An Integrated Strategy for Nutraceuticals from Haematoccus pluvialis: From Cultivation to Extraction.

The aim of this study was to develop an effective integrated cultivation system for Haematococcus pluvialis as a source of bioactive compounds such as astaxanthin, lutein, proteins, and fatty acids (FAs). The Chlorophyta H. pluvialis was cultivated in a vertical bubble column photobioreactor (VBC-PBR) under batch mode, allowing switching from green to red phase for astaxanthin induction. The combined effect of light intensity and nutrients on bioactive compound formation was investigated. Results showed that growth under lower nutrients availability and light intensity led to a higher concentration of biomass. Growth under high light intensity with an appropriate concentration of nitrate, sulfate, phosphate and magnesium led to ~85% and ~58% higher production of total carotenoids and fatty acids, respectively. Under high stress conditions, ~90% nitrate and phosphate consumption were observed.

Effectiveness of Dunaliella salina Extracts against Bacillus subtilis and Bacterial Plant Pathogens.

Several bacteria pathogens are responsible for plant diseases causing significant economic losses. The antibacterial activity of Dunaliella salina microalgae extracts were investigated in vitro and in vivo. First, biomass composition was chemically characterized and subjected to extraction using polar/non-polar solvents. The highest extraction yield was obtained using chloroform:methanol (1:1 v/v) equal to 170 mg g-1 followed by ethanol (88 mg g-1) and hexane (61 mg g-1). In vitro examination of hexane extracts of Dunaliella salina demonstrated antibacterial activity against all tested bacteria. The hexane extract showed the highest amount of ß-carotene with respect to the others, so it was selected for subsequent analyses. In vivo studies were also carried out using hexane extracts of D. salina against Pseudomonas syringae pv. tomato and Pectobacterium carotovorum subsp. carotovorum on young tomato plants and fruits of tomato and zucchini, respectively. The treated young tomato plants exhibited a reduction of 65.7% incidence and 77.0% severity of bacterial speck spot disease. Similarly, a reduction of soft rot symptoms was observed in treated tomato and zucchini fruits with a disease incidence of 5.3% and 12.6% with respect to 90.6% and 100%, respectively, for the positive control.

The Use of Exogenous Lung Surfactant (Poractant Alfa) in Acute Respiratory Failure by Drowning.

Drowning is an acute respiratory failure as a result from immersion or submersion of the airways in a liquid medium (predominantly water). Inhalation of water causes severe lung damage due to the destruction of pulmonary surfactant, resulting in decreased lung elasticity, alveolar collapse, alteration of ventilation-perfusion ratio, intrapulmonary blood shunting, hypoxia, acute lung injury, and Acute Respiratory Distress Syndrome (ARDS). Poractant alfa (Curosurf®), a natural surfactant effective in the treatment of newborn respiratory distress, has been used in various forms of ARDS, but in drowning syndromes, experience is still poor. We describe a series of nine clinical cases of drowning, six adults and three children, treated in our Intensive Care Unit (ICU) with endobronchial administration of poractant alfa. After 24 and 48 hours of administration in all cases, there was an improvement in arterial blood gas analysis (ABG) parameters and imaging. All patients were discharged without clinical consequences.

Enhancing Biomass and Lutein Production From Scenedesmus almeriensis: Effect of Carbon Dioxide Concentration and Culture Medium Reuse.

The main purpose of this study is to investigate the effects of operative parameters and bioprocess strategies on the photo-autotrophic cultivation of the microalgae Scenedesmus almeriensis for lutein production. S. almeriensis was cultivated in a vertical bubble column photobioreactor (VBC-PBR) in batch mode and the bioactive compounds were extracted by accelerated solvent extraction with ethanol at 67°C and 10 MPa. The cultivation with a volume fraction of CO2 in the range 0-3.0%v/v showed that the highest biomass and lutein concentrations - 3.7 g/L and 5.71 mg/g, respectively - were measured at the highest CO2 concentration and using fresh growth medium. Recycling the cultivation medium from harvested microalgae resulted in decreased biomass and lutein content. The nutrient chemical composition analysis showed the highest consumption rates for nitrogen and phosphorus, with values higher than 80%, while sulfate and chloride were less consumed.

