animal and human supplements. From a biorefinery point of view, it is important to take advantage of all the qualities and benefits that microalgae have by combining their great capacity to treat wastewater and exploit the produced biomass, analysing its composition for subsequent valorisation, for example. In this study, Chlorella vulgaris was used to treat aquaculture wastewater from a trout farm aquaculture facility, and the treatment efficiency was evaluated. To valorise the resulting biomass, its composition was also assessed. C. vulgaris successfully grew in the effluent with growth rates of 0.260 +/- 0.014 d(-1) and with average productivity of 32.9 +/- 1.6 mg L-1 d(-1). The achieved removal efficiencies were 93.5 +/- 2.1% for total nitrogen, 98.0 +/- 0.1% for nitrate-nitrogen and 92.7 +/- 0.1% for phosphate-phosphorus. Concerning biomass composition, the lipids (15.82 +/- 0.15%), carbohydrates (48.64 +/- 0.83%), and pigment contents (0.99 +/- 0.04% for chlorophyll a + b and 0.21 +/- 0.04% for carotenoids) were similar to the values of similar studies. However, the protein content obtained (17.93 +/- 1.21%) was lower than the ones mentioned in the literature

Expression of histone methyltransferases as novel biomarkers for renal cell tumor diagnosis and prognostication

Renal cell tumors (RCTs) are the most lethal of the common urological cancers. The widespread use of imaging entailed an increased detection of small renal masses, emphasizing the need for accurate distinction between benign and malignant RCTs, which is critical for adequate therapeutic management. Histone methylation has been implicated in renal tumorigenesis, but its potential clinical value as RCT biomarker remains mostly unexplored. Hence, the main goal of this study was to identify differentially expressed histone methyltransferases (HMTs) and histone demethylases (HDMs) that might prove useful for RCT diagnosis and prognostication, emphasizing the discrimination between oncocytoma (a benign tumor) and renal cell carcinoma (RCC), especially the chromophobe subtype (chRCC). We found that the expression levels of three genes-SMYD2, SETD3, and NO66-was significantly altered in a set of RCTs, which was further validated in a large independent cohort. Higher expression levels were found in RCTs compared to normal renal tissues (RNTs) and in chRCCs comparatively to oncocytomas. SMYD2 and SETD3 mRNA levels correlated with protein expression assessed by immunohistochemistry. SMYD2 transcript levels discriminated RCTs from RNT, with 82.1% sensitivity and 100% specificity (AUC=0.959), and distinguished chRCCs from oncocytomas, with 71.0% sensitivity and 73.3% specificity (AUC: 0.784). Low expression levels of SMYD2, SETD3, and NO66 were significantly associated with shorter disease-specific and disease-free survival, especially in patients with non-organ confined tumors. We conclude that expression of selected HMTs and HDMs might constitute novel biomarkers to assist in RCT diagnosis and assessment of tumor aggressiveness.This study was funded by research grants from Research Center of Portuguese Oncology Institute – Porto (CI-IPOP 4-2012) and European Community’s Seventh Framework Program – Grant number FP7-HEALTH-F5-2009-241783. ASP-L and FQV are and were supported by FCT-Fundação para a Ciência e a Tecnologia grants (SFRH/SINTD/94217/2013 and SFRH/ BD/70564/2010, respectively)