Smart Method for Carotenoids Characterization in Haematococcus pluvialis red phase and Evaluation of Astaxanthin Thermal Stability.

Haematococcus pluvialis microalgae is a promising source of astaxanthin, an excellent antioxidant carotenoid. H. pluvialis, as well as other species, could find more extensive applications as healthy food for a variegated carotenoids composition in addition to astaxanthin. Official method has not currently been used for this purpose. The objective of this work was to propose a method to characterize carotenoids in H. pluvialis after the comparison between spectrophotometric and liquid chromatography analysis. In addition, in order to improve the use of astaxanthin in the food industry, thermal stability was investigated. In this context, the effect of temperature at 40-80 °C, over a 16 h storage period was tested on astaxanthin produced by H. pluvialis. A further test was carried out at room temperature (20 °C) for seven days. A decrease in the astaxanthin concentration was observed at all tested temperatures with a decrease >50% of all-trans isomer at 80 °C after 16 h and an increase of 9-cis and 13-cis isomers. In conclusion, the obtained results showed the importance of evaluating the degradation effect of temperature on astaxanthin used as a food additive for a future greater enhancement of this bioproduct in the food field.

Use of Ceftolozane/Tazobactam in a Case of Septic Shock by Puerperal Sepsis.

Multiresistant bacteria infections cause widespread morbidity and mortality and lead to an increase in expenses for hospital stays and complications. We describe the case of a 27-year-old patient with puerperal sepsis after cesarean section due to Escherichia coli complicated by multiresistant Klebsiella ESBL-producing superinfection with septic shock and multiple organ dysfunction syndrome, successfully treated with ceftolozane/tazobactam.

Selective Extraction of ω-3 Fatty Acids from Nannochloropsis sp. Using Supercritical CO2 Extraction.

In this article, microalgae Nannochloropsis sp. was used for fatty acid (FA) extraction, using a supercritical fluid-carbon dioxide (SF-CO2) extraction method. This study investigated the influence of different pre-treatment conditions by varying the grinding speed (200-600 rpm), pre-treatment time (2.5-10 min), and mixing ratio of diatomaceous earth (DE) and Nannochloropsis sp. biomass (0.5-2.0 DE/biomass) on FAs extraction. In addition, the effect of different operating conditions, such as pressure (100-550 bar), temperature (50-75 °C), and CO2 flow rate (7.24 and 14.48 g/min) on eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) recovery, was analyzed. Experimental data evidenced that, keeping constant the extraction conditions, the pre-treatment step enhanced the FAs extraction yield up to 3.4 fold, thereby the maximum extracted amount of FAs (61.19 mg/g) was attained with the pre-treatment with a ratio of DE/biomass of 1 at 600 rpm for 5 min. Moreover, by increasing both SF-CO2 pressure and temperature, the selectivity towards EPA was enhanced, while intermediate pressure and lower pressure promoted DHA recovery. The highest amount of extracted EPA, i.e., 5.69 mg/g, corresponding to 15.59%, was obtained at 75 °C and 550 bar with a CO2 flow rate of 14.48 g/min, while the maximum amount of extracted DHA, i.e., ~0.12 mg/g, equal to 79.63%, was registered at 50 °C and 400 bar with a CO2 flow rate of 14.48 g/min. Moreover, the increased CO2 flow rate from 7.24 to 14.48 g/min enhanced both EPA and DHA recovery.

Supercritical Fluid Extraction of Lutein from Scenedesmus almeriensis.

Lutein has several benefits for human health, playing an important role in the prevention of age-related macular degeneration (AMD), cataracts, amelioration of the first stages of atherosclerosis, and some types of cancer. In this work, the Scenedesmus almeriensis microalga was used as a natural source for the supercritical fluid (SF) extraction of lutein. For this purpose, the optimization of the main parameters affecting the extraction, such as biomass pre-treatment, temperature, pressure, and carbon dioxide (CO2) flow rate, was performed. In the first stage, the effect of mechanical pre-treatment (diatomaceous earth (DE) and biomass mixing in the range 0.25-1 DE/biomass; grinding speed varying between 0 and 600 rpm, and pre-treatment time changing from 2.5 to 10 min), was evaluated on lutein extraction efficiency. In the second stage, the influence of SF-CO2 extraction parameters such as pressure (25-55 MPa), temperature (50 and 65 °C), and CO2 flow rate (7.24 and 14.48 g/min) on lutein recovery and purity was investigated. The results demonstrated that by increasing temperature, pressure, and CO2 flow rate lutein recovery and purity were improved. The maximum lutein recovery (~98%) with purity of ~34% was achieved operating at 65 °C and 55 MPa with a CO2 flow rate of 14.48 g/min. Therefore, optimum conditions could be useful in food industries for lutein supplementation in food products.

Extraction of Bioactive Compounds Using Supercritical Carbon Dioxide.

Microalgae Dunaliella salina contains useful molecules such as ß-carotene and fatty acids (FAs), which are considered high value-added compounds. To extract these molecules, supercritical carbon dioxide was used at different operative conditions. The effects of mechanical pre-treatment (grinding speed at 0⁻600 rpm; pre-treatment time of 2.5⁻7.5 min) and operating parameters for extraction, such as biomass loading (2.45 and 7.53 g), pressure (100⁻550 bars), temperature (50⁻75 °C) and CO2 flow rate (7.24 and 14.48 g/min) by varying the extraction times (30⁻110 min) were evaluated. Results showed that the maximum cumulative recovery (25.48%) of ß-carotene was achieved at 400 bars and 65 °C with a CO2 flow rate of 14.48 g/min, while the highest purity for stage (55.40%) was attained at 550 bars and 65 °C with a CO2 flow rate of 14.48 g/min. The maximum recovery of FAs, equal to 8.47 mg/g, was achieved at 550 bars and 75 °C with a CO2 flow rate of 14.48 g/min. Moreover, the lowest biomass loading (2.45 g) and the first extraction cycle (30 min) allowed the maximum extraction of ß-carotene and FAs.

Extraction of Astaxanthin and Lutein from Microalga Haematococcus pluvialis in the Red Phase Using CO2 Supercritical Fluid Extraction Technology with Ethanol as Co-Solvent.

Astaxanthin and lutein, antioxidants used in nutraceutics and cosmetics, can be extracted from several microalgal species. In this work, investigations on astaxanthin and lutein extraction from Haematococcus pluvialis (H. pluvialis) in the red phase were carried out by means of the supercritical fluid extraction (SFE) technique, in which CO2 supercritical fluid was used as the extracting solvent with ethanol as the co-solvent. The experimental activity was performed using a bench-scale reactor in semi-batch configuration with varying extraction times (20, 40, 60, and 80 min), temperatures (50, 65, and 80 °C) and pressures (100, 400, and 550 bar). Moreover, the performance of CO2 SFE with ethanol was compared to that without ethanol. The results show that the highest astaxanthin and lutein recoveries were found at 65 °C and 550 bar, with ~18.5 mg/g dry weight (~92%) astaxanthin and ~7.15 mg/g dry weight (~93%) lutein. The highest astaxanthin purity and the highest lutein purity were found at 80 °C and 400 bar, and at 65 °C and 550 bar, respectively.

Microalgae Characterization for Consolidated and New Application in Human Food, Animal Feed and Nutraceuticals.

The exploration of new food sources and natural products is the result of the increase in world population as well as the need for a healthier diet; in this context, microalgae are undoubtedly an interesting solution. With the intent to enhance their value in new commercial applications, this paper aims to characterize microalgae that have already been recognized as safe or authorized as additives for humans and animals (Chlorella vulgaris, Arthrospira platensis, Haematococcus pluvialis, Dunaliella salina) as well as those that have not yet been marketed (Scenedesmus almeriensis and Nannocholoropsis sp.). In this scope, the content of proteins, carbohydrates, lipids, total dietary fiber, humidity, ash, and carotenoids has been measured via standard methods. In addition, individual carotenoids (beta-carotene, astaxanthin, and lutein) as well as individual saturated, monounsaturated, and polyunsaturated fatty acids have been identified and quantified chromatographically. The results confirm the prerogative of some species to produce certain products such as carotenoids, polyunsaturated fatty acids, and proteins, but also show how their cellular content is rich and diverse. H. pluvialis green and red phases, and Nannochloropsis sp., in addition to producing astaxanthin and omega-3, contain about 25⁻33% w/w proteins on a dry basis. D. salina is rich in beta-carotene (3.45% w/w on a dry basis), S. Almeriensis is a source of lutein (0.30% w/w on a dry basis), and the C. vulgaris species is a protein-based microalgae (45% w/w on a dry basis). All, however, can also produce important fatty acids such as palmitic acid, γ-linolenic acid, and oleic acid. Considering their varied composition, these microalgae can find applications in multiple sectors. This is true for microalgae already on the market as well as for promising new sources of bioproducts such as S. almeriensis and Nannochloropsis sp.

Supercritical Carbon Dioxide Extraction of Astaxanthin, Lutein, and Fatty Acids from Haematococcus pluvialis Microalgae.

Haematococcus pluvialis microalgae in the red phase can produce significant amounts of astaxanthin, lutein, and fatty acids (FAs), which are valuable antioxidants in nutraceutics and cosmetics. Extraction of astaxanthin, lutein, and FAs from disrupted biomass of the H. pluvialis red phase using carbon dioxide (CO2) in supercritical fluid extraction (SFE) conditions was investigated using a bench-scale reactor in a semi-batch configuration. In particular, the effect of extraction time (20, 40, 60, 80, and 120 min), CO2 flow rate (3.62 and 14.48 g/min) temperature (50, 65, and 80 °C), and pressure (100, 400, and 550 bar.) was explored. The results show the maximum recovery of astaxanthin and lutein achieved were 98.6% and 52.3%, respectively, at 50 °C and 550 bars, while the maximum recovery of FAs attained was 93.2% at 65 °C and 550 bars.

Extraction of astaxanthin from microalga Haematococcus pluvialis in red phase by using generally recognized as safe solvents and accelerated extraction.

Solvent Extraction was tested to extract astaxanthin from Haematococcus pluvialis in red phase (HPR), by investigating effects of solvents, extraction pressure and temperature. Astaxanthin isomers were identified and quantified in the extract. The performances of acetone and ethanol, Generally Recognized As Safe (GRAS) solvents, were explored. Negligible effect of pressure was found, while with increasing extraction temperature astaxanthin recovery increased till a maximum value, beyond which thermal degradation seemed to be greater than the positive effect of temperature on extraction. Furthermore, to maximize the extraction yield of astaxanthin, mechanical pre-treatment of HPR biomass was carried out and several extraction runs were consecutively performed. Experimental results showed that after the mechanical pre-treatment the astaxanthin recovery strongly increased while a single extraction run of 20 min was sufficient to extract more than 99% of total astaxanthin extracted. After pre-treatment, maximum recovery of about 87% was found for acetone (pressure = 100 bar; temperature = 40 °C; total time = 60 min).

Evaluation of the treatability of a winery distillery (vinasse) wastewater by UASB, anoxic-aerobic UF-MBR and chemical precipitation/adsorption.

A multi-stage pilot-scale treatment cycle consisting of an Upflow Anaerobic Sludge Blanket reactor (UASB) followed by an anoxic-aerobic Ultra Filtration Membrane Bio Reactor (UF-MBR) and a post treatment based on chemical precipitation with lime or adsorption on Granular Activated Carbons (GAC), was applied in order to evaluate the treatment feasibility of a real winery distillery wastewater at laboratory and bench scale. The wastewater was classified as high strength with acidic pH (3.8), and concentrations of 44,600, 254, 604 and 660 mg/l for CODtot, total nitrogen, total phosphorous and phenols, respectively. The UASB reactor was operated at Organic Loading Rates (OLR) in the range 3.0-11.5 kgCODtot/m3/d achieving treatment efficiency up to 97%, with an observed methane production of 340 L of CH4/kgCOD. The MBR system was operated with an organic load in the range 0.070-0.185 kgCOD/kgVSS/d, achieving a removal up to 48%, 67% and 65% of the influent COD, total nitrogen and phenols, respectively. The combination of UASB and UF-MBR treatment units was not effective in phosphate and colour removal assigning to further chemical precipitation and adsorption processes, respectively, their complete removal in order to comply with legal standards for wastewater discharge. Subsequently, the optimization of the investigated treatment chain was assessed by applying a chemical precipitation step upstream and downstream the UASB reactor, and a related treatment unit cost assessment is presented in view of a further technological scale-up.

The Use of Coupled Plasma Filtration Adsorption in Traumatic Rhabdomyolysis.

Severe musculoskeletal injuries induce the release of sarcoplasmic elements such as muscle enzymes, potassium, and myoglobin in the systemic circulation. The circulating myoglobin damages the glomerulus and renal tubules. Conventional haemodialysis is not able to remove myoglobin, due to its high molecular weight (17,8 kilodaltons [kDa]). We treated four traumatic rhabdomyolysis patients with Coupled Plasma Filtration Adsorption (CPFA) in order to remove myoglobin followed by 14 hours of Continuous Veno-Venous Hemofiltration (CVVH). During the treatment, all patients showed clinical improvement with a decrease in muscular (creatine kinase [CK] and myoglobin) and renal (creatinine and potassium) damage indices. One patient, in spite of full renal recovery, died of cerebral haemorrhage on the 26th day of hospital stay.

Microbial responses to polycyclic aromatic hydrocarbon contamination in temporary river sediments: Experimental insights.

Temporary rivers are characterized by dry-wet phases and represent an important water resource in semi-arid regions worldwide. The fate and effect of contaminants have not been firmly established in temporary rivers such as in other aquatic environments. In this study, we assessed the effects of sediment amendment with Polycyclic Aromatic Hydrocarbons (PAHs) on benthic microbial communities. Experimental microcosms containing natural (Control) and amended sediments (2 and 20 mg PAHs kg(-1) were incubated for 28 days. The PAH concentrations in sediments were monitored weekly together with microbial community structural (biomass and phylogenetic composition by TGGE and CARD-FISH) and functional parameters (ATP concentration, community respiration rate, bacterial carbon production rate, extracellular enzyme activities). The concentration of the PAH isomers did not change significantly with the exception of phenanthrene. No changes were observed in the TGGE profiles, whereas the occurrence of Alpha- and Beta-Proteobacteria was significantly affected by the treatments. In the amended sediments, the rates of carbon production were stimulated together with aminopeptidase enzyme activity. The community respiration rates showed values significantly lower than the Control after 1 day from the amendment then recovering the Control values during the incubation. A negative trend between the respiration rates and ATP concentration was observed only in the amended sediments. This result indicates a potential toxic effect on the oxidative phosphorylation processes. The impoverishment of the energetic resources that follows the PAH impact may act as a domino on the flux of energy from prokaryotes to the upper level of the trophic chain, with the potential to alter the temporary river functioning.

An integrated approach for monitoring efficiency and investments of activated sludge-based wastewater treatment plants at large spatial scale.

WISE, the Water Information System for Europe, is the web-portal of the European Commission (EU) that disseminates the quality state of the receiving water bodies and the efficiency of the municipal wastewater treatment plants (WWTPs) in order to monitor advances in the application of both the Water Framework Directive (WFD) as well as the Urban Wastewater Treatment Directive (UWWTD). With the intention to develop WISE applications, the aim of the work was to define and apply an integrated approach capable of monitoring the efficiency and investments of activated sludge-based WWTPs located in a large spatial area, providing the following outcomes useful to the decision-makers: (i) the identification of critical facilities and their critical processes by means of a Performance Assessment System (PAS), (ii) the choice of the most suitable upgrading actions, through a scenario analysis. (iii) the assessment of the investment costs to upgrade the critical WWTPs and (iv) the prioritization of the critical facilities by means of a multi-criteria approach which includes the stakeholders involvement, along with the integration of some technical, environmental, economic and health aspects. The implementation of the proposed approach to a high number of municipal WWTPs highlighted how the PAS developed was able to identify critical processes with a particular effectiveness in identifying the critical nutrient removal ones. In addition, a simplified approach that considers the cost related to a basic-configuration and those for the WWTP integration, allowed to link the critical processes identified and the investment costs. Finally, the questionnaire for the acquisition of data such as that provided by the Italian Institute of Statistics, the PAS defined and the database on the costs, if properly adapted, may allow for the extension of the integrated approach on an EU-scale by providing useful information to water utilities as well as institutions.

Organic priority substances and microbial processes in river sediments subject to contrasting hydrological conditions.

Flood and drought events of higher intensity and frequency are expected to increase in arid and semi-arid regions, in which temporary rivers represent both a water resource and an aquatic ecosystem to be preserved. In this study, we explored the variation of two classes of hazardous substances (Polycyclic Aromatic Hydrocarbons and Nonylphenols) and the functioning of the microbial community in river sediments subject to hydrological fluctuations (Candelaro river basin, Italy). Overall, the concentration of pollutants (∑PAHs range 8-275ngg(-1); ∑NPs range 299-4858ngg(-1)) suggests a moderate degree of contamination. The conditions in which the sediments were tested, flow (high/low) and no flow (wet/dry/arid), were associated to significant differences in the chemical and microbial properties. The total organic carbon contribution decreased together with the stream flow reduction, while the contribution of C-PAHs and C-NPs tended to increase. NPs were relatively more concentrated in sediments under high flow, while the more hydrophobic PAHs accumulated under low and no flow conditions. Passing from high to no flow conditions, a gradual reduction of microbial processes was observed, to reach the lowest specific bacterial carbon production rates (0.06fmolCh(-1)cell(-1)), extracellular enzyme activities, and the highest doubling time (40h) in arid sediments. In conclusion, different scenarios for the mobilization of pollutants and microbial processes can be identified under contrasting hydrological conditions: (i) the mobilization of pollutants under high flow and a relatively higher probability for biodegradation; (ii) the accumulation of pollutants during low flow and lower probability for biodegradation; (iii) the drastic reduction of pollutant concentrations under dry and arid conditions, probably independently from the microbial activity (abiotic processes). Our findings let us infer that a multiple approach has to be considered for an appropriate water resource exploitation and a more realistic prevision of the impact of pollutants in temporary waters.

Response of bacterial isolates from Antarctic shallow sediments towards heavy metals, antibiotics and polychlorinated biphenyls.

The response of bacterial isolates from Antarctic sediments to polychlorinated biphenyls (Aroclor 1242 mixture), heavy metal salts (cadmium, copper, mercury and zinc) and antibiotics (ampicillin, chloramphenicol, kanamycin and streptomycin) was investigated. Overall, the ability to growth in the presence of Aroclor 1242 as a sole carbon source was observed for 22 isolates that mainly belonged to Psychrobacter spp. Tolerance to the heavy metals assayed in this study was in the order of Cd > Cu > Zn > Hg and appeared to be strictly related to the metal concentrations, as determined during previous chemical surveys in the same area. With regards to antibiotic assays, the response of the isolates to the tested antibiotics ranged from complete resistance to total susceptibility. In particular, resistances to ampicillin and chloramphenicol were very pronounced in the majority of isolates. Our isolates differently responded to the presence of toxic compounds primarily based on their phylogenetic affiliation and secondarily at strain level. Moreover, the high incidence of resistance either to metal or antibiotics, in addition to the capability to grow on PCBs, confirm that bacteria are able to cope and/or adapt to the occurrence pollutants even in low human-impacted environments